A, B, D cells and a fourth cell type in long-term cultures of fetal rat pancreas. Whole pancreases from fetal rats of 13 days and 18 days gestation were explanted onto rayon grids and grown in organ culture. Cultures were fixed in Bouin's fluid, sectioned and stained with the fluorescent antibody techniques for glucagon and insulin, aldehyde fuchsin for B cells, pseudoisocyanin for D cells and a silver technique for the fourth cell type. The 13-day explants were fixed after 10 days in culture. A, B and D and the fourth cell type were seen, indicating that precursors of all four endocrine cell types must be present in the fetal pancreas shortly after the formation of the pancreatic bud (11 days). Further, the presence of these four cell types in the walls of tubules in these cultures indicates the tubules as the site of origin of all the endocrine tissue. The 18-day explants were collected every other day of culture from 2 to 30 days in a long-term experiment. A number of large islets with well granulated B cells was still present after 30 days of culture. The relative abundance of cell types at different stages was estimated as follows: 18-day fetal controls, A greater than B=4 greater than D; after 2 to 10 days in culture, B greater than A greater than or equal to D; after 18 to 30 days in culture, B greater than D greater than A greater than 4.


A unique property of fetal bovine serum: high levels of protein-glutathione mixed disulfides. Fetal bovine serum has been reported to delay or inhibit "spontaneous" neoplastic transformation in vitro as compared with all other sera tested. The present results indicate that fetal bovine serum is also unique in containing high levels of protein-glutathione mixed disulfides (3 to 7 microng glutathione as mixed disulfide per ml serum). The level of mixed disulfide appears to vary in accordance with the period of gestation of the fetal calves used to prepare the serum, decreasing below detectable levels (less than 0.2 microng per ml) with near-term fetal calves. Calf, adult bovine, fetal horse, and swine sera did not contain detectable levels of this type of mixed disulfide.


Associative control of the immune response to cell surface antigens. The immune response to cell surface antigens is regulated by the activity of helper and suppressor cells, probably of TH and possibly of TCS types. For this to operate, recognition must take place of two or more antigenic determinants, which may be carried on the same (intramoblecular help) or different (intermolecular, intrastructural help) molecules. Regulation of this type has been shown to operate in the response to the murine allo-antigens H-2K, H-2D, Thy-1, and H-minors.


Neutralization of Salmonella toxin-induced elongation of Chinese hamster ovary cells by cholera antitoxin. A partially purified preparation of the delayed skin permeability factor from Salmonella typhimurium caused Chinese hamster ovary cells to elongate. The elongation effect and the skin test activity were blocked by monospecific rabbit antisera against cholera toxin and against the B fragment of cholera toxin,


Effectiveness of parenteral and oral typhoid vaccination in mice challenged with a Salmonella typhi-Salmonella typhimurium hybrid. Live Salmonella typhi administered intraperitoneally, acetone-killed S. typhi administered intraperitoneally, and live S. typhi given orally, with their effectiveness decreasing in that order, protected Swiss white mice against death from challenge with a virulent Salmonella typhimurium hybrid expressing S. typhi antigens.


Immunological properties of ampicillin polymers. The immunologic effects of ampicillin polymers were studied in mice and rabbits. Polymerized ampicillin--in contrast to monomers--had a suppressive effect on the antibody response to penicilloylated proteins. Unfractionated ampicillin polymer, and high molecular weight fractions were found to have an immunogenic effect in mice, when tested by a haemolytic plaque assay. The immunogenic effect of variously sized polymers did not correspond to their antigenic activity as reflected by passive cutaneous anaphylaxis experiments.


Optimal antibody-receptor ratio in anaphylactic-sensitization. The antigen-induced release of histamine from guinea pig and monkey lung fragments sensitized in vitro with hyperimmune rabbit antibody and human reaginic serum, respectively, was studied. When the amounts of whole antiserum, gamma-globulin fraction, or purified antibody were increased over the optimum in passive anaphylactic sensitization, the subsequent histamine release was reduced in spite of the fact that proportionately greater amounts of antigen were added to the system. The observed inhibition by higher antibody concentrations might be due to a reduction in cell-stimulating capacity of the antigen-antibody-receptor complex caused by inappropriate ratios among any of these three components.


Detoxified hymenoptera venoms: preliminary studies of in vitro cytotoxicity and antigenicity of Apis mellifera 'venomoid'. The preparation of detoxified venom (venomoid) of Apis mellifera is described. The venomoid obtained by formaldehyde detoxification retained significant antigenicity. Radioallergosorbent test (RAST) inhibition studies indicated a loss of the affinity of the venomoid for the venom-specific human IgE antibody. In rabbits the venomoid was capable of producing precipitating antibodies which cross-reacted with the original venom. Cytotoxic activity of venomoid as measured by nonimmunological histamine release and hemolytic activity with human cells was not detectable in vitro. Further studies seem warranted for the establishment of a venomoid as a diagnostic and/or therapeutic agent in the management of the hypersensitivity reactions to venom.


C3 and C5-cleaving properdin enzymes formed on zymosan incubated with human serum: the decay and the regeneration of the enzymes. Incubation os zymosan (Z) with normal human serum led to the formation of ZXhu, which had the abilities to cleave C3 and C5. Kinetic studies on ZXhu formation revealed that the amount of inactivated C5 by ZXhu, when expressed in site-forming unit (SFU), was much greater than that of inactivated C3. When ZXhu having limited C3 and C5-cleaving was incubated at 37 degrees C it decayed with a first order reaction completely in 120 min. At any stage of decay of ZXhu, the activities could be restored by the addition of both B and D, but not by the addition of B or D alone. Not D but B could be bound to the completely decayed ZXhu, and activation of bound B by D led to the regeneration of ZXhu.


Characterization of the in vitro cytotoxic assay of immune lymphoid cells in experimental allergic encephalomyelitis (EAE). Complement, antilymphocyte globulin, antiencephalitogenic antigen serum, actinomycin D and mitomycin C inhibited the cytotoxicty against syngenic fibroblasts of spleen cells more than lymph node cells. The results strongly suggest that the immune cell populations in EAE in the spleen andlymph nodes are qualitatively and/or quantitatively different. Cytotoxicity was dependent upon the number of immnue cells in the test. There was evidence of antigenic competition between encephalitogenic antigen and Myocobacterium tuberculosis in the induction of EAE.


Failure of the putative IgE pentapeptide to compete with IgE for receptors on basophils and masts cells. We have examined the claim that a pentapeptide resembling a portion of epsilon-chain of IgE can inhibit allergic reactions by competing with IgE for basophil-mast cell IgE receptors. Four laboratories with expertise in different areas collaborated. The following was found. (1) There was no evidence, by autoradiography, that the pentapeptide could block IgE fixation to basophils or displace IgE from these cells. (2) There was no evidence, by histamine release studies that the pentapeptide could block basophil sensitization or remove IgE from actively sensitized cells. The pentapeptide, also, did not impair antigen-induced histamine release. (3) There was no evidence that the pentapeptide could block the skin test response in Prausnitz-Küstner testing with the antigen injected intradermally or orally administered. Neither the correct pentapeptide sequence from the epsilon-chain nor the Fc" fragment of IgE impaired the P-K test. In each series of experiments, IgE myeloma protein, as a positive control, blocked the allergic response. The failure of the pentapeptide to block these reactions was observed at molar ratios of pentapeptide/IgE of 10(6) to 10(7). Our results, therefore, provide no evidence for competition between the pentapeptide and IgE. Even if there should be some weak affinity between the pentapeptide and IgE receptors on basophils or mast cells this would, by virtue of quantitative considerations, be of no clinical relevance.


Comparison of human stratum corneum, callus and psoriatic scales by means of serological methods. Homogenated stratum coreum, callus and psoriatic scales were extracted with (1) phenol water (PW) and (2) the combined use of trypsin digestion and phenol water extraction (TPW). The serological properties of the various preparations obtained were compared, using indirect haemagglutination, absorption and inhibition tests. The PW and TPW water phases contained two different antigens which were common to all three tissues. In addition, the periodate-treated TPW water phase of stratum corneum contained an erythrocyte-sensitizing antigenic determinant. This, however, cross-reacted with the untreated and periodate-treated preparation of psoriatic scales, whereas callus lacked the determinant present after treatment with periodate. Apparently callus and psoriatic scales lacked some components present in stratum corneum, but determinants specific for callus or psoriatic scales were not detected.


Failure of increased intracranial pressure to affect rapid axonal transport at the optic nerve head. In owl monkeys, elevation of intracranial pressure to 500 mm. saline for 4 to 7 hours failed to cause an accumulation at the optic nerve head of protein carried by rapid axonal transport. This suggests that the block of rapid transport observed by others during papilledema may be the result of axon swelling, not its cause. Alternatively, more than 8 hours may be required for intracranial pressure to show an effect, only the slow transport may be affected initially, or other factors than simple hydrostatic pressure may be operative in papilledema. In addition, the elevated intracranial pressure did not prevent the block of axonal transport at the lamina cribrosa produced by elevated intraocular pressure, even though the elevated intracranial pressure reduced the pressure gradient to which the axon is subjected as it crosses the lamina cribrosa. Perhaps the block produced by intraocular pressure is not due to a simple mechanical or hydrostatic mechanism.


Orthograde and retrograde axoplasmic transport during acute ocular hypertension in the monkey. Orthograde and retrograde axoplasmic transport have been studied in the optic nerve heads of 37 Macaca fascicularis eyes with normal or elevated intraocular pressure (IOP) produced by cannulation of the anterior chamber. Orthograde transport was labeled by 3H-amino acids injected intravitreally and incorporated into retinal ganglion cell proteins. Retrograde transport was studied in the same eyes by injecting horseradish peroxidase (HRP) into one or both optic tracts and dorsal lateral geniculate nuclei (dLGN). Both tracers accumulated in the lamina scleralis (LS) of eyes maintained at pressures of 25 to 150 mm. Hg for 12 to 28 hours (pressure in normal controls = 10 to 14 mm. Hg) but the HRP technique was markedly more sensitive. The degree of retrograde transport obstruction in the LS appeared to be directly proportional to both the height and the duration of elevated IOP. In one experiment, the blockades of orthograde and retrograde transport induced at 50 mm. Hg were demonstrated to be reversible. Serial reconstructions of radioautographs and peroxidase-reacted sections of the optic nerve heads demonstrated that the orthograde and retrograde transport obstructions were coincidental anatomically by light microscopy in the LS and occurred most prominently in the temporal quadrants of the nerve head. These transport obstructions occurred at moderate elevations of IOP (25 TO 50 mm. Hg) despite (1) elevated arterial PO2 levels during inhalation of 100 percent oxygen and (2) intact nerve head capillary circulation, as demonstrated by perfusion with nucleated avian erythrocytes.


An animal model for the growth of human tumor cell lines. Newborn rats were thymectomized at less than 24 hr of age and received antilymphocyte serum and heterologous human tumor cells. This model allows the growth of large primary tumors with a high metastatic percentage. This model may be applicable to the study of a wide variety of genitourinary tumors.


Bladder wall tension in benign prostatic hypertrophy. Two hundred forty-three synchronous urinary flow and intravesical pressure recordings were carried out on 51 men all of whom were over 50 years of age. They consisted of 12 normal subjects and 39 patients with benign prostatic hypertrophy in various stages of bladder outlet obstruction. Bladder wall tension was determined from each pressure and flow recording. Bladder wall tension at the commencement of micturition was found to be of value for the diagnosis of bladder outlet obstruction attributable to benign prostatic hypertrophy. A technique for direct recording of bladder wall tension is desirable.


An immunofluorescence technique with counterstain on fixed cells for the detection of antibodies to human herpesviruses; antibody patterns in patients with Hodgkin's disease and nasopharyngeal carcinoma. An indirect immunofluorescence (IF) test on fixed cells with Evans' blue counterstain is described for all four human herpesviruses, i.e., herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2), varicella-zoster virus (VZV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Comparison with immunodiffusion (ID) for HSV-2 and with ID and complement fixation (CF) for VZV and CMV demonstrated the specificity and high sensitivity of the IF test. Also introduced is a modification of the anti-complement immunofluorescence (ACIF) test for EBV-determined nuclear antigen (EBNA), permitting simultaneous titration of antibodies to this nuclear antigen and of the anti-nuclear factor (ANF). Seroepidemiological studies of these viruses in patients with Hodgkin's disease (HD) in the Netherlands revealed the following pattern: (1) in nodular sclerosing (NS) HD there is a 4-fold (significant) elevation in antibody titer to EBV-VAC, but no elevation to EBV-EA and EBNA; (2) in mixed cellularity (MC) HD a 10-fold (significant) elevation to both EBV-VCA and EA, but no elevation to EBNA is found compared to the control groups. These patterns in NS and MC HD are different from the pattern in nasopharyngeal carcinoma (NPC) which manifests elevations in antibody titers to EBV-VCA and EA as well as to EBNA. Antibody titers to HSV, VZV and CMV are not significantly elevated in either HD or NPC.


Mixed infection with two tobamoviruses: the formation of particles containing the coat protein messenger RNAs of either virus. Plants mixedly infected with the U2 strain of tobacco mosaic virus (T2MV) and sunnhemp mosaic virus (SHMV) and grown at 35 degrees, yield particles of the same modal lengths (300 and 40 nm) as those found in plants singly infected with SHMV, but not in plants infected with T2MV, which yield only the long particles. At least some of the particles produced in mixedly infected plants contain coat proteins of both viruses. When RNAs from these particles are translated in vitro the coat proteins of both viruses are produced, although when a mixture of RNAs from particles of SHMV and T2MV, grown separately, are translated in vitro only SHMV protein is produced. These and other results suggest that the short particles produced in mixedly infected plants contain both coat protein messengers.


Characterization of two new serotypes of San Miguel sea lion virus. Two new virus isolates, one from a California sea lion (Zalophus californianus californianus) and the other from a northern fur seal (Callorhinus ursinus) were partially characterized. Their physicochemical characteristics were similar to those of vesicular exanthema of swine virus (VESV) and San Miguel sea lion virus (SMSV). The virion morphology was in both instances typically calicivirus. On the basis of this and the serum cross-neutralization testing, these isolates were classed as two new types of SMSV and were designated serotypes SMSV-4 and SMSV-5.


Biological and immunological studies on two substrains, c-1 and c-3, derived from the Nakayama-NIH strain of Japanese encephalitis virus. A comparative analysis was performed on two substrains, c-1 and c-3, derived from the Nakayama-NIH strain of Japanese encephalitis virus. The infectivity of the two substrains was inactivated at similar rates by treatment with Tween 80 and Tween 20 but not by treatment with Tween 60. However, hemagglutinating activity as measured at pH 6.8 of the c-3 substrain treated with Tween 80 was more labile than that of the c-1 substrain treated with Tween 80. Also, the immunogenicity (neutralizing and hemagglutination-inhibiting antibodies) was different between the two substrains in sensitivity to low pH (6.8) and to Tween 80. With the c-1 substrain a combination of these treatments reduced the immunogenicity, but each treatment applied singly did not materially reduce it. The immunogenicity of the c-3 substrain was reduced by single as well as combined applications of the two treatments. Evidence was obtained which suggested that the part of the antigen sensitive to these treatments was strain-specific, whereas the resistant antigen was common among the Japanese encephalitis-St. Louis encephalitis complex members.


Isolation and assay of rabies serogroup viruses in CER cells. Infection of CER cell cultures with field strains of rabies virus, ranging from 0 to 5 mouse brain passages, was detected by immunoflurescence within 2-4 days after infection. A fluorescent focus assay for measuring infectivity of seven rabies serogroup viruses was rapid and reproducible. Rabies field strians and other rabies serogroup viruses also induced cytopathic effect, usually on initial passage. The hemadsorption-negative (HAD-) plaque test in BSC-1 cells was successfully applied to laboratory-adapted rabies strains. HAD- test attempts were unsuccessful with CER cells and with field isolates of rabies virus in both cell lines. CER cells are heteroploid and are antigenically related to BHK-21 cells by fluorsecent antibody tests.


Physicochemical characterization of two serologically unrelated equine rhinoviruses. The physicochemical properties of two serologically distinct equine rhinoviruses have been examined. Each virus sedimented at approximately 160S but co-centrifugation of the two viruses in a sucrose gradient revealed a small difference in their sedimentation coefficients. The two viruses also have different buoyant densities in cesium chloride. The equine rhinovirus type 1 equilibrated as a sharp peak at 1.45 g/ml whereas the type 2 virus equilibrated as a heterogeneous band with a peak at 1.44 g/ml but ranging in density from 1.41 to 1.45 g/ml. The relative sedimentation coefficients of the two virus RNAs were 35S for rhinovirus 1 and 37S for rhinovirus 2. A limited number of base composition analyses also showed differences between the two virus RNAs. The polypeptide profile of each serotype in polyacrylamide gels was generally similar to those of other picornaviruses but the two serotypes could be distinguished readily from each other.


Specificity of the Tumor-specific transplantation antigen induced by JC virus, a human polyomavirus. The specificity of the tumor-specific transplantation antigen (TSTA) induced by JC virus was investigated by cross protection tests in weanling hamsters. Hamsters immunized with JC virus, BK virus, or SV40 were challenged 5 weeks later with known numbers of JC virus- or SV40-induced hamster tumor cells. Both the JC virus-immune and the SV40-immune hamsters showed resistance to challenge with homologous but not heterologous tumor cells, and the BK virus-immune hamsters were not resistant to either heterologous tumor cell. The TSTA induced by JC virus did not cross-react with that induced by SV40.


Homogeneity of lipopolysaccharides from Acholeplasma. Five methods were employed to determine the heterogeneity or homogeneity of lipopolysaccharides from four acholeplasmal species, Acholeplasma axanthum, A. granularum, A. laidlawii, and A. modicum. A axanthum lipopolysaccharide behaved as a single component in all tests. A. granularum exhibited two components of identical composition and antigenic specificity. A. modicum lipopolysaccharide behaved as three components in two tests, but all three were similar in composition and identical serologically. The separable components of lipopolysaccharides from A. granularum and A. modicum probably represent size differences only. A. laidlwii lipopolysaccharide contained two distinct components by all methods. One was identified as the previously reported amino sugar polymer, whereas the other was a lipopolysaccharide containing both neutral and amino sugars.


Regulation of ribonucleoside diphosphate reductase synthesis in Escherichia coli: increased enzyme synthesis as a result of inhibition of deoxyribonucleic acid synthesis. Inhibition of deoxyribonucleic acid (DNA) synthesis in Escherichia coli by chemical inhibitors or by shifting cultures of temperature-sensitive elongation (dnaE and dnaB) or initiation (dnaA) mutants to nonpermissive conditions led to greatly increased synthesis of the enzyme ribonucleoside diphosphate reductase, which catalyzes the first reaction unique to the pathway leading to DNA replication. In contrast to the Gudas and Pardee proposed model for control of the synthesis of DNA repair enzymes, in which both DNA inhibition and DNA degradation are involved, DNA synthesis inhibition in recA, recB, recC, or lex strains results in increased synthesis of ribonucleotide reductase, which suggests that DNA degradation is not required. We propose that inhibition of DNA synthesis causes a cell to accumulate an unknown compound that stimulates the initiation of a new round of DNA replication, and that this same signal is used to induce ribonucleotide reductase synthesis.


Inhibition of germ tube formation in Neurospora. Phenethyl alcohol, m-cresol, and related compounds cause inhibition of germ tube formation in conidia of Neurospora crassa. Conidia continue to swell and form large spherical cells that are capable of multiple germ tube formation upon removal of inhibitor.


Isolation of chicken hemoglobin mRNA and synthesis of complementary DNA. Chicken globin mRNA has been purified and partially characterized. Globin-specific sequences are found primarily as 9 S RNA, but also are found with ribosomal RNA, preferentially the 28 S moiety. The chicken globin mRNA preparation has been translated in the wheat germ and Krebs ascites cell-free systems. The products have been identified by sodium dodecyl sulfate-gel electrophoresis as the alpha- and beta-globin polypeptides. The globin mRNA is resolved into two asymmetric peaks by polyacrylamide gel electrophoresis in 98% formamide. The minor rapidly migrating peak consists primarily of alpha message while the major slowly migrating peak contains a mixture of alpha and beta messages. The synthesis of cDNA has been optimized and the products analyzed by polyacrylamide gel electrophoresis in 98% formamide. The products consist primarily of full copy transcripts that can be resolved into three discrete species.


Genetic regulation of UDP-glucuronosyltransferase induction by polycyclic aromatic compounds in mice. Co-segregation with aryl hydrocarbon (benzo(alpha)pyrene) hydroxylase induction. Induction of hepatic 4-methylumbelliferone UDP-glucuronosyltransferase (EC 2.4.1.17) by polycyclic aromatic compounds, such as 3-methylcholanthrene or beta-naphthoflavone, occurs in C57BL/6N, A/J, PL/J, C3HeB/FeJ, and BALB/cJ but not in DBA/2N, AU/SsJ, AKR/J, or RF/J inbred strains of mice. This pattern of five responsive and five nonresponsive mouse strains parallels that of the Ah locus, which controls the induction of aryl hydrocarbon (benzo[alpha]pyrene) hydroxylase (EC 1.14.14.2). Induction of the transferase is maximal in C57BL/6N mice with 200 mg of 3-methylcholanthrene/kg body weight; no induction occurs in nonresponsive DBA/2N mice even at a dose of 400 mg/kg. The rise of inducible transferase activity lags 1 or more days behind the rise of inducible hydroxylase activity and peaks 5 days after a single dose of 3-methylcholanthrene. In offspring from the appropriate backcrosses and intercross between C57BL/6N and DBA/2N parent strains, the genetic expression of 3-methylcholanthrene-inducible transferase activity is inherited as an additive (co-dominant) trait. This expression differs distinctly from that of the inducible hydroxylase activity, which is inherited almost exclusively as a single autosomal dominant trait in these same animals. The more potent inducer 2,3,7,8-tetrachlorodibenzo-p-dioxin induces the transferase more than 3-fold in C57BL/6N mice and less than 2-fold in DBA/2N mice, whereas the hydroxylase is induced equally (about 8-fold) in both strains. A dose of 3-methylcholanthrene given 3 days after 2,3,7,8-tetrachlorodibenzo-p-dioxin, at a time when hydroxylase induction in both strains is very high, does not enhance the rise in inducible transferase activity seen in C57BL/6N or DBA/2N mice which have received 2,3,7,8-tetrachlorodibenzo-p-dioxin alone. These data indicate that (a) the inducibility of two metabolically coordinated membrane-bound enzyme activities may be regulated by a single genetic locus, and (b) although the hydroxylase can be fully induced in the nonresponsive DBA/2N strain by 2,3,7,8-tetrachlorodibenzo-p-dioxin prior to 3-methylcholanthrene treatment, metabolites of the 3-methylcholanthrene treatment, metabolites of the 3-methylcholanthrene treatment, metabolites of the 3-methylcholanthrene, presumably present in the liver, are incapable of inducing further the transferase activity. The difference in sensitivity between 3-methylcholanthrene and the more potent inducer 2,3,7,8-tetrachlorodibenzo-p-dioxin for both the hydroxylase and the transferase activities suggests the possibility of a common receptor in regulating both enzyme induction processes.


The restoration of the articular surfaces overlying Replamineform porous biomaterials. Replamineform porous implants (4 mm X 4 mm diameter) were placed into full-thickness cartilage and bone defects of the weight-bearing surface of the lateral femoral condyles of adult male white rabbits. These were analyzed at 1 day, 1 week, 6 weeks, 3 months, and 6 months for 1) ingrowth of tissue within the implants and 2) restoration of the articular surface overlying them. Appropriate unfilled, but similar, control defects were also studied. Mineralized bone was seen within the substance of both the TiO2 and hydroxyapatite implants at 1 week; this extremely rapid response was present in every specimen studied and was not seen with alphaAl2O3 or control animals. With the passage of time, maturation of this bone ingrowth occurred so that by 3 months, they were all incorporated into the surrounding bone. Only the hydroxyapatite implants showed consistent regenerative healing of hyaline articular cartilage from the margins of the defects with the passage of time; this occurred whenever the subchondral bone adjacent to the defect proliferated in a "creeping" fashion over the articular aspect of the implant, and the undamaged cartilage then followed it. Fibrocartilage, and not hyaline cartilage, formed the articular surface over the TiO2 and alphaAl2O3 implants and in the controls.


Supramolecular structure of polymorphic collagen fibrils. Reconstituted cartilage collagen fibrils with an oblique banding pattern or with two types of symmetrical patterns, and reconstituted rattail tendon fibrils with a third type of symmetrical pattern were examined by electron microscopy and found to consist of narrow subfibrils having native-type cross-striations. Analysis of the four types of patterns by a graphic method of specific band matching revealed the orientation and axial relation of individual subfibrils and their component molecules. In fibrils with an oblique pattern, subfibrils have the same orientation and a regular 100A axial displacement. Observations on staining characteristics, folded fibrils, and transverse sections of embedded fibrils suggest that the obliquely banded fibrils are ribbonlike or layered structures. In the three types of fibrils with a symmetrical pattern, adjacent subfibrils are oppositely oriented and aligned within a 119-A segment of the 670-A major period. Considered together, the observations suggest that interaction sites on the surface of subfibrils (and perhaps on the surface of native collagen fibrils) occur in various patterns that are manifested accouding to the nature of the environment during fibril formation, and that such patterns can be mapped on the surface of subfibrils by noting the arrangement of subfibrils in polymorphic forms.


Defects in the cartilaginous growth plates of brachymorphic mice. Homozygous brachymorphic (bm/bm) mice are characterized by disproportionately short stature. Newborn bm/bm epiphyseal cartilages are shorter than normal although the cells in the different zones of growth are relatively well organized. The extracellular matrix reacts poorly with stains specific for sulfated glycosaminoglycans. The ultrastructural appearance of the cartilage matrix indicates normal collagen fibrils; however, proteoglycan aggregate granules are smaller than normal and are present in reduced numbers, particularly in the columnar and hypertrophic zones of the growth plate. In addition, a prominent network of fine filaments, which are extractable in 4 M guanidine hydrochloride, are present in the bm/bm cartilage matrix. These findings suggest that a defect affecting the proteoglycan component of cartilage occurs in bm/bm mice.


Patterns of DNA synthesis during pollen embryogenesis in henbane. Continued DNA synthesis in the generative cell nucleus, followed by mitosis and cytokinesis, results in the formation of pollen embryoids in cultured anthers of H. niger. In contrast, the nucleus of the vegetative cell undergoes no DNA synthesis after it is cut off, or synthesizes DNA only during a limited number of cell cycles. DNA synthetic patterns in the generative and vegetative cell nuclei confirm the ontogeny of embryoids described in this plant.


Microfluorometric analysis of cellular DNA following incorporation of bromodeoxyuridine. Bromodeoxyuridine (BrdU) incorporation into cellular DNA has a differential effect on the cell-associated fluorescence of several DNA-specific dyes. After cells were treated with BrdU, flow microfluorometry was used to study the relative increase or decrease influorescence of stained cells. Bromodeoxyuridine incorporation into CHO cells increased the fluorescence of mithramycin-, olivomycin-, or chromomycin-stained cells, decreased that of propidium iodide-stained cells, and had little, if any, effect on the fluorescence of acriflavine Feulgen-stained cells. Changes in relative fluorescence of cell associated dyes are due to changes in the amounts of dye bound to cells with BrdU-substituted DNA. Colorimetric and absorbance measurement of DNA content showed that BrdU does not alter the diploid DNA content of CHO cells; however, BrdU induces perturbations in the distribution of cells about the cell cycle which cause an increase in average DNA content.


Rapid purification of an RNA tumor virus and proteins by high-performance steric exclusion chromatography on porous glass bead columns. High-performance steric exclusion chromatography on a 1250-A pore size polyethylene glycol-treated glass bead column was used to purify avian myeloblastosis virus and hamster melanoma virus from plasma protein and tissue culture media. The purified hamster melanoma virus was still infectious and the avian myeloblastosis virus-associated RNA-directed DNA polymerase showed a 1100-fold purification of the virus from one column treatment. Electron microscopy of the purified virus showed intact particles, with surface projections evident. The time required for column purification of the virus was 5 min.


[The quantitative detection of estrogens and antithyroid drugs by thin-layer and high performance thin-layer chromatography in animal tissue (author's transl)]. Methods of thin-layer chromatography (TLC) and high-performance thin-layer chromatography (HPTLC) were developed for the determination of estrogens and antithyroid drugs in extracts of animal tissues. TLC proved to be suitable for quantities in the range of 200-2000 ng, with a detection limit of 50-200 ng, HPTLC in the range of 10-200 ng, showing calibration curves of good linearity even in extracts. By HPTLC better detection limits, better separation and faster ascending could be achieved than by TLC.


Gram stain evaluation of the quality of sputum specimens for mycobacterial culture. A group of 34 mycobacteria, consisting of 25 Mycobacterium tuberculosis and nine strains of three other species, was isolated from 400 expectorated sputum specimens submitted on 148 patients from county-wide sources. Eight strains (24% of the total) were isolated from specimens evaluated by Gram stain to be oropharyngeal fluids. The remaining 26 strains were isolated from ungradable specimens and those primarily of lower respiratory origin. It was concluded that the random examination of sputum by Gram stain to determine the specimen's quality for mycobacterial isolation is not necessary.


Counterimmunoelectrophoresis in the diagnosis of bacterial meningitis. The aim of the present study was to investigate whether counterimmunoelectrophoresis (CIE) would facilitate the rapid, etiological diagnosis of bacterial meningitis when used in parallel with other routine methods in a medical bacteriological laboratory. Of 3,674 consecutive specimens of cerebrospinal fluid (CSF) received at the Department of Diagnostic Bacteriology, Statens Seruminstitu, 283 specimens (each representing one patient) were selected for examination by CIE on the basis of the following criteria: bacteria or pleocytosis or both by microscopy or positive culture or both. CIE was performed with antisera to Neisseria meningitidis (groups A, B and C), Streptococcus pneumoniae (omni-serum and pools A to 1), and Haemophilus influenzae type b. Antigen was detected in 57% (72/126) of specimens in which cultures revealed these three kinds of microorganisms in CSF and in 12% (17/139) of the culture-negative specimens. CSF specimens from 21 patients with bacterial meningitis caused by other species were all negative in CIE, except four, three of which contained Escherichia coli antigen reacting with antiserum to N. meningitidis group B and one E. coli antigen reacting with antiserum to H. influenzae type b. Specific diagnosis was achieved in 60% (170/283) of the specimens studied and could be extablished within 1 h in 85% (145/170) by the combined results of microscopy and CIE. Ten specimens, nine of which showed a reaction with antiserum to N. meningitidis group A, were positive by CIE only.


Sucrose-negative variants of Candida tropicalis. Four cultures of a Candida sp. that lacked alpha-glucosidase activity were isolated from clinical specimens. Physiological, morphological, and serological characterizations of the yeasts and deoxyribonucleic acid reassociation studies supported their classification as a variant of C. tropicalis.


Ultrasonic evaluation of radiation therapy ports. Radiation therapy is useful in the palliative treatment of large, unresectable abdominal and pelvic malignancies. The purpose of this study is to evaluate the effectiveness of B mode ultrasound scanning as an adjunct in radiation treatment planning of these patients. Fifty-one patients with large abdominal or pelvic masses had radiation therapy ports established using findings from surgery, radiographs, and physical examination. All the patients then had B mode ultrasound examinations. The position of the therapy ports was marked on the echogram. Thirty-one of the 51 patients studied (61%) had evidence of extension of tumor beyond the therapy ports. This was often due to "iceberging", i.e. when the deep portion of the tumor was considerably larger than the palpable margins of the superficial portion. B mode ultrasound scanning is a valuable adjunct in planning palliative radiation therapy of patients with large, unresectable, abdominal and pelvic malignancies.


A comparison of the erythrocyte sedimentation rate and plasma viscosity in detecting changes in plasma proteins. The Westergren erythrocyte sedimentation rate (ESR) and plasma viscosity were compared in 114 patients and their correlations with total and differential plasma protein fractions were analysed. There is a linear correlation between these two screening tests. Higher correlation coefficients were obtained between the plasma viscosity and fibrinogen and alpha and gamma globulins than with the ESR. Albumin affected the two tests in opposite directions. The ESR was falsely increased by a fall in haemoglobulin even within two standard deviations from the mean. Both tests gave an appreciable number of incorrect values--the plasma viscosity in 21 cases and the ESR in 33. The cause for these is discussed. It is concluded that the plasma viscosity is the more sensitive and reliable measure of changes in acute phase protein reactants and more useful for monitoring clinical progress.


Cyclic AMP in dental and periodontal tissues during tooth eruption in kittens. Cyclic adenosine 3',5'-monophosphate (cAMP) concentrations and cellular distribution were studied in dental and periodontal tissues during tooth eruption in kittens. Although the mean levels of cAMP around developing teeth were similar in all the tissue samples, there were marked differences in cAMP stainability of tissues apical and occlusal to the erupting teeth.


Self-instruction in dentistry: a critique. Recent activity and interest in self-instruction and programmed self-instruction emerged from studies which demonstrated the central importance of learner response with prompt consequences. Self-instructional procedures in formal education, e.g., teaching machines, were initially designed to be an improvement over conventional show-and-tell instruction in the arrangement of systematic student response, prompt consequences, and student self-pacing. A minority of self-instructional procedures in dentistry reflect that intent. The evidence discussed suggests that dental education may be the worse for that omission. Suggestions for the improvement of self-instructional in dentistry are presented.


A comparison of the analgesic effect of oxymorphone by rectal suppository and intramuscular injection in patients with postoperative pain. The relative analgesic potency of oxymorphone by rectal suppository and intramuscular injection was evaluated in a double-blind, twin-crossover comparison of graded single doses in 136 patients with postoperative pain. The time-effect curves of the two routes of administration differed substantially; rectal resulted in lower and more delayed peak analgesia and a longer duration of action than intramuscular administration. When both duration and intensity of analgesia are considered (total effect), rectal oxymorphone was 1/10 as potent as the intramuscular form; in peak effect, it was only 1/16 to 1/20 as potent. However, because intramuscular oxymorphone is nine to ten times as potent as intramuscular morphine, 5 to 10 mg oxymorphone by suppository provides analgesia comparable to that provided by the usually used doses of parenteral narcotics. Rectal oxymorphone produced no more, and perhaps somewhat fewer, side effects than doses of intramuscular oxymorphone producing equivalent total analgesic effect. None of the patients objected to the rectal route of analgesic administration. This study demonstrates the feasibility of well-controlled analgesic assays employing the double-dummy technique to compare suppositoreis with oral or parenteral analgesic dosage forms. Our observations also suggest that the rectal route is an acceptable and practical way of administering potent analgesics and is probably being underutilized by physicians in the control of moderate to severe pain.


Rapid chemical dehydration of biologic samples for scanning electron microscopy using 2,2-dimethoxypropane. Acidified 2,2-dimethoxypropane (DMP) has been used as a rapid chemical dehydrating agent en route to plastic embedding and ulthrathin sectioning for conventional electron miscroscopy (J Histochem Cytochem 23:107, 1975). We have used DMP to dehydrate biologic specimens prior to critical point drying and metal coating for scanning electron microscopy. There is no difference in either the gross architecture or the fine surface structure of mouse small intestine and trachea, rat trachea and kidney, and cultured fibroblasts, between samples dehydrated in DMP for 5 min to 30 days and those conventionally dehydrated in ethanol or acetone. DMP dehydration is advantageous in speed, economy and apparent completeness.


Scanning Feulgen-deoxyribonucleic acid cytophotometry of Papanicolaou destained preparations. Feulgen deoxyribonucleic acid cytophotometry of Papanicolaou destained specimens revealed a differential loss in Feulgen reactivity among human buccal and cervical smears, cultured embryonic lung fibroblasts and invasive cervical carcinoma cells. Loss in Feulgen reactivity in Papanicolaou destained fibroblasts and polyploid nuclei of malignant lesions was observed to result in underestimates of relative Feulgen deoxyribonucleic acid and nuclear area values using scanning integrating microdensitometry. Thus, Papanicolaou stained preparations may not be suitable for deoxyribonucleic acid quantification of high ploidy lesions since distributional absorption error is unpredictably influenced by such factors as ploidy level, nuclear size, chromatin dispersion and differential aldehyde loss during destaining. Feulgen deoxyribonucleic acid cytophotometry of Papanicolaou stained preparations can be useful for differentiating benign from malignant lesions if extent of aneuploidy (as reflected in abnormal deoxyribonucleic acid frequency distribution profile) is used as a diagnostic indicator.


Experimental infection with Ancylostoma caninum larvae in mice. IV. Serum protein pattern during primary infection. Serum protein of Swiss albino mice infected with Ancylostoma caninum larvae, were analysed electrophoretically. Twenty mice were infected with a dose of 1000 larvae and the alterations in serum protein during an infection period of 30 days were recorded and compared with uninfected controls. The result showed a significant decrease in albumin and gamma globulin with an increase in beta globulin. There were no significant changes in the alpha-1 and alpha-2 globulins. The total globulins increased and A/G ratio decreased. These changes appeared to be remarkable on the 9th day after infection.


Tumor growth and antibodies after RSV-challenge in normal chickens and in chickens congenitally infected with avian leukosis virus. Normal chickens and chickens congenitally infected with an avian leukosis virus (ALV) of antigenic subgroup A were challenged with strains of Rous sarcoma virus (RSV) of two different antigenic subgroups (B and C) and tumor induction and growth as well as humoral antibody to viral envelope antigen (VEA) and tumor-specific surface antigen (TSSA) were measured. There was no effect of congenital ALV infection on RSV tumor incidence or latent period but the growth rate and size of the tumors were much higher in congenitally infected birds as compared to controls. Whereas most tumors in the RSV-challenged normal birds regressed, tumors in ALV-infected birds grew progressively. There were no striking differences in the number of birds in either group in the incidence of anti-TSSA or anti-VEA antibodies nor did the presence of either type of antibody reflect the tumor status of the host.


Systemic expression of cutaneous basophil hypersensitivity. Guinea pigs primed for cutaneous basophil hypersensitivity (CBH) with several soluble proteins or with sheep erythrocytes developed a systemic, delayed-onset, maculopapular rash when challenged parenterally with specific antigen. The rash was most readily induced 5 to 7 days after immunization, at a time when local CBH skin test reactivity was also optimal. Miscroscopically, the rash resembled local CBH skin test reactions, being comprised of a papillary dermal infiltrate of basophils and lymphocytes and a striking dilatation and compaction of superficial venules. In addition to the systemic rash, animals expressing systemic CBH (SCBH) exhibited a striking eosinophilia at 24 hr which gave way to basophilia at 48 hr. Focal collections of eosinophils, and of smaller numbers of basophils, were found in the lungs and spleen; both eosinophils and basophils infiltrated the medulla of the thymus. Thus, basophil-rich infiltrations are favored in the skin even after systemic challenge with antigen and occur only to a much smaller extent in other organs where eosinophils may predominate. These differences in the response of various organs to challenge with parenteral antigen suggest that as yet unidentified local factors play a determinative role in regulating the inflammatory response. The pathogenesis of SCBH is not yet established, but it shares many of the properties of local CBH: histology, carrier specificity, development early after sensitization in the absence of detectable antibodies. Passive transfer has not been accomplished with serum alone but has been achieved irregularly with cells plus serum. SCBH may serve as a useful model for several disease states in man characterized by a systemic rash and eosinophilia, including certain types of drug reaction.


Cross-reacting antigens in chemically induced sarcomas are fetal determinants. Serologic cross-reactivity has been demonstrated among three MCA-induced sarcomas in C57BL/6N female mice (MCA-2, MCA-3, and MCA-12) with a microcytotoxicity assay. Serum from mice bearing MCA-3 tumor was cytotoxic to both MCA-2 and MCA-3 tumor cells at a titer of 1:8. Sequential absorptions of this serum with syngeneic embryo cells completely eliminate cytotoxicity against MCA-2 cells without affecting the cytotoxic titer against MCA-3 cells. Serum hyperimmune to the MCA-3 tumor reacted with MCA-3, MCA-2, and MCA-12 tumors. Absorption of this serum with embryo cells eliminated cytotoxicity against MCA-2 and MCA-12 cells, but was incapable of lowering the titer against MCA-3 cells below 1:40. Similarly, serum hyperimmune to MCA-2 tumor was lytic to MCA-2, MCA-3, and MCA-12 before absorption, but was lytic only to MCA-2 cells after absorption with sygeneic embryo cells. Thus, the in vitro cross-reactivity between MCA-induced sarcomas is due to a common fetal antigen(s), which is distinct from the individual tumor-specific antigens of each tumor. Since these tumors do not exhibit cross-reactivity in in vivo challenge experiments, it appears that this fetal antigen is not responsible for in vivo immune protection.


Age-related changes in T cell function.

Age-related changes in T cell function. A comparison was made of the abilities of carrier (BGG)-primed T cell populations from young (4-month old), middle-aged (14- and 19-month old) and old (31- and 34-month old) mice to collaborate with hapten (DNP)-primed B cells from young mice in a cell-transfer system. The plaque-forming cell responses to 2,4-dinitrophenol (DNP) were measured by a modification of the Jerne plaque assay. The DNP-specific antibody-forming cell responses of old T cell/young B cell combinations were significantly lower than those of young T cell/young B cell combinations, both in the number of T cells needed for peak response and in the size of that response. These data indicate that the primed T cell populations of old mice are deficient by a factor of 6 in their ability to initiate B cell proliferation and differentiation into antibody-forming cells.


Murine plasma cells secreting more than one class of immunoglobulin heavy chain. II. SAMM 368--a plasmacytoma secreting IgG2b-kappa and IgA-kappa immunoglobulins which do not share idiotypic determinants. SAMM 368 is a BALB/c plasmacytoma which secretes IgG2b-kappa and IgA-kappa paraproteins. Immunofluorescence studies of ascites cells from the tumor with purified, heavy chain class-specific antiglobulins demonstrate that single cells contain both IgG2 and IgA heavy chains. Idiotypic antisera prepared in mice and rabbits indicate that the two paraproteins produced by the tumor do not share idiotypic determinants. Analysis of the purified paraproteins for allotype markers showed that SAMM 368 IgA bears the BALB/c A12,13,14 determinants. SAMM 368 IgG2b does not carry any detectable allotypic determinants in spite of the fact that heterologous antisera identify the paraprotein as IgG2b.


In vitro induction of polyclonal killer T cells with 2-mercaptoethanol and the essential role of macrophages in this process. Killer T cells against allogeneic and syngeneic tumor cells were generated in vitro by the addition of 2-mercaptoethanol (2-ME) to the murine spleen cell culture in the absence of any antigenic stimulation. The maximum activity of T cell-mediated cytotoxicity (CMC) induced with 2-ME was observed on day 4 of culture and the induction of CMC was completely inhibited by the addition of inhibitor of DNA synthesis, hydroxyurea, or cytosin arabinoside. CMC induced with 2-ME was specifically inhibited by the addition of unlabeled target cells to the 51Cr-release assay system. These results indicated that killer T cells were generated in the presence of 2-ME as a result of nonspecific polyclonal activation of precursors into cytotoxic effector cells and that they recognized target cells with antigen-specific recognition receptors. Spleen cells deprived of adherent cells showed impaired induction of CMC with 2-ME. The addition of peritoneal exudate macrophages to splenic T cells restored this response. The result indicated that macrophages were essential for the induction of CMC with 2-ME. The possibility that the function of macrophages was mediated by soluble factor(s) released from macrophages was demonstrated by the separate culture of splenic T cells and macrophages in double-chambered, Marbrook-type vessels and by the addition of supernatants from macrophage cultures to splenic T cells. 2-ME and soluble factor(s) released from macrophages seemed to be required for the activation of precursors into killer cells.


Histamine-induced suppressor factor (HSF): effect on migration inhibitory factor (MIF) production and proliferation. Histamine added in vitro to cultures of sensitized lymphocytes suppresses antigen-induced production of migration inhibitory factor (MIF) and proliferation by these cells. Recent studies have suggested that lymphocytes bearing histamine type-2 receptors play a regulatory role in these in vitro responses. The present studies were undertaken to determine if suppressor function by cells having histamine receptors was mediated through a soluble product. It was found that lymph node cells from nonimmune or immune strain 2 guinea pigs elaborate a nondialyzable factor into the culture supernatant when incubated with 10(-3) to 10(-5) M histamine (histamine-induced suppressor factor of HSF). HSF, when cocultured with sensitized lymphocytes, suppressed their MIF and proliferative responses to antigen. HSF was made by lymphocytes but not macrophages. Its production could be blocked by an H2 receptor antagonist (burimamide) but not an H1 receptor antagonist (chlorpheniramine). Furthermore, the inhibitory effect of HSF was reversible as lymphocytes washed free of the factor after 24 hr and recultured with fresh medium and antigen were able to produce MIF. Gel filtration by Sephadex G-100 chromatography indicated that HSF had an approximate m.w. of 23,000 to 40,000. These results suggest that the release of histamine at the sites of immediate hypersensitivity reactions, possibly by generating HSF activity, may play a regulatory role in the subsequent development of cellular-immune reactions at the same site.


Specificity of rat xenogeneic cell-mediated cytolysis for the products of the K and D loci of the mouse H-2 complex. Lewis rat lymph node (LN) cells, when cultured with irradiated mouse spleen cells, develop cytotoxic effectors that were shown to be specific for the sensitizing mouse strain. Alloantisera directed to the products of the K and D loci of the target inhibited cytolysis whereas an antiserum to the I region gene products did not inhibit cytolysis even though LPS-induced spleen blasts were used as targets. Thus, it appears that even when effector cells from another species are generated to mouse spleen cells, the gene products of the K and D loci continue to play a predominant role.


The class of surface immunoglobulin on virgin and memory B lymphocytes. The class of surface immunoglobulin receptors for antigen on B cell precursors of different classes of antibody-forming cells was determined by utilizing a technique of class-specific antigen suicide. Spleen cells are first treated with a class-specific antiserum under conditions that result in the stripping of that class from the cell surface. The cells are then permitted to bind a highly radioactive trinitrophenyl (TNP)-conjugated protein, which leads to lethal irradiation of all TNP-specific B cells except those whose TNP receptors had been removed by the class-specific stripping of surface immunoglobulin. In this way, the class of antibody-forming cells resulting from TNP stimulation of B cells with different classes of surface immunoglobulin can be examined. It was found that the virgin B cell precursors of IgM-producing cells are two types: cells bearing IgM receptors only and those bearing both IgM and IgD receptors. All virgin B cells that gave rise to IgG1 antibody-forming cells had both IgM and IgD on their surfaces, demonstrating that an antigen-dependent switch from IgM and IgD to IgG1 production is a common feature of B cell maturation. In contrast, memory B cell precursors of IgG1 antibody-forming cells had predominantly IgG1 as their surface antigen receptor. The implications of these findings on current models of B cell maturation are analyzed.


Variability of the lytic susceptibility of Neisseria gonorrhoeae to human sera. The bactericidal activity of human sera for Neisseria gonorhoeae was studied. Sera were obtained from a group of patients with gonococcal infections who had acute urethritis, acute pelvic inflammatory disease, disseminated gonococcal infection, or who were asymptomatic carriers. The homologous and heterologous strains were tested with these sera. The development of serum bactericidal antibodies was not a universal event. With few exceptions, the susceptibility of a particular strain to human antibody and complement appeared to be largely independent of the particular person from whom the serum was obtained and was due instead to antigenic properties intrinsic to each individual strain. Lipopolysaccharide appeared to be the predominant antigen against which bactericidal antibodies were directed. The principal bactericidal antibody class was IgM. Blocking antibodies were not found to account for the lack of lytic activity. A correlation of bactericidal antibodies with protection from developing gonococcal infection could not be demonstrated in three pateints.


Modulation of lipopolysaccharide (LPS)-mediated function by structural differences of two physically distinct fractions of Escherichia coli K235 LPS. Lipopolysaccharide (LPS), extracted from Escherichia coli K235 by the butanol water technique, was fractionated by gel filtration chromatography into high m.w. (LPS I) and low m.w. (LPS II) fractions. These two forms of LPS were characterized by different densities and chemical compositions. Chemical analysis provided evidence for greater amounts of lipid A and Lipd A-associated protein (LAP) per unit weight associated with LPS II. The biologic activity of the two LPS preparations was compared over a spectrum of different parameters. LPS II was shown to be a more potent mitogen and toxin than LPS I, whereas the two preparations were demonstrated to be of equal activity as polyclonal B cell activators, immunogens, and adjuvants. A modulatory role for the polysaccharide component of the LPS molecule is discussed.


Immunologic tolerance to HGG in mice. I. Suppression of the HGG response in normal mice with spleen cells or a spleen cell lysate from tolerant mice. Adoptive transfer of spleen cells or spleen cell lysates from mice tolerant to human-gamma-globulin (HGG) specifically suppressed the response of normal syngeneic recipients to HGG. The suppressive activity could be transferred for over 100 days after tolerance induction. The suppression induced by both spleen cells and spleen cell lysate was found to be specific as evidenced by a normal response to a challenge with turkey-gamma-globulin or goat erythrocytes. The activity of the suppressive lysate could be removed by passing the material through an HGG immunoadsorbent column but not by passing it through an anti-HGG column or a BSA column. These results indicated that the factor had antigen specificity and was probably not antigen-antibody complexes. That this suppression was not due to a shifting of the kinetics of the antibody response has also been demonstrated. The antigen-specific suppressor factor in the tolerant spleen cell lysates was a protein with a m.w. of approximately 45,000 daltons. The kinetics of the appearance of both suppressor cells and suppressor factor were consistent with a mechanism of active suppression functioning in the maintenance of tolerance to HGG.


Temperature dependence of antigen-specific rosette formation by lymphocytes from immunised mice. Spleen cell suspensions from mice undergoing a secondary response to sheep erythrocytes (SRBC) contained about one tenth as many specific antigen-binding, rosette-forming cells (RFC) when they had been washed at 37 degrees C instead of 4 degrees C before rosetting. This difference was correlated with the presence of IgG anti-SRBC antibody in the serum, and the 37 degrees C washings of immunised spleen cells could passively allergise non-immune spleen cells at 4 degrees C for specific rosette formation which was inhibitable by anti-mouse F(ab)2 serum. The RFC from actively immunised mice were lymphocytes and not macrophages by morphological and cytochemical criteria. It is suggested that the 37 degrees C-labile RFC are lymphocytes to which IgG antibodies bind in the cold. These data indicate that in the use of antigen-binding cell assays to monitor immunological responses, it is necessary to wash lymphocytes at 37 degrees C before testing.


Studies on H-2 specificities on mouse tumour cells by a new microradioassay. Seven mouse tumour cell lines of different aetiology and origin were tested for the expression of surface alloantigens using twenty-four well defined H-2 alloantisera and anti-Thy 1.2. We used a new radioassay that involves antibody-complement treatment of the tumour target cells followed by postlabeling the surviving tumour cells with 14C-thymidine. With a relatively high frequency the anti-H-2 sera were reacting differently with the tumour cells than with respective syngeneic lymphoid cells. Thirty-six anomalous reactions out of 129 investigated were detected. Absorbtion experiments performed with H-2 antigen positive or negative lymphoid cells revealed a striking similarity between these extra-specificities and H-2 specificities of foreign haplotypes. The results are discussed in relation to the biological importance of the extra-specificities, both with regard to origin (derepression) and function (transplantation antigen properties).


Molecular relationship between H-2 and ML antigens on leukaemia L-1210/v cells. The molecular relationship between H-2 antigens and mammary leukaemia (ML) antigen at the surface of leukaemic L-1210/v cells was studied by the 'lyso-strip' method. Molecules carrying the private specificities of the H-2d haplotype are independent entities in the membrane of the L-1210/v leukaemic cells and are not linked to molecules carrying the 'ML' antigen.


Experimental production of rabbit anti-guinea-pig epidermal cell sera. Comparison to pemphigus antibodies. Anti-epidermal cell sera (AES) were obtained by immunizing rabbits with enzymatically dispersed, viable guinea-pig epidermal cells followed by absorption with guinea-pig red blood cells, spleen and thymus cells, and liver powders. Complement-mediated cytotoxicity and immunofluorescence demonstrated that AES were towards cell surface antigen specific for stratified squamous epithelia of guinea pigs, and cross reacted with the corresponding tissues of humans and monkeys. Immunofluorescence revealed that AES reacted with Hassall's corpuscles and surrounding epithelial cells of the guinea-pig thympus which seemed to share common antigens with the epidermis; AES gave no reaction with other organs. While antigens reactive with pemphigus antibodies (PA) were demonstrated by membrane immunofluorescence to be present on the epidermal cell surface, PA showed no cytotoxicity to guinea-pig and human epidermal cells. Re-treatment of isolated epidermal cells with trypsin showed that antigens ractive with PA were more susceptible to the enzyme than those reactive with AES. These findings suggest that the cell surface antigens binding AES are different from the antigens which bind PA.


Developmental changes in the protein profile of cornifying lingual epithelium. Epithelia from the tongue dorsum of 14- to 21-day embryos, 21-day embryos, 3-week-old, and adult rats were separated from their connective tissues by incubation in balanced salt solution containing EDTA. Aliquots from total extracts of these tissues were electrophoresed on sodium dodecyl sulfate-polyacrylamide gels. Scans of gels stained with fast green (FG) revealed more than 20 peaks. Ten major peaks ranging from apparent molecular weights (MW) of 120,000 to 14,000 daltons comprised about 70% of the total protein on each gel. This report focuses primarily on two pairs of peaks, arbitrarily numbered 2 and 3 (MWs 71,000 and 69,000) and 9 and 10 (MWs 17,500 and 14,000). Peaks 9 and 10 predominated in the 15-day embryos where they comprised about 30% of the total protein. As development proceeded, there was a gradual shift in the protein profile in favor of peaks 2 and 3 until on the 20th day the relative amounts of these peaks reached a maximum and peaks 9 and 10 decreased in relative amounts. The protein profile on the 20th fetal day resembled that of the 3-week-old rats and the adults. The rise in the relative amounts of peaks 2 and 3 coincided with the morphologic appearance of large numbers of tonofilaments and the onset of cornification. When the gel was stained by a procedure specific for sulfhydryl groups, peaks 9 and 10 were especially reactive after the 18th day; plainmetric analysis revealed that these had twice the relative affinity for this stain than for FG whereas other peaks had equal or less affinity. The incorporation of [3H]cystine into peaks 9 and 10 was relatively greater than into the other proteins.


Gamma benzene hexachloride in guinea-pig brain after topical application. Blood and brain levels of gamma benzene hexachloride had been measured following topical application in guinea pigs. Levels increased with number of application, and the concentration in brain was at least 10 times greater than in blood.


Immunochemical analyses of a major antigen of Mycobacterium szulgai. Unheated culture filtrate of Mycobacterium szulgai and a homologous goat antiserum were prepared. Immunoelectrophoretic analysis demonstrated a dominant anodal antigen in the culture filtrate. By the use of diethylaminoethyl-cellulose chromatography, a fraction designated MSP, which was rich in this anodal antigen, was recovered. The major antigen of MSP was demonstrated to have partial identity with reference mycobacterial antigen 6, and evidence was obtained for separate shared and specific antigenic determinants. MSP was found to be a potent, delayed skin-test antigen of considerable specificity when used in sensitized guinea pigs. Arthus reactions were also observed and were not specific.


Effect of combinations of penicillin and aminoglycosides on Streptococcus faecalis: a comparative study of seven aminoglycoside antibiotics. The 50% inhibitory concentrations (IC50) of benzylpenicillin, streptomycin, sisomicin, gentamicin, tobramycin, kanamycin, amikacin, and butirosin were determined for 58 clinical isolates of Streptococcus faecalis, 28 of which were recovered from cultures of blood samples from patients with endocarditis. The IC50 of streptomycin was less than 100 microng/ml for 42 strains, 192-10,000 microng/ml for eight, and larger than or equal to 10,000 micron/ml for eight. One isolate that was highly resistant to streptomycin was also highly resistant to kanamycin and butirosin. Extraordinarily high resistance to the other aminoglycosides was not observed. The bactericidal effects of combinations of penicillin and aminoglycosides were studied in 20 strains of S. faecalis that represented different levels of resistance to streptomycin. Significant enhancement of the effect of the combination of penicillin and streptomycin was found only in strains with an IC50 of smaller than or equal to 190 microng/ml. Combinations of penicillin and sisomicin, gentamicin, or tobramycin were effective even against strains that were highly resistant to streptomycin and kanamycin.


Plasma cell antigens in human multiple myeloma. Tumor-specific antigens capable of eliciting a response from autologous lymphocytes have been described in mouse plasmacytoma systems. This paper presents evidence of similar antigens in human myeloma. Myeloma plasma cells isolated from the bone marrow of 27 patients stimulated autologous and allogeneic peripheral blood lymphocytes (PBL) from myeloma patients in mixed leukocyte culture. Plasma cells isolated fromt  the bone marrow of normal patients or patients with benign plasmacytosis failed to stimulate PBL in mixed leukocyte culture. Similar results were found in passive cytotoxicity assays with the use of chinken red blood cells (CRBC) coated with 3M KC1 plasma cell extracts from myeloma patients. PBL from myeloma patients caused 30 to 80 percent chromium-51 release from tanned chromium-labeled plasma cell extract-coated CRBC targets, whereas PBL from normal patients or patients with benign plasmacytosis caused only 10 to 25 percent chromium-51 release. This study indicates the presence of material resembling tumor-specific antigens on human myeloma plasma cells. Immune response to suce antigens is elicited in autologous and allogeneic myeloma patients.


Plasma levels of human chorionic somatomammotrophin, progesterone, unconjugated oestradiol and oestriol, and alpha-foetoprotein in patients with hydatidiform moles. Human chorionic somatomammotrophin (HCS), progesterone, and unconjugated oestradiol and oestriol were measured in the plasma of 13 patients with intact hydatidiform moles from week 9 to 19 of pregnancy and in 89 normal women from week 5 to 20 of pregnancy. Plasma alpha-foetoprotein (AFP) was also measured in 9 out of 13 patients and in 23 of the normal women from week 13 to 20 of pregnancy. All the compounds were measured by radioimmunoassay. The plasma HCS concentration in 35 samples from 13 patients with hydatidiform moles ranged from 10 to 910 ng/ml; this was lower than that in normal pregnancy of corresponding duration in eight patients; within the normal range in four patients and high in one patient. The plasma progesterone concentration ranged from 17-5 to 79-2 ng/ml; it was higher than that in normal pregnancy in eight patients and within the normal range in five patients. The plasma unconjugated oestradiol concentration ranged from 1-82 to 8-10 ng/ml; it was higher than normal in six patients and within the normal range in seven patients. The plasma unconjugated oestriol concentration ranged from 0-168 to 1-37 ng/ml, the levels at 15-19 weeks of gestation being significantly lower than those in normal pregnancy at this time (P less than 0-005). Plasma AFP was not detectable in the nine patients (less than 10 ng/ml) whereas it ranged from 10 to 80 ng/ml in 18 out of 23 women in week 13-20 of normal pregnancy. The present results suggest that both plasma oestriol and AFP could be helpful in the diagnosis of hydatidiform mole at about 12-14 weeks though diagnosis could not be made with absolute certainty.


Ia antigenic specificities are oligosaccharide in nature: hapten-inhibition studies. We have previously reported that the Ia specificities, coded for by the I region within the H-2 complex, appear to consist predominantly of carbohydrate. This conclusion was reached by examining low molecular weight Ia-bearing oligosacharides isolated from mouse serum. We now report hapten-inhibition studies which indicate that the binding of both allogeneic and xenogeneic anti-Ia antibodies to the Ia glycoproteins found predominantly on B lymphocytes can be specifically inhibited by certain free sugars. Both inhibition assays revealed that the specificity for the following Ia antigens resides predominantly in the following sugars: (a) Ia.1: N-acetyl-D-mannosamine or related sugars; (b) Ia.3: alpha-D-galactose and related sugars; (c) Ia.7: L-fucose;  and (d) Ia.15: N-acetyl-D-glucosamine. It seems likely that these sugars are found at the terminal nonreducing ends of the carbohydrate portion of the Ia-bearing glycoproteins present in the lymphocyte membrane. In contrast, several public and private H-2 antigenic specificities did not appear to be sugar defined. These studies imply that at least some of the Ia genes from both the I-A and I-C subregions of the I region code for glycosyl transferases which modify oligosaccharide structure and impart specificity to the Ia antigens by alteration of their terminal sugar residues.


Immune mechanisms in leukemia role of the Ia antigens. We have shown that the selective removal of cells possessing Ia determinants coded by the I-A, I-B, and I-J regions of the H-2 gene complex completely abrogates the protective capacity of nylon-wool-purified T lymphocytes against leukemic challenge. This suggests that the Ia antigen bearing T cells play an important role in tumor immunity.


The capsule of the gonococcus.

The capsule of the gonococcus. 16 strains of Neisseria gonorrhoeae were subjected to several established techniques for capsule demonstration by light microscopy. In all strains examined, encapsulation of the gonococcus was demonstrated. Although the capsules were somewhat more easily seen in strains recently isolated from clinical material, organisms that had been passaged for several years also were encapsulated as were all the colony types within these strains. The gonococcal capsule is easily disrupted by mechanical forces.


Histocompatibility antigens and genetic control of the immune response in guinea pigs IV. Specific inhibition of lymphocyte proliferation by auto-anti-idiotypic antibodies. The in vitro T-cell proliferation induced by penicilloylated bovine IgG (BPO-BGG) in sensitized strain 2 guinea pigs could be specifically blocked by the serum of guinea pig (*305) which had been repeatedly immunized with BPO-BGG over a period of 9 mo. The antibodies which appeared in the serum of this animal (*305) were functionally similar to the strain 2 anti-idiotypic antibodies (a strain 2 BPO-BGG) raised by immunizing strain 2 guinea pigs with immuno-adsorbent column-purified BPO-BGG. Animal *305 had no detectable antibodies to BPO-BGG and failed to give a delayed hypersensitivity skin response when challenged with BPO-BGG.


T-T interactions in the induction of suppressor and helper T cells: analysis of membrane phenotype of precursor and amplifier cells. The Ly and Ia phenotypes of T lymphocytes involved in the in vitro generation of helper and suppressor cells were identified. The precursors of both cells are found in adult thymectomized spleen. Helper precursors are Ly-1+2-3-Ia-, while suppressor precursors are Ly-1-2+3+Ia-, although the suppressor effector is Ia+. In both cases a second 'amplifier' cell is required for differentiation of precursors to occur. This cell is found in anti-lymphocyte serum-treated spleen and has the phenotype Ly-1+2+3+Ia-.


Genetic control of alpha-fetoprotein synthesis in the mouse. To approach the genetic mechanism that turns off the synthesis of alpha-fetoprotein (AFP) after birth, we assumed that a change in this mechanism might affect the low basal level of AFP that can be detected in the adult organism. The concentration of AFP was therefore determined for serum from adult mice of 27 different inbred strains. With one exception, this basal level was between 34 and 173 ng/ml, which is about 10(5)-fold less than the serum concentration at birth. In one strain, BALB/c/J, the AFP level was found to be considerably increased; it was about 10-fold higher than in other strains at 9-10 wk of age. Two other substrains of BALF/c mice showed normally low AFP levels. Kinetic studies show that the rate with which AFP disappears from serum after birth is reduced in BALB/c/J mice as compared to other strains. The increased AFP level of BALB/c/J mice appears to be due to an increased rate of synthesis of AFP, since the rate of catabolism of AFP was found to be normal in these mice. Genetic analysis was performed by crossing BALB/c/J mice with mice having an ordinary AFP level, followed by determination of AFP levels in mice of the F1 and F2 generations as well as in back-cross mice. The results clearly indicate that the increased AFP level in BALB/c/J mice is controlled by a single recessive Mendelian gene, which has been named Raf (for regulation of alphafetoprotein). The Raf gene could be directly involved in the regulation of AFP synthesis, but it may also control AFP levels only indirectly, e.g., by regulating the synthesis of a hormone that controls AFP synthesis.


Immunosuppressive factor(s) extracted from lymphoid cells of nonresponder mice primed with L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) III. Immunochemical properties of the GAT-specific suppressive factor. The GAT-specific suppressor T-cell factor (GAT-TsF) extracted from lymphoid cells from GAT-primed, nonresponder DBA/1 mice has been partially characterized. It is a protein that has affinity for GAT and determinants encoded by the I region of the H-2 complex. On the basis of specificity and avidity, GAT-TsF resembles anti-GAT-MBSA antibodies produced by DBA/1 mice in spite of the fact that it is too small to be classical antibody and has no constant-region determinants of heavy or light chains. Further, GAT or a fragment of GAT is associated with the GAT-TsF. GAT-TsF has been partially purified from the crude extract by absorption to GAT-Sepharose and elution with 0.4 to 0.6 KCl. GAT-TsF purified on the basis of its affinity for GAT bears I-region determinants but not detectable GAT or GAT fragment.


The monoclonal anti-phosphorylcholine antibody response in several murine strains: genetic implications of a diverse repertoire. The idiotypic identification of monoclonal antibodies has been used to define and enumerate clonotypes within the murine repertoire of B cells specific for phosphorylcholine (PC). The response in the BALB/c strain is dominated by a single antibody specificity which is identical to TEPC 15 protein; however, antibody without the TEPC 15 idiotype appears heterogeneous by idiotypic cross-reactivity and hapten inhibition of binding to antigen. Dissection of the PC-specific repertoire in the AKR, A/He, and C3H strains has indicated that some monoclonal antibodies share binding-site idiotypic determinants with TEPC 15, although these clones represent a minority of the precursor cells. In addition to providing insights into the heterogeneity and expression of the murine B-cell repertoire, these studies emphasize structural relationships between PC-specific clonotypes. Within the BALB/c strain, some antibodies share combining-site-related idiotypic specificities with TEPC 15, but differ in other variable region determinants. Among allotypically distinct strains, there exists a remarkable similarity of variable region determinants in at least a minority of antibodies.


Enhancement of IgE-mediated histamine release from human basophils by viruses: role of interferon. Human leukocytes maintained in culture are induced to release histamine when exposed to ragweed antigen E or anti-IgE. Leukocyte cultures incubated with virus (i.e. HSV-1, Influenza A, and Adeno-1) but not exposed to ragweed antigen E or anti-IgE fail to release histamine. If, however, leukocyte cultures are first exposed to virus and then to ragweed antigen E or anti-IgE, significant enhancement of histamine release occurs. Both infectious and inactivated virus enhance histamine release and the degree of enhancement is related to the concentration of virus and the length of the incubation. Tissue culture fluid harvested 8 h after exposure of leukocytes to virus contains a soluble factor which is capable of enhancing histamine release when added to fresh leukocyte cultures. This factor has all the properties of interferon including species specificity and cannot be dissociated from the antiviral activity of interferon. Moreover, both known inducers of interferon (poly I:poly C) and standard preparations of interferon are capable of enhancing histamine release. The enhancement of histamine release by interferon represents a new biological role for interferon.


The inheritance of antibody V regions in the rabbit: linkage of an H-chain-specific idiotype to immunoglobulin allotypes. Anti-idiotype antibodies specific for the H chain of an homogeneous antistreptococcal antibody (4135 Ab) were prepared by injection of recombinant molecules consisting of the H chains from 4135 Ab and L chains isolated from the injected rabbit. The antibodies prepared in this fashion (anti-HId) were specific for the VH region of 4135 Ab. Using this preparation in an inhibition of binding assay, sera from rabbits related and unrelated to 4135 were screened  for the presence of the 4135 HId. It was found that about 45% of the related rabbits, when immunized with streptococcal Group C vaccine, produced antibodies with a cross-reactive idiotype, while less than 10% of similarly immunized unrelated rabbits produced molecules bearing HId. The expression of HId was linked to the a3 allotype present in the H chain allogroup J. Antibody molecules bearing the HId determinant were isolated from heterozygous (a1a3 and a2a3) rabbits and shown to express the a3 allotype. One rabbit lacking the a3 allotype produced significant amounts of antibodies expressing HId. These antibodies were found to express both VH and CH  allo-types characteristic of the J allogroup, although neither allotype was found in a preimmune IgG sample from this rabbit.


Differences in the mechanism of tolerance to dinitrophenylated bovine gamma globulin when induced in normal adult mice or in reconstituted irradiated mice: dependence of the mechanism of tolerance on the structural organization of the lymphoid system. Tolerance can be induced in adult mice by a single intravenous injection of 0.5 mg dinitrophenylated bovine gamma globulin. The cellular mechanism of the unresponsive state is different depending upon whether the tolerance is induced in normal intact adult mice or in reconstituted, irradiated mice. The tolerant state induced in intact mice is characterized by a high avidity of the residual antibody-forming cells in partially tolerant animals and a prompt reversibility on cell transfer. The overall properties of this unresponsive state are consistent with the hypothesis that it is mediated by the production of small amounts of high affinity antibody in response to the tolerance-inducing injection of antigen. In contrast, the unresponsiveness induced in reconstituted, irradiated mice by the same procedure was characterized by a low avidity of the residual antibody-forming cells in partially tolerant animals and stability on transfer of spleen cells from unresponsive into irradiated recipients. No suppressor cell activity was detected and mixed cell transfer studies were consitent with the view that this unresponsive state represented a B-lymphocyte clonal deletion. The presence or absence of T lymphocytes in the population of cells used for reconstituting the irradiated recipients did not effect the ease of tolernace induction or the cellular mechanism of the tolerant state which was produced. If irradiated mice reconstituted with B and T lymphocytes were rested for 2 wk before tolerance induction then a reversible "high affinity"-type tolerance is obtained such as is typical of normal intact animals. Restorationof a "normal" response to the tolerance-inducing injection of antigen is dependent upon the presence of thymus cells in the population of cells used for reconstitution. It is suggested that the structural integrity of the lymphoid tissue is critical in determining whether B cell will be rendered tolerant after exposure to antigen in vivo.


The release of an endogenous pyrogen from guinea pig leukocytes in vitro: a new model for investigating the role of lymphocytes in fevers induced by antigen in hosts with delayed hypersensitivity. Guinea pig periotoneal exudate (PE) cells incubated overnight in vitro with heat-killed Staphylococci released an endogenous pyrogen (EP) that could be assayed by intravenous injection in rabbits. The febrile responses were linearly related to the dosage of EP over an eightfold range. PE cells derived from guinea pigs with delayed hypersensitivity (DH) to bovine gamma globulin (BGG), also released EP when incubated with antigen in vitro. This reaction was specific and did not occur withe PE cells from normal or complete Freund's adjuvant-sensitized guinea pigs. Studies indicated that monos and/or polymorphonuclear leukocytes rather than lymphocytes were the source of EP. However, when incubated with BGG and sufficient dosages of BGG-sensitized lymphocytes, normal PE cells released EP over a 42 h period. These results suggest that antigen stimulates specifically sensitized lymphocytes to release an agent (perhaps a lymphokine) that activates phagocytic cells to release EP. This model offers unique advantages for investigating in vitro the role of the lymphocyte in antigen-induced fever in DH as well as the relationship of this lymphocyte-induced activity to other known biologic activities mediated by antigen stimulated lymphocytes.


H-2 restriction of cell-mediated immunity to an intracellular bacterium: effector T cells are specific for Listeria antigen in association with H-21 region-coded self-markers. The protective activity of anti-Listeria-immune T cells assayed in an adoptive transfer system in H-2 restricted. As shown in the present studies, the demonstration of the restriction is directly dependent on the dose and the relative protective activity of spleen cells. In addition, some H-2-unrestricted protection is conferred predominantly by other than immunoglobulin-negative spleen cells. Thus, the activity of Listeria-immune T cells appears to be 'absolutely' restricted and is in this respect comparable to in vivo T-cell-mediated anti-viral protection. The predominant genetic region of H-2 coding for the structures which are mainly involved in this restriction in T-cell immunity to this prototype intracellular bacterium is the I region. The specificity of Listeria-immune T cells is determined by the H-2 haplotype of the donor. Thus, F1 hybrids seem to possess at least two separable sets of T cells, each specific for one parental haplotype. As is true in the virus model, the results cannot distinguish between an altered-self or a dual recognition model of T-cell recognition to explain H-2 restriction. They are, however, compatible with the idea and I-coded cell surface structures may serve as receptors for cell-specific differentiation signals, which trigger direct or lymphokin-mediated activation of macrophages to manifest increased bactericidal capacity. The interesting parallels in self-marker recognition of T cells in the virus and intracellular bacterium systems, respectively, appear to be reasonably explained by the different types of signals transmitted by T cells to various target cells via the distinctly different self-markers employed (i.e., K or D vs I).


In vitro tolerance induction of neonatal murine B cells as a probe for the study of B-cell diversification. The susceptibility to in vitro tolerance induction has been implicated as a characteristic of B cells early in their development, since DNP-reactive B cells are tolerizable only during the first days after birth, and 25% of adult bone marrow cells are tolerizable. In the present study, a modification of the in vitro splenic focus technique was utilized to determine if PC-specific B cells, by virtue of their late expression (approximately 1 wk post-parturition), also display susceptibility to tolerance induction. The results demonstrate that at 7-10 days after birth, when over 90% of the DNP-specific splenic B cells are resistant to tolerance induction, the majority of PC-specific B cells are tolerizable. These results re-emphasize tolerance susceptibility as a characteristic of developing clones, confirm the late acquisition of PC-specific B cells, and support the contention that the acquisition of the specificity repertoire is a highly ordered, specifically predetermined process which is independent of antigen-driven events.


Cytotoxic T lymphocytes specific for I region determinants do not require interactions with H-2K or D gene products. Gene products coded for by the major hisocompatibility complex (MHC) can serve as target antigens for cytotoxic T lymphocytes (CTL) (1). A variety of test systems are available which have yielded information consistently reinforcing the importance of this complex of genes in the generation and effector phases of the cytotoxic immune response. Originally, it was shown that allogeneically-induced CTL had specificity primarily for the products of the K and D loci of the mouse H-2 complex (2). More recently this has also been found to be the case for xenogeneic immunizations (3,4). Additional examples of T cell-mediated lysis have been reported involving viral-infected or chemically- modified syngeneic stimulating and target cells in which homology at H-2K or H-2D was required between the responding and target cells for appreciable lysis to occur (5-7). Moreover, CTL specific for minor histocompatability antigens are able to lyse only target cells bearing these membrane antigens and sharing a common H-2K or H2-D gene product with the effector (8,9). Two hypotheses have been proposed to explain the requirement for H-2 identity between effector and targets in these systems. CTL may recognize new antigenic determinants created by the interaction of the modifier with syngeneic K and D gene products. Alternately, a dual recognition system my exist, requiring an antigen-specific receptor as well as a second receptor with specificity for homologous H-2K or H-2D determinants (5). Neither model can be excluded at this time. The I region also contains genes coding for histocompatibility loci since animals differing at the I-A or I-C regions of the H-2 complex reject skin grafts (10-12), though less rapidly than mice differing at the H-2K or H-2D regions, Also CTL can be generated to I region determinants but less efficiently than CTL specific for H-2K or H-2D gene products (12-14). The question can therefore be raised, whether the I region minor histocompatibility loci function independently from the H-2K or H-2D loci or whether I region-specific cytolysis requires the participation of H-2K or H-2D gene products of the target cell. This communication illustrates the generation of CTL showing specificity for I region determinants in primary mixed lymphocyte cultures. Further, we demonstrate by genetic analysis and byt eh use of speficit alloantisera that CTL directed to Ia determinants (a) do not see these antigens as modifications of H-2K or H-2D gene products but as independent gene products coded for by the I region, and (b) they do not require interaction with target cells bearing the same H-2K or H-2D gene product as the effect CTL.


Evidence for the dissimilarity of acrosin and pancreatic trypsin as revealed by a sensitive immunoassay. Common antigenic determinants between two species of pancreatic trypsin and twelve species of acrosin were studied using a highly sensitive, enzyme-linked immunoassay. No common antigenic determinants between trypsin and acrosin could be detected.


Characterization of three antigenic particles of swine vesicular disease virus. Three distinct particles were isolated from cell culture harvests of swine vesicular disease virus (SVDV) by sucrose and CsCl gradient centrifugation. Virions (148S), RNA-free empty capsids (81S), and a third particle (49S) also free of RNA showed immune reactivity with SVDV antiserum. The 81S and 49S particles had polypeptides typical of naturally occurring empty capsids. Injection of purified antigens into guinea pigs produced antisera which distinguished empty capsids from virions on immunodiffusion; the 49S antigen appeared similar to virions. Antisera produced to freshly prepared virus antigen grown in brains of baby mice distinguished SVDV from the serologically related Coxsackie B-5 virus but did not distinguish the individual S particle antigens. Partly purified virus preparations degraded to empty capsids when incubated in guinea pig serum. The possible origin of empty capsids and 49S particles and their relationship to antigenicity of virus preparations are discussed.


Characterization of human papovavirus RFV: comparison with SV40 and BKV. Human papovavirus, RFV, isolated from urine of a renal transplant patient was compared with two strains of SV40 and with the prototype human papovavirus, BKV. Neutralization tests showed that RFV and BKV are indistinguishable, while large-plaque (LP) and small-plaque(SP) isolates of SV40 gave a low but significant level of cross-reaction with rabbit or human antisera against RFV. DNA reassociation saturation tests using 125I-labelled RFV DNA show that BKV has 88% homology, and SP-SV40 has 29% homology to RFV. We conclude that RFV and BKV are nearly, if not totally, identical and are not SV40 variants.


Fast axonal transport: effect of antimitotic drugs and inhibitors of energy metabolism on the rate and amount of transported protein in frog sciatic nerves. Mitosis inhibitors, drugs affecting the energy metabolism, heavy water, and ouabain were used to partially inhibit fast axonal transport in frog sciatic nerves. Effects on the rate and on the amount of pulse labeled protein could be separated. The pulse of labeled protein, released after a cold-block, rapidly reached a maximum height which indicated that the transport system was saturated in the nerve segment occupied by the pulse. Both the rate and the amount were reduced by the mitosis inhibitors colchicine, vinblastine, and griseofulvin. Colchicine had a differential effect and reduced the rate of material migrating in the advancing front of the pulse less than the rate of that moving in the peak. Preincubation at low temperature potentiated the effects of colchicine. Two inhibitors of energy metabolism, NaCN and IAA, reduced the amount of labeled material in the pulse. The slope of the pulse was markedly reduced and multiple peaks appeared. The distance covered by the migrating pulse was largely unaffected, but some retardation of late components might have occurred. In contrast, 2.4-DNP reduced the rate without any effects on the amount of migrating material. Heavy water uniformly reduced the rate of the migrating pulse, whereas the main effect of ouabain was a diminished amount and multiple peaks as with NaCN and IAA. All drugs were tested for their effects on the electrical activity of sciatic nerves. The compound action potential was not affected by the mitosis inhibitors and heavy water, but was depressed by the inhibitors of energy metabolism and abolished by ouabain. The results indicate that the effects of various transport inhibitory drugs can be differentiated if both the rate and the amount are considered.


Lead neuropathy. 1) Morphometry, nerve conduction, and choline acetyltransferase transport: new finding of endoneurial edema associated with segmental demyelination. Morphometric and pathologic studies along the length of the peripheral nervous system were obtained in groups of rats fed 4% lead carbonate for 3 and 6 months and in match-fed controls. The number and diameter histograms of L6 cytons of spinal ganglia and of myelinated fibers of proximal and distal portions of peroneal and sural nerve were not significantly different from the control groups. On the other hand, segmental demyelination occurred approximately as frequently in proximal as in distal parts of nerves. At 3 months approximately 1/3 of teased myelinated fibers showed changes of segmental demyelination (Condition C), or of remyelination after segmental demyelination (Condition F) or of both segmental demyelination and of remyelination (Condition D), while at 6 months more than 4/5ths of fibers showed these changes. As expected, regression lines of axonal area on number of lamellae of myelin, were less steep in nerves of rats fed on lead for 6 months as compared to controls. Axonal transport of choline acetyltransferase in lead neuropathy did not differ from that in control rats. As expected from the studies of others, conduction velocity of myelinated fibers of caudal nerve were low. A new finding was the often quite striking increase of transverse fascicular area of peripheral nerves. This was due to edema which appeared to develop at about the time of onset os segmental demyelination. Although the edema may be an epiphenomenon, it could be an important observation bearing on the development of lead neuropathy. It would be important to know next whether or not the blood nerve barrier is altered in lead neuropathy.


Electrophysiological investigations in a case of cephalic tetanus. In a case of cephalic tetanus the amplitudes and the latencies of the compound action potentials recorded from the facial muscles after stimulation of the facial nerve at the mastoid were not significantly different on the two sides. With repetitive stimulation at 30/sec a pathological facilitation was observed on both sides. The blink reflex bilaterally had normal latencies but the amplitudes of the reflex potentials were always significantly lower on the paretic side. It is concluded that facial palsy in cephalic tetanus is mainly due to a functional block of conduction in the course of the peripheral nerve whereas the disturbance of neuromuscular transmission probably has little importance in these cases.


The regional curare test in myasthenia gravis. 30 subjects without disturbance of neuromuscular transmission and 18 patients with myasthenia gravis were used in conducting the regional curare test. The adductor pollicis and the hypothenar muscles were studied with the 3/sec stimulation test. With three different dosages of curare one could find no reliable border between "normal" and "pathological". In the patients with myasthenia no definite relation could be found between the findings with the regional curare test and the clinical picture. The curare concentration reaching the muscle is probably quite variable from case to case as regards diffusion and volume in the tissue. The 3/sec stimulation test with registration from the deltoid muscle, and in certain cases the systemic curare test, appear more suitable than the regional curare test for routine diagnosis as well as indication for thymectomy. But for cases of ocular myasthenia showing no further weakness by the systemic curare test, the regional curare test can be put to use. The advantage lies in the higher concentration of curare which can thereby be brought to the muscle. The precautionary measures should be similar to those taken with the systemic curare test.


Electromyogram and nerve conduction in patients with acute intermittent porphyria. Diminished activity of uroporphyrinogen I-synthetase in the liver and other tissues may be regarded to be the primary genetic deficiency of acute intermittent porphyria (AIP). Increased production and renal excretion of delta-aminolevulinic acid (ALA) und porphobilinogen (PBG) are secondary phenomena. The neuropsychiatric symptomatology of AIP consists of neuropathy, vegetative crises and exogenous psychoses. In this study electromyographic and neurographic investigations were performed on 20 persons with AIP. 16 patients had experienced attacks of AIP, 10 of them including neuropathy. 4 persons showed the biochemical findings of AIP but had not yet had symptoms. In cases with persistent pareses following porphyric neuropathy denervation signs or sequelae were still present. In patients without clinical symptoms and in latent cases there were normal or borderline findings. Motor nerve conduction velocity was mostly decreased in combination with denervation signs and in a range that indicated a primarily axonal nerve lesion and consequent myelin damage rather than primary demyelinization. The mean motor conduction velocity of n. tibialis was somewhat lower in patients with porphyric crises without neuropathy than in latent cases without any clinical crises. The differences were not significant in other nerves. The findings are discussed under consideration of the electrodiagnostic results of other investigations and of neuropathological and clinical data.


Retrograde changes in motor and sensory conduction velocity after nerve injury. Nerve section is followed by a reduction of motor and sensory conduction velocity in the proximal segment of the injured nerve. This reduction of velocity is associated with retrograde changes in fiber size. If reinnervation does not occur within the next 1 1/2--2 years, retrograde degeneration of nerve fibers results, and the amplitude of the evoked nerve potential in the proximal segment of the injured nerve decreases. This retrograde degeneration is probably significant in view of the poor results frequently obtained after nerve transplantation which is carried out too late.