This proposal will define a training program that will allow Dr. Subia to redevelop and strengthen her current knowledge of molecular biology and immunology and to redevelop research skills that will ultimately allow her to become established as an independent investigator. Dr. Subia's long term research goals are to gain more knowledge about the pathogenesis of Neisseria gonorrhoeae. Phase I of this proposal will allow Dr. Subia to learn the most current molecular biological concepts and techniques in order to address the Specific Aims. Specific research plans for Phase II will be a continuum based on findings during Phase I and will involve the use of techniques acquired during Phase I. Dr. Subia is interested in studying the pathogenesis of Neisseria gonorrhoeae. This organism is a major cause of sexually transmitted disease and is a significant cause of morbidity and infertility. The mechanism of pathogenesis of Neisseria is complex. A major factor in pathogenesis is the organism's ability to invade eucaryotic cells. There is now increasing evidence to suggest that lipooligosaccharide of N. gonorrhoeae may initiate the process of invasion into eucaryotic cells. The terminal Ga1beta1-4GlcNac moiety recognized by monoclonal antibody 3F11 appears to be specifically involved as a component of the gonococcal-eukaryotic cell interaction. The underlying hypothesis of the research component of this grant is that a surface receptor on eukaryotic cells binds the LOS thru the 3F11 epitope and that anti-idiotypes to 3F11 will permit us to isolate this receptor from eukaryotic cells. This receptor probably is in the family of the Galactose binding proteins which have been described on the surface of a number of human cell types. For the purposes of this proposal we will refer to the receptor which binds to the LOS as the Ga1beta(1-4)GlcNAc receptor. The hypothesis stated above will be demonstrated through the following Specific Aims: 1) The generation of a hybridoma(s) producing an anti-idiotype antibody (Ab2beta) to the binding site of MAb 3F11 (MAb 3F11 binds specifically to Ga1beta1-4GlcNAc moiety of lipooligosaccharide), 2) the isolation, from either Hec1B or Hep2G cells, of the receptor which binds to the Ga1beta(1-4)GlcNAc LOS epitope by use of affinity chromatography with 3F11 anti-idiotype as ligand, 3) the generation of monoclonal antibodies to the Ga1beta1-4GlcNAc binding receptor and determine the N-terminal sequence of this protein, 4) the use of these reagents generated in Aim 3 to clone and sequence Ga1beta1-4GlcNAc receptor, 5) the study of the role of the gonococcus in inducing human cells to modulate expression of the receptor using the in vitro cell culture systems. In addition, I plan to determine the cell type(s) upon which this receptor resides in normal human cervical, endometrial and fallopian tube tissue.