matrix cohorts, according to the protocol plan. We were able to administer the maximum permitted dose of 108 pfu without any serious adverse events by vaccination of the first of 18 patients. These initial 18 patients were followed throughout the one-year period after injection. An additional 14 patients were treated at the MTD dose level , as planned in the protocol and confirmed by a letter to the FDA. The purpose of the additional patients was to have sufficient numbers of patients in the groups to statistically evaluate the anti -PSA immune responses induced by the Ad/PSA vaccine. In summary, 32 patients were treated in the study followed through the one-year period. Two additional patients were enrol led in the study (total number of enrolled patients, 34) but never received the vaccine and were therefore not evaluable. One patient chose to have radiation therapy instead of participating in the trial and the second was diagnosed with a second malignancy (melanoma) shortly after his enrollment in the phase I study. 1.4.1 Phase I study results The median age of the patients was 70.2 years (range, 52 to 89). The vaccine was administered as an aqueous suspension or in a collagen (Gelfoam) matrix to 32 patients . Sixteen (16/32) or 50% of the patients exhibited grade 1 vaccine-related adverse events (AE), 1/32 (3.1%) that exhibited a grade 2 AE, and one patient exhibited a grade 3 AE which was a decrease in neutrophil count. There were no vaccine-related grades 4 or 5 AEs. The following Table 2 is a listing of all AEs, sorted by relationship to the vaccine injection. Figure 2 . Effect of the presence of anti-adenovirus Ab on Ad5- PSA immunization with and without Gelfoam. Mice were injected i.p. with 10 9 PFU of Ad5- lacZ or PBS 2 wk before immunization with 10 9 PFU Ad5- PSA delivered s.c. in the fluid phase (PBS) or Gelfoam. CTL were tested against RM-11psa and RM-11neo to demonstrate PSA specificity. Cytolytic activity of each group against RM-11neo targets was <10% for all E:T ratios. Lubaroff Ad/PSA Phase II, protocol #2, page 8 Table 2 Ad/PSA Phase I Trial Adverse Events – by Vaccine Relationship Patient System Day of onset* Event Vaccine - related AP005 Neural 21 Agitation No AP006 Musculoskeletal 13 Left hip and thigh pain No AP007 Musculoskeletal 22 Left back pain No AP007 Neural 14 Situational depression No AP009 GI 21 Constipation No AP009 Musculoskeletal 14 Joint aches, secondary to fall No AP009 Neural 11 Fall, causing head lacerations & loss of consciousness No AP010 GU 21 Proteinuria No AP010 Skin 21 Edema No AP014 Constitutional 22 Fatigue No AP014 Musculoskeletal 14 Leg pain (bone – femur) No AP015 Respiratory 1 Cough No AP016 GU 172 Left neph rolithiasis No AP016 GU 183 New primary tumor – papillary bladder No AP016 Immune 8 Flu-like symptoms No AP016 Musculoskeletal 4 Bone pain – left leg No AP016 Musculoskeletal 21 Decreased left leg strength No AP018 Musculoskeletal 8 Bilateral rib pain No AP018 Neural 8 Insomnia No AP020 Neural 168 cord compression No AP022 GI 2 constipation No AP022 Hematologic 14 decreased lymphocyte count No AP022 Hematologic 63 decreased WBC & neutrophils No AP022 Musculoskeletal 5 increased bone pain No AP02 2 Neural 16 depression No AP023 Musculoskeletal 20 pain in left buttocks No AP025 Musculoskeletal 16 bone pain No AP026 Cardiovascular 14 Lower extremity edema No AP027 GI 1 GI cramping No AP027 GI 1 constipation No AP034 Respiratory prior Edema, whe ezing (change in inhaler in same time period) No AP035 Constitutional 64 Weight loss No AP035 GI 50 Constipation No AP035 GU 22 Proteinuria No AP035 Hematologic 15 Lymphopenia No AP035 Musculoskeletal 22 Increased alkaline phosphatase No AP037 GI 73 Nausea No AP018 Skin 1 Injection site tenderness Possible AP019 Cardiovascular 1 Hypotension Possible AP019 Constitutional 1 fever Possible AP019 Constitutional 14 fatigue Possible AP019 GU 1 Proteinuria Possible AP020 GI 21 increased alkaline phosp hatase Possible AP020 GU 21 ketonuria Possible Lubaroff Ad/PSA Phase II, protocol #2, page 9 AP020 Metabolic 14 hyponatremia Possible AP020 Metabolic 14 hyperglycemia Possible AP020 Respiratory 9 viral symptoms Possible AP005 Musculoskeletal 1 Left inguinal pain Possibly AP008 GU 21 Proteinuria Possibly AP008 Respiratory 3 Cold symptoms Possibly AP010 Hematologic 1 Decrease in absolute neutrophils count Possibly AP027 Liver 21 increased AST Possibly AP036 Hematologic 1 Anemia Possibly AP036 Hematologic 63 Anemia Possibly AP037 Hematologic 1 Lymphopenia Possibly AP002 Musculoskeletal 3 Groin pain Unlikely AP002 Neural 3 headache Unlikely AP002 Skin 1 Itching Unlikely AP003 Pulmonary 21 Pleural effusion Unlikely AP006 Constitutional 14 Chills Unlikely AP006 Neural 14 Migraine headache Unlikely AP012 Cardiovascular 21 Hypotension Unlikely AP012 GI 0 Heartburn Unlikely AP012 Neural 21 Dizziness Unlikely AP014 Hematologic 82 Anemia Unlikely AP015 Cardiovascular 137 Myocardial infarction Unlikely AP018 Respiratory 43 DOE? Unlikely AP02 1 Hematologic 1 decreased lymphocytes Unlikely AP021 Hematologic 63 thrombocytopenia Unlikely AP023 Immune 16 urinary tract infection Unlikely AP023 Liver 21 elevated AST Unlikely AP024 Hematologic 14 anemia Unlikely AP024 Hematologic 1 anemia Unlikel y AP025 GI 85 abdominal distention, constipation, apparently related to metastatic disease in the periaortic lymph nodes and periprostatic tumor mass Unlikely AP025 Hematologic 14 thrombocytopenia Unlikely AP025 Neural 122 Headache Unlikely AP027 Immun e 21 Infection Unlikely AP034 Cardiovascular 14 Hypotension/dizziness Unlikely AP034 Musculoskeletal 14 Back pain Unlikely AP036 Cardiovascular 63 Edema – bilateral ankles Unlikely AP002 Skin 0 hematoma at injection site Yes AP003 Skin 0 Bruising at i njection site Yes AP004 Skin 0 Ecchymosis at injection site Yes AP005 Skin 0 Ecchymosis and erythema at injection site Yes AP008 Skin 0 Ecchymosis at injection site Yes AP009 Skin 0 Ecchymosis at injection site Yes AP018 Skin 0 Ecchymosis Yes AP025 Skin 0 Ecchymosis at injection site Yes AP026 Skin 0 Erythema at injection site Yes AP034 Skin 0 Pain at injection site Yes * day 0 = day of injection; day 1 = day after injection Lubaroff Ad/PSA Phase II, protocol #2, page 10 We measured the anti -PSA immune responses, both antibody and T cell, in all patients enrolled in the study. Antibody responses to PSA were measured by the binding to PSA -secreting cell lines using the method adapted from Cavacini, et al.