Oral vaccination of white-tailed deer using a recombinant Bacillus Calmette-Guérin vaccine expressing the Borrelia burgdorferi outer surface protein A: prospects for immunocontraception.

PROBLEM: Reduction of excess numbers of white-tailed deer (Odocoileus virginianus) is a prime example of a potential use for immunocontraception as a means of wildlife population management. Oral vaccination appears to be the most pragmatic way to deliver immunocontraceptive vaccines to free-roaming populations of deer, but there was little, if any, prior evidence that oral vaccination is a viable concept in deer. METHOD OF STUDY: We used live Bacillus Calmette-Guérin (BCG) in a recombinant form (rBCG), which expressed Borrelia burgdorferi outer surface protein A, to test whether deer vaccinated orally with a specific antigen expressed in a live vector produce detectable antibody titers. RESULTS: The data indicate that oral vaccination of deer with an expressed antigen is feasible, as demonstrated by peak antibody titers to the expressed antigen. Also, peak titers measured by enzyme-linked immunosorbent assay were highest in orally vaccinated deer: 1600 in deer vaccinated by injection and 6400 in those vaccinated orally. CONCLUSIONS: The results of this study demonstrate that it is feasible to vaccinate deer orally with a live vector.

Comparative efficacy of two immunocontraceptive vaccines.

As part of our research program to develop immunocontraception as a wildlife management tool, we compared the physiological responses of wild Norway rats (Rattus norvegicus) to two immunocontraceptive vaccines; one involved mouse zona pellucida peptide (MZPP); the other involved gonadotropin releasing hormone (GnRH). Efficacy was monitored by immune, hormonal, and natality responses. Both vaccines were effective, but GnRH was much more effective (100% sterility of both sexes vs. 50% sterility of MZPP-treated females). Breeding success of control rats was 88% with litters of 5-9 pups; breeding success of MZPP rats was 50% with litters of 2-8; GnRH rats produced no young. In GnRH-treated male rats monitored for up to 17 months, testosterone was nondetectable and testes were atrophied to about 10% of their original volume for 10-13 months. There were no notable differences in mortality or body weights among groups, and, with the exception of testicular regression, there were no changes in general appearance. The GnRH vaccine is potentially a good rat reproductive control agent that may be effective over the normal lifespan of a rat under natural conditions in the wild.

T-cell receptor biases and clonal proliferations in blood and pleural effusions of patients with lung cancer.

We sought evidence that pulmonary carcinomas mediate a cellular immunologic response by analyzing T-cell antigen receptor beta-chain variable gene (TCRBV) repertoires of lymphocytes from peripheral blood (PBL) and malignant pleural effusions (PEL) of five lung cancer patients. Expression levels of 27 TCRBV were quantitated by multiprobe RNase protection assay (RPA), and clonal expansions were identified by sequence enrichment nuclease assay (SENA) and junctional region sequencing. Abnormal TCRBV expansions were identified in all subjects by RPA (mean 6.9 +/- 1.7/patient), and their number closely correlated with elapsed time since initial diagnosis (r = 0.97). SENA, performed in specimens from three patients, confirmed the presence of mono or oligoclonality in 48% of abnormal RPA expansions, and further identified T-cell clones among TCRBV with normal expression levels. The majority of clonal expansions were among PEL, and were nearly equally divided between CD4 and CD8. These data show that T-cell repertoires of lung cancer patients are characterized by marked abnormalities and frequent clonal expansions, most likely representing responses to unique, tumor-specific antigens (TSA). Moreover, this process appears exaggerated among PEL, further suggesting that malignant effusions include local proliferations of tumor reactive T cells. These findings imply the presence of lung cancer TSA capable of eliciting cellular immune responses and raise the possibility that selective immunotherapies can ultimately be developed.

T cell receptor biases and clonal proliferations among lung transplant recipients with obliterative bronchiolitis.

Obliterative bronchiolitis (OB) is the most serious late complication of lung transplantation, but the pathogenesis of this disorder has not been elucidated. We sought evidence that OB is mediated by a cellular immunologic response by characterizing T cell antigen receptor beta-chain variable gene (TCRBV) repertoires in lung allograft recipients. Expression levels of 27 TCRBV among recipients were determined by multiprobe RNase protection assay after PCR amplification. In comparison to recipients with no evidence of rejection (n = 9), the PBL TCRBV repertoires of OB subjects (n = 16) exhibited more frequent expansions (16 vs. 9% of all measured TCRBV, P < 0.02), and the magnitudes of these abnormalities were greater (8.2 +/- 0.8 vs. 4.5 +/- 0.3 SD from mean normal values, P < 0.01). TCRBV sequencing showed these expansions were composed of clonal or oligoclonal populations. Thus, T cell responses in the recipients are marked by highly selective clonal expansions, presumably driven by indirect recognition of a limited number of immunodominant alloantigens. These processes are exaggerated among allograft recipients with OB, implying that cognate immune mechanisms are important in the pathogenesis of the disorder. Furthermore, the prominence of finite, distinct TCR phenotypes raise possibilities for development of novel diagnostic modalities and targeted immunotherapies for OB and other manifestations of chronic allograft rejection.

Monoclonal antibody KS1/4-methotrexate immunoconjugate studies in non-small cell lung carcinoma.

The antigen reactive with murine monoclonal antibody (MAb) KS1/4 is expressed on epithelial malignancies and some normal epithelial tissues. Studies were undertaken to evaluate KS1/4-methotrexate (KS1/4-MTX) immunoconjugate in patients with advanced non-small cell carcinoma of the lung. Eleven patients in two different groups received KS1/4-MTX in two different escalating dose infusion schedules with a maximal tolerated dose of 1,750 mg/M2 and a cumulative dose of MTX of 40 mg/M2. Toxicities were similar in both groups and included fever, anorexia, nausea, vomiting, diarrhea, abdominal pain, guaiac positive stool, and hypoalbuminemia. Two patients had an associated aseptic meningitis. One patient had a 50% decrease in two lung nodules without a change in lymphangitic infiltrates. This patient received a second course of treatment and developed an immune complex-mediated arthritis and serum sickness. Four patients mounted a human antimouse antibody response. Post-treatment tumor biopsies documented binding of MAb KS1/4. These studies document the feasibility and potential usefulness of a MAb directed against tumor-associated antigens with the targeting of chemotherapeutic drugs in patients with non-small cell lung carcinoma.

Phase I clinical comparative study of monoclonal antibody KS1/4 and KS1/4-methotrexate immunconjugate in patients with non-small cell lung carcinoma.

A Phase Ia clinical trial was undertaken to evaluate and compare murine monoclonal antibody KS1/4 and KS1/4-methotrexate immunoconjugate in patients with Stage IIIB or IV non-small cell carcinoma of the lung. Six patients received KS1/4 alone and five patients received KS1/4-methotrexate conjugate. The maximal total dose received per patient in both groups was 1661 mg. Mild to moderate side effects in both groups included fever, chills, anorexia, nausea, vomiting, diarrhea, anemia, and brief transaminasemia. One patient who received antibody alone had an apparent acute immune complex-mediated reaction. Ten of 11 patients had a human anti-mouse response. Posttreatment carcinoma biopsies revealed binding of monoclonal antibody KS1/4 and deposition of C3d and C4c complement fragments. Monoclonal antibody binding and complement deposition correlated with increasing doses of infused antibody. There was one possible clinical response.

Isolated canine mastocytoma cells: propagation and characterization of two cell lines.

Five different dog mastocytoma tumors were successfully transplanted and propagated in BALB/c nude mice. Cells from two of these tumors were passaged serially through at least four generations of mice without morphological or functional change. The average yield from a 2-cm tumor harvested from a mouse was 1.2 +/- 2.8 X 10(9) mast cells with greater than 90% viability. Cells of one line were larger and more heavily granulated than the other, and contained 1.29 +/- 0.74 pg histamine/cell (mean +/- SD). Calcium ionophore A23187 and compound 48/80 caused dose dependent histamine release with no significant difference in release from generation to generation. The smaller cells contained 0.06 +/- 0.06 pg histamine/cell. Histamine release after calcium ionophore or compound 48/80 was dose dependent and unchanged through serial passages. Following passive sensitization antigen caused dose-dependent histamine release confirming the presence of IgE receptors on these cells. In both cell lines histamine release was inhibited by terbutaline, dibutyryl adenosine 3',5'-cyclic monophosphate, or isobutylmethylxanthine. These methods provide a morphologically and functionally stable population of nearly pure canine mast cells for biochemical and physiological studies.

Alkyl chain length and acute oral toxicity of p-aminophenones.

There exists a structure-toxicity relationship between the length of the alkyl chain of p-aminopropiophenone homologues and their oral LD50's in albino mice and rats. The acute oral toxicity decreased in both species with the removal of one carbon atom from the alkyl group, increased with the addition of one and two carbon atoms, and decreased again with the addition of three carbon atoms to the alkyl group.

Taste preferences of the common vampire bat (Desmodus rotundus).

Taste preference tests, with simultaneous presentation of treated and untreated food, were administered to 24 common vampire bats (Desmodus rotundus). The bats received brief exposures to four different stimuli representing sweet, salty, sour, and bitter tastes, each at four different concentrations. Despite a strong location bias, the bats significantly (P < 0.01) avoided the highest concentrations of the salty, sour, and bitter tastes. Consumption of the sweet stimulus at all concentrations was similar to that of the untreated standard. Vampires evidently can discriminate based on taste, although their ability is apparently poorly developed when compared with some euryphagous species such as the rat. Hence, taste is probably not a factor in host selection by the vampire.