Identification of antigenic determinants by polyclonal and hybridoma antibodies induced during the course of infection by vaccinia virus

In order to extend the understanding of determinants involved in the humoral response in the infected host, mice were subjected to an immunization regimen using both active and uv-killed vaccinia virus. The spectrum of antibody specificity in hyperimmune sera was followed by Western blotting. Comparable studies involving Western blotting and immunofluorescence were conducted with a panel of monoclonal antibodies derived from hybridomas selected from similarly immunized animals. Hyperimmune sera contained circulating antibodies primarily against three polypeptides of 28K, 35K, and 62K. These antigens were shown to be located both at the surface and within the virion. The repertoire of monoclonal antibodies included some that reacted with the 28K and 35K antigens and others that recognized a 32K core complex component or a nonvirion cell surface component, corresponding to the viral hemagglutinin. Within the panel of monoclonal antibodies was a large group which reacted with a 32K antigen found in the IHD-J virion but absent from the IHD-W strain. This finding correlates with the absence of a 32K polypeptide from the IHD-W particle. Overall, the current findings reveal the absence of any particular correlation between the incidence of polyclonal antibodies in the circulation of the immune host and the frequency of selected hybridomas against vaccinia antigens. Application of this type of immunological analysis should provide useful data concerning the detection and mapping of the antigens and their epitopes which are significant for humoral immunity.     

Human male infertility caused by degeneration and death of sperm in the epididymis

Four patients with persistent oligospermia and necrospermia were found to have severely degenerated sperm in the ejaculate. However, in those examined, testicular sperm were ultrastructurally normal, indicating that sperm degeneration and death was occurring during epididymal passage or storage or both or upon mixing with the seminal plasma at ejaculation. Seminal plasma was found to be nontoxic to normal donor sperm. In three patients, frequent ejaculation (two ejaculates per day for 4 or 5 days) was used to deplete epididymal sperm reserves and reduce the period spent in the epididymis. This resulted in a threefold to sevenfold increase in percentage of motile sperm in the ejaculate and a similar increase in sperm motility index. The authors propose the term "epididymal necrospermia" to describe this previously undefined type of male infertility.     

Comparison of total alkaline phosphatase and three assays for bone-specific alkaline phosphatase in childhood and adolescence

We compared serum levels of total alkaline phosphatase (TAP) and bone-specific alkaline phosphatase (BAP) as determined by three different assays (lectin affinity electrophoresis, immunoradiometric assay, enzyme-linked immunosorbent assay) in subjects aged 5–20 years suffering from X-linked hypophosphatemic rickets ( n = 14), chronic renal failure ( n = 10) and chronic cholestatic liver disease ( n = 16). Results were compared to controls of the same age and were expressed as standard deviation scores (SDS). TAP correlated significantly with BAP ( r > 0.9 for each assay; p <0.001) in controls. In children with cholestatic diseases, TAP (median SDS + 2.0) was elevated, but BAP, as measured by the electrophoretic assay, was within the reference range for most patients (median SDS: -0.4; p = 0.003 for the difference between the median SDS of TAP and BAP). In contrast, results for BAP as determined by the two immunoassays were not significantly different from TAP in any of the three patient groups ( p > 0.05 in each group for both assays). In this study, the two immunoassays did not have a detectable advantage over lectin affinity electrophoresis in the determination of BAP.     

Fanconi's anemia treated by allogeneic marrow transplantation

Eight patients with Fanconi's anemia were given cyclophosphamide alone (seven patients) or combined with procarbazine and antithymocyte globulin (one patient) followed by marrow grafts from HLA-identical siblings. All patients had engraftment. Seven developed acute and three chronic graft-versus-host disease (GVHD). Three patients died with GVHD and infectious complications (days 19, 56, and 82) and one with an intracerebral hemorrhage (day 540). Four patients are surviving 647- 3435 days after grafting, two are well, and two have chronic GVHD that is improving. These results show that Fanconi's anemia can be treated successfully by allogeneic marrow transplantation.