PubMed · 6951353
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D O White. 1981. Recent developments in virology.. https://pubmed.ncbi.nlm.nih.gov/6951353/
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Induction protocols for pediatric renal transplant recipients commonly utilize polyclonal or monoclonal agents in sequence with additional immunosuppression. Polyclonal agents Minnesota antilymphoblast globulin (MALG), anti-thymocyte globulin (ATGAM) effect both T and B cells while monoclonal agents (OKT3) are T-cell specific. Flow cytometric analysis of T-cell subsets has become the marker of adequacy of immunosuppression during OKT3 therapy, yet to date no marker exists to measure the adequacy of immunosuppression with polyclonal agents. At the University of Michigan, flow cytometric analysis in pediatric renal transplant recipients undergoing polyclonal induction reveals that CD3, CD4, and CD8 suppression initially occurs. As opposed to OKT3, a rebound of CD3 cells occurs despite daily use of a polyclonal agent in sequence with additional immunosuppressives. During these analyses, a single kidney was lost which flow cytometry failed to predict. No significant difference in flow cytometric patterns occurred when comparing ATGAM to MALG induction. Flow cytometry utilized during polyclonal induction in pediatric renal transplant recipients revealed a variety of T-cell suppression patterns but failed to demonstrate persistence of suppression. Despite this lack of suppression as indicated by flow cytometry, a 97% 1 year allograft survival rate exists in the pediatric transplant program at the University of Michigan Medical Center. Therefore, the role of flow cytometry during polyclonal induction has yet to be well defined.
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The present experiments were initiated to see if cells capable of binding antigens could make polyreactive antibodies. Fluorescein isothiocyanate-labeled self and non-self antigens were incubated with B cells from normal individuals. Antigen-binding cells were separated from non-antigen-binding cells by flow cytometry, immortalized with Epstein-Barr virus and analyzed at the clonal level for their capacity to make polyreactive antibodies. Four to six times more cells making polyreactive antibodies were found in the B cell subset that bound antigens than in the B cell subset that did not bind antigens. The majority of the polyreactive antibodies were of the immunoglobulin (Ig)M isotype. Immunoflow cytometry revealed that cell lines making polyreactive antibodies bound a variety of antigens (e.g., insulin, IgGFc and beta-galactosidase), whereas cell lines making monoreactive antibodies bound only a single antigen. The binding of antigens to B cell lines that made polyreactive antibodies could be inhibited (range, 28%-57%) by both homogeneous and heterogeneous antigens. Both CD5+ and CD5- antigen-binding B cells made polyreactive antibodies, but the frequency was slightly higher in the CD5+ antigen-binding (85%) as compared to the CD5- antigen-binding (50%) population. Comparison of CD5+ B cells that bound antigens with CD5+ B cells that did not bind antigens showed that approximately 86% of the former, but only 15% of the latter, made polyreactive antibodies. It is concluded that cells capable of binding a variety of different antigens can make polyreactive antibodies and that antigen binding is a good marker for identifying polyreactive antibody-producing cells.