Biomedical subjects
Publications and source records attributed to L N Lemeneva.
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RhD immunisation which follows pregnancy can be prevented by administration to the rhesus-negative mother of 20 micrograms anti-D immunoglobulin per 1 ml of D-positive fetal red cells in the maternal circulation. With the aim of substitution of polyclonal anti-D with monoclonal one we developed human-mouse cell lines producing anti-RhD IgG1 by fusing EBV-transformed human immune lymphocytes with murine myeloma. After several clonings and passages the EBV genome was eliminated from the cell lines. The antibodies were purified from culture supernatants using protein A affinity chromatography and tested for sterility, virus contamination, pyrogenecity, toxicity and DNA content. The monoclonals were compared with the standard polyclonal anti-RhD in in vitro and in vivo assays. In antibody-dependent cell cytotoxicity (ADCC) 2 of 4 studied monoclonals promoted greater RBS lysis than polyclonal anti-D at equivalent concentrations. Detection of binding site number of monoclonals anti-D revealed about 10,000/RBS D-determinants on DCe/dce erythrocyte which agrees with the data for polyclonal anti-D. Best in ADCC monoclonal anti-D sharply increased human autologous 51Cr-labelled rbc sensitised in vitro as well as accelerated clearance of RhD RBS from circulation of Rh-negative volunteers injected with 150 micrograms monoclonal anti-D. After clinical trial using of monoclonal anti-D is permitted in Russia.
Human B-lymphoblastoid cell lines producing specific anti-D antibodies against Rh0(D) antigen have been established by EBV-transformation. The strong influence of the donor's immune status (serum titer of anti-D antibodies, intravenous booster injection prior to bleeding) is shown. The cell lines cloned by limiting dilution continuously produce anti-D antibodies in culture for 7 months or longer which is sufficient to support large-scale production of anti-D antibodies based on the cultivation of cryopreserved cells. Anti-D IgGl monoclonal antibodies obtained react with Rh-positive and Du cells but not with Rh-negative cells in serological tests. These human monoclonal antibodies could be useful in the prevention of Rh hemolytic disease of newborns.
The protocol of Epstein-Barr virus (EBV)-induced transformation of B-lymphocytes from reripheral blood of normal humans has been described. The requirements for mononuclear EBV-infection and for selection, cultivation, and cloning of EBV-transformed cell lines have been investigated. Successful establishment of stable human immunoglobulin-producing cell lines from lymphocytes of immunized donors could be achieved following the protocol presented.
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Defective growth of parental bone marrow in an F1 hybrid is associated with delay of the exponential growth phase of injected hemopoietic stem cells rather than with their rejection. This is demonstrated both by parental hemopoietic stem cell kinetics in the irradiated hybrid and by the increase in the number of spleen colonies with time after hemopoietic cell injection. After passage through an F1 hybrid the parental hemopoietic stem cells acquire ability for better growth in the same F1 host. This phenomenon, which we designated "adaptive modification of hemopoietic stem cells," is associated with the appearance, on their surface, of histocompatibility molecules carrying H-2 determinants of the recipient. Treatment of the modified cells with antiserum against the second parental strain abrogates the state of adaptive modification.
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Injection of various parental strain hemic cells reduces the ability of the F1 hybrid to resist growth of grafted hemopoietic tissue from the same parent strain. Such an effect can be exerted by cells from lymph nodes, spleen, bone marrow, or thymus, the peritoneal macrophages, and the peripheral blood leukocytes. Embryonic and adult liver cells or blood erythrocytes do not inhibit hybrid resistance. It is assumed that hemic cells inhibit hybrid resistance through an effect on the hemopoietic microenvironment. The mechanism of this effect cannot be considered the result of a graft-versus-host reaction since lymphoid cells which are "immune" or "tolerant" to the hybrid antigens possess inhibitory activity similar to that of intact lymphoid cells. The hybrid resistance inhibitory cells maintain the inability of "parent-in-F1 hybrid" type chimeras to express hybrid resistance for at least 2 months.
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