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Biomedical subjects

M Belvedere

Publications and source records attributed to M Belvedere.

11 recordsLinked to original sources

Sera from volunteers immunized by planned blood transfusions as source of DR cytotoxic typing reagents.

Sera from volunteers immunized with planned blood transfusions were tested for anti DR cytotoxic antibodies with a panel of HLA typed cultured human lymphoid cells using a variety of serological techniques. The majority of sera contained DR cytotoxic antibodies. The specificity of DR antibodies in seven sera was determined by testing them with a panel of B peripheral lymphocytes typed with DR alloantisera submitted to the 7th International Histocompatibility Workshop. The temporal evolution of DR and HLA-A and B cytotoxic antibodies was determined in two subjects by testing serial bleedings with B lymphoid cells, coated with Fab2 fragments from anti beta 2 mu and anti DR xenoantisera. Results indicated a parallel evolution of DR and HLA-A and B cytotoxic antibodies.

Antibody Specificity

Appearance and evolution of anti-Da (B cell-specific) antibodies after planned immunizations.

Appearance and evolution of anti-Da antibodies has been followed in eight volunteers immunized by whole blood transfusions or leukocyte intradermal injections form a single donor incompatible for HLA--A,--B,--C and--D specificities. Several unabsorbed bleedings from each recipient were studied against the specific immunizer with three different complement-dependent lymphocytotoxicity (CdL) techniques: (1) standard NIH CdL on total peripheral blood lymphocytes (PBL); (2) VII Workshop standard CdL technique on B cell-enriched suspensions; (3) beta2 microglobulin blanketing test ("bb" test) on B cells. Results obtained with the "bb" test were confirmed with platelet-absorbed sera. The "bb" and the absorbed sera allow discrimination between anti-Da and anti-HLA--A,--B,--C antibodies. Stage of appearance and evolution are rather similar for an anti-HLA--A,--B,--C and anti-Da. An early appearance of antibodies positive only against B cells is due to weak anti-HLA--A,--B antibodies which react better with B cells than with total PBL. Immunogenicity of Da antigens seems to be of the same order as HLA--A, and--B. In fact, Da reactivity was present in all eight recipients studied. These reactivities always segregated in familes with these HLA haplotypes. On a small panel of unrelated D-typed donors, three sera showed a significant positive association with D alleles.

Antibody Specificity

Serologic and immunochemical characterization of HLA-A9 xenoantisera.

The specificity of A9 antiserum produced in rabbits immunized with high density lipoprotein-associated HLA-A9 from serum was investigated by using serologic and immunochemical approaches. After suitable absorption, the xenoantiserum No. 2958 is monospecific for A9 antigens in direct cytotoxic test. Data from lysostrip experiments and from blocking of cytotoxicity with Fab2 fragments indicate that the A9 xenoantiserum No. 2958 react with the same or closely associated antigenic structures recognized by A9 alloantisera. Immunoprecipitates obtained with the specific xenoantiserum No. 2958 and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis contained only proteins of the size of detergent-solubilized HLA and beta2-microglobulin (45,000 and 12,000 m.w.). Immunodepletion experiments indicated that the precipitation was mediated by antibodies directed to the heavy chain, and not the light chain of the complex. These results indicate that monospecific HLA-A9 xenoantisera have the same discriminatory capacity of alloantisera and can be useful for the serologic and immunochemical characterization of HLA antigens.

Absorption

Antigenic determinants of the W28 molecule different from but spatially close to W28 determinant.

The existence of two allelic determinants, subtypic to W28 (named TO54 and TO55), and of an antibody cross-reacting W28 times HL-A10, is demonstrated. These new determinants have been studied by means of population and family studies and with the following serological techniques: absorption, elution, lysostrip and blocking by F(ab)-2 fragment. The results suggest that determinants TO54 and TO55 are very similar each other and are spatially close to the supertypic factor.

Adsorption