PubMed Health⌕ Search

Biomedical subjects

A E Bussard

Publications and source records attributed to A E Bussard.

At least 19 recordsLinked to original sources
PubMed1985

Monoclonal autoantibodies against mouse red blood cells: a family of structurally restricted molecules.

Cultured mouse peritoneal cells from unstimulated mice developed plaque-forming activity against isologous bromelain-treated erythrocytes. Several IgM monoclonal autoantibodies obtained by fusion of peritoneal cells from NZB or CBA origin with BALB/c myeloma cells were purified by affinity chromatography on trimethyl ammonium (TMA) column on the basis of their cross-reactivity with TMA, phosphorylcholine (PC) or choline haptens. Binding affinity for PC hapten was of the order of 10(3) M-1. Idiotypic studies with a polyclonal rabbit anti-idiotypic reagent revealed strong cross-reactions with all hybridoma autoantibodies thus far tested. In addition, the rabbit anti-idiotypic serum detected idiotypes or cross-reactive idiotypes in the sera of NZB and CBA as well as BALB/c mice. N-terminal amino acid sequence analyses of three hybridoma autoantibodies from NZB mice and one from CBA mice were carried out. The sequences of the first 32 residues of the four heavy chains showed that three were identical while one had one amino acid interchange; they belong to the VHIII-subgroup. The light chains were identical in the first 35 residues with the exception of a substitution at position 3 in two light chains and are members of the VK-9-subgroup. These results entirely support the idiotypic data. These monoclonal autoantibodies from NZB and CBA mice although isolated and eluted from PC-related haptens do not have any apparent structural nor idiotypic relationship to PC-specific antibodies. Idiotypic and V-region N-terminal sequence data suggest that these autoantibodies constitute a highly restricted family of molecules likely to be encoded by unique germ-line genes which may be expressed as such or as somatic variants in different mouse strains.

Aging↗
PubMed1982

Relationship between choline derivatives and mouse erythrocyte membrane antigens revealed by mouse monoclonal antibodies. I. Anticholine activity of anti-mouse erythrocyte monoclonal antibodies.

Different clones of mouse hybridomas, derived from the fusion of unstimulated mouse peritoneal cells with mouse myeloma cells, producing IgM monoclonal antibodies directed against the membrane of bromelain-treated mouse erythrocytes (MRBC(Br)) have been previously established. We have recently shown that one of these hybridomas produce, in ascites, antibodies cross-reacting with phosphorylcholine derivatives (trimethylammonium (TMA) derivatives). In this work the cross-reactivity for TMA derivatives of the monoclonal antibodies produced by 4 anti-MRBC(Br) hybridomas have been studied at the cell level (plaque-forming cells). Phosphorylcholine, choline bromide and p-aminophenyl-trimethylammonium were found to be potent specific inhibitors of plaque formation (anti MRBC(Br)). The hemolytic activities of ascites and tissue culture supernatants were studied and their inhibition by TMA derivatives was determined. Immunoglobulins from ascites purified on TMA immunoadsorbent column were analyzed by two-dimensional gel electrophoresis, their spectrotype was compared to the spectrotype of immunoglobulins from tissue culture supernatants from the same hybridoma radioactively tagged by internal incorporation of [14C]leucine. It could be shown without ambiguity that the PTMA column retained an IgM with the same characteristics as the IgM secreted in vitro.

Animals↗
PubMed1981

The participation of trimethylammonium in the mouse erythrocyte epitope recognized by monoclonal autoantibodies.

Monoclonal IgM anti-erythrocyte autoantibody produced by a NZB-derived hybridoma has been found to react with trimethylammonium-containing compounds. Such compounds are able to prevent the lysis of bromelain-treated mouse erythrocytes (BrMRBC) by those autoantibodies. Using a column of insolubilized betaine hydrazine (BH) the monoclonal anti-erythrocyte antibody has been specifically retained. Elution of this antibody was accomplished by 0.15 M choline (Ch).

Animals↗
PubMed1976

The relationship between storage and secretion of specific antibody by immune lymphoid cells: ultrastructural localization of anti-peroxidase antibodies in plaque-forming cells of the rabbit popliteal lymph node.

The ultrastructure of antibody-forming cells (AFC) has been studied in the lymph node cell population from rabbits locally immunized with horseradish peroxydase (PO) incorporated in complete Freund's adjuvant, and the kinetics of AFC development followed from day 7 to day 18 after one injection of PO. Identification of the AFC was done by local hemolysis assay, using carboxymethyl cellulose solidifying medium and PO-coated sheep erythrocytes. AFC were thereafter transferred by micromanipulation into a Beem capsule, fixed, treated by PO for fine ultrastructure detection of anti-PO antibodies, included, sectioned and studied by electron microscopy. It was found that the AFC were essentially of three categories: lymphocytes, proplasmacytes and plasmacytes, with (+), or without (-), intracellular antibody. The proportion of these categories varies with the time elapsed since the injection of antigen and with the plaque-forming activity of the population: lymphocytes (-) are relatively more numerous (over 30%) at the early stages of immunization (day 7). The number of plasmacytes increases with immunization. If most of them contain intracellular antibody at the early stages (up to day 9), the proportion of plasmacytes (+) decreases markedly afterwards. The cell type distribution is compatible with the idea that the lymphocytes are the precursors of plasmacytes, proplasmacytes being transitional forms, but no direct filiation scheme can actually be deduced from these experiments.

Animals↗
PubMed1975

Formation of plaques by adherent spleen cells from nonimmunized mice, incubated in vitro with the nonadherent fraction of immune allogeneic cells.

Twenty-four-hour incubation of adherent spleen cells from normal C57BL mice with purified nonadherent spleen cells from CBA mice, in the peak of their primary response against sheep red blood cells, resulted in the appearance of plaque-forming cells in the adherent cell population. The immunological nature of these plaques was demonstrated by their complement dependency and the inhibitory effect of specific anti-mouse IgM serum. Direct, but not indirect plaque formation was found to be associated with the adherent cell population for which direct cell to cell contact is necessary. Experiments with cytotoxic specific anti-strain sera revealed that in this system, the adherent plaque-forming cells are the nonimmune adherent C57BL cells.

Animals↗