The diversity of antigen-specific antibodies in humans and in two xenochimeric SCID mouse models.
We applied two-dimensional gel electrophoresis (2-DE) to study the repertoire of tetanus toxoid (TT)-specific antibodies produced after TT immunization in healthy humans and in severe combined immunodeficient mice xenotransplanted with either human peripheral blood leukocytes (PBLe) or with human adult tonsil (hu-ton) pieces. Specific anti-TT antibodies, as well as total immunoglobulins (Ig), were purified by affinity chromatography on TT-Sepharose or Protein G-Sepharose, respectively. 2-DE unambiguously allowed us to differentiate between the specific humoral responses produced either by humans or by the two xenochimeric mouse models. Anti-TT antibodies produced by humans were polyclonal with a superimposed oligoclonality that was donor-dependent and that did not change upon time. By contrast, immunized hu-PBLe-SCID mice exhibited an evident clonal restriction of the Ig, which increased with time after boosting. Hu-ton-SCID mice showed a clonal diversity which was intermediate between those observed in humans and in hu-PBLe-SCID mice, and which was stable over time. In addition, information was gained by 2-DE, correlating with data obtained by enzyme-linked immunosorbent assay (ELISA), on the isotype composition of the anti-TT IgM response. Altogether, our results clearly demonstrated that the clonal diversity of monospecific antibodies can be appreciated by 2-DE, and that the largest diversity was found in humans when compared to that in xenochimeric models. In addition, mice implanted with pieces of lymphoid organs had the broadest anti-TT Ig diversity, an observation supporting the use of this model for the generation of antibodies with restricted specificity.
Service régional vaudois de transfusion sanguine, Lausanne, Switzerland.
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A
Layer
J D
JD
Tissot
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P
Schneider
M A
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Duchosal
D000818:Animals; D000916:Antibody Diversity; D017690:Cell Transplantation; D002678:Chimera; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005260:Female; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology / Q000378:metabolism*; D007132:Immunoglobulin Isotypes; D007147:Immunoglobulin Light Chains / Q000276:immunology / Q000378:metabolism*; D007136:Immunoglobulins / Q000145:classification / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D051379:Mice; D016513:Mice, SCID; D014066:Palatine Tonsil / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology*; D016378:Tissue Transplantation; D014183:Transplantation, Heterologous
D007143:Immunoglobulin Heavy Chains; D007132:Immunoglobulin Isotypes; D007147:Immunoglobulin Light Chains; D007136:Immunoglobulins; D013745:Tetanus Toxoid