Demonstration of T4, T8, M1 and B7 determinants on human T cells with a rosette test: implications for the assay specificity of monoclonal antibodies.
Under optimal test conditions significantly more freshly isolated human T cells reacted with OKT4, OKT8, OKM1 and OKB7 monoclonal antibodies (Mabs) in the indirect antiglobulin rosetting reaction (IARR) than by indirect immunofluorescence. Rabbit erythrocytes (E) coated with anti-mouse immunoglobulin were more sensitive indicator cells in the IARR than similarly coated sheep E. Treatment of T cells with neuraminidase further enhanced T cell reactivity in the IARR with each Mab so that an average of 60% or more of T cells were T4+, T8+ and M1+ and at least 40% had the T4+ T8+ phenotype. The various findings suggest that the rosette assay detects determinants on T cells that are expressed below the detection threshold of immunofluorescence. Moreover, these findings indicate that the cellular specificities of a particular Mab may change when one assay system is substituted for another or when the protocol of a particular assay is altered.
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000939:Epitopes / Q000276:immunology; D005069:Evaluation Studies as Topic; D005455:Fluorescent Antibody Technique; D006801:Humans; D009439:Neuraminidase / Q000494:pharmacology; D011817:Rabbits / Q000097:blood; D012397:Rosette Formation; D012756:Sheep / Q000097:blood; D013601:T-Lymphocytes / Q000145:classification* / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009439:Neuraminidase