Demonstration of T4, T8, M1 and B7 determinants on human T cells with a rosette test: implications for the assay specificity of monoclonal antibodies |
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Authors: | D G Haegert |
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Affiliation: | Department of Pathology, Montreal General Hospital and Montreal General Hospital Research Institute, Montreal, Quebec, Canada |
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Abstract: | 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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Keywords: | T cells monoclonal antibody rosette test immunofluorescence surface markers Mab, monoclonal antibody E, erythrocyte Ig, immunoglobulin IARR, indirect antiglobulin rosetting reaction IIF, indirect immunofluorescence |
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