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Rosette Formation of Human Lymphoid Cells with Monkey Red Blood Cells¹

From the Departments of Molecular Immunology and of Immunopathology, Scripps Clinic and Research Foundation, 476 Prospect Street, La Jolla, California 92037

Abstract

Studies of human peripheral lymphocytes (HPL) from healthy subjects and of human lymphoblastoid cell lines (HLC) revealed spontaneous formation of rosettes with Macaca speciosa monkey red blood cells (MRBC). The percentage of HPL forming rosettes with MRBC paralleled the percentage of HPL exhibiting markers ascribed to human B cells. The MRBC-rosetting cells were positive for membrane-bound immunoglobulin (MBIg). Furthermore, by employing a mixed rosette method, lymphocytes carrying both EAC3d^mo and FITC-MRBC were encountered. In contrast, no lymphocytes carrying both SRBC and FITC-MRBC were observed. Ninety-six per cent of purified B cells obtained at the interface after interaction of HPL with SRBC and Ficoll-Isopaque centrifugation formed rosettes with MRBC. In contrast, only 8% of an enriched population of T cells obtained at the interface after interaction with EAC3d^mo and Ficoll-Isopaque centrifugation formed MRBC-rosettes. Incubation of HPL with MRBC and subsequent centrifugation on Ficoll-Isopaque resulted in a highly enriched T cell population at the interface. Several lymphoblastoid cell lines with B cell characteristics, but not two cell lines with T cell characteristics, formed spontaneous rosettes with MRBC. Inhibition experiments showed that the structure on the lymphoid cell surfaces mediating the adherence of MRBC was independent of the MBIg as well as the receptors for IgGFc, C3b, and C3d. It appears that rosetting with MRBC will provide a new detection marker for human cells classified as B type and a technique for obtaining an enriched T cell population.

Footnotes

¹ This is Publication No. 983 from the Departments of Molecular Immunology and of Immunopathology. This investigation was supported by United States Public Health Service Research Grants CA 16071, AI 10180, and AI13154 from the National Institutes of Health, the California Division of the American Cancer Society Senior Fellowship D-221, and the United States Department of the Army Grant DADA 17-73-C-3137.

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