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Detection of Cell-Bound Immunoglobulins by a Radioisotopic Micro-Mixed Hemadsorption Reaction with Technetium-99m-Labeled Erythrocytes^1,2,

From the Department of Pathology and Oncology, University of Kansas Medical Center, College of Health Sciences and Hospital, Kansas City, Kansas 66103

Abstract

The mixed hemadsorption (MHA) reaction detects antibodies reactive with cell surface antigens by means of antiglobulin-coated indicator erythrocytes. We have developed a radioisotopic modification which employs sheep erythrocytes (SRBC) that have been prelabeled with technetium-99m (^99mTc), a high specific activity metastable -emitter of short half life. The ^99mTc MHA reaction was performed on human and murine cells cultured in Microtest II plates with six replicate wells per serum dilution. Antibody activity in species-specific xenoantisera and mono- and polyspecific alloantisera was detected in high titer. The sensitivity of ^99mTc micro-mixed hemadsorption was 2 times that of the visual assessment of mixed hemadsorption, 100 to 200 times that of the ¹²⁵I-mixed antiglobulin reaction and 500 to 1000 times more sensitive than indirect immunofluorescence. The assay system was applied successfully to confirm the species of origin of a panel of previously karyotyped human and mouse cell lines. Our results indicate that the ^99mTc micro-mixed hemadsorption method is a rapid, sensitive, quantitative test for the detection of cell surface antigens and membrane reactive antibodies.

Footnotes

¹ Presented in part at the 57th Annual Meeting of the Federation of American Societies for Experimental Biology, Atlantic City, New Jersey, April 15 to 20, 1973.

² This investigation was supported by Public Health Service General Research Grant 5 SO4 RRO6147, Grants AI 09947 and CA 13190 from the National Institutes of Health, and DRG 1075 B, Damon Runyon Memorial Fund.

³ Postdoctoral Fellow, United States Public Health Service, 1972–1973.