An Indirect Anti-immunoglobulin Rosetting Reaction to detect alloantibodies to human lymphocytes

A test is reported which detects alloantibody, absorbed onto the surface of human lymphocytes from multiparous antisera, by means of a red cell rosette assay. The red cells, trypsinised ox, are coupled with anti-immunoglobulin using chromic chloride. The antiglobulin used is rabbit anti-human IgG (Fc), chosen to avoid reaction with the surface immunoglobulin naturally present on human B lymphocytes. The reaction is termed the Indirect Anti-immunoglobulin Rosetting Reaction (IARR). The IARR is shown to be specific in the following ways: anti-immunoglobulin coupled ox cells do not react with normal human lymphocytes nor with lymphocytes treated with non-reactive serum. Red cells coupled with normal rabbit IgG do not react with normal or alloantibody coated lymphocytes. Multiparous sera (reactive with other individuals) do not react with cells of the serum donor in the IARR. Finally, the coupled red cells do not usually react with lymphocytes which have absorbed immune complexes onto their Fc receptors. The IARR is shown to be more sensitive than a standard cytotoxic test for detection of alloantibody. Several possible applications of the IARR are discussed.