Spectrophotometric determination of lymphocyte mediated sheep red blood cell hemolysis in vitro

Simpson, M. A.; Gozzo, J. J.

An assay was developed in which the degree of in vitro spleen cell-mediated sheep red blood cell (SRBC) hemolysis was quantitated spectrophotometrically (QHS). Immune mouse spleen cells, SRBC and guinea pig complement were incubated at 37.degree. C for 1 h in buffer, centrifuged and the supernatant solutions read at 413 nm. The greatest degree of cell-mediated SRBC hemolysis was observed in day +5 harvested spleens following a day 0, 5% SRBC i.p. injection. A stimulator SRBC dose of 0.2% resulted in maximal stimulation with little non-specific hemolysis. This assay was presented as an alternative to the well established plaque forming and rosette assays.