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

An assay has been developed in which the degree of in vitro spleen cell-mediated sheep red blood cell (SRBC) hemolysis is quantitated spectrophotometrically (QHS). Immune mouse spleen cells, SRBC and guinea pig complement were incubated at 37 degrees 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. In the assay system, it was found that a stimulator SRBC dose of 0.2% resulted in maximal stimulation with little non-specific hemolysis. This assay is presented as an alternative to the well established plaque forming and rosette assays.