A SIMPLE METHOD FOR EVALUATING THE REJECTION OF GRAFTED SPLEEN-CELLS BY FLOW-CYTOMETRY AND TRACING ADOPTIVELY TRANSFERRED CELLS BY LIGHT-MICROSCOPY

In this report we describe a simple method using the vital dye 5- (and 6-) carboxyfluorescein diacetate succinimidyl ester (CFSE) to follow splenic graft rejection by flow cytometry. CFSE-labelled spleen cell s uspensions were injected intravenously into various recipients and blo od samples were taken at different time points to follow the transferr ed cells. We found that the labelled cells could be readily detected b y flow cytometry for up to eleven weeks. The loss of these labelled ce lls in various transfusion experiments with different major and minor histocompatibility differences followed the rejection kinetics previou sly described for skin transplants. Thus, this method offers a simple tool to test the histocompatibility of donor cells/grafts with the hos t in adoptive/transplantation experiments in which donor and host are not completely syngeneic. Furthermore, we developed a method to trace adoptively transferred fluorescent CFSE-labelled cells by light micros copy by converting in situ immunofluorescence staining into immunoenzy me staining. (C) 1997 Elsevier Science B.V.