Description and comparison of three in vitro methods for the detection of guinea pig MHC antigens

Two methods (an immunoenzymatic and a cell dot immunobinding assay) are described and compared with a reference protein A rosetting assay, for the detection of guinea pig class I and class II histocompatibility antigens. The main requirement of these three assays for obtaining good results is the use of whole fresh target cells since all tested experimental conditions for fixing cellular antigens (glutaraldehyde, formaldehyde, paraformaldehyde), as well as for obtaining cell monolayers (adherence, desiccation, poly-L-lysine) presented several drawbacks and were unsuccessful in our hands. The comparison between the three methods shows that the immunoenzymatic assay gives good results but that it is less reproducible than the other two. The cell dot immunobinding is highly sensitive, reproducible and economical. It can be used for genetic typing and alloantigen analysis of guinea-pig lines, instead of the rosetting method, which is highly reliable but time consuming.