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A Biochemical Analysis of the Membrane-Associated Gene Products of Major Histocompatibility Complex of the Rat¹

From the Wistar Institute of Anatomy and Biology, 36th Street at Spruce, Philadelphia, Pennsylvania 19104

Abstract

Thus far, the rat major histocompatibility complex (MHC), AgB or RTI, have been divided by recombinational analysis into two regions, AgB.A (RTI.A) coding for transplantation antigens (class I gene products) and AgB.B (RTI.B) controlling the humoral immune response (Ir genes) and the proliferative response to allogeneic cells (MLR, class II genes). In this study, an attempt was made by biochemical analysis to determine whether there exists more than one AgB.A (class I) gene product, as has been found in the mouse (H-2K and H-2D) and in the human (HLA.A and HLA.B antigens). NP40-solubilized ³H-fucose-labeled rat MHC membrane products were immunoprecipitated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Firstly, with a congenic anti-erythrocyte serum, only membrane structures with an approximate m.w. of 45,000 (AgB.A) were precipitated. A congenic anti-lymphocyte serum absorbed with erythrocytes precipitated antigens with an apparent m.w. of 33,000 and 28,000 (AgB.B) whereas the unabsorbed serum precipitated in addition a 45,000-dalton component (AgB.A). Secondly, sequential treatment of a DA (AgB4, RTI^a) extract with a strongly cross-reactive serum followed by an (Lewis x BN.B2)F₁ anti-AgB4-specific serum demonstrated that the AgB4 haplotype codes for at least two AgB.A and at least two AgB.B gene products.

Footnotes

¹ This work was supported by United States Public Health Service Research Grants CA-07973, CA-10097, and CA-10815 from the National Cancer Institute and Grant RR-05540 from the Division of Research Resources.