Serological and Immunogenic Activity of Soluble Mouse Transplantation Antigens Controlled by the H -2 Locus

Abstract

The solubilization and partial purification of mouse transplantation antigens were monitored by (1) an in vitro assay for alloantigenic specificities and (2) an in vivo assay for transplantation antigens controlled by the H-2 histocompatibility locus. Antigens from A/J mice were solubilized by papain and fractionated on a Sephadex G-150 column. The eluate showed a 280 nm absorbance peak (F(1)) in the excluded volume and two peaks (F(2) and F(3)) in the included volume. H-2 specificities 1, 3, 4, 5, 11, 23, and 28 were confined to a single peak in the F(2) fraction. Fractions F(1), F(2), and F(3), were tested for their ability to accelerate skin graft rejection in noncoisogenic strains which differ at both H-2 and non-H-2 loci, and coisogenic strains which differ only at the H-2 locus. All fractions produced significant acceleration of graft rejection in the noncoisogenic strains, but only fraction F(2) produced significant acceleration in the coisogenic strains. These findings indicate that H-2 transplantation antigens detected by our in vivo assay, and H-2 alloantigenic specificities detected by our in vitro assay are solubilized by papain and are eluted in the same peak during Sephadex fractionation.