Kaufman J F, Flajnik M F, Du Pasquier L
J Immunol. 1985 May;134(5):3258-64.
The class II antigens from four inbred strains of Xenopus laevis (r, f, g, and j haplotypes) and six gynogenetic LG clones (two Xenopus laevis, two Xenopus gilli haplotypes) with functionally well-defined MHC types have been immunoprecipitated with the rabbit anti-human class II beta-chain serum anti-p29boost and analyzed by two-dimensional gel electrophoresis. The glycosylated material from 15-hr biosynthetically labeled cells runs as a broad fuzzy band around 33kD that, upon removal of N-linked glycans by Endo F, resolves into upper beta-chain bands and lower alpha-chain bands. Both the glycosylated and deglycosylated class II antigens give rise to multiple IEF spots in evenly spaced arrays (alpha-chain: two to eight spots in one to three arrays, beta-chain: two to 12 spots in one to five arrays). Both chains are polymorphic and both map to the functionally defined MHC. The large number of spots argues for multiple class II antigens; by radioactive N-terminal sequencing, two homologous alpha-chains and five beta-chains are present in the f haplotype. By comparison with MHC-linked alloantisera, anti-p29boost recognizes all major polymorphic class II molecules in Xenopus laevis. A selection of outbred animals were typed by using an IEF procedure requiring only a million PHA-stimulated blood cells.
来自非洲爪蟾四个近交系（r、f、g和j单倍型）以及六个雌核发育LG克隆（两个非洲爪蟾、两个非洲姬蛙单倍型）且具有功能明确的MHC类型的II类抗原，已用兔抗人II类β链血清抗-p29boost进行免疫沉淀，并通过二维凝胶电泳进行分析。来自15小时生物合成标记细胞的糖基化物质在33kD左右呈现为一条宽的模糊带，在用内切糖苷酶F去除N-连接聚糖后，可分解为上部的β链带和下部的α链带。糖基化和去糖基化的II类抗原在均匀间隔的阵列中都产生多个IEF斑点（α链：在一到三个阵列中有两到八个斑点，β链：在一到五个阵列中有两到十二个斑点）。两条链都是多态性的，并且都定位于功能明确的MHC。大量的斑点表明存在多种II类抗原；通过放射性N端测序，f单倍型中存在两条同源α链和五条β链。通过与MHC连锁的同种抗血清比较，抗-p29boost识别非洲爪蟾中所有主要的多态性II类分子。通过使用仅需要一百万个PHA刺激的血细胞的IEF程序对一组远交动物进行分型。
