Strober S, Appella E, Law L W
Proc Natl Acad Sci U S A. 1970 Oct;67(2):765-72. doi: 10.1073/pnas.67.2.765.
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.
通过(1)同种抗原特异性的体外测定和(2)由H-2组织相容性基因座控制的移植抗原的体内测定,监测小鼠移植抗原的溶解和部分纯化。来自A/J小鼠的抗原用木瓜蛋白酶溶解,并在葡聚糖G-150柱上分级分离。洗脱液在排阻体积中显示出280nm吸光度峰(F(1)),在包含体积中有两个峰(F(2)和F(3))。H-2特异性1、3、4、5、11、23和28局限于F(2)级分中的单个峰。测试了F(1)、F(2)和F(3)级分在H-2和非H-2基因座均不同的非同基因品系以及仅在H-2基因座不同的同基因品系中加速皮肤移植排斥的能力。所有级分在非同基因品系中均显著加速了移植排斥,但只有F(2)级分在同基因品系中显著加速了移植排斥。这些发现表明,我们的体内测定检测到的H-2移植抗原和我们的体外测定检测到的H-2同种抗原特异性被木瓜蛋白酶溶解,并在葡聚糖分级分离过程中在同一峰中洗脱。