Timpl R, Glanville R W, Wick G, Martin G R
Immunology. 1979 Sep;38(1):109-16.
Basement membrane (type IV) collagens were extracted from a mouse tumour with acetic acid and from human placenta after limited enzymatic digestion. Antisera were produced against both collagens in rabbits and guinea-pigs and examined by various assays. These antisera were found to be specific for basement membrane collagen and showed little or no cross-reactions with the interstitial collagens, types I, II and III or with human placenta collagen consisting of alpha A and alpha B chains. Varying degrees of cross-reaction were observed between antisera to human and mouse type IV collagen. Immunochemical analyses demonstrated the presence of three distinct determinants in the tumour type IV collagen. Rabbit antisera against this antigen reacted with either collagenase-resistant segments or with a collagenous, disulphide-bonded segment (P3). Guinea-pig antisera recognized primarily antigenic determinants in the P3 segment. Antisera to placenta type IV collagen reacted with another collagenous, pepsin fragment (P1) which lacks disulphide bonds. These antisera showed complete cross-reaction with collagenous alpha 1 (IV) chains prepared from pepsin-digests of human placenta and bovine lens capsule.
用乙酸从小鼠肿瘤中提取基底膜(IV型)胶原蛋白,并在有限的酶消化后从人胎盘中提取。用兔和豚鼠制备针对这两种胶原蛋白的抗血清,并通过各种检测方法进行检测。发现这些抗血清对基底膜胶原蛋白具有特异性,与I、II和III型间质胶原蛋白或由αA和αB链组成的人胎盘胶原蛋白几乎没有交叉反应。在针对人IV型和小鼠IV型胶原蛋白的抗血清之间观察到不同程度的交叉反应。免疫化学分析表明肿瘤IV型胶原蛋白中存在三种不同的决定簇。针对该抗原的兔抗血清与抗胶原酶片段或与胶原性二硫键结合片段(P3)反应。豚鼠抗血清主要识别P3片段中的抗原决定簇。针对胎盘IV型胶原蛋白的抗血清与另一个缺乏二硫键的胶原性胃蛋白酶片段(P1)反应。这些抗血清与从人胎盘和牛晶状体囊的胃蛋白酶消化物中制备的胶原性α1(IV)链完全交叉反应。