Wolf H, Hála K, Boyd R L, Wick G
Eur J Immunol. 1984 Sep;14(9):831-9. doi: 10.1002/eji.1830140912.
Surface antigens on chicken thymus and bursa cells were analyzed by immunoprecipitation using polyclonal and monoclonal antisera raised against (and specific for) thymus (ATS) or bursa (ABS) cells, respectively. The antigens identified were compared with those governed by the B-F, B-L and B-G regions of the chicken major histocompatibility complex (B complex). Four proteins were precipitated from thymus cells by 2 polyclonal ATS: both antisera recognized molecules of apparent molecular mass of 172-182, 132-135, 75-76 kDa, and one antiserum in addition recognized a protein of 102 kDa. The 172-182 and 102-kDa peaks were still demonstrable under reducing conditions indicating that they are composed of a single polypeptide chain, the other 2 were lost under reducing conditions, therefore, must be composed of smaller subunits. Of the 2 monoclonal ATS tested, one identified a single protein of 186 kDa and the other a 135-kDa protein (in addition to 2 smaller molecules); whether these are the same as those precipitated by the polyclonal antisera remains to be determined as they behaved differently under reducing conditions. Proteins of 162 and 78-84 kDa were revealed by 2 polyclonal ABS under nonreducing conditions but the former may in one case be a polymer (it disappeared under reducing conditions) and in the other a single molecule. In addition molecules of 182 kDa were identified by one antiserum and of 84 and 60 kDa by the other under nonreducing conditions. Of the 4 monoclonal ABS only one identified a 200-kDa protein: molecules of 115-125, 90-100, 48-52 and 40-43 kDa were also precipitated, all of which were reduced to smaller molecules. With 2 specific anti-B-F alloantisera we were able to precipitate the "conventional" B-F antigen from red blood cell lysates of CB-strain chickens resolving into a 40-kDa peak and a light chain of about 12 kDa corresponding to beta 2 microglobulin. Precipitates from peripheral blood lymphocytes, bursa and thymus cells revealed an additional protein of 22 kDa. With 2 specific B-L alloantisera two peaks of 33 kDa and 31 kDa were obtained from peripheral blood lymphocytes. Using anti-B-G alloantisera a double band corresponding to 47 and 42 kDa was seen under reducing conditions. There is no evidence from these data to indicate that the polyclonal and monoclonal antibodies are directed towards major histocompatibility complex antigens.
分别使用针对胸腺细胞（ATS）或法氏囊细胞（ABS）产生的多克隆和单克隆抗血清，通过免疫沉淀法分析鸡胸腺和法氏囊细胞上的表面抗原。将鉴定出的抗原与鸡主要组织相容性复合体（B复合体）的B-F、B-L和B-G区域所控制的抗原进行比较。两种多克隆ATS从胸腺细胞中沉淀出四种蛋白质：两种抗血清均识别表观分子量为172 - 182、132 - 135、75 - 76 kDa的分子，另外一种抗血清还识别一种102 kDa的蛋白质。在还原条件下，172 - 182和102 kDa的峰仍可检测到，表明它们由单条多肽链组成，另外两种在还原条件下消失，因此必定由较小的亚基组成。在所测试的两种单克隆ATS中，一种鉴定出一种186 kDa的单一蛋白质，另一种鉴定出一种135 kDa的蛋白质（以及另外两种较小的分子）；由于它们在还原条件下表现不同，这些蛋白质是否与多克隆抗血清沉淀出的蛋白质相同仍有待确定。两种多克隆ABS在非还原条件下揭示出162和78 - 84 kDa的蛋白质，但前者在一种情况下可能是聚合物（在还原条件下消失），在另一种情况下是单分子。此外，一种抗血清在非还原条件下鉴定出182 kDa的分子，另一种抗血清鉴定出84和60 kDa的分子。在四种单克隆ABS中，只有一种鉴定出一种200 kDa的蛋白质：还沉淀出115 - 125、90 - 100、48 - 52和40 - 43 kDa的分子，所有这些分子在还原条件下都还原为较小的分子。使用两种特异性抗B-F同种异体抗血清，我们能够从CB品系鸡的红细胞裂解物中沉淀出“常规”的B-F抗原，分解为一个40 kDa的峰和一条约12 kDa的轻链，对应于β2微球蛋白。外周血淋巴细胞、法氏囊和胸腺细胞的沉淀物显示出一种额外的22 kDa蛋白质。使用两种特异性B-L同种异体抗血清，从外周血淋巴细胞中获得了33 kDa和31 kDa的两个峰。使用抗B-G同种异体抗血清，在还原条件下观察到对应于47和42 kDa的两条带。这些数据没有证据表明多克隆和单克隆抗体针对主要组织相容性复合体抗原。
