MITOGENIC LECTINS BIND TO THE ANTIGEN RECEPTOR ON HUMAN-LYMPHOCYTES
MITOGENIC LECTINS BIND TO THE ANTIGEN RECEPTOR ON HUMAN-LYMPHOCYTES
复制标题
DOI：
10.1002/eji.1830190225
复制
发表时间：
1989-02-01
影响因子：
5.4
通讯作者：
KELLYCHILSON, AE
中科院分区：
文献类型：
--
作者：
CHILSON, OP;KELLYCHILSON, AE
探讨了有丝分裂凝集素和非有丝分裂凝集素与二硫键连接的细胞表面受体在人淋巴细胞上相互作用的特异性。用凝集素-琼脂糖衍生物(或牛血清白蛋白琼脂糖凝胶)吸附表面放射性碘标记的扁桃体淋巴细胞和T淋巴母细胞(HPB-ALL)的裂解物(NONIDT-P40)，或用适当的单抗进行免疫沉淀。用十二烷基硫酸钠-聚丙烯酰胺双向(未还原/还原)凝胶电泳法分析凝集素洗脱物和增溶免疫沉淀物。放射性标记多肽通过放射自显影显影。在各种凝集素结合多肽中，豌豆凝集素、刀豆凝集素A和扁豆凝集素分别与豌豆凝集素、刀豆蛋白A和小扁豆凝集素结合，而L的白血球凝集素和菜豆的红血球凝集素又与菜豆的植物血凝素结合。商陆凝集素只识别II和III。这些分子与花生凝集素、大豆凝集素、紫云英凝集素-I、扁豆凝集素、苦参凝集素有弱结合，但不与Helix pomatia凝集素或BSA-琼脂糖结合。杂二聚体II(82-88 kDa)由50-55 kDa和40-43 kDa亚基组成，可能代表α/β亚基。T细胞抗原受体(TCRα/β)。异源二聚体III(-72 kDa)由41 kDa和37 kDa亚基组成，可能代表TcR.同源二聚体I(120-130 kDa)和IV(55-61 kDa)分别由55-60 kDa和30 kDa多肽组成，显然以前没有描述过。针对HPB-ALL细胞上抗原受体的单抗H1-2D4和E-PHA与共同分子相互作用的证据包括：(A)从N-乙酰半乳糖胺洗脱的不溶凝集素的糖肽部分中与H1-2D4免疫沉淀TCR；以及(B)E-PHA-琼脂糖从溶解的H1-2D4免疫沉淀物中吸附TCR。E-PHA对CD3的识别是通过UCHT1从E-PHA特异性洗脱的糖肽部分免疫沉淀CD3蛋白来指示的。这一结果与有丝分裂凝集素与人淋巴细胞上某些二硫键连接分子相互作用的观点是一致的，包括TCRα/β。也许还有TcR。虽然一些非促有丝分裂的凝集素也能识别这些受体，但这种相互作用的亲和力很低。
The specificity of interactions between mitogenic and non-mitogenic lectins and disulfide-linked cell surface receptors on human lymphocytes was explored. Lysates (Nonidet-P40) of surface-radioiodinated tonsil lymphocytes and T lymphoblastoid cells (HPB-ALL) wer absorbed with lectin-agarose derivatives (or bovine serum albumin, BSA-agarose) or immunoprecipitated with appropriate monoclonal antibodies (mAb). Lectin eluates and solubilized immunoprecipitates were analyzed by two-dimensional (nonreduced/reduced) sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Radiolabeled polypeptides were visualized by autoradiography. Among the various lectin-binding polypeptides, two disulfide-linked heterodimers (II and III) and two apparent homodimers (I and IV) are bound by pea lectin, concanavalin A and lentil lectin on tonsil lymphocytes; II, III and IV were bound both leukoagglutinating (L)- and erythroagglutinating (E)-phytohemagglutinins from Phaseolus vulgaris (PHA). Pokeweed mitogen recognize only II and III. These molecules are weakly bound by peanut agglutinin, soybean agglutinin; Ulex europaeus agglutinin-I, Dolichos biflorus agglutinin, Vicia villosa agglutinin and Sophora japonica agglutinin, but are not bound by Helix pomatia agglutinin or BSA-agarose. Heterodimer II (82-88 kDa), comprised of 50-55-kDa and 40-43 kDa subunits, probably represents the .alpha./.beta. T cell antigen receptor (TcR .alpha./.beta.). Heterodimer III (64-72 kDa), comprised of 41-kDa and 37-kDa subunits, may represent TcR.gamma.. The homodimers, I (120-130 kDa) and IV (55-61 kDa), comprised of 55-60 kDa and 30-kDa polypeptides, respectively, have apparently not been previously described. Evidence that H1-2D4, a mAb directed against the antigen receptor on HPB-ALL cells, and E-PHA interact with a common molecule includes; (a) immunoprecipitation of TcR with H1-2D4 from the glycopeptide fraction specifically eluted from insolubilized lectin with N-acetylgalactosamine; and (b) adsorption of TcR from a solubilized H1-2D4 immunoprecipitate by E-PHA-agarose. Recognition of CD3 by E-PHA is indicated by immunoprecipitation of CD3 protein by UCHT1 from the glycopeptide fraction specifically eluted from E-PHA. The results are consistent with the view that mitogenic lectins interact with certain disulfide-linked molecules on human lymphocytes, including the TcR .alpha./.beta. and perhaps TcR.gamma.; while some nonmitogenic lectins also recognize these receptors, the interactions is of low affinity.