Lillehoj H S, Kim S, Lillehoj E P, Bacon L D
USDA, Protozoan Diseases Laboratory, Beltsville, Maryland 20705.
Poult Sci. 1988 Nov;67(11):1525-35. doi: 10.3382/ps.0671525.
A monoclonal antibody (MAb), designated P2M11, that detects a monomorphic determinant of chicken class II antigens was produced from the fusion of P3X63 myeloma cells with spleen cells from BALB/c mice immunized with chicken splenic lymphocytes. Flow cytometric analyses of lymphocytes from the SC and FP strains of chickens showed 30 to 50% staining of bursa cells, 15 to 20% staining cells, and less than 5% staining of thymus cells. Addition of MAb P2M11 to splenic of T cell cultures stimulated with allogeneic cells or concanavalin A resulted in a significant inhibition of the T cell proliferation responses. Immunoprecipitation of 35S-methionine-labeled spleen cell extracts using MAb P2M11 identified molecules with apparent molecular weights of approximately 28,000, 30,000, and 32,000 by sodium dodecyl-polyacryl-amide gel electrophoresis. Taken together, these data indicate that the antigens detected by MAb P2M11 are similar in cell distribution and structure to chicken Ia antigens encoded by B-L genes. Using this MAb, a strain difference was demonstrated in the tissue distribution of Ia antigen positive lymphocytes in the spleens but not the thymuses of 15I5-B congenic and inbred strains of chickens. This difference may be due to the genes associated with B-complex genes.
一种名为P2M11的单克隆抗体,它能检测鸡Ⅱ类抗原的一个单态决定簇,是通过将P3X63骨髓瘤细胞与用鸡脾淋巴细胞免疫的BALB/c小鼠的脾细胞融合产生的。对鸡的SC和FP品系淋巴细胞进行的流式细胞术分析显示,法氏囊细胞有30%至50%被染色,染色细胞为15%至20%,胸腺细胞染色不到5%。将单克隆抗体P2M11添加到用同种异体细胞或伴刀豆球蛋白A刺激的脾T细胞培养物中,会导致T细胞增殖反应受到显著抑制。使用单克隆抗体P2M11对35S-甲硫氨酸标记的脾细胞提取物进行免疫沉淀,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳鉴定出表观分子量约为28,000、30,000和32,000的分子。综合这些数据表明,单克隆抗体P2M11检测到的抗原在细胞分布和结构上与由B-L基因编码的鸡Ia抗原相似。使用这种单克隆抗体,在15I5-B同源近交系鸡的脾脏而非胸腺中,Ia抗原阳性淋巴细胞的组织分布存在品系差异。这种差异可能归因于与B复合体基因相关的基因。