Koumans-van Diepen J C, Egberts E, Peixoto B R, Taverne N, Rombout J H
Department of Experimental Animal Morphology and Cell Biology, Agricultural University, Wageningen, The Netherlands.
Dev Comp Immunol. 1995 Jan-Feb;19(1):97-108. doi: 10.1016/0145-305x(94)00061-j.
B cell and immunoglobulin (Ig) heterogeneity was demonstrated in carp, Cyprinus carpio L., using two monoclonal antibodies (MAbs; WC14, WCI12) produced against carp serum Ig. Immunochemical results showed that both WCI4 and WCI12 react with a protein determinant on the heavy chain of Ig (relative molecular mass approximately 70,000). Immunofluorescence microscopic and flow cytometric analyses of lymphoid cells suggest three distinct subpopulations of B cells and plasma cells: WCI4+12- cells, WCI4-12+ cells, and WCI4+12+ cells. WCI4-12+ and WCI4+12+ anti-DNP antibody-secreting cells were also demonstrated with the ELISPOT assay in pronephros and spleen cell suspensions from primary immunised carp. Affinity chromatography of carp serum and sequential immunoprecipitation of 125I-labelled peripheral blood leucocyte (PBL) membrane proteins only indicated the presence of two antigenically different Ig molecules, i.e., WCI4-12+ and WCI4+12+ molecules. WCI4+12- molecules could not be detected by affinity chromatography or immunoprecipitation. During ontogeny, a shift in percentages of WCI4+12- and WCI4-12+ cells was found in the spleen and the pronephros. In these organs, WCI4+12- cells formed the majority of B cells at 2 weeks of age, but the percentages of this cell type decreased during ontogeny. On the other hand, the percentages of WCI4-12+ cells increased during development, and these cells became the major population of B cells from 13 weeks onward. The proportion of WCI4+12+ cells remained almost constant during ontogeny. The distribution of B cell subpopulations in blood was more or less stable at all ages. The functional significance of Ig heterogeneity in fish and in particular carp is discussed.
利用两种针对鲤鱼血清免疫球蛋白（Ig）产生的单克隆抗体（MAbs；WC14、WCI12），在鲤鱼（Cyprinus carpio L.）中证实了B细胞和免疫球蛋白（Ig）的异质性。免疫化学结果显示，WC14和WCI12均与Ig重链上的一个蛋白质决定簇发生反应（相对分子质量约为70,000）。对淋巴细胞进行免疫荧光显微镜和流式细胞术分析，提示B细胞和浆细胞存在三个不同的亚群：WC14+12-细胞、WC14-12+细胞和WC14+12+细胞。用ELISPOT分析法在初次免疫的鲤鱼的前肾和脾细胞悬液中也证实了WC14-12+和WC14+12+抗二硝基苯酚（DNP）抗体分泌细胞的存在。鲤鱼血清的亲和层析和125I标记的外周血白细胞（PBL）膜蛋白的顺序免疫沉淀仅表明存在两种抗原性不同的Ig分子，即WC14-12+和WC14+12+分子。通过亲和层析或免疫沉淀无法检测到WC14+12-分子。在个体发育过程中，在脾脏和前肾中发现WC14+12-和WC14-12+细胞的百分比发生了变化。在这些器官中，WC14+12-细胞在2周龄时构成了B细胞的大多数，但这种细胞类型的百分比在个体发育过程中下降。另一方面，WC14-12+细胞的百分比在发育过程中增加，并且从13周龄起这些细胞成为B细胞的主要群体。WC14+12+细胞的比例在个体发育过程中几乎保持不变。B细胞亚群在血液中的分布在所有年龄段都或多或少是稳定的。文中讨论了鱼类尤其是鲤鱼中Ig异质性的功能意义。
