Rodrigues P N, Dixon B, Roelofs J, Rombout J H, Egberts E, Pohajdak B, Stet R J
Department of Animal Sciences, Cell Biology and Immunology Group, Wageningen Agricultural University, The Netherlands.
Dev Immunol. 1998;5(4):263-75. doi: 10.1155/1998/15984.
Expression of beta2-microglobulin (beta2m) in the common carp was studied using a polyclonal antibody raised against a recombinant protein obtained from eukaryotic expression of the Cyca-B2m gene. Beta2m is expressed on peripheral blood Ig+ and Ig lymphocytes, but not on erythrocytes and thrombocytes. In spleen and pronephros, dull- and bright-positive populations could be identified correlating with the presence of erythrocytes, thrombocytes, and mature leucocytes or immature and mature cells from the lympho-myeloid lineage, respectively. Thymocytes were shown to be comprised of a single bright-positive population. The Cyca-B2m polyclonal antiserum was used in conjunction with a similarly produced polyclonal antiserum to an MHC class I (Cyca-UA) alpha chain to investigate the expression of class I molecules on peripheral blood leucocytes (PBL) at different permissive temperatures. At 12 degrees C, a temporary downregulation of class I molecules was demonstrated, which recovered to normal levels within 3 days. However, at 6 degrees C, a lasting absence of class I cell-surface expression was observed, which could be restored slowly by transfer to 12 degrees C. The expression of immunoglobulin molecules on B cells was unaffected by temperature changes. The absence of the class I cell-surface expression was shown to be the result of a lack of sufficient Cyca-B2m gene transcription, although Cyca-UA mRNA was present at comparable levels at all temperatures. This suggests that class I expression is regulated by a temperature-sensitive transcription of the Cyca-B2m gene.
利用针对从鲤鱼(Cyca)-B2m基因真核表达获得的重组蛋白产生的多克隆抗体,研究了β2-微球蛋白(β2m)在鲤鱼中的表达。β2m在外周血Ig +和Ig淋巴细胞上表达,但在红细胞和血小板上不表达。在脾脏和前肾中,可以分别鉴定出与红细胞、血小板以及成熟白细胞或淋巴-髓系谱系的未成熟和成熟细胞的存在相关的暗淡阳性和明亮阳性群体。胸腺细胞显示由单一的明亮阳性群体组成。将鲤鱼(Cyca)-B2m多克隆抗血清与同样制备的针对MHC I类(Cyca-UA)α链的多克隆抗血清结合使用,以研究在不同允许温度下外周血白细胞(PBL)上I类分子的表达。在12℃时,I类分子出现暂时下调,3天内恢复到正常水平。然而,在6℃时,观察到I类细胞表面表达持续缺失,转移到12℃后可缓慢恢复。B细胞上免疫球蛋白分子的表达不受温度变化影响。尽管在所有温度下Cyca-UA mRNA水平相当,但I类细胞表面表达的缺失被证明是由于缺乏足够的鲤鱼(Cyca)-B2m基因转录所致。这表明I类表达受鲤鱼(Cyca)-B2m基因温度敏感转录的调节。