Expression and temperature-dependent regulation of the beta2-microglobulin (Cyca-B2m) gene in a cold-blooded vertebrate, the common carp (Cyprinus carpio L.).

P N Rodrigues, B Dixon, J Roelofs, J H Rombout, E Egberts, B Pohajdak, R J Stet
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引用次数: 49

Abstract

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.

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冷血脊椎动物鲤鱼β -微球蛋白(Cyca-B2m)基因的表达和温度依赖性调控。
用一种针对Cyca-B2m基因真核表达重组蛋白的多克隆抗体研究了β -微球蛋白(beta2m)在鲤鱼体内的表达。Beta2m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。在脾脏和原肾中,暗阳性和亮阳性人群分别与红细胞、血小板、成熟白细胞或淋巴-髓系未成熟和成熟细胞的存在相关。胸腺细胞是由一个单一的亮阳性群体组成的。Cyca-B2m多克隆抗血清与类似制备的MHC I类(Cyca-UA) α链多克隆抗血清联合使用,研究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类细胞表面持续缺乏表达,转移到12℃后可缓慢恢复。免疫球蛋白分子在B细胞上的表达不受温度变化的影响。尽管在所有温度下Cyca-UA mRNA的水平相当,但I类细胞表面表达的缺失被证明是缺乏足够的Cyca-B2m基因转录的结果。这表明I类表达受Cyca-B2m基因的温度敏感转录调控。
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