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人类CD45RC特异性。不同成熟和激活阶段T细胞的新型标志物。

Human CD45RC specificity. A novel marker for T cells at different maturation and activation stages.

作者信息

Zapata J M, Pulido R, Acevedo A, Sánchez-Madrid F, de Landázuri M O

机构信息

Servicio de Immunologia, Hospital de la Princess, Universidad Autónoma de Madrid, Spain.

出版信息

J Immunol. 1994 Apr 15;152(8):3852-61.

PMID:8144955
Abstract

The different isoforms of CD45 seem to play an important role in thymocyte maturation and T cell activation and function. To investigate the particular contribution of CD45-exon C (exon 6) to human T cell development, we first investigated the expression of the CD45-exon C isoforms on different human lymphoid cell populations. A wide panel of mAbs against the different isoforms of CD45 was generated by immunizing mice with purified human CD45. Two of these mAbs (RP1/12 and RP2/19) selectively recognized mouse 300-19 pre-B cells that had been transfected with CD45 cDNA that contained exon C, suggesting that epitopes recognized by these mAbs are dependent on the expression of exon C. Immunofluorescence analysis of PBLs showed that CD45RC is expressed at high levels on B, NK, and CD8+ T cells, although a subset of CD4+ T cells was found to be negative. Activation of PBMCs and purified NK cells resulted in down-regulation of CD45RC, which took place in a coordinate manner involving both down-regulation of the expression of CD45RA and CD45RB and up-regulation of CD45R0. Two-color immunofluorescence analyses showed that thymocytes expressing CD45RC were CD1-to lo, CD3hi and HLA class Ihi, typical markers of mature thymocytes. Accordingly, immunohistochemical analysis of a normal thymus showed that whereas the anti-CD45RA mAb stained only a subset of medullary thymocytes, the anti-CD45RC mAb stained the majority of medullary thymocytes. Thymocytes that bear CD45RA also express CD45RC, but do not express CD45RO; a subset of CD45 RA- RO+ thymocytes was also detected. Immunoprecipitation of CD45 molecules from both PBMC and thymocyte cell lysates with mAbs against the different specificities of this molecule revealed that the RC determinants are found on CD45 polypeptides of 220 and 205 kDa that also bear RB or both RB and RA determinants, respectively. Together these results support both the existence of a tight regulation of the expression of CD45 specificities during lymphocyte activation and thymocyte maturation and also that CD45RC is selectively expressed by mature medullary thymocytes during T cell ontogeny.

摘要

CD45的不同同工型似乎在胸腺细胞成熟、T细胞激活及功能中发挥重要作用。为研究CD45外显子C(外显子6)对人类T细胞发育的特殊贡献,我们首先研究了CD45外显子C同工型在不同人类淋巴细胞群体上的表达。通过用纯化的人类CD45免疫小鼠,产生了一系列针对CD45不同同工型的单克隆抗体。其中两种单克隆抗体(RP1/12和RP2/19)选择性识别已转染含外显子C的CD45 cDNA的小鼠300-19前B细胞,这表明这些单克隆抗体识别的表位依赖于外显子C的表达。外周血淋巴细胞的免疫荧光分析显示,CD45RC在B细胞、NK细胞和CD8+ T细胞上高水平表达,尽管发现一部分CD4+ T细胞为阴性。外周血单个核细胞和纯化的NK细胞的激活导致CD45RC下调,这是以一种协同方式发生的,涉及CD45RA和CD45RB表达的下调以及CD45R0表达的上调。双色免疫荧光分析显示,表达CD45RC的胸腺细胞为CD1低至阴性、CD3高表达且HLA I类高表达,这些是成熟胸腺细胞的典型标志物。因此,对正常胸腺的免疫组织化学分析显示,抗CD45RA单克隆抗体仅染色髓质胸腺细胞的一部分,而抗CD45RC单克隆抗体染色大多数髓质胸腺细胞。携带CD45RA的胸腺细胞也表达CD45RC,但不表达CD45RO;还检测到一部分CD45 RA- RO+胸腺细胞。用针对该分子不同特异性的单克隆抗体对来自外周血单个核细胞和胸腺细胞裂解物的CD45分子进行免疫沉淀,结果显示,RC决定簇存在于分别也携带RB或同时携带RB和RA决定簇的220 kDa和205 kDa的CD45多肽上。这些结果共同支持在淋巴细胞激活和胸腺细胞成熟过程中存在对CD45特异性表达的严格调控,以及在T细胞个体发育过程中成熟髓质胸腺细胞选择性表达CD45RC。

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