Goto-Koshino Yuko, Tomiyasu Hirotaka, Suzuki Hiyoshi, Tamamoto Takashi, Mizutani Noriyuki, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime
Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Vet Immunol Immunopathol. 2014 Jul 15;160(1-2):118-22. doi: 10.1016/j.vetimm.2014.03.011. Epub 2014 Mar 28.
CD45 is one of the most abundant molecules expressed on the white blood cell surface in various mammals. In this study, we investigated the differential expression of CD45 isoforms in normal canine white blood cells. It has been shown that all canine nucleated blood cells express CD45. We characterized two major isoforms of canine CD45 derived from alternative splicing: a higher molecular weight isoform, CD45RA, and a lower molecular weight isoform, CD45RO. The nucleotide sequences of the two isoforms were identical, except for the region corresponding to a part in the extracellular domain. Flow cytometry analysis using an antibody that recognizes CD45RA, but not CD45RO, revealed that granulocytes did not express CD45RA, and monocytes express low levels of CD45RA. We further analyzed the expression levels of CD45RA in each lymphocyte subpopulation and found that the expression of CD45RA on CD21+ B cells was uniform. On the other hand, expression of CD45RA on CD3+ T cells was variable. Upon stimulation of lymphocytes with Con A, the CD45RA+ fraction increased, indicating that not only the phenotypes but also the activation status influences the isoform expression pattern of CD45. Our finding provides a basic knowledge of the expression of canine CD45, which could be a tool to study lymphocytes with various phenotypes, developmental stages, and activation status.
CD45是各种哺乳动物白细胞表面表达最为丰富的分子之一。在本研究中,我们调查了正常犬类白细胞中CD45异构体的差异表达。已表明所有犬类有核血细胞均表达CD45。我们鉴定了源自可变剪接的犬类CD45的两种主要异构体:一种分子量较高的异构体CD45RA,以及一种分子量较低的异构体CD45RO。除了对应于细胞外结构域一部分的区域外,这两种异构体的核苷酸序列相同。使用识别CD45RA但不识别CD45RO的抗体进行的流式细胞术分析显示,粒细胞不表达CD45RA,单核细胞表达低水平的CD45RA。我们进一步分析了每个淋巴细胞亚群中CD45RA的表达水平,发现CD21+B细胞上CD45RA的表达是均匀的。另一方面,CD3+T细胞上CD45RA的表达是可变的。用刀豆蛋白A刺激淋巴细胞后,CD45RA+部分增加,表明不仅表型而且激活状态都会影响CD45的异构体表达模式。我们的发现提供了犬类CD45表达的基础知识,这可能是研究具有各种表型、发育阶段和激活状态的淋巴细胞的一种工具。