Tsumura H, Kimura S, Kawano M, Tsurudome M, Shimura K, Ito Y
Department of Microbiology, Mie University School of Medicine, Mie-Ken, Japan.
Cell Immunol. 1998 Mar 15;184(2):153-60. doi: 10.1006/cimm.1998.1271.
Murine FRP-1 (mFRP-1) and Ly10 antigens are alloantigens. Anti-Ly10.1, monoclonal antibody (mAb) reacts to HeLa cells stably expressing mFRP-1.1 (HeLa cells/mFRP-1.1), but does not react to HeLa cells/ mFRP-1.2. On the other hand, anti-Ly10.2 mAb reacts to HeLa/mFRP-1.2 cells, but does not react to HeLa cells/mFRP-1.1. These findings indicate that Ly10.1 and Ly10.2 antigens are identical to the heavy chains of mFRP-1.1 and mFRP-1.2 molecules, respectively. Thymocytes and fibroblast cells obtained from CDF1 and BDF1 mice showed reactivity to both anti-Ly10.1 and anti-Ly10.2/FRP-2 antibodies, indicating that mFRP-1/Ly10 alloantigens are codominantly expressed in the F1 mice. Intriguingly, IMC carcinoma cells, derived from CDF1 mice, express mFRP-1.2, but do not express mFRP-1.1. Surprisingly, DBT cells, derived from CDF1 mice, consist of two different cell populations: one expresses both mFRP-1.1 and mFRP-1.2 antigens and the other expresses only mFRP-1.1 antigen. Consequently, we tried to isolate cloned DBT cell lines by limiting dilution. Finally, six cloned DBT cell lines were obtained, and three clones of these cloned DBT cells expressed both mFRP-1.1 and mFRP-1.2, and another three clones of these cloned DBT cells expressed both mFRP-1.1 and mFRP-1.2, and another three clones including clone 5 cells expressed only mFRP-1.1 antigen. No induction of mFRP-1.2/Ly10.2 by 5-azacytidine and sodium n-butyrate could be detected in DBT clone 5 cells. Neither mFRP-1.2 mRNA nor the genomic cDNA clone encoding mFRP-1.2 could be detected in DBT clone 5 cells, nor could the genomic cDNA encoding mFRP-1.1 be detected in IMC cells, indicating that aberrant expression of mFRP-1/Ly10 allotypes in CDF1 mice-derived cells is due to the gene deletion.
小鼠FRP-1(mFRP-1)和Ly10抗原是同种异体抗原。抗Ly10.1单克隆抗体(mAb)与稳定表达mFRP-1.1的HeLa细胞(HeLa细胞/mFRP-1.1)发生反应,但不与HeLa细胞/mFRP-1.2发生反应。另一方面,抗Ly10.2 mAb与HeLa/mFRP-1.2细胞发生反应,但不与HeLa细胞/mFRP-1.1发生反应。这些发现表明,Ly10.1和Ly10.2抗原分别与mFRP-1.1和mFRP-1.2分子的重链相同。从CDF1和BDF1小鼠获得的胸腺细胞和成纤维细胞对抗Ly10.1和抗Ly10.2/FRP-2抗体均有反应,表明mFRP-1/Ly10同种异体抗原在F1小鼠中是共显性表达的。有趣的是,源自CDF1小鼠的IMC癌细胞表达mFRP-1.2,但不表达mFRP-1.1。令人惊讶的是,源自CDF1小鼠的DBT细胞由两种不同的细胞群体组成:一种表达mFRP-1.1和mFRP-1.2抗原,另一种仅表达mFRP-1.1抗原。因此,我们试图通过有限稀释法分离克隆的DBT细胞系。最终,获得了六个克隆的DBT细胞系,其中三个克隆的DBT细胞表达mFRP-1.1和mFRP-1.2,另外三个克隆,包括克隆5细胞,仅表达mFRP-1.1抗原。在DBT克隆5细胞中未检测到5-氮杂胞苷和正丁酸钠对mFRP-1.2/Ly10.2的诱导作用。在DBT克隆5细胞中未检测到mFRP-1.2 mRNA或编码mFRP-1.2的基因组cDNA克隆,在IMC细胞中也未检测到编码mFRP-1.1的基因组cDNA,这表明CDF1小鼠来源的细胞中mFRP-1/Ly10同种异型的异常表达是由于基因缺失所致。