Detection of CD45iota mRNA in murine Th1 but not Th2 clones.
CD45, a prototype of the receptor-like protein tyrosine phosphatase (PTPase) family, is one of the essential molecules in signal transduction through T cell receptors. Because at least 8 types of CD45 isoforms can potentially be produced by alternative mRNA splicing of exons 4, 5, and 6, the analyses at the transcription and protein levels of CD45 during the development and differentiation of T cells have been performed using RT-PCR and isoform-specific monoclonal antibodies, respectively. We report here that the ninth and smallest isoform of CD45, designated as CD45iota (CD45t), which is alternatively spiced from exons 4, 5, and 6 as well as exon 7, is present in the fetal thymus and splenic T cells of mice, and in murine Th1 clones, but not in Th2 clones. The expression of full-length CD45t mRNA as the functional CD45 PTPase was confirmed by RT-PCR analysis. Furthermore, the expression vector of CD45t was constructed, and its expression was detected in combination with anti-pan CD45 mAb and our newly established anti-LAR/CD45 PTPase domain mAb. These results suggested that CD45t might be an important isoform of CD45 for differentiation signaling of Th cells, and might be used as a marker to distinguish between Th1 and Th2 cells.
Tsujikawa
K
K
Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Japan. tujikawa@phs.osaka-u.ac.jp
Uchino
Y
Y
Ichijo
T
T
Furukawa
T
T
Yamamoto
H
H
D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002478:Cells, Cultured; D005333:Fetus; D005822:Genetic Vectors; D017493:Leukocyte Common Antigens / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D020033:Protein Isoforms / Q000096:biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology*; D018418:Th2 Cells / Q000276:immunology*; D013950:Thymus Gland / Q000276:immunology*; D014162:Transfection
D020033:Protein Isoforms; D012333:RNA, Messenger; D011994:Recombinant Proteins; D017493:Leukocyte Common Antigens