DNA microarray gene expression profile of T cells with the splice variants of TCRζ mRNA observed in systemic lupus erythematosus

Kensei Tsuzaka, Kyoko Nozaki, Chika Kumazawa, Kiyono Shiraishi, Yumiko Setoyama, Keiko Yoshimoto, Katsuya Suzuki, Tohru Abe, Tsutomu Takeuchi

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15 Citations (Scopus)

Abstract

We have reported that the TCRζ mRNA with alternatively spliced 3′ UTR (ζ mRNA/as-3′-untranslated region (UTR)) and ζ mRNA lacking exon 7 (ζ mRNA/exon 7-) observed in systemic lupus erythematosus patient T cells can lead to down-regulation of both ζ and TCR/CD3 complexes. To determine whether these T cells expressing decreased ζ exhibit differential transcription patterns, we transfected retrovirus vectors containing wild-type ζ cDNA, ζ cDNA/as-3′ UTR, and ζ cDNA/exon 7- into murine T cell hybridoma MA5.8 cells which lack ζ expression to construct the MA5.8 mutants WT, AS3′ UTR, and EX7-, respectively. FACS analyses demonstrated reduced cell surface expression of ζ and TCR/CD3 complexes on the AS3′ UTR mutant and the EX7- mutant in comparison to that on the WT mutant. Total RNA was collected after stimulating the MA5.8 mutants with anti-CD3 Ab. Reverse-transcribed cDNA was applied to the mouse cDNA microarray containing 8691 genes, and the results were confirmed by real-time PCR. The results showed that 36 genes encoding cytokines and chemokines, including IL-2, IL-15, IL-18, and TGF-β2, were down-regulated in both the AS3′ UTR mutant and the EX7- mutant. Another 16 genes were up-regulated in both, and included genes associated with membranous proteins and cell damage granules, including the genes encoding poliovirus receptor-related 2, syndecan-1, and granzyme A. Increased protein expression of these genes was confirmed by Western blot and FACS analyses. Identification of these responsive genes in T cells in which the ζ and TCR/CD3 complexes were down-regulated may help to better understand the pathogenesis of systemic lupus erythematosus.

Original languageEnglish
Pages (from-to)949-956
Number of pages8
JournalJournal of Immunology
Volume176
Issue number2
Publication statusPublished - 2006 Jan 15
Externally publishedYes

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ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Tsuzaka, K., Nozaki, K., Kumazawa, C., Shiraishi, K., Setoyama, Y., Yoshimoto, K., Suzuki, K., Abe, T., & Takeuchi, T. (2006). DNA microarray gene expression profile of T cells with the splice variants of TCRζ mRNA observed in systemic lupus erythematosus. Journal of Immunology, 176(2), 949-956.