Characterization of alternative splice variants of human interleukin-2

V. N. Tsytsikov, V. V. Yurovskv, S. P. Atamas, B. White

Research output: Contribution to journalArticle

Abstract

We used reverse transcription-PCR to analyse IL-2 mRNA transcripts obtained from activated human peripheral blood mononuclear cells of healthy donors. We have identified and characterized two alternatively spliced mRNAs, called IL-262 and PL-283. As shown by DNA sequence analysis, IL-2S2 and IL-283 lack exon 2 or exon 3, respectively. To test functions of IL-282 and FL-263, we expressed IL282, IL-283, and wild type IL-2 in GS115 PicHa pastoris expression system. Recombinant proteins were identified by SDS-PAGE and Western blot analysis with anti-IL-2 antibody. Each recombinant protein had the expected molecular weight. We tested whether IL-2, IL-282 and IL-283 proteins have different functions. In contrast to IL-2, IL-282 and IL-283 had no costimulatory effect on T-cell proliferation. Preliminary data show that IL-282 and IL-283 inhibit FL-2-induced T cell proliferation in vitro. This inhibition is not mediated through toxicity. We hypothesize that IL-282 and IL-283 are competitive inhibitors of IL-2 binding to IL-2 receptors. Additional studies are in progress to test this hypothesis.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996
Externally publishedYes

Fingerprint

interleukin-2
Interleukin-2
T-cells
Cell proliferation
Recombinant Proteins
Exons
Cell Proliferation
recombinant proteins
T-Lymphocytes
exons
Messenger RNA
cell proliferation
Interleukin-2 Receptors
DNA sequences
T-lymphocytes
Transcription
DNA Sequence Analysis
Reverse Transcription
Toxicity
Polyacrylamide Gel Electrophoresis

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Tsytsikov, V. N., Yurovskv, V. V., Atamas, S. P., & White, B. (1996). Characterization of alternative splice variants of human interleukin-2. FASEB Journal, 10(6).

Characterization of alternative splice variants of human interleukin-2. / Tsytsikov, V. N.; Yurovskv, V. V.; Atamas, S. P.; White, B.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

Tsytsikov, VN, Yurovskv, VV, Atamas, SP & White, B 1996, 'Characterization of alternative splice variants of human interleukin-2', FASEB Journal, vol. 10, no. 6.
Tsytsikov VN, Yurovskv VV, Atamas SP, White B. Characterization of alternative splice variants of human interleukin-2. FASEB Journal. 1996;10(6).
Tsytsikov, V. N. ; Yurovskv, V. V. ; Atamas, S. P. ; White, B. / Characterization of alternative splice variants of human interleukin-2. In: FASEB Journal. 1996 ; Vol. 10, No. 6.
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N2 - We used reverse transcription-PCR to analyse IL-2 mRNA transcripts obtained from activated human peripheral blood mononuclear cells of healthy donors. We have identified and characterized two alternatively spliced mRNAs, called IL-262 and PL-283. As shown by DNA sequence analysis, IL-2S2 and IL-283 lack exon 2 or exon 3, respectively. To test functions of IL-282 and FL-263, we expressed IL282, IL-283, and wild type IL-2 in GS115 PicHa pastoris expression system. Recombinant proteins were identified by SDS-PAGE and Western blot analysis with anti-IL-2 antibody. Each recombinant protein had the expected molecular weight. We tested whether IL-2, IL-282 and IL-283 proteins have different functions. In contrast to IL-2, IL-282 and IL-283 had no costimulatory effect on T-cell proliferation. Preliminary data show that IL-282 and IL-283 inhibit FL-2-induced T cell proliferation in vitro. This inhibition is not mediated through toxicity. We hypothesize that IL-282 and IL-283 are competitive inhibitors of IL-2 binding to IL-2 receptors. Additional studies are in progress to test this hypothesis.

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