Alternative TitleSignificance of CD244 expression in functional development of CD8+ cytotoxic T lymphocytes
Periodical title平成17(2005)年度 科学研究費補助金 基盤研究(C) 研究成果報告書 = 2005 Fiscal Year Final Research Report
Note (General)(1)新生児CD8+T細胞はCD244陰性であるのに対し,成人CD8+T細胞は約40-50%がCD244陽性であり,加齢に伴いCD244陽性比率は増加した.CD28,CD62L発現を3重解析するとCD244陰性細胞は全てCD28陽性CD62L陽性であるのに対し,CD244陽性細胞はCD28,CD62L陽性,陰性の亜群が存在した.伝染性単核球症(IM)や麻疹のCD8+T細胞ではCD244+細胞の増加が顕かで,特にCD28,CD62L陽性CD244陽性CD8+T細胞の増加が著明であった.(2)CD244陽性,陰性CD8+T細胞亜群における表面抗原,細胞内傷害性蛋白を解析した.CD244-細胞はどの表面分子発現からもナイーブ細胞として表面形質を示し,パーフォリン,グランザイムB, TIA-1発現も認めなかった.一方,CD244陽性細胞は,メモリー細胞表面形質を呈した.(3)Naive T細胞ではTCRレパートワは多様性に富むがmemory-T細胞では限られたT細胞レパートワであることが予想され,TCR CDR3 spectrum解析をCD244陽性、陰性各細胞亜群において解析した.CD244陰性細胞は全てのTCRVβでガウス分布を呈したが,CD244陽性分画は多くのTCRVβでskewedパターンを示した.また,CD28陽性細胞より陰性細胞で,CD62L陽性細胞より陰性細胞で複雑指数は低下していた。(4)CD244陽性CD8+T細胞への生体内でのCD244陰性細胞から陽性細胞への転換をin vitroにてCD244-CD8+T細胞を刺激培養して確認した.CD244発現はCD3刺激にて培養早期から増強した.一方,CD28,CD62L-分画への転換は2-3週間かかり緩除に進行した.(5)機能的分化における細胞分裂の程度を定量化するため、各細胞分画におけるTREC含有量を測定したところ、CD244陰性細胞は最も多くTREC量が多く、CD28CD62L陽性CD244陽性細胞、CD28/CD62L陰性CD244陽性細胞分画の順にTREC含有量は減少した。
(1) About 40-50% adult CD8+ T cell were CD244 positive and CD244 positive fraction increased with aging whereas almost of newborn infant CD8+ T cell was CD 244 negative. CD28/CD62L positive, or negative two subgroup were present in CD244 positive cells when they analyzed three-color flow cytometry, whereas all CD244 negative CD8+ T cells were CD28 positive CD62L positive. Increase of CD244+ cells have been found in patients with infectious mononucleosis (IM) and measles., compared it with a normal subject, and in particular, and increase of CD28,CD62L positive CD244 positive CD8+ T cell were prominent.(2) We analyzed the difference of expression of surface antigen and intracellular cytotoxic granular proteins in CD244-positive, and CD244-negative CD8+ T cell subgroup. CD244-negative cells showed surface phenotype almost same as naive cell from all surface molecule expression and did not show perform, granzyme B, TIA-1 expression either. On the other hand, CD244-positive cells showed me mory cell surface phenotype.(3) Naive T cell have TCR repertoire with full of diversity, but T cell repertoire diversity was expected to be limited in memory-T cells, and TCR CDR spectrum were analyzed in each CD244-positive, CD244-negative cell subgroup. CD244 negative cells presented Gaussian distribution in all TCRVβ, but CD244-positive demarcation showed a skewed pattern in many TCRVβ. In addition, complexity index of CDR3 spectra in CD28-negative cell or CD62L negative cell decreased more than that in CD28 positive cells, or CD62L positive cell.(4) CD244-negative CD8+ T cell separated from normal adult CD8+T cells were stimulated and incubated, in vitro. We confirmed conversion from CD244-negative to CD244-positive phenotypte in CD8+ T cells. CD244 expression were increased with CD3 stimulation at early stage of culture. On the other hand, switch to CD28,CD62L- phenotype took two or three weeks and progressed in slowly.(5) We measured TREC content in each cell fractionation to quantify degree of mitosis in functional differentiation phase.CD 244 negative cells showed abundant TREC content, and the TREC content decreased in order of CD28CD62L positive CD244-positive cells, CD28/CD62L negative CD244-positive cell populations.
研究課題/領域番号:16591013, 研究期間(年度):2004-2005
出典：「CD244抗原発現とCD8^+細胞傷害性T細胞の機能的分化」研究成果報告書 課題番号16591013 (KAKEN：科学研究費助成事業データベース（国立情報学研究所）) 本文データは著者版報告書より作成
Related Materialhttps://kaken.nii.ac.jp/search/?qm=30204366
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16591013/
https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-16591013/165910132005kenkyu_seika_hokoku_gaiyo/
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