优势化Ⅱ型T细胞表达Ly49C和Ly49A及其功能

Ly49A and Ly49C Expression and Their Functions of Polarized Type Ⅱ T Cells in Mice

探讨ＫＩＲ分子参与优势化的Ｔｈ２、Ｔｃ２细胞活化抑制和免疫耐受的可能机制．①体 外经ＩＬ－４优势化诱导Ⅱ型Ｔ细胞； ②ＡＢＣ法检测Ｌｙ４９Ａ表达； ③优势化Ⅱ型Ｔ淋巴细胞的 Ｌｙ４９ＣｍＲＮＡ的原位分子杂交；④ ３Ｈ－ＴｄＲ掺入法测定单向和双向 ＭＬＲ中 Ｂ６（Ｈ－２ｂ）小鼠优势 化Ⅱ型Ｔ淋巴细胞增殖；⑤Ｌｙ４９Ｃ阻断后活化Ⅱ型Ｔ细胞对ＥＬ９６ｌｌ细胞的杀伤活性测试．结 果有：①在ｍＩＬ－４、ＣｏｎＡ和Ｉｏｎｍｙｃｉｎ环境下体外培养４天即可使静息淋巴细胞ｍＩＬ－４的分泌 水平由低于２０ｕｇ／ｍＬ提高到８３３．±２８．７３ｕｇ／ｍＬ（Ｐ＜０．０１）；②优势化的Ⅱ型Ｔ淋巴细胞的 Ｌｙ４９ＣｍＲＮＡ表达阳性率为（２５．６±６．２％（静息Ｔ细胞的质量分数＜３％），Ｌｙ４９Ａ的膜表达为 （２．５±４．９）％（静息Ｔ细胞的质量分数＜５％）ＦＣＭ检测结果显示：Ｌｙ４９ Ｃ在Ⅱ型Ｔ细胞表 达阳性率（２９．８±４．３）％，其中 ＣＤ４＋细胞表达阳性率为（１０．２２±０．９）％、 ＣＤ８＋细胞阳性表达 率为（１７．８ ± １．３）％抗Ｌｙ４９Ｃ单克隆抗体（５Ｅ６）可以使Ｃ５７ＢＬ／６Ｈ－２ｂ－ＢＡ；Ｂ／ｃＨ－２ｄ之间单向 ＭＬＲ和双向ＭＬＲ的增殖?

To explore the possible mechanism of killer cell inhibitory receptors (KIR) in immunosuppression and immunity tolerance of the polarized Th2 Tc2 cells. ① T cells of C57BL/6 mice were induced to turn toward Th2, Tc2 cells (type Ⅱ cells) by culture T cells with mIl, ionomycine and ConA In vitro; ② T cells producing IL-4 were confirmed by ELISA and FCM methods; ③ Ly49A on T cells were essayed by ABC method; ④ Ly49CmRNA in type Ⅱ cells were detected by in situ hybridization hitochewhsty; ⑤ Prolierative activity of type Ⅱ cells in B6H-２b mice was examined by mix 3H-TdR into DNA; ⑥ Cytoxic activity of type Ⅱ T cell ligated Ly49C were measured by detecting the killingactivity on EL9611 cells. Stationary T lymphoxcytes can be induced to turn toward class Ⅱ cytokines producing cells after 4 days culturing with mLL-4 ConA and Ionomycin in vitro. The mIL-4 se crcating level rises from 20ug/mLto 833 .33 ± 28 .73 ug/mL. The rate of Ly49A mRNA and Ly49A protein expressing cells in type Ⅱ cells were (25 .6 ± 6. 2) % and (22. 5 ± 4. 9) % respectively (P < 3% and P < 5% respectively in stationion T cells). The results of FCM inspecting show that the rate of Ly49C expressing cells in type Ⅱ cells was (29.8 ± 4.3) %, among of which, the rates ｏf CD8+ and CD4+ cells were (10.2 ±0.9) % and (17. 8 ± 1. 3) % respectively. The proliferative activity of T cells ligated Ly49C in directional and bi-directional MLR between C57BI/6H-2b and C57BL/6H-2d were elevated by (21 .5 ± 3.4)% and (13.8 ± l.6)% respectively after ligating Ly49A with. with mAb 5E6, Cytoxic activity of T cells in C57BL/6 mice was heightened by (47. 2 ± 8. 5) % when Ly49C was ligated by mAh 5E6. Conclusion: The Ly49A, Ly49C expressing increased in T cells while T cells were induced into type fi cells by culture with M and T cell activating reagents. Inhibitory signal transfer pass ways of T cells have been established by KIR on T cells conjunct with MHC-I expressed on target cells.