肺癌患者外周血及癌组织中白细胞介素-21＋、CD8＋、γ-干扰素＋T细胞的变化及相互作用

The change and interaction of interleukin- 21＋ and CD8＋ interferon -γ＋ T cell in perigheral blood and lung lesion of lung cancer patients

目的观察白细胞介素-21（IL-21）＋、CD8＋、γ-干扰素＋T细胞（Tcl）在非小细胞肺癌（NSCLC）患者外周血、肺癌组织及癌旁组织中的变化，探讨其在NSCLC中的作用及相互关系。方法收集NSCLC患者外周血、癌组织及癌旁组织50例。同期收集健康对照组外周血30例。酶联免疫吸附试验（ELISA）法检测血清、癌组织及癌旁组织中IL-21的浓度；流式细胞术检测上述标本中CD8＋IL-21R＋和Tcl细胞比例；磁珠分选纯化患者外周血CD8＋T细胞，重组人白细胞介素-21（rhIL-21）与纯化的CD8＋T细胞分别共培养24、48h，设对照组和共培养组，流式细胞术检测其Tel细胞比例。结果NSCLC血清中IL-21浓度高于对照组[（23．13±3．92）ng／L比（3．46±1．19）ng／L，P〈0．05]，外周血CD8＋IL-21R＋和Tcl细胞比例均低于对照组[（1．56±0．35）％比（3．11±1．00）％。（11．34±2．62）％比（33．94±8．38）％，P〈0．05]。NSCLC癌组织中IL-21浓度高于癌旁组织[（37．82±7．24）ng／L比（15．87±3．82）ng／L，P〈0．05]，CD8＋IL-21R＋、Tcl细胞比例均高于癌旁组织[（1．62±O．25）％比（1．18±0．24）％，（5．70±1．10）％比（3．85±O．52）％，P〈0．05]。IL-21浓度和CD8＋IL-21R＋、Tcl细胞比例在不同TNM分期外周血、癌组织中有差异，随临床分期的进展有下降的趋势。rhlL-21与患者外周血CD8＋T细胞在培养24h和48h后，培养组中Tcl细胞比例均高于对照组[（18．89±3．48）％比（13．03±2．22）％，（24．62±3．78）％比（15．14±2．76）％，P〈0．05]。结论在NSCLC外周血及肺癌组织中，IL-21浓度升高，并随临床分期的进展而下调；CD8＋IL-21R＋及Tcl细胞在患者外周血中下降，而在肺癌组织处聚集，并随临床分期的进展而下调；IL-21可能通过与CD8＋IL-21R结合，刺激和活化Tcl的产生，促进γ-干扰素（IFN-γ）生成，杀伤肿瘤细胞。

Objective To explore the change and interaction of interleukin - 21 （ IL - 21 ） ＋ and CD8 ＋ interferon （IFN） - γ ＋ T （ Tcl ） cell in perigheral blood （ PB ） and lung lesion of the non small cell lung cancer （NSCLC） patients. Methods The samples of PB, lung lesion and para - cancer tissues was collected from 50 NSCLC inpatients, and the PB of 30 healthy people was control group （CG）. The con- centration of IL-21 in this samples was tested by enzyme linked immunosorbent assay （ELISA） kit and the proportion of CD8 ＋ IL -21R ＋ and Tcl cell was determined by flow cytometry; CD8 ＋T cells were ob- tained from PB of the patients using human T - cell isolation kit, purified CD8 ＋ T cells were cultured with recombinant human interleukin 21 （ rhIL - 21 ） or without, co - cultured group and control group. After 24 h and 48 h of culture, the percentage of Tcl cell was calculated by flow cytometry. Results The con- centration of IL -21 in the serum from patients was significantly higher as compared to the CG [ （23.13 ± 3.92） ng/L vs. （3.46 ± 1.19） ng/L （P 〈0. 05） ], the proportion of CD8 ＋ IL -21R＋ and Tcl cell from patients PB was significantly lower compared with the CG, respectively [ （ 1.56 ± 0. 35 ） % vs. （3.11±1.00）% （P〈0.05）, （11.34±2.62）% vs. （33.94±8.38）% （P〈0.05）1. As well as the concen- tration of IL -21 from tissue homogenates [ （37.82 ±7.24） ng/L vs. （15.87 ±3.82） ng/L （P〈 0. 05 ）] , the percentage of CD8 ＋ IL- 21R＋ and Tel cell from lesion was significantly increase compared with the para - lung cancer tissues, respectively [ （ 1.62 ± 0. 25 ） % vs. （ 1.18 ± 0. 24） % （ P 〈 0.05 ） , （5.70 ± 1.10） % vs. （3.85 ± 0. 52 ） % （ P 〈 0. 05 ） ]. The concentration of IL - 21 in serum and lesion from patients, and the frequency of Tel , CD8 ＋ IL- 21R ＋ cell in patients PB and lung cancer tissues both [lad tile differentiation in different TNM phase, and had the downtrend with the development of the disease. After 24 h and 48h of culture with rhIL - 21, the frequencies of Tcl from co - cultured group were both higher than control group [ （18.89 ±3.48）% vs. （13.03 ±2.22）% （P〈0.05）, （24.62±3.78）% vs. （ 15. 14 ± 2. 76） % （P 〈 O. 05） 3. Conclusion In the PB and lung lesion of NSCLC, the content of IL- 21 was increasing and downtrend with the developing of the disease; the CD8 ＋ IL- 21R ＋ and Tcl cell in PB was down regulation and recruitment into lung lesion, and both had downtrend with the advance of tile disease; IL -21 may combine to CD8 ＋ IL- 21R, then promote CD8 ＋T cell secret the IFN -γ and im- plcunent the anti -tumor effect.