自然杀伤T淋巴细胞对黑色素瘤细胞的免疫调节及杀伤功能实验研究

目的探讨自然杀伤T淋巴细胞(NKT细胞)在细胞免疫调节及杀伤功能中的作用。方法建立混合淋巴细胞培养(MLC)体系,B16F10-luc-G5细胞作为靶细胞,以总淋巴细胞为效应细胞。(1)调节效应实验以NKT细胞或CD4~+CD25~+T淋巴细胞为调节细胞,分为3组:NKT组、CD4~+CD25~+T组、靶细胞空白对照组;另设有1640空白对照组(RPMI1640溶液)。(2)杀伤效应实验以NKT或自然杀伤(NK)细胞为效应细胞,分为3组:NKT组、NK组、靶细胞空白对照组。混合培养24、48、72h后,通过活体成像系统检测该培养系统的靶细胞生物发光,以监测NKT细胞的调节及杀伤效应。结果 (1)调节效应实验:NKT组及CD4~+CD25~+T组测定光子数分别与两空白对照组比较,以及NKT组内培养24h与72h时测得光子数比较,差异均有统计学意义(P0.05)。(2)杀伤效应实验:NKT组及NK组与靶细胞空白对照组所测光子数比较,差异均有统计学意义(P0.05);且培养24、72h时NKT组与NK组光子数比较,差异均有统计学意义(P0.05)。结论 NKT细胞具有抑制总淋巴细胞对靶细胞的杀伤效应,且抑制效应具有时间性,其调节作用较CD4~+CD25~+T淋巴细胞强。NKT细胞具有对靶细胞的杀伤效应,但较NK细胞弱。

Study on the immunoregulatory and lethal effects of natural killer T cells on melanoma cells in vitro

Objective To explore the immunoregulatory and lethal effects of natural killer T lymphocytes(NKTs) in vitro .Meth‐ods The mixed lymphocyte cultured(MLC) system was established ,in which the B16F10‐luc‐G5 cells were set as target cells ,the total lymphocyte cells were set as effector cells .(1)In the experiment on immunoregulatory effects ,NKT lymphocytes or CD4+CD25+ T lymphocytes were set as regulating cells ,there was three groups ,including the NKT group ,CD4+CD25+ T group and pure target cell control group .Otherewise ,the 1640 blank control group was set by only adding RPMI1640 solution .(2)In the ex‐periment on antitumor effects ,the NKT or natural killer(NK) lymphocytes were set as killer cells ,there was three groups ,inclu‐ding the NKT group ,NK group and pure target cell control group .Mixed culturing 24 ,48 and 72 hours ,the bioluminescence of target cells in MCL system was detected by using the in vivo imaging system .Results (1)In the experiment on immunoregulatory effects ,there were statistically significant differences in measured average photon numbers between NKT group ,CD4+ CD25+ T group and the two control groups(P<0 .05) .The statistically significant differences were also found in the NKT group between 24 hours and 72 hours (P<0 .05) .(2)In the experiment on antitumor effects ,there were statistically significant differences in meas‐ured average photon numbers ,when the NKT group and NK group were compared to the pure target cell control group(P<0 .05) . After culturing 24 and 72 hours ,statistically significant differences were found between NKT group and NK group(P<0 .05) .Con‐clusion The NKT cells could inhibit the lethal effects of lymphocyte cells on target cells ,and the inhibitory effects are changed by the length of culturing .Compared with the CD4+CD25+ T lymphocytes ,NKT lymphocytes have strongger regulatory effects .Addi‐tionally ,the NKT cells have lethal effects on target cells ,which might be weaker than that of NK cells .