淋巴瘤患者外周血TCRVα24^+Vβ11^+自然杀伤T细胞体外活化特性研究  

Expansion capacity and cytokine releasing characteristics of TCRVα24^+Vβ11^+ NKT cells in peripheral blood of patients with lymphoma

作　　者：王迎雪[1] 李丽珍[1] 窦爱霞[2] 孙俊华[2] 郭成山[2] 张锑[1] 彭军[1] 徐从高[1] 

机构地区：[1]山东大学齐鲁医院血液学研究室,山东济南250012 [2]山东大学第二医院血液科,山东济南250033

出　　处：《中国病理生理杂志》2008年第6期1129-1133,共5页Chinese Journal of Pathophysiology

摘　　要：目的:研究淋巴瘤患者外周血TCRVα24+Vβ11+自然杀伤T(NKT)细胞的数量以及体外活化后的功能状态,与正常人外周血NKT细胞的数量及功能状态进行比较。方法:制备30例淋巴瘤患者和30例年龄及性别匹配的正常对照外周血单个核细胞(PBMNCs),以流式细胞术(FACS)检测TCRVα24+Vβ11+NKT细胞数量,以α-半乳糖神经酰胺(α-Galcer)及白细胞介素-2(IL-2)从PBMNCs中扩增活化NKT细胞,采用细胞内细胞因子流式细胞术检测手段,测定NKT细胞中胞内白细胞介素-4(IL-4)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)阳性细胞的比例。结果:淋巴瘤患者与正常对照PBMNCs中TCRVα24+Vβ11+NKT细胞的细胞比率分别为0.17%±0.10%、0.28%±0.18%(P<0.05)。PBMNCs培养体系中加入α-Galcer及IL-2,将NKT细胞扩增活化7d后,淋巴瘤患者与正常对照的NKT细胞的扩增倍数分别为101.37±44.61、129.66±56.31(P<0.05)。扩增活化后,淋巴瘤患者与正常对照的NKT细胞中胞内细胞因子IFN-γ阳性细胞的比例分别为41.96%±15.06%、52.48%±18.85%(P<0.05);TNF-α阳性细胞的比例分别为46.30%±16.03%、71.37%±17.28%(P<0.05);IL-4阳性细胞的比例分别为36.19%±11.74%、33.12%±12.95%(P>0.05)。不同病理分型及分期的淋巴瘤患者之间上述各指标无显著差异。结论:淋巴瘤患者外周血TCRVα24+Vβ11+NKT细胞数量较正常对照明显减少,经α-Galcer扩增活化后扩增倍数较正常对照降低,分泌细胞因子IFN-γ、TNF-α的功能较正常降低,此数量及功能的降低与淋巴瘤的分型及分期无关。但其仍保持有对α-Galcer刺激后的扩增活化反应能力。AIM： To analyze the quantity of TCRVa24^＋ Vβ11^＋ natural killer T （NKT） cells and cytokines production induced by a- galactosylceramide （ a- Galcer） in vitro in lymphoma patients. METHODS ： Flow cytometry was utilized to enumerate TCRVa24^＋ Vβ11^＋ NKT cells in peripheral blood mononuclear cells （PBMNCs） in 30 cases of lymphoma patients and 30 cases of age - and gender - matched healthy controls. NKT cells were activated with a- Galcer and interleukin - 2 （ IL - 2） after expansion in vitro. The percentages of positive NKT cells which expressed intracellular intedeukin - 4 （ IL - 4）, interferon - γ（ IFN -γ） and tumor necrosis factor - ct （ TNF - a） were then determined by flow cytometry. RESULTS： The rates of NKT ceils in PBMNCs were 0. 17% ±0. 10% and 0.28%±0. 18% （P 〈0.05） in lymphoma patients and controls, respectively. Seven days after expansion and activation with a- Galcer and IL - 2, the fold of expansion of NKT cells in two groups was 101.37±44. 61 and 129.66±56. 31 （P 〈0.05）, respectively. The ratio of TCRVa24^＋ NKT cells that secreted IFN -γ or TNF -a in lymphoma patients was significantly lower than that in controls （41.96% ±15.06% vs52.48% ±18.85%, P〈0.05; 46.30%±16.03% vs71.37% ±17.28%, P〈0.05）. While the ratio of TCRVa24^＋ NKT cells that secreted IL - 4 was not significantly different between the two groups （ 36. 19% ±11.74% vs 33. 12% ±12. 95%, P 〉 0. 05 ）. There was no significant difference in above parameters among different groups of lymphoma patients subdivided by pathology and clinical stage. CONCLUSION： The quantity of NKT cells in PBMNCs in lymphoma patients is lower than that in controls. The expansion capacity and the function of producing cyto- kines IFN - γ and TNF - a of NKT cells stimulated with et - Galcer are decreased. This decrease is independent of lympho- ma pathology type or clinical stage.

关 键 词：杀伤细胞 天然 半乳糖神经酰胺类 细胞因子类 淋巴瘤 

分 类 号：R551.2[医药卫生—血液循环系统疾病]