利妥昔单抗对弥漫大B细胞淋巴瘤患者Th17细胞及相关细胞因子影响的体外实验

目的 探讨利妥昔单抗对弥漫大B细胞淋巴瘤(DLBCL)患者Th17细胞及相关细胞因子体外的影响及其意义.方法 DLBCL初治患者(DLBCL组)20例和健康对照者(健康对照组)20名,每个对象分别采集4份外周血标本,分离外周血单个核细胞(PBMC),按照培养条件的不同分成4个亚组:空白组(A亚组)、加入利妥昔单抗组(B亚组)、加入利妥昔单抗和血清组(C亚组)和极化组(D亚组)[加白细胞介素6(IL-6)和转化生长因子(TGF-β)].培养后分别采用流式细胞术检测各亚组外周血PBMC中Th17细胞比例,酶联免疫吸附法(ELISA)检测上述培养液中白细胞介素17(IL-17)的表达水平.结果 健康对照组中,D亚组的Th17细胞比例和IL-17水平分别为(17.12±4.90)％、(45.735±10.012)pg/ml,均高于A、B、C亚组(P＜0.05),A、B、C亚组之间比较差异均无统计学意义(P＞0.05).DLBCL组A亚组的Th17细胞比例及IL-17水平为(0.69±0.24)％、(6.012±1.312)pg/ml,均低于健康对照组A亚组的(2.43±0.61)％、(8.217± 1.681)pg/ml(P＜ 0.05);在与利妥昔单抗一起体外培养后,DLBCL组中B、C和D亚组的Th17细胞比例分别为(2.34±0.48)％、(2.31±0.53)％和(16.92±4.81)％,IL-17水平分别为(7.944±1.538) pg/ml、(7.957±1.533) pg/ml和(44.417±9.881) pg/ml,均高于A亚组,且D亚组明显高于B、C亚组(P＜0.05).B、C亚组之间比较差异无统计学意义(P＞0.05).结论 体外实验证实,DLBCL患者外周血PBMC中Th17细胞比例低于健康人,利妥昔单抗作用于DLBCL患者外周血PBMC后可升高Th17细胞比例.

Rituximab impacts Th17 cells and related cytokines in patients with diffuse large B-cell lymphoma in vitro

Objective To explore the impact of rituximab on Th17 cells and related cytokines in patients with diffuse large B-cell lymphoma (DLBCL) in vitro and its significance.Methods 20 cases of DLBCL untreated patients and 20 healthy subjects were enrolled in the name of DLBCL group and health control group, respectively.4 peripheral blood samples were collected from every case to separate peripheral blood mononuclear cells (PBMCs), which were assigned to 4 subgroups according to different culture conditions: blank subgroup(subgroup A), rituximab subgroup (subgroup B), rituximab and serum subgroup (subgroup C) and polarization subgroup (subgroup D) (added IL-6 and TGF-β).After cultured in vitro, the percentage of Th17 cells in each subgroup was tested by flow cytometry, and the cytokine IL-17 in the abovementioned culture fluid was measured by enzyme-linked immunosorbent assay (ELISA).Results In health control group, the percentage of Th17 cells and the level of IL-17 in subgroup D [(17.12 ± 4.90) ％ and (45.735±10.012) pg/ml] were significantly higher than those in subgroup A, B, C (P ＜ 0.05), and there was no difference in each other subgroup A, B, C (P ＞ 0.05).The percentage of Th17 cells and the level of IL-17 in the DLBCL subgroup A were significantly lower than those in health control subgroup A [(0.69±0.24) ％ and (6.012±1.312) pg/ml vs (2.43±0.61) ％ and (8.217±1.681) pg/ml (P ＜ 0.05)].In DLBCL group, after cultured with rituximab, the percentages of Th17 cells in subgroup B, C, D were (2.34±0.48) ％, (2.31±0.53) ％ and (16.92±4.81) ％, and the levels of IL-17 were (7.944±1.538) pg/ml, (7.957±1.533) pg/ml and (44.417±9.881) pg/ml, respectively, which were all significantly higher than those in subgroup A.Besides, the percentage of Th17 cells and the level of IL-17 in DLBCL subgroup D were significantly higher than those in subgroup B, C (P ＜ 0.05), while there was no difference between subgroup B and subgroup C.Conclusion Experiments in vitro confirmed that the percentage of Th17 cells in PBMCs of DLBCL patients was lower than that in healthy persons, and rituximab could elevate the percentage of Th17 cells in PBMCs of DLBCL patients.