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

Abstract

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 above-mentioned 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). 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. Key words: Lymphoma, large B-cell, diffuse; Rituximab; Th17 cells; Interleukin-17; In vitro experiment