CAJ | 학술논문

目的研究小鼠骨髓来源未成熟树突状细胞(imDC)诱导同种幼稚T淋巴细胞分化为调节性T淋巴细胞(Treg细胞)及辅助性T淋巴细胞(TH)1、TH2和TH 17的能力.方法培养Balb/c小鼠骨髓来源树突状细胞(DC),加入脂多糖刺激DC成熟.分别将imDC与成熟DC(mDC)与同种C57BL/6小鼠幼稚T淋巴细胞共同培养,用酶联免疫斑点法检测TH 1类细胞因子[γ干扰素(IFN-γ)和白细胞介素(IL)-2]、TH2类细胞因子(IL-4和IL-10)、TH17类细胞因子(IL-17)的分泌情况.imDC和mDC分别与同种幼稚T淋巴细胞在加入IL-2及转化生长因子β1的条件下共同培养,用流式细胞术检测T淋巴细胞中Treg细胞比例.结果分别与imDC或mDC共同培养后,T淋巴细胞中分泌IFN-γ细胞的数量分别为(11.67±2.18)/2×105个细胞和(182.00±23.71)/2×105个细胞(P＜0.01),分泌.IL-2细胞的数量分别为(26.67±2.96)/2×105个细胞和(318.30±18.62)/2×105个细胞(P＜0.01),分泌IL-4细胞的数量分别为(17.00±3.78)/2×105个细胞和(45.33±3.48)/2×105个细胞(P＜0.01),分泌IL-10细胞的数量分别为(7.00±1.00)/2×105个细胞和(158.70±10.90)/2×105个细胞和(P＜0.001),分泌IL-17细胞的数量为(0.66±0.33)/2×105个细胞和(238.30±24.39)/2×105个细胞(P＜0.001).imDC诱导产生的Treg细胞占(22.70±1.53)％,高于mDC诱导产生的(5.42±1.27)％ (P＜0.001).结论 imDC诱导同种幼稚T淋巴细胞较多分化为Treg细胞,而较少分化为TH1、TH2和TH17细胞.
목적 연구소서골수래원미성숙수돌상세포(imDC)유도동충유치T림파세포분화위조절성T림파세포(Treg세포)급보조성T림파세포(TH)1、TH2화TH 17적능력.방법 배양Balb/c소서골수래원수돌상세포(DC),가입지다당자격DC성숙.분별장imDC여성숙DC(mDC)여동충C57BL/6소서유치T림파세포공동배양,용매련면역반점법검측TH 1류세포인자[γ간우소(IFN-γ)화백세포개소(IL)-2]、TH2류세포인자(IL-4화IL-10)、TH17류세포인자(IL-17)적분비정황.imDC화mDC분별여동충유치T림파세포재가입IL-2급전화생장인자β1적조건하공동배양,용류식세포술검측T림파세포중Treg세포비례.결과 분별여imDC혹mDC공동배양후,T림파세포중분비IFN-γ세포적수량분별위(11.67±2.18)/2×105개세포화(182.00±23.71)/2×105개세포(P＜0.01),분비.IL-2세포적수량분별위(26.67±2.96)/2×105개세포화(318.30±18.62)/2×105개세포(P＜0.01),분비IL-4세포적수량분별위(17.00±3.78)/2×105개세포화(45.33±3.48)/2×105개세포(P＜0.01),분비IL-10세포적수량분별위(7.00±1.00)/2×105개세포화(158.70±10.90)/2×105개세포화(P＜0.001),분비IL-17세포적수량위(0.66±0.33)/2×105개세포화(238.30±24.39)/2×105개세포(P＜0.001).imDC유도산생적Treg세포점(22.70±1.53)％,고우mDC유도산생적(5.42±1.27)％ (P＜0.001).결론 imDC유도동충유치T림파세포교다분화위Treg세포,이교소분화위TH1、TH2화TH17세포.
Objective To explore the differentiation of allogeneic naive T cells to regulatory T cells (Tregs) and T helper (TH) 1/2/17 cells by coculture with bone marrow-derivedimmature dendritic cells (irnDC).Method Bone marrow-derived imDC were cultivated from Balb/c mice.Lipopolysaccharide-stimulated DC were harvested as mature dendritic cells (mDC) and unstimulated cells were collected as imDC.Then irnDC or mDC were cocultured with allogeniec naive T cells,respectively.TH1 cytokines [interferon-γ (IFN-γ) and interleukin (IL)-2],TH2 cytokines (IL-4 and IL-10),and TH17 cytokine (IL-17) of co-cultured cells were detected by enzyme linked immunospot assay.CD4+ Forkhead box p3 (FoxP3) + Treg proportion in CD4+ cells in the co-cultured system with IL-2 and transforming growth factor-β1 (TGF-β1) was analyzed by flow cytometry.Result As compared with mDC,na(i)ve T cells cocultured with imDC secreted much less IFN-γ (11.67 ± 2.18 vs.182.00±23.71,P＜0.01),IL-2 (26.67±2.96 vs.318.30± 18.62,P＜0.01),IL-4 (17.00±3.78 vs.45.33±3.48,P＜0.01),IL-10(7.00±1.00vs.158.70±10.90,P＜0.001) and IL-17 (0.66 ± 0.33 vs.238.30 ± 24.39,P＜0.001).Furthermore,imDC induced more CD4+ FoxP3+ Tregs than mDC after adding IL-2 and TGF-β1 in the coculture system for 7 days (22.70 ± 1.53 ％ vs.5.42 ± 1.27％,P＜0.01).Conclusion imDC are more effective to induce na ve T cells to Tregs,but not differentiate to TH 1/TH 2/TH 17 cells.These findings provide in vitro experimental evidence for induction of transplant tolerance by adoptive transfer of imDC.