刺激原诱导的华支睾吸虫感染大鼠淋巴细胞体外增殖和细胞因子的产生

目的　研究感染华支睾吸虫的大鼠T淋巴细胞对寄生虫抗原或促有丝分裂原植物血凝素刺激后的体外增殖及细胞因子产生的免疫效应。　方法　观察感染华支睾吸虫的大鼠脾淋巴细胞和肠系膜淋巴结细胞 ,在促有丝分裂原PHA ,华支睾吸虫分泌排泄抗原 ,华支睾吸虫成虫粗抗原和异尖线虫幼虫粗抗原刺激下 ,体外淋巴细胞增殖及细胞因子的水平 ,包括IFN γ ,IL 2 ,IL 4,IL 1 0。　结果　肠系膜淋巴结的淋巴细胞增殖高于脾淋巴细胞。淋巴细胞浓度为 3× 1 0 6或 9× 1 0 6 细胞/孔时 ,脾脏和肠系膜淋巴细胞增殖均显著高于对照组 (P <0 .0 1 )。淋巴细胞数为 5× 1 0 6 细胞 /孔 ,刺激原浓度为 5或 1 0 μg/ml时 ,淋巴细胞活化明显。肠系膜淋巴结细胞的细胞因子 ,IFN γ和IL 1 0的产生水平显著增加。　 结论　淋巴细胞浓度 5×1 0 6 细胞/孔和刺激原浓度 1 0 μg/ml的条件为激活T淋巴细胞增殖和细胞因子产生的最适培养条件。华支睾吸虫感染大鼠的成虫分泌排泄抗原在体内可能刺激淋巴细胞增殖和细胞因子产生

Proliferation and Cytokine Production of Lymphocytes from Clonorchis sinensis-infected Rats in Response to Stimulators in vitro

OBJECTIVE: To study the rat lymphocyte proliferation, differentiation and cytokine production in response to Clonorchis sinensis infection. METHODS: The lymphocyte proliferation and cytokine production (IFN-gamma, IL-2, IL-4, IL-10) in response to mitogen phytohaemagglutinin (PHA), C. sinensis excretory-secretory antigen (ES Ag), C. sinensis crude antigen (crude Ag) and Anisakis larvae antigen were detected in vitro from splenic lymphocytes (SLC) and mesenteric lymph node cells (MLNC) of rats infected with C. sinensis. Statistical analysis was performed by Sigma Plot System. RESULTS: Lymphocyte proliferations in MLNC were higher than that in SLC. At concentrations of 3 x 10(6) or 9 x 10(6) cells/well, lymphocyte proliferations were significantly higher in both SLS and MLNC than in the control with cell alone (P < 0.01). At the lymphocyte concentration of 5 x 10(6) cells/well and stimulator concentration of 5 or 10 micrograms/ml, significant lymphocyte activation was observed. Under the same culture condition (5 x 10(6) cells/ well with 10 micrograms/ml stimulator), cytokine IFN-gamma and IL-10 production in vitro increased significantly in MLNC. CONCLUSION: Concentrations of 5 x 10(6) lymphocytes/well and 10 micrograms/ml stimulator were selected as the optimal culture condition for activation of lymphocyte proliferation and cytokine production. Since the production of Th1-type cytokine IFN-gamma and Th2-type cytokine IL-10 was much enhanced from MLNC of C. sinensis-infected rats, it is considered that C. sinensis ES Ag may stimulate lymphocyte proliferation and cytokine production in C. sinensis-infected rats in vivo.