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目的利用三氯乙烯(TCE)染毒的小鼠模型,研究T、B淋巴细胞及细胞因子在TCE诱发过敏反应中的作用。方法以皮肤接触同时结合吸入对12只小鼠进行致敏处理,6周后耳部涂抹TCE进行激发。以耳肿胀系数作为评价小鼠过敏反应的指标;分离脾细胞体外培养,采用噻唑蓝(MTT)法测定脾淋巴细胞抗原特异性增殖反应;酶联免疫吸附测定(ELISA)方法测定细胞培养上清液中IgG和白介素-4(IL-4)、γ型干扰素(IFN-γ)水平;应用流式细胞技术进行淋巴细胞亚群分析。结果TCE致敏组小鼠脾淋巴细胞加入TCE体外培养后,细胞存活率明显高于溶剂对照组[(79±10)%vs(63±11)%,P<0·05];细胞培养上清液中IgG水平(与二甲基亚砜对照孔比较的相对值)在两组间比较差异有显著性[(70±5%)vs(53±6)%,P<0·01]。流式细胞分析结果显示,TCE致敏组脾淋巴细胞与TCE共培养后,CD3+T细胞占总淋巴细胞的比例[(41·6±4·4)%]及CD4+/CD8+(2·1±0·6)与对照组比较[分别为(39·4±4·0)%和(2·3±0·9)],差异无显著性(P>0·05)。TCE致敏组小鼠激发时未见耳肿胀,然而TCE致敏组小鼠IFN-γ/IL-4比值(0·54±0·12)明显低于溶剂对照组(0·90±0·22,P<0·01)。结论TCE能够诱导T淋巴细胞的增殖活化,分泌Th2型细胞因子,并刺激B淋巴细胞分泌抗原特异性的IgG抗体。
Objective To study the role of T, B lymphocytes and cytokines in the TCE-induced allergic reaction in mice induced by trichlorethylene (TCE). Methods Twelve mice were sensitized with skin contact and inhalation. TCE was applied to the ear after 6 weeks. Ear swelling index was used as an index to evaluate the anaphylaxis in mice. Splenocytes were isolated and cultured in vitro. Antigen-specific proliferative responses of splenic lymphocytes were measured by MTT assay. ELISA assay was used to determine the cell- IgG, IL-4 and IFN-γ levels in the blood were measured. Lymphocyte subsets were analyzed by flow cytometry. Results TCE sensitized group of mice spleen lymphocytes added TCE cultured in vitro, the cell survival rate was significantly higher than the solvent control group (79 ± 10) vs (63 ± 11)%, P <0 · 05]; cell culture Serum IgG levels (relative to dimethylsulfoxide control) were significantly different between the two groups (70 ± 5% vs 53 ± 6%, P <0.01). Flow cytometry analysis showed that the ratio of CD3 + T cells to total lymphocytes (41.6 ± 4.4%) and CD4 + / CD8 + (-2.1) in TCE sensitized group after co-cultured with TCE ± 0 · 6) compared with the control group [(39.4 ± 4.0)% and (2.3 ± 0.9)] respectively, with no significant difference (P> 0.05). TCE sensitized mice did not have ear swelling when stimulated, however, the ratio of IFN-γ / IL-4 in TCE-sensitized mice was significantly lower than that in the solvent control group (0 · 90 ± 0 · 22, P <0.01). Conclusion TCE can induce the proliferation and activation of T lymphocytes, secrete Th2-type cytokines, and stimulate B lymphocytes to secrete antigen-specific IgG antibodies.