免疫毒性评价中不同淋巴细胞激活剂致大鼠外周全血产生细胞因子作用的评价

Evaluation on Cytokine Production in Rat Peripheral Whole Blood Induced by Different Lymphocyte Stimulants in Immunotoxicity Bioassay

淋巴细胞激活剂可有效活化免疫细胞,进而促进细胞因子的产生和分泌,但不同激活剂及激活方式的选择直接影响细胞因子产生的种类和水平。为评价不同激活剂促细胞因子产生的作用,筛选一种快速有效的激活方法用于细胞因子检测,本研究采用不同浓度佛波酯(PMA)/钙离子载体、植物血凝素(PHA)、刀豆蛋白A(Con A)、脂多糖(LPS)和美洲商陆素(PWM)体外激活大鼠外周全血0、2、4、6、8、10 h,对各样本中10种细胞因子(白细胞介素2(IL-2)、白细胞介素4(IL-4)、白细胞介素6(IL-6)、白细胞介素10(IL-10)、白细胞介素13(IL-13)、干扰素γ(IFN-γ)、肿瘤坏死因子α(TNF-α)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)、受激活调节正常T细胞表达和分泌因子(RANTES)和转化生长因子β(TGF-β))含量进行检测,并评价了环孢素A和硫唑嘌呤2种免疫抑制剂对激活剂诱导产生细胞因子的抑制作用。结果表明25 ng·mL-1PMA和1μg·mL-1钙离子载体体外诱导6 h可以显著升高大鼠外周全血IL-2、IFN-γ、TNF-α、RANTES和TGF-β含量,是一种作用广泛的激活方法。其诱导产生细胞因子作用的变化可反映机体免疫系统和功能的损伤,可有效用于外源化学物免疫毒性评价。

Lymphocyte stimulants can effectively promote the production and release of several cytokines byactivating the immune cells, but the types and levels of cytokines depend on the kinds of stimulants. To stud-y the activation effects of different lymphocyte stimulants on cytokine production, the authors analyzed theproduction of interleukin-2 （IL-2）, interleukin-4 （IL-4）, interleukin-6 （IL-6）, interleukin-10 （IL-10）, interleukin-13（IL-13）, interferon-γ（IFN-γ）, tumor necrosis factor α （TNF-α）, granulocyte-macrophage colony-stimulating fac-tor （GM-CSF）, regulated upon activation normal T cell expressed and secreted factor （RANTES） and transfor-ming growth factor β （TGF-β） in undiluted rat peripheral whole blood after in vitro culture （0, 2, 4, 6, 8, 10 h） with different concentrations of phorbol-12-myristate-13-acetate （PMA）/Ionomycin, phytohaemagglutinin （PHA）, concanavalin A （Con A）, lipopolysaccharide （LPS） and pokeweed-mitogen （PWM）. Besides, the effects of immunosuppressants （cyclosporine A and azathioprine） on cytokines production induced by the above lymphocyte stimulants were also studied. The results showed that a rapid increase of cytokine （including IL-2, IFN-γ, TNF-α, RANTES and TGF-β） secretion within 6 h after stimulation with 25 ng· mL-1 PMA and 1 μg. mL-1 ionomycin. The inhibition of these cytokine profiles reflected the effects of xenobiotics on immune sys- tem. Therefore, simultaneous detection of cytokine pr^fdes in undiluted whole blood stimulated with PMA/ Ionomycin is a powerful tool for immunotoxicity assessment.