Wang Yafei, Ghali Wa-el, Pingle Pavani, Traboulsi Amar, Dalal Tajas, O'Rourke James, Cone Robert E
Department of Pathology, Vision Immunology Center, University of Connecticut Health Center, Farmington, CT, USA.
Ocul Immunol Inflamm. 2003 Mar;11(1):39-52. doi: 10.1076/ocii.11.1.39.15578.
To characterize the immunoregulatory mechanisms in vitro of spleen cells that are activated by intracameral injection of antigen (AC-spleen cells).
AC-spleen cell regulation of in-vitro antigen-induced proliferation and interferon-gamma production by lymph node cells from TNP-BSA-immunized mice was quantified by co-culture of the lymph node cells with TNP-BSA and AC-spleen cells induced by intracameral TNP-BSA. Cytokine production was quantified by ELISA.
AC-spleen cells produced significantly more IL-4 and IL-10 than spleen cells from TNP-BSA-immunized mice or naive spleen cells; unlike spleen cells from immunized mice, AC-spleen cells did not produce IFN-gamma. AC-splenic CD4(+), CD8(+), CD4(-)/CD8( -) (DN) T cells differentially suppressed antigen-induced proliferation and IFN-gamma production by immunized lymph node cells by a mechanism dependent on IL-10 and antigen. Cultures of lymph node cells, antigen, and AC-splenic T cells contained increased amounts of IL-10 and/or TGFbeta2.
The differential, cytokine-dependent immunoregulatory effects of CD4( +) and CD8(+) AC-spleen cells observed in vitro parallel their effects in vivo. We suggest that the suppression of antigen-induced lymphocyte proliferation and IFN-gamma production by AC-spleen cells provides a useful in-vitro assay of the immunoregulatory activity of cell populations that are induced by the injection of antigen into the anterior chamber.
表征经前房内注射抗原激活的脾细胞(AC-脾细胞)在体外的免疫调节机制。
通过将来自TNP-BSA免疫小鼠的淋巴结细胞与TNP-BSA和经前房内注射TNP-BSA诱导的AC-脾细胞共培养,定量分析AC-脾细胞对体外抗原诱导的增殖以及干扰素-γ产生的调节作用。通过酶联免疫吸附测定法(ELISA)对细胞因子的产生进行定量分析。
与来自TNP-BSA免疫小鼠的脾细胞或未免疫的脾细胞相比,AC-脾细胞产生的白细胞介素-4(IL-4)和白细胞介素-10(IL-10)显著更多;与免疫小鼠的脾细胞不同,AC-脾细胞不产生干扰素-γ(IFN-γ)。AC-脾细胞中的CD4(+)、CD8(+)、CD4(-)/CD8(-)(双阴性,DN)T细胞通过一种依赖于IL-10和抗原的机制,差异性地抑制免疫淋巴结细胞的抗原诱导增殖和IFN-γ产生。淋巴结细胞、抗原和AC-脾T细胞的培养物中白细胞介素-10和/或转化生长因子β2(TGFβ2)的含量增加。
在体外观察到的CD4(+)和CD8(+)AC-脾细胞的差异性、细胞因子依赖性免疫调节作用与其在体内的作用相似。我们认为,AC-脾细胞对抗原诱导的淋巴细胞增殖和IFN-γ产生的抑制作用为注射抗原至前房诱导的细胞群体免疫调节活性提供了一种有用的体外检测方法。