Sage Peter T, Sharpe Arlene H
Department of Microbiology and Immunobiology, Harvard Medical School, NRB, 77 Avenue Louis Pasteur, Boston, MA, 02115, USA.
Methods Mol Biol. 2015;1291:151-60. doi: 10.1007/978-1-4939-2498-1_13.
T follicular helper (T(FH)) cells stimulate, whereas T follicular regulatory (T(FR)) cells inhibit, B cell responses. Despite the potent immunoregulatory roles for TFR cells in controlling the magnitude of antibody production, the precise mechanisms by which T(FR) cells exert their suppressive effects are not yet clear. The lack of specific assays to assess T(FR) cell function separately from differentiation has hindered progress in elucidating T(FR) cell function. This is due, in part, to difficulty in separating T(FR) cells from phenotypically similar, but functionally different, T(FH) cells. Here we describe an in vitro approach for sensitively and quantitatively assessing the capacity of T(FR) cells to suppress T(FH)-mediated B cell antibody production utilizing both ELISA and flow cytometry to measure B cell responses. Beyond assessing T(FR) function, this assay system can also be used to sensitively measure T(FH) stimulatory capacity as well as B cell function.
滤泡辅助性T（T(FH)）细胞可刺激B细胞反应，而滤泡调节性T（T(FR)）细胞则抑制B细胞反应。尽管TFR细胞在控制抗体产生量方面具有强大的免疫调节作用，但其发挥抑制作用的确切机制尚不清楚。缺乏能够独立于分化来评估T(FR)细胞功能的特异性检测方法，阻碍了对T(FR)细胞功能的阐明。部分原因在于，难以将T(FR)细胞与表型相似但功能不同的T(FH)细胞分离。在此，我们描述了一种体外方法，利用酶联免疫吸附测定（ELISA）和流式细胞术来测量B细胞反应，灵敏且定量地评估T(FR)细胞抑制T(FH)介导的B细胞抗体产生的能力。除了评估T(FR)功能外，该检测系统还可用于灵敏地测量T(FH)的刺激能力以及B细胞功能。
