School of Nursing, Anhui Medical University, Hefei, Anhui, China.
Department of Epidemiology and Biostatistics, School of Public Health, Anhui Medical University, Hefei, Anhui, China.
Clin Rheumatol. 2024 Nov;43(11):3515-3523. doi: 10.1007/s10067-024-07124-x. Epub 2024 Sep 5.
This work aims to investigate whether RIP2 silencing in naive CD4 T cells from lupus-prone mice impacts Th17 cell activity or differentiation in vitro.
Naive CD4 T cells isolation from MRL/lpr mice's spleens. Three RNA interference target sequences of RIP2 were packaged with lentivirus and transfected into naive CD4 T cells. The shRIP2 with the highest interference efficiency was selected and transfected into naive CD4 T cells. Naive CD4 T cells were cultured under conventional (TGF-β1 and IL-6) and pathogenic (IL-6, IL-23, IL-1β) differentiation environments, respectively. Then, RT-qPCR, Western blot or Flow Cytometry were used for measuring the amounts of RIP2 and IL-17 and the differentiation of Th17 cells in two settings.
Under the conventional Th17 (cTh17) cell differentiation environment (TGF-β1 and IL-6), RIP2 deficiency is linked to decreased IL-17A levels (1.00 ± 0.03 vs 0.80 ± 0.03) and attenuated cTh17 cell (2.46 ± 0.08 vs 0.78 ± 0.03) differentiation (all, P < 0.05). Under the pathogenic Th17 (pTh17) cell environment (IL-1β, IL-23, IL-6), RIP2 deficiency is linked to elevated IL-17A levels (1.03 ± 0.05 vs 1.63 ± 0.07) and enhanced pTh17 cell (3.69 ± 0.19 vs 5.49 ± 0.10) differentiation (all, P < 0.05).
Our data suggest that RIP2 inhibition induces preferential differentiation of naive CD4 T cells to pathogenic Th17 cells, while being able to upregulate IL-17A levels in the context of pTh17 cell differentiation. Our study opens up new research areas to reveal the underlying mechanisms and potential therapeutic targets for the prevention and treatment of SLE patients. Key Points • Silencing of RIP2 in naive CD4 T cells from lupus-prone mice promotes pathogenic Th17 (pTh17) cell differentiation and IL-17A production under pTh17 cell (IL-1β, IL-23, and IL-6) conditions. • RIP2 deficiency in naive CD4 T cells reduces conventional Th17 (cTh17) cell differentiation and IL-17A production under cTh17 cell (TGF-β1 and IL-6) conditions. • RIP2-deficient naive CD4 T cells preferentially differentiate towards pTh17 cells rather than cTh17 cells in vitro. • Inhibition of RIP2 may be involved in the development of SLE via effects on Th17/IL-17.
本研究旨在探讨在狼疮易感小鼠的初始 CD4 T 细胞中沉默 RIP2 是否会影响体外 Th17 细胞的活性或分化。
从 MRL/lpr 小鼠的脾脏中分离初始 CD4 T 细胞。将 RIP2 的三个 RNA 干扰靶序列包装到慢病毒中，并转染到初始 CD4 T 细胞中。选择干扰效率最高的 shRIP2 并转染到初始 CD4 T 细胞中。在常规（TGF-β1 和 IL-6）和致病性（IL-6、IL-23、IL-1β）分化环境下培养初始 CD4 T 细胞。然后，使用 RT-qPCR、Western blot 或流式细胞术分别测量两种环境下 RIP2 和 IL-17 的量以及 Th17 细胞的分化。
在常规 Th17（cTh17）细胞分化环境（TGF-β1 和 IL-6）下，RIP2 缺陷与 IL-17A 水平降低（1.00±0.03 对 0.80±0.03）和 cTh17 细胞分化减弱（2.46±0.08 对 0.78±0.03）相关（均，P<0.05）。在致病性 Th17（pTh17）细胞环境（IL-1β、IL-23、IL-6）下，RIP2 缺陷与 IL-17A 水平升高（1.03±0.05 对 1.63±0.07）和 pTh17 细胞分化增强（3.69±0.19 对 5.49±0.10）相关（均，P<0.05）。
我们的数据表明，RIP2 抑制诱导初始 CD4 T 细胞向致病性 Th17 细胞优先分化，同时能够在 pTh17 细胞分化的情况下上调 IL-17A 水平。我们的研究开辟了新的研究领域，以揭示 SLE 患者预防和治疗的潜在机制和治疗靶点。
狼疮易感小鼠的初始 CD4 T 细胞中 RIP2 的沉默促进了致病性 Th17（pTh17）细胞在 pTh17 细胞（IL-1β、IL-23 和 IL-6）条件下的分化和 IL-17A 的产生。
初始 CD4 T 细胞中 RIP2 的缺失减少了常规 Th17（cTh17）细胞在 cTh17 细胞（TGF-β1 和 IL-6）条件下的分化和 IL-17A 的产生。
在体外，RIP2 缺陷的初始 CD4 T 细胞优先向 pTh17 细胞而不是 cTh17 细胞分化。
RIP2 抑制可能通过对 Th17/IL-17 的影响参与 SLE 的发生。
