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Title: [Regulatory effect of lactate on peripheral blood CD4⁺ T cell subsets in patients with rheumatoid arthritis].
Author: Huang H, Zhao J, Zhao X, Bai Z, Li X, Wang G.
Journal: Beijing Da Xue Xue Bao Yi Xue Ban; 2024 Jun 18; 56(3):519-525. PubMed ID: 38864139.
Abstract:
OBJECTIVE: To investigate the serum lactate level in patients with rheumatoid arthritis (RA) and its relationship with disease activity, and to analyze the effect of sodium lactate on the activation of CD4⁺ T cells, the ability of secreting cytokines and CD4⁺T cell subsets in peripheral blood of the RA patients. METHODS: The peripheral blood of healthy controls (HC) and RA patients was collected, and the content of lactate in the supernatant was detected by lactate detection kit, the correlation between the content of lactate and the disease score of the RA patients was analyzed; the activation level of CD4⁺ T cells, the proportion of CD4⁺ T cell subsets and the cytokines secreted by CD4⁺ T cells in peripheral blood of all the RA patients were detected by flow cytometry after being stimulated with sodium lactate. RESULTS: The serum lactate level in the RA patients (n=66) was significantly higher than that in the HC (n=60, P < 0.001), and there was a certain correlation with disease activity score in 28 joints (DAS28)-C-reactive protein (CRP) (r=0.273, P=0.029), The levels of rheumatoid factor [RF, 197.50 (26.03, 783.00) IU/mL vs. 29.30 (0.00, 102.60) IU/mL, P < 0.01], CRP [37.40 (11.30, 72.60) mg/L vs. 5.83 (2.36, 12.45) mg/L, P < 0.001], were increased in patients with the lactate concentration greater than 5 mmol/L were significantly higher than those in patients with the lactate concentration less than or equal 5 mmol/L, however, there was no significant difference in the expression of erythrocyte sedimentation rate [ESR, 42.00 (19.00, 77.00) mm/h vs. 25.00 (12.50, 45.50) mm/h, P>0.05] and anti-cyclic citrullinated peptied (CCP) antibody [82.35 (17.70, 137.00) RU/mL vs. 68.60 (25.95, 119.70) RU/mL, P>0.05]. Compared with the control group, the expression of PD-1 (46.15%±8.54% vs. 41.67%±9.98%, P < 0.001), inducible costimulatory molecule (ICOS, 5.77%±8.60% vs. 18.65%±7.94%, P < 0.01) and CD25 (25.89%±5.80% vs. 22.25%±4.59%, P < 0.01) on the surface of CD4⁺ T cells in the RA patients treated with sodium lactate was significantly increased. Compared with the control group, the proportion of Th17 (4.62%±1.74% vs. 2.93%±1.92%, P < 0.05) and Tph (28.02%±6.28% vs. 20.32%±5.82%, P < 0.01) cells in CD4⁺T cells of the RA patients in the sodium lactate treatment group increased. Compared with the control group, the expression of IL-21 (5.73%±1.59% vs. 4.75%±1.71%, P < 0.05) in CD4⁺T cells was up-regulated in the RA patients treated with sodium lactate. CONCLUSION: The level of serum lactate in RA patients is increased, which promotes the activation of CD4⁺T cells and the secretion of IL-21, and up-regulates the proportion of Th17 and Tph cells in the RA patients. 目的: 研究类风湿关节炎(rheumatoid arthritis, RA)患者血清乳酸水平及其与疾病活动度的关系，分析乳酸对RA患者外周血中CD4⁺T细胞的活化、分泌细胞因子能力以及对CD4⁺T细胞亚群的作用。方法: 选择大连医科大学附属第二医院风湿免疫科2019年10月至2021年9月连续收治的RA患者作为病例组，同期与RA患者性别年龄相匹配的大连医科大学附属第二医院体检中心健康人(healthy control, HC)作为对照组，收集两组外周血，乳酸检测试剂盒检测上清液中乳酸含量，并分析其与RA患者疾病评分的相关性。用乳酸刺激后，通过流式细胞术检测所有病例组RA患者外周血中CD4⁺T细胞的活化水平、CD4⁺T细胞亚群的比例以及CD4⁺T细胞分泌的细胞因子。结果: RA患者(n=66)的血清乳酸水平[(5.13±1.01) mmol/L vs. (1.72±0.91) mmol/L, P＜0.001]明显高于HC(n=60)，且与RA患者DAS28(disease activity score in 28 joints)-C反应蛋白(C-reactive protein，CRP)评分有相关性(r=0.273, P=0.029)，乳酸浓度>5 mmol/L较浓度≤5 mmol/L的类风湿因子(rheumatoid factor, RF)[197.50 (26.03, 783.00) IU/mL vs. 29.30 (0.00, 102.60) IU/mL, P＜0.01]和CRP [37.40 (11.30, 72.60) mg/L vs. 5.83 (2.36, 12.45) mg/L, P＜0.001]的表达均升高，而红细胞沉降率(erythrocyte sedimentation rate, ESR)[42.00 (19.00, 77.00) mm/h vs. 25.00 (12.50, 45.50) mm/h, P>0.05]和抗环瓜氨酸多肽(cyclic citrullinated peptied, CCP)抗体[82.35 (17.70, 137.00) RU/mL vs. 68.60 (25.95, 119.70) RU/mL, P>0.05]的表达差异无统计学意义。与对照组HC相比，乳酸处理组的RA患者CD4⁺T细胞表面PD-1 (46.15%±8.54% vs. 41.67%±9.98%, P < 0.001)、诱导共刺激分子(inducible costimulatory molecule, ICOS)(25.77%±8.60% vs. 18.65%±7.94%, P < 0.01)和CD25 (25.89%±5.80% vs. 22.25%±4.59%, P < 0.01)的表达明显升高；乳酸处理组的RA患者CD4⁺T细胞中Th17 (4.62%±1.74% vs. 2.93%±1.92%, P < 0.05)、外周辅助性T (peripheral helper T, Tph)细胞(28.02%±6.28% vs. 20.32%±5.82%, P < 0.01)的比例升高；乳酸处理组的RA患者CD4⁺T细胞中IL-21(5.73%±1.59% vs. 4.75%±1.71%, P < 0.05)的表达上调。结论: RA患者血清乳酸水平升高，促进了RA患者CD4⁺T细胞活化和分泌IL-21，以及上调了RA患者Th17、Tph细胞的比例。

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