补体调节蛋白CD55和CD59在Stevens-Johnson综合征/中毒性表皮坏死松解症患者外周血的表达水平
Expression levels of complement regulatory proteins CD55 and CD59 in the peripheral blood of patients with Stevens-Johnson syndrome/toxic epidermal necrolysis
目的:检测Stevens-Johnson综合征(SJS)和中毒性表皮坏死松解症(TEN)患者外周血补体调节蛋白CD55和CD59的表达水平,初步分析其在发病中的可能作用。方法:收集2017年12月至2022年12月在安徽医科大学第一附属医院和安徽医科大学第二附属医院皮肤科住院治疗的SJS和TEN患者(SJS/TEN组),同时收集发疹型药疹患者(轻症对照)和健康对照。采用流式细胞仪检测外周血单个核细胞(PBMC)中CD4 + T淋巴细胞和CD8 + T淋巴细胞的比例。通过流式液相多重蛋白定量技术检测炎症相关细胞因子肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-17、IL-10、γ干扰素(IFN-γ)、IL-2和IL-4的表达水平。采用实时荧光定量PCR检测PBMC中CD55和CD59 mRNA水平。采用流式细胞术检测CD8 + T淋巴细胞表面CD55和CD59蛋白的表达情况。采用单因素方差分析和Tukey检验进行统计学分析。 结果:共收集13例SJS/TEN患者,27例发疹型药疹患者,40例健康对照。SJS/TEN患者中,男8例,女5例,18 ~ 84(47.15 ± 19.99)岁,病程(7.74 ± 2.63) d,3组性别和年龄的差异均无统计学意义( P > 0.05)。3组CD4 + T淋巴细胞比例差异无统计学意义( F = 3.84, P = 0.051)。SJS/TEN患者外周血CD8 + T淋巴细胞比例(25.60% ± 4.57%)较健康对照(16.20% ± 6.28%)更高( q = 4.59, P = 0.018)。SJS/TEN患者细胞因子TNF-α、IL-6、IL-10、IL-17和IFN-γ的表达水平高于健康对照和轻症对照组(均 P < 0.001)。此外,SJS/TEN患者PBMC中CD55( F = 9.46, P < 0.001)和CD59 mRNA表达( F = 15.14, P < 0.001)均低于健康对照和轻症对照组。SJS/TEN患者CD8 + T淋巴细胞表面CD55( F = 51.51, P < 0.001)和CD59蛋白( F = 31.59, P < 0.001)亦明显低于其他两组或健康对照组。 结论:补体调节蛋白CD55和CD59在SJS/TEN患者中表达降低,可能与SJS/TEN患者CD8 + T淋巴细胞激活以及体内过度的炎症反应相关。
更多Objective:To detect expression levels of complement regulatory proteins CD55 and CD59 in the peripheral blood of patients with Stevens-Johnson syndrome (SJS) /toxic epidermal necrolysis (TEN), and to preliminarily analyze their potential roles in the occurrence of SJS/TEN.Methods:Hospitalized patients with SJS/TEN (SJS/TEN group) were collected from the Department of Dermatology of the First Affiliated Hospital of Anhui Medical University and the Second Affiliated Hospital of Anhui Medical University from December 2017 to December 2022. Meanwhile, patients with maculopapular exanthema (MPE) and healthy physical examinees were also collected and served as the mild group and healthy control group, respectively. Flow cytometry was performed to determine the proportions of CD4 + T lymphocytes and CD8 + T lymphocytes in peripheral blood mononuclear cells (PBMCs). The expression levels of inflammatory cytokines tumor necrosis factor (TNF) -α, interleukin (IL) -6, IL-17, IL-10, interferon (IFN) -γ, IL-2, and IL-4 were detected using flow cytometric bead array technology. The mRNA expression levels of CD55 and CD59 in PBMCs were detected by real-time fluorescence-based quantitative PCR (qRT-PCR). Flow cytometry was also performed to determine the protein expression of CD55 and CD59 on the surface of CD8 + T lymphocytes. Statistical analyses were carried out using one-way analysis of variance and Tukey's test. Results:Totally, 13 patients with SJS/TEN, 27 patients with MPE, and 40 healthy controls were collected. Among the SJS/TEN patients, there were 8 males and 5 females, with their age being 18 to 84 (47.15 ± 19.99) years, and disease duration being 7.74 ± 2.63 days. No significant differences were observed in the gender distribution or age among the 3 groups (both P > 0.05). The proportions of CD4 + T lymphocytes did not differ among the 3 groups ( F = 3.84, P = 0.051). The proportions of CD8 + T lymphocytes in the peripheral blood were significantly higher in the SJS/TEN group (25.60% ± 4.57%) than in the healthy control group (16.20% ± 6.28%; q = 4.59, P = 0.018). The expression levels of inflammatory cytokines TNF-α, IL-6, IL-10, IL-17, and IFN-γ were significantly higher in the SJS/TEN group than in the healthy control group and mild group (all P < 0.001). In addition, the mRNA expression of CD55 ( F = 9.46, P < 0.001) and CD59 in PBMCs ( F = 15.14, P < 0.001) was significantly lower in the SJS/TEN group than in the mild group and healthy control group. The protein expression levels of CD55 ( F = 51.51, P < 0.001) and CD59 ( F = 31.59, P < 0.001) on the surface of CD8 + T lymphocytes were also significantly lower in the SJS/TEN group than in the other two groups and the healthy control group, respectively. Conclusion:Complement regulatory proteins CD55 and CD59 were downregulated in SJS/TEN patients, which may be associated with the activation of CD8 + T lymphocytes and excessive inflammatory responses.
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