Low T cell production of TNFalpha and IFNgamma in ankylosing spondylitis: its relation to HLA-B27 and influence of the TNF-308 gene polymorphism.
Low T cell production of TNFalpha and IFNgamma in ankylosing spondylitis: its relation to HLA-B27 and influence of the TNF-308 gene polymorphism.
复制标题
强直性脊柱炎中 T 细胞产生的 TNFα 和 IFNγ 低：与 HLA-B27 的关系以及 TNF-308 基因多态性的影响。
DOI：
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发表时间：
2001
影响因子：
27.4
通讯作者：
J. Braun
中科院分区：
文献类型：
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作者：
M. Rudwaleit;S. Siegert;Z. Yin;J. Eick;A. Thiel;Andreas Radbruch;J. Sieper;J. Braun
客观化
为验证强直性脊柱炎(AS)是一种辅助性T细胞2型极化疾病的假说，在单个细胞水平上对AS患者T细胞细胞因子干扰素-γ(IFNGamma)、白介素4(IL4)、肿瘤坏死因子α(TNFpha)和IL10进行定量，并与正常对照组和正常对照组进行比较。
方法
用佛波酯/离子霉素刺激65例受试者(25例活动期AS患者、18例正常对照和22例阴性对照)外周血单个核细胞，表面CD3和CD8染色，细胞内细胞因子干扰素-γ、肿瘤坏死因子α、白介素4和白介素10染色，用流式细胞仪进行分析。肿瘤坏死因子α的产生与肿瘤坏死因子α启动子的-308和-238位基因多态有关。
结果
在人类白细胞抗原B27阳性的AS患者外周血中，肿瘤坏死因子α+T细胞的百分比(中位数为5.1%)显著低于健康对照组(中位数9.5%；p=0.008)。令人惊讶的是，在健康的人类白细胞抗原-B27阳性的对照组中，肿瘤坏死因子α+T细胞的百分比(中位数7.48%)也显著低于健康的人类白细胞抗原-B27阴性的对照组(p=0.034)。此外，强直性脊柱炎患者和HLAB27阳性对照组的IFNγ+T细胞百分率均低于HLAB27阴性对照组(p=0.005和p=0.003)。强直性脊柱炎患者外周血中IL10+/CD8+T细胞百分率明显高于正常对照组。在HLAB27阳性受试者中，TnF1/2杂合子位于-308(n=6)与TnF1/1纯合子T细胞的百分比(n=25；中位数9.97%vs5.11%；p=0.017)相关。相反，在HLA-B27阴性对照组(n=18)中，没有这种基因/表型相关性(中位数9.4%vs10.6%)。
结论
人类白细胞抗原B27阳性的AS患者和正常对照在单个细胞水平表现出较低的T细胞分泌TNFα和IFNGamma，这可能是导致人类白细胞抗原B27阳性患者发生AS的易感性增加的原因之一。初步的基因型-表型相关性表明，在人类白细胞抗原-B27阳性的受试者中，位于-308位的TNF2或一个连锁基因会导致较高的TNFα产量，因此，可能是保护性单倍型的标志。
OBJECTIVE
To test the hypothesis that ankylosing spondylitis (AS) is a T helper cell type 2 polarised disease by quantifying the T cell cytokines interferon gamma (IFNgamma), interleukin 4 (IL4), tumour necrosis factor alpha (TNFalpha), and IL10 at the single cell level in patients with AS in comparison with healthy HLA-B27 negative and HLA-B27 positive controls.
METHODS
Peripheral blood mononuclear cells from 65 subjects (25 HLA-B27 positive patients with active AS, 18 healthy HLA-B27 positive controls, and 22 healthy HLA-B27 negative controls) were stimulated with phorbol myristate acetate/ionomycin for six hours, surface stained for CD3 and CD8, intracellularly stained for the cytokines IFNgamma, TNFalpha, IL4, and IL10, and analysed by flow cytometry. TNFalpha production was related to the genotype of the TNFalpha promoter at the -308 and -238 polymorphisms.
RESULTS
In peripheral blood the percentage of TNFalpha+ T cells was significantly lower in HLA-B27 positive patients with AS (median 5.1% for CD4+ T cells) than in healthy HLA-B27 negative controls (median 9.5%; p=0.008). Surprisingly, the percentage of TNFalpha+ T cells was also significantly lower in healthy HLA-B27 positive controls (median 7.48%) than in healthy HLA-B27 negative controls (p=0.034). Furthermore, the percentage of IFNgamma+ T cells was lower in patients with AS and in healthy HLA-B27 positive controls than in healthy HLA-B27 negative controls (p=0.005 and p=0.003, respectively). The percentage of IL10+/CD8+ T cells was higher in patients with AS than in both control groups. In HLA-B27 positive subjects, TNF1/2 heterozygosity at -308 (n=6) was associated with a higher percentage of TNFalpha+ T cells than TNF1/1 homozygosity (n=25; median 9.97% v 5.11% for CD4+ T cells; p=0.017). In contrast, in HLA-B27 negative controls (n=18) there was no such genotype/phenotype correlation (median 9.4% v 10.6%).
CONCLUSIONS
The lower T cell production of TNFalpha and IFNgamma shown at the single cell level in HLA-B27 positive patients with AS and healthy HLA-B27 positive controls may contribute to the increased susceptibility of HLA-B27 positive subjects to develop AS. Preliminary genotype-phenotype correlations suggest that in HLA-B27 positive subjects TNF2 at -308 or a linked gene results in higher TNFalpha production and, therefore, might be a marker for a protective haplotype.