CAJ | 학술논문

目的探讨寻常型天疱疮(pemphigus vulgaris,PV)患者抗原特异性Th1和Th2细胞在不同疾病阶段的变化,进一步了解自身反应性T细胞在疾病中的作用.方法收集24例PV患者资料,合成桥粒芯糖蛋白抗原肽段DG3(96～112).体外用该肽段分别刺激患者的外周血单一核细胞(PBMC),用酶联免疫斑点(ELISPOT)方法检测干扰素(IFN)-γ+ Th1细胞和白介素(IL)-4+ Th2细胞的数量,以及记忆性B细胞的数量.用t检验或单因素方差分析(one-way ANOVA)对各组数值进行比较,用Pearson相关系数对Th细胞、记忆性B细胞及抗桥粒芯糖蛋白3(Dsg3)抗体滴度进行相关性分析.结果 24例PV患者男女比例1.67:1,平均年龄(56.59±14.66)岁.5×105个PBMC中,PV患者特异性IFN-γ+ Th1细胞绝对数为420.18±350.29,高于健康对照145.12±86.56,差异有统计学意义(P＜0.05).PV患者特异性IL-4+ Th2的细胞数(366.76±192.44)与健康对照(335.88±164.96)之间差异无统计学意义.PV患者外周血中特异性IL-4+ Th2占总Th2细胞的百分率为37.03％±23.44％,特异性IFN-γ+ Th1细胞为23.62％±16.77％;7例患者进行了治疗前后的自身对照比较.特异性IL-4+ Th2细胞的数量在治疗后(241.68±160.60)较治疗前(452.82±199.29)明显下降,差异有统计学意义(t=2.48,P＜ 0.05).特异性Th细胞、记忆性B细胞及抗Dsg3抗体滴度之间无统计学相关性(均P＞0.05).结论抗原肽段DG3(96～112)含有被PV患者特异性Th细胞识别的致病性抗原表位;特异性IFN-γ+ Th1细胞和IL-4+ Th2细胞均在疾病中发挥一定的作用;特异性IL-4+ Th2细胞在疾病活动中可能起到了更重要的致病作用.
목적 탐토심상형천포창(pemphigus vulgaris,PV)환자항원특이성Th1화Th2세포재불동질병계단적변화,진일보료해자신반응성T세포재질병중적작용.방법 수집24례PV환자자료,합성교립심당단백항원태단DG3(96～112).체외용해태단분별자격환자적외주혈단일핵세포(PBMC),용매련면역반점(ELISPOT)방법검측간우소(IFN)-γ+ Th1세포화백개소(IL)-4+ Th2세포적수량,이급기억성B세포적수량.용t검험혹단인소방차분석(one-way ANOVA)대각조수치진행비교,용Pearson상관계수대Th세포、기억성B세포급항교립심당단백3(Dsg3)항체적도진행상관성분석.결과 24례PV환자남녀비례1.67:1,평균년령(56.59±14.66)세.5×105개PBMC중,PV환자특이성IFN-γ+ Th1세포절대수위420.18±350.29,고우건강대조145.12±86.56,차이유통계학의의(P＜0.05).PV환자특이성IL-4+ Th2적세포수(366.76±192.44)여건강대조(335.88±164.96)지간차이무통계학의의.PV환자외주혈중특이성IL-4+ Th2점총Th2세포적백분솔위37.03％±23.44％,특이성IFN-γ+ Th1세포위23.62％±16.77％;7례환자진행료치료전후적자신대조비교.특이성IL-4+ Th2세포적수량재치료후(241.68±160.60)교치료전(452.82±199.29)명현하강,차이유통계학의의(t=2.48,P＜ 0.05).특이성Th세포、기억성B세포급항Dsg3항체적도지간무통계학상관성(균P＞0.05).결론 항원태단DG3(96～112)함유피PV환자특이성Th세포식별적치병성항원표위;특이성IFN-γ+ Th1세포화IL-4+ Th2세포균재질병중발휘일정적작용;특이성IL-4+ Th2세포재질병활동중가능기도료경중요적치병작용.
Objective To study the changes of antigen-specific T helper type 1 (Th1) cells and Th2 cells in patients with pemphigus vulgaris (PV) at different stages,so as to better understand the roles of autoreactive T cells in PV.Methods The DG3 (96-112) peptide was synthesized.Twenty-four patients with PV and 10 health checkup examinees were included in this study.Peripheral blood mononuclear cells (PBMCs) were obtained from the health checkup examinees and all the patients before treatment and seven patients about one month after treatment,and stimulated with the DG3 peptide of 10 mg/L for different durations.Then,enzyme-linked immunospot (ELISPOT) assay was performed to count the number of DG3-specific IFN-γ+ Th1 cells,IL-4+ Th2 cells and memory B cells.One-way analysis of variance (ANOVA) and t test were done to compare the number of cells between different groups,and Pearson correlation coefficient was used to evaluate the correlation among Th cells,memory B cells and anti-desmoglein 3 (Dsg3) antibody titers.Results In these patients,the male to female ratio was 1.67 ∶ 1,and the average age was (56.59 ± 14.66) years.Compared with the health check-up examinees,the patients with PV showed a higher absolute number of DG3-specific IFN-γ+ Th1 cells (420.18 ± 350.29 vs.145.12 ± 86.56,t =3.25,P ＜ 0.05),but a similar absolute number of specific IL-4+ Th2 cells (366.76 ± 192.44 vs.335.88 ± 164.96,P＞ 0.05) per 5 × 105 PBMCs.The percentage of DG3-specific IL-4+ Th2 cells in Th2 cells was 37.03％ ± 23.44％,and the percentage of IFN-γ+ Th1 cells was 23.62％ ± 16.77％ in peripheral blood of patients with PV.The number of DG3-specific IL-4+ Th2 cells per 5 × 105 PBMCs significantly decreased from 452.82 ± 199.29 before treatment to 241.68 ± 160.60 after treatment in seven patients (t =2.48,P ＜ 0.05).There was no significant correlation between specific Th cells,memory B cells and anti-Dsg3 antibody titers (all P ＞ 0.05).Conclusions The peptide DG3 (96-112) has pathogenic epitopes which could be recognized by specific Th cells of patients with PV.Both antigen-specific IFN-γ+ Th1 cells and IL-4+ Th2 cells play certain roles in the pathogenesis of pemphigns vulgaris,and IL-4+ Th2 cells appear to be more important.