自体全血注射对自体血清皮肤试验阳性的难治性慢性自发性荨麻疹患者嗜碱性粒细胞FcεRⅠ与CD63表达的影响

Effect of autologous whole blood injections on expression of FcεRⅠ and CD63 on basophils in patients with refractory chronic spontaneous urticaria and positive autologous serum skin test

目的评估自体全血注射(AWBI)联合抗组胺药治疗对自体血清皮肤试验(ASST)阳性的难治性慢性自发性荨麻疹患者的临床疗效,并检测其对嗜碱性粒细胞高亲和力IgE受体(FcεRⅠ)以及CD63表达水平的影响,探讨AWBI治疗ASST阳性慢性荨麻疹的机制。方法2017年11月至2018年6月在陆军军医大学第一附属医院皮肤科门诊收集ASST阳性的慢性难治性荨麻疹患者80例,随机数字表法分成AWBI组(40例)及对照组(40例),两组均常规口服氯雷他定和依巴斯汀治疗,AWBI组同时接受AWBI治疗每周1次,共12次。治疗前及12周后评估两组周荨麻疹活动程度评分(UAS7)以及皮肤病生活质量指数(DLQI)。AWBI组30例在治疗前及第4、8、12周用流式细胞仪检测外周血嗜碱性粒细胞FcεRⅠ、CD63表达水平。采用GraphPad Prism7.00软件进行统计分析,两组UAS7和DLQI比较采用t检验,FcεRⅠα表达水平比较采用Mann-Whitney U检验,不同时点指标两两比较采用Wilcoxon's配对秩和检验,指标相关性采用Spearman相关分析。结果AWBI组在治疗前及治疗后第12周UAS7分别为27.15±4.53、14.25±7.56,DLQI分别为16.88±6.01、8.48±4.15;对照组UAS7分别为26.90±5.22、19.93±6.32,DLQI分别为17.08±6.79、13.93±5.43。两组治疗前UAS7与DLQI评分差异均无统计学意义;治疗后两组UAS7与DLQI均较治疗前明显降低(均P<0.01),且AWBI组UAS7及DLQI评分显著低于对照组(均P<0.01)。AWBI组患者治疗前和第4、8、12周时嗜碱性粒细胞FcεRⅠα荧光强度中位数(P25,P75)分别为22532(16740,29220)、16911(10240,21816)、13282(7600,16848)与11466(7161,14578),ASST阳性血清诱导的CD63阳性嗜碱性粒细胞比例中位数(P25,P75)分别为35.25%(26.75%,49.13%)、25.95%(19.37%,37.54%)、13.57%(7.79%,19.57%)与9.87%(6.43%,16.52%),治疗第4周时嗜碱性粒细胞FcεRⅠα与CD63表达水平均较基线显著降低,且第8周较第4周亦显著降低(均P<0.01)。AWBI治疗前至第4周、第4~8周以及第8~12周FcεRⅠα的变化值与ASST阳性血清诱导的CD63变化值均呈不同程度正相关,r值分别为0.364、0.422、0.455,均P<0.05。结论AWBI联合抗组胺药有助于改善ASST阳性难治性慢性自发性荨麻疹的临床症状,并且可能通过影响嗜碱性粒细胞FcεRⅠ以及CD63表达水平发挥作用。

Objective To evaluate the clinical efficacy of autologous whole blood injections(AWBI)combined with antihistamines for the treatment of patients with refractory chronic spontaneous urticaria and positive autologous serum skin test(ASST),to evaluate its effect on the expression of the high-affinity IgE receptor(FcεRⅠ)and CD63 on basophils,and to analyze the possible mechanism underlying the treatment of ASST-positive chronic urticaria with AWBI.Methods Eighty patients with ASST-positive chronic intractable urticaria were enrolled from Department of Dermatology,The First Hospital Affiliated to Army Medical University between November 2017 and June 2018,and randomly and equally divided into two groups by a random number table:AWBI group and control group were both conventionally treated with oral loratadine and ebastine,and AWBI group were additionally treated with AWBI once a week for 12 sessions.Before the treatment and after 12-week treatment,urticaria activity score of 7 days(UAS7)and dermatology life quality index(DLQI)in the two groups were evaluated.Among 30 patients in the AWBI group,flow cytometry was performed to determine the expression of FcεRⅠ and CD63 on the basophils in the peripheral blood at the baseline,weeks 4,8 and 12 after the initial treatment.Statistical analysis was carried out with GraphPad Prism 7.00 software by t test for the comparison of UAS7 or DLQI scores,Mann-Whitney U test for the comparison of FcεRⅠα expression,paired Wilcoxon signed rank test for comparing FcεRⅠα or CD63 expression between two different time points,and Spearman correlation analysis for analyzing the correlation between FcεRⅠα and CD63 expression.Results Before the treatment,no significant differences in UAS7 or DLQI scores were observed between the AWBI group and control group(UAS7:27.15±4.53 vs.26.90±5.22;DLQI:16.88±6.01 vs.17.08±6.79;both P > 0.05).After 12-week treatment,UAS7 and DLQI scores both significantly decreased in the two groups compared with those before the treatment(all P<0.01),and were significantly lower in the AWBI group than in the control group(UAS7:14.25±7.56 vs.19.93±6.32;DLQI:8.48±4.15 vs.13.93±5.43;both P<0.01).At the baseline,weeks 4,8 and 12 after the initial treatment,the fluorescence intensities of FcεRⅠα on basophils(M [P25,P75])in the AWBI group were 22 532(16 740,29 220),16 911(10 240,21 816),13 282(7 600,16 848)and 11 466(7 161,14 578)respectively,and the proportions of CD63^+ basophils induced by ASST-positive serum(M [P25,P75])in the AWBI group were 35.25%(26.75%,49.13%),25.95%(19.37%,37.54%),13.57%(7.79%,19.57%)and 9.87%(6.43%,16.52%)respectively.At week 4 after the initial treatment,the expression of FcεRⅠα and CD63 on basophils in the AWBI group both significantly decreased compared with those at the baseline(both P<0.01),but significantly increased compared with those at week 8(both P<0.01).The changes in FcεRⅠα expression from baseline to week 4,from week 4 to week 8,and from week 8 to week 12 were positively correlated with the changes in CD63 expression induced by ASST-positive serum(r=0.364,0.422,0.455,respectively,all P<0.05).Conclusion AWBI combined with antihistamines can improve the clinical symptoms of ASST-positive refractory chronic urticaria,likely by affecting the expression of FcεRⅠand CD63 on basophils.