摘要目的 探讨氨基酮戊酸光动力(ALA-PDT)治疗尖锐湿疣的局部免疫反应.方法 分别采用体外细胞共培养体系和在体免疫组化方法进行研究.将前期建立的稳定表达人乳头瘤病毒16E7蛋白HaCaT细胞株即HPV 16E7/HaCaT细胞作为HPV感染角质形成细胞模型,将ALA-PDT处理后的HPV 16E7/HaCaT细胞与外周血单一核细胞(PBMC)经Transwell小室共培养,3h后观察PBMC的趋化情况;同时采用免疫组化方法研究10例尖锐湿疣患者光动力前及光动力后1、2、3、48 h时,皮损中CD4+T细胞、CD8+T细胞、CD68+巨噬细胞的数量及CD4+/CD8+T细胞数比值变化情况.结果 与ALA-PDT处理后的HPV 16E7/HaCaT细胞共培养3h后,PBMC有明显趋化现象.临床标本免疫组化结果显示,48 h时CD4+T细胞、CD8+T细胞数量及其比值明显升高(P＜0.05),而CD68+巨噬细胞在3h及48 h均升高(P＜0.05).结论 ALA-PDT治疗尖锐湿疣的过程中,可诱导机体产生局部抗病毒免疫反应.

abstractsObjective To investigate the local immune response in condyloma acuminatum treated with aminolevulinic acid-photodynamic therapy (ALA-PDT).Methods In vitro and in vivo studies were performed.A previously established keratinocyte cell line human papilloma virus (HPV) 16E7/HaCaT which stably expresses HPV16E7 protein was used in this study.Peripheral blood mononuclear cells (PBMCs) were separated from 10 healthy volunteers.After pretreatment with ALA-PDT,HPV16E7/HaCaT cells were cocultured with the PBMCs for 3 hours in a Transwell chamber followed by the observation of chemotactic migration of PBMCs.Tissue samples were obtained from the lesions of 10 patients with condyloma acuminatum before,and at 1,2,3 and 48 hours after the first session of ALA-PDT.Immunohistochemistry was conducted to determine the number of CD4+ T cells,CD8+ T cells and CD68+ macrophages as well as CD4/CD8 T-cell ratio in the tissue samples.Results After 3-hour coculture with HPV16E7/HaCaT cells pretreated by ALA-PDT,PBMCs showed apparent chemotactic migration.Immunohistochemistry revealed a statistical increase in the number of CD4+ T cells,CD8+ T cells and CD4/CD8 T-cell ratio at 48 hours (all P ＜ 0.05),as well as in the number of CD68+ macrophages at 3 hours and 48 hours (both P ＜ 0.05) after the first session of ALA-PDT.Conclusion ALA-PDT may induce local antiviral immune response in condyloma acuminatum.