Immunovirology of Gingival Grevicular Fluid in HIV Patients

目的：探讨艾滋病相关牙周病变局部免疫反应的特点及其与临床和免疫生物学指标的关系，提出HIV相关牙周炎(HIV-P)的诊断标准。方法：采用流式细胞术(FCM)对CDC IV期AIDS患者牙周液中P24+细胞进行检测。外周血(PB)白细胞作为系统对照。对数值数据集进行主成分分析。结果：在91名筛查的患者中，44名患者患有HIV-P(48.35%)。23例患者的三色FCM结果显示，GCF中CD68+/p24+和CD14+/CD68+/p24+细胞比例明显高于PB，CD14+/p24+比例低于PB，CD14+/p24+细胞中p24的荧光强度(F1)高于PB。主成分分析显示有3个因子可解释75%以上的变异：因子1受GI、LA、BL和%CD68/p24调节，因子2受%CD68/p24、BL和T4调节，其中%CD68/p24与另外两个参数负相关；因子3受年龄、T4、BL控制。结论：GCF中存在HIV感染的p24+/CD68+巨噬细胞，CD16表达下调提示其失活。P24+单核细胞从血液中被动员到HIV-P损伤处，然后在原位分化为巨噬细胞。HIV-P的特征是侵袭性的组织破坏和p24+巨噬细胞的渗透。PCA提示，在疾病过程中，BL和p24+巨噬细胞的动员可能是不同步发生的。

Objectives : To characterize the local immune response in AIDS-associated periodontal lesions and to assess the relationship between clinical and immunobiological parameters to propose diagnostic criteria of HIV-related periodontitis (HIV-P).Methods : P24+ cell from Gingival crevicular fluid (GCF) leukocytes of CDC stage IV AIDS patients were characterized by flow cytometry (FCM). Peripheral blood (PB) leukocytes served as systemic controls. Numerical datasets were subjected to a principal component analysis (PCA).Results : Of the 91 patients screened, 44 had HIV-P (48.35%). Two-color FCM of GCF cells from 15 patients disclosed a high p24+ macrophage fraction and a low CD16+CD11b+ neutrophil rate in GCF.Three-color FCM of 23 patients showed that CD68+/p24+ and CD14+/CD68+/p24+ fractions were significantly higher in GCF than in PB.CD14+/p24+ fraction is lower in GCF than in PB.The fluorescence intensities (F1) for p24 in CD14+ and CD14+/CD68+ cells were higher in GCF.The p24 FI of CD68+ macrophages in GCF inversely correlated with CD4+ lymphocytes count in PB.P24 FI levels of CD14+ monocytes in GCF and PB significantly correlated. PCA revealed 3 factors explaining over 75% of the variance : Factor 1 regulated by GI, LA and BL and % CD68/p24 ; Factor 2, regulated by % CD68/p24, BL and T4, where % CD68/p24 negatively correlated with the two other parameters ; Factor 3 was dominated by age, T4, BL.Conclusion : Putatively HIV-infected p24+/CD68+ macrophages were detected from GCF.Downregulated CD16 on GCF neutrophil implied their de-activation. P24+ monocytes appeared mobilised from blood to the HIV-P lesions, then differentiate in situ into macrophages. HIV-P can be characterized by an aggressive tissue destruction and an infiltration of p24+ macrophages. PCA suggested that BL and p24+ macrophage mobilization may asynchronously occur over the disease course.