UKY DENTAL COBRE: ORAL INFECTIONS AND HIV RECRUDESCENCE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 HAART方案(高效抗逆转录病毒疗法)的引入显著改变了艾滋病毒的病程，提高了艾滋病毒感染者的存活率和生活质量。然而，用现有的抗逆转录病毒疗法不能完全根除艾滋病毒感染。这主要是由于潜伏感染的CD4+T细胞、巨噬细胞和其他宿主细胞(例如，树突状细胞、肥大细胞)在急性艾滋病毒感染的最早阶段，持续极长的半衰期。HIV-1在包括巨噬细胞在内的各种宿主免疫细胞内的持久性构成了控制HIV-1感染的主要障碍。我们正在使用模型T细胞和巨噬细胞系统来评估口腔微生物挑战重新激活HIV的能力。我们的初步研究使用了BF24细胞系，这是一种感染了HIV LTR启动子的人巨噬细胞，驱动着氯霉素乙酰转移酶(CAT)基因。具体目的1是检测口腔细菌(PG、Sm、Cr、FN)或超声波对这些细胞的影响，以评估HIV启动子激活随种类、剂量和时间的变化。我们还评估了多菌挑战在这种激活中协同作用的能力。结果用CAT-ELISA法测量。不同的口腔细菌和细菌超声波激活HIV的能力存在差异，支持牙周炎患者口腔感染的特点可以不同地影响HIV的重新激活。我们还注意到，与单独使用任何一种细菌相比，多细菌对BF24细胞的挑战表现出相加的重新激活。具体目标2侧重于宿主诱导的分子重新激活HIV的能力。在这些研究中，我们用不同的细菌超声波挑战了人牙龈成纤维细胞(HGF)的培养。然后用产生的上清液刺激BF24细胞中的HIV重新激活。结果表明，HGF上清液对BF24细胞的刺激存在剂量和时间效应。然而，这些反应似乎比细菌对巨噬细胞的直接刺激要小得多。最后，我们启动了一些共培养实验，其中肝细胞生长因子、巨噬细胞和细菌挑战在相同的环境中进行。初步结果表明，这些培养物中的因素可能实际上调节了激活HIV启动子的能力。因此，这些数据表明，口腔细菌有能力激活潜伏感染的巨噬细胞中的艾滋病毒。这种激活的幅度表现出一定的特异性，这些感染细胞的多细菌攻击可以增强HIV的重新激活。这些结果表明，慢性口腔感染对艾滋病毒感染患者的治疗成功构成了风险。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The introduction of HAART regimens (Highly Active Antiretroviral Therapy) has significantly modified the course of HIV disease, with longer survival rates and improvement of life quality in HIV-infected individuals. However, complete eradication of HIV infection cannot be achieved with currently available antiretroviral regimens. This primarily results from the establishment of a pool of latently infected CD4+ T cells, macrophages, and other host cells (eg. dendritic cells, mast cells) during the very earliest stages of acute HIV infection that persists with an extremely long half-life. This persistence of HIV-1 within various host immune cells, including macrophages constitutes a major obstacle in the control of HIV-1 infection. We are using model T cell and macrophage systems to evaluate the capacity of the oral microbial challenges to reactivate HIV. Our initial studies have used the BF24 cell line, a human macrophage transfected with the HIV LTR promoter driving a chloramphenicol acetyltransferase (CAT) gene. Specific Aim 1 was to examine the alteration of these cells were challenged with oral bacteria (Pg, Sm, Cr, Fn) or sonicates to assess variations in HIV promoter activation related to species, dose, and time. We also evaluated the ability of a polymicrobial challenge to synergize in this activation. The outcome was measured using a CAT ELISA. Differences were noted in the ability of the various oral bacteria and bacterial sonicates to activate HIV, supporting that the characteristics of oral infection in periodontitis patients could variably impact reactivation of HIV. We also noted that a polybacterial challenge of the BF24 cells demonstrated an additive reactivation when compared to either bacterium alone. Specific Aim 2 focuses on the ability of host induced molecules to reactivate HIV. In these studies, we have challenged human gingival fibroblast (HGF) cultures with various bacterial sonicates. The resulting supernatants were then used to stimulate HIV reactivation in the BF24 cells. The findings indicated a dose and time responsiveness for the HGF supernatants to stimulate the BF24 cells. However, these reactions appeared substantially less that direct bacterial stimulation of the macrophages. Finally, we have initiated some co-culture experiments in which the HGF, macrophages and bacterial challenge take place in the same milieu. Initial results suggest that factors in these cultures may actually modulate the ability to activate the HIV promoter. Thus, the data demonstrate that oral bacteria have the capacity to activate HIV in latently infected macrophages. There appeared some specificity to the magnitude of this activation and a polybacterial challenge of these infected cells can enhance HIV reactivation. The results suggest chronic oral infections provide a risk to treatment success in HIV infected patients.