Long-lived plasma cells in HIV infection and vaccination

1背景/理由：这笔赠款的前提是尽管抗体在人类免疫系统中的潜在重要性 2预防和控制艾滋病毒感染，在抗艾滋病毒gp120反应中缺乏持久性是一个问题 3艾滋病毒疫苗的开发威胁到其潜在的实用性。 4. 5目标：我们的假设是长期存活的浆细胞之间的数量和质量上的差异 6产生抗HIV Env抗体的人与不产生抗HIV Env抗体的人相比。我们有最新和详细的数据来自 7我们从其获得详细骨髓库数据的艾滋病毒感染者的数量， 8鉴定和分离循环中的主要抗体。知道哪些抗体构成了 9谱系使我们能够识别和跟踪不同浆细胞室中的抗体分泌细胞。 10通过这样做，我们已经确定了产生抗gp120的浆细胞，包括长寿的浆细胞 11慢性HIV感染患者骨髓中确实存在抗体。这样做的具体目的是 12个建议是1)比较骨髓长寿浆细胞分泌抗-DNA抗体的转录图谱 13个针对非gp120长寿浆细胞的gp120抗体；2)比较不同骨骼的表型差异 14个骨髓长寿浆细胞分泌抗非gp120长寿浆细胞抗体；3) 15测定CART治疗对骨髓中分泌抗Env抗体浆细胞的影响。 16年 17方法：我们计划研究几组不同个体的浆细胞：慢性HIV 18名感染者、手推车上的艾滋病毒感染者和接种了IHV01艾滋病毒疫苗的正常志愿者 19项研究。骨髓样本将根据CD19和CD138的表达被分成四个群体。这个 第一步是确定抗gp120、抗gp41、抗p24和抗白喉的骨髓谱系。 21名捐赠者(6名慢性感染者接种车前后和6名接受艾滋病毒疫苗接种的捐赠者)。此后，细胞签名 22(转录组)将在不同的抗原特异性浆细胞之间进行比较， 23个浆细胞亚群和队列。其次，表型差异(克隆频率、ELISPOT、分泌率) 24人将被测量，然后在组之间进行比较，如上所述。使用购物车前后的配对样本 25治疗后，我们将研究CART对浆细胞转录组和表型的影响。 26个子集。我们还将确定在HIV感染中看到的抗gp120长寿命浆细胞是否是“暂时的”。 27(与购物车一起消失)，以及是否可以在艾滋病毒感染或疫苗接种中看到“真正的”长寿人口。 28 29影响：我们建议上述研究将阐明抗病毒抗体寿命缺陷的性质。 30 HIV-1 gp120，这将对艾滋病毒疫苗设计产生直接影响。

1 Background/Rationale: The premise of this grant is that despite the potential importance of antibodies in the 2 prevention and control of HIV infection, the lack of durability in the anti-HIV gp120 response is a problem for 3 HIV vaccine development that threatens their potential utility. 4 5 Objectives: Our hypothesis is that quantitative and qualitative differences between the long-lived plasma cells 6 that make anti-HIV Env antibodies compared to those that do not. We have recent and detailed data from a 7 number of HIV-infected individuals from whom we have obtained detailed bone marrow repertoire data, 8 identifying and isolating the predominant antibodies in circulation. Knowing which antibodies constitute the 9 repertoire allows us to identify and track the antibody secreting cells in the various plasma cell compartments. 10 Doing this, we have identified that plasma cells, including long-lived plasma cells, that produce anti-gp120 11 antibody can indeed be found in the bone marrow patients with chronic HIV infection. The specific aims of this 12 proposal are 1) Compare the transcriptional profile of bone marrow long-lived plasma cells secreting anti- 13 gp120 antibodies to non-gp120 long-lived plasma cells; 2) Compare phenotypic differences between bone 14 marrow long-lived plasma cells secreting anti-gp120 antibodies to non-gp120 long-lived plasma cells; 3) 15 Determine the effect of cART treatment on anti-Env antibody secreting plasma cells in the bone marrow. 16 17 Methods: We plan on studying the plasma cells from several different groups of individuals: chronic HIV 18 infected, HIV-infected on cART, and normal volunteers who have been vaccinated in the IHV01 HIV vaccine 19 studies. Bone marrow samples will be sorted into four populations based on CD19 and CD138 expression. The 20 first step is to define the anti-gp120, anti-gp41, anti-p24, and anti-diphtheria bone marrow repertoire in the 21 donors (6 chronically infected pre and post-cART and 6 HIV vaccinee donors). Thereafter the cell signatures 22 (transcriptome) amongst the various antigen-specific plasma cells will be compared with each other, across 23 plasma cell subsets, and cohorts. Next, the phenotypic differences (clonal frequency, Elispot, secretion rates) 24 will be measured and then compared between groups, as above. Using paired samples before and after cART 25 therapy, we will study the effects of cART on the transcriptome and phenotypic profile of the plasma cell 26 subsets. We will also determine if the anti-gp120 long-lived plasma cells seen in HIV infection are “transient” 27 (disappear with cART), and whether a “true” long-lived population can be seen in HIV infection or vaccination. 28 29 Impact: We propose that the studies above will elucidate the nature of the antibody longevity defect against 30 HIV-1 gp120, which will have direct implications for HIV vaccine design.