Martinez-Meza Samuel, Premeaux Thomas A, Cirigliano Stefano M, Friday Courtney M, Michael Stephanie, Mediouni Sonia, Valente Susana T, Ndhlovu Lishomwa C, Fine Howard A, Furler O'Brien Robert L, Nixon Douglas F
Institute of Translational Research, Feinstein Institutes for Medical Research, Northwell Health, Manhasset, NY, USA.
Division of Infectious Diseases, Department of Medicine, Weill Cornell Medicine, New York, NY, USA.
J Neuroinflammation. 2025 Mar 5;22(1):66. doi: 10.1186/s12974-025-03375-w.
HIV-1-associated neurocognitive impairment (HIV-1-NCI) is marked by ongoing and chronic neuroinflammation with loss and decline in neuronal function even when antiretroviral drug therapy (ART) successfully suppresses viral replication. Microglia, the primary reservoirs of HIV-1 in the central nervous system (CNS), play a significant role in maintaining this neuroinflammatory state. However, understanding how chronic neuroinflammation is generated and sustained by HIV-1, or impacted by ART, is difficult due to limited access to human CNS tissue.
We generated an in vitro model of admixed hematopoietic progenitor cell (HPC) derived microglia embedded into embryonic stem cell (ESC) derived Brain Organoids (BO). Microglia were infected with HIV-1 prior to co-culture. Infected microglia were co-cultured with brain organoids BOs to infiltrate the BOs and establish a model for HIV-1 infection, "HIV-1 M-BO". HIV-1 M-BOs were treated with ART for variable directions. HIV-1 infection was monitored with p24 ELISA and by digital droplet PCR (ddPCR). Inflammation was measured by cytokine or p-NF-kB levels using multiplex ELISA, flow cytometry and confocal microscopy.
HIV-1 infected microglia could be co-cultured with BOs to create a model for "brain" HIV-1 infection. Although HIV-1 infected microglia were the initial source of pro-inflammatory cytokines, astrocytes, neurons and neural stem cells also had increased p-NF-kB levels, along with elevated CCL2 levels in the supernatant of HIV-1 M-BOs compared to Uninfected M-BOs. ART suppressed the virus to levels below the limit of detection but did not decrease neuroinflammation.
These findings indicate that HIV-1 infected microglia are pro-inflammatory. Although ART significantly suppressed HIV-1 levels, neuronal inflammation persisted in ART-treated HIV-1 M-BOs. Together, these findings indicate that HIV-1 infection of microglia infiltrated into BOs provides a robust in vitro model to understand the impact of HIV-1 and ART on neuroinflammation.
即使抗逆转录病毒药物疗法（ART）成功抑制病毒复制，HIV-1相关神经认知障碍（HIV-1-NCI）仍以持续的慢性神经炎症以及神经元功能丧失和衰退为特征。小胶质细胞是HIV-1在中枢神经系统（CNS）中的主要储存库，在维持这种神经炎症状态中起重要作用。然而，由于获取人类CNS组织的机会有限，了解HIV-1如何产生和维持慢性神经炎症或受ART影响具有一定难度。
我们构建了一种体外模型，将造血祖细胞（HPC）来源的小胶质细胞与胚胎干细胞（ESC）来源的脑类器官（BO）混合培养。在共培养之前，小胶质细胞先感染HIV-1。将感染的小胶质细胞与脑类器官BO共培养，使其浸润到BO中，建立HIV-1感染模型“HIV-1 M-BO”。对HIV-1 M-BO进行不同方向的ART处理。通过p24 ELISA和数字液滴PCR（ddPCR）监测HIV-1感染情况。使用多重ELISA、流式细胞术和共聚焦显微镜通过细胞因子或p-NF-kB水平测量炎症。
HIV-1感染的小胶质细胞可与BO共培养，以创建“脑”HIV-1感染模型。尽管HIV-1感染的小胶质细胞是促炎细胞因子的最初来源，但与未感染的M-BO相比，HIV-1 M-BO上清液中的星形胶质细胞、神经元和神经干细胞的p-NF-kB水平也有所升高，CCL2水平也升高。ART将病毒抑制到检测限以下，但并未减轻神经炎症。
这些发现表明HIV-1感染的小胶质细胞具有促炎作用。尽管ART显著抑制了HIV-1水平，但在接受ART治疗的HIV-1 M-BO中神经元炎症仍然存在。总之，这些发现表明浸润到BO中的小胶质细胞的HIV-1感染提供了一个强大的体外模型，以了解HIV-1和ART对神经炎症的影响。
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