Nanodepots Encapsulating a Latency Reversing Agent and Broadly Neutralizing Antibody Enhance Natural Killer Cell Cytotoxicity Against an in vitro Model of Latent HIV.
Nanodepots Encapsulating a Latency Reversing Agent and Broadly Neutralizing Antibody Enhance Natural Killer Cell Cytotoxicity Against an in vitro Model of Latent HIV.
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DOI：
10.2147/ijn.s401304
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发表时间：
2023
影响因子：
8
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中科院分区：
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目前针对人类免疫缺陷病毒（HIV）的抗逆转录病毒疗法（ART）无法治愈，因为病毒存在于潜伏库中，需要终身坚持ART并增加合并症的风险。“休克和杀死”的方法，以重新激活艾滋病毒从潜伏的水库，然后管理的抗艾滋病毒药物是一个有希望的战略，消除潜伏的艾滋病毒。为了实现有效的休克和杀伤，我们描述了一种消除HIV储库的策略，该策略结合了潜伏期逆转剂（LRA），广泛中和抗体（bnAb）和自然杀伤（NK）细胞。该策略利用共封装LRA和bnAb的聚合物纳米贮库（ND）来重新激活潜伏感染并引起来自共施用的NK细胞的增强的细胞毒性。采用纳米共沉淀法制备了LRA（TNF-α）和bnAb（3BNC 117）共包封的聚乳酸-羟基乙酸共聚物（PLGA）NDs（TNF-α-3BNC117-NDs）。ACH-2细胞用作潜伏HIV感染的细胞模型。将经遗传修饰以组成型表达Fc受体CD 16的NK 92亚系与TNF-α-3BNC117-ND联合给予ACH-2细胞。分别通过流式细胞术和ELISA测量ACH-2细胞死亡和细胞外p24。稳定的PLGA ND以高效率共包封TNF-α和3BNC 117，并在生理条件下释放这些试剂。在TNF-α-3BNC117-ND存在下，NK 92表型保持相似。ND释放的TNF-α有效地重新激活ACH-2细胞中的HIV，通过细胞内p24阳性细胞频率增加3.0倍来测量。释放的3BNC 117中和并结合再活化的病毒，靶向总ACH-2细胞的57.5%。重要的是，TNF-α-3BNC117-ND显著增强了NK 92细胞介导的ACH-2细胞杀伤（1.9倍），并将细胞外p24水平降至基线水平。这些研究结果表明，我们的新的ND为基础的三方策略的治疗潜力，重新激活潜伏感染的细胞的HIV，产生一个HIV特异性位点的bnAb结合，并增强NK 92细胞的重新激活的HIV感染的靶细胞的杀伤。
Current antiretroviral therapies (ART) for human immunodeficiency virus (HIV) are not curative, as the virus persists in latent reservoirs, requiring lifelong adherence to ART and increasing the risk of co-morbidities. “Shock and kill” approaches to reactivate HIV from latent reservoirs followed by administration of anti-HIV drugs represent a promising strategy for eradicating latent HIV. To achieve effective shock and kill, we describe a strategy to eradicate the HIV reservoir that combines latency reversing agents (LRAs), broadly neutralizing antibodies (bnAbs), and natural killer (NK) cells. This strategy utilizes a polymer nanodepot (ND) that co-encapsulates the LRA and bnAb to reactivate latent infection and elicit enhanced cytotoxicity from co-administered NK cells. Poly(lactic-co-glycolic acid) (PLGA) NDs were synthesized using the nanoprecipitation method to co-encapsulate an LRA (TNF-α) and a bnAb (3BNC117) (TNF-α-3BNC117-NDs). ACH-2 cells were used as a cellular model of latent HIV infection. An NK92 subline, genetically modified to constitutively express the Fc receptor CD16, was administered to ACH-2 cells in combination with TNF-α-3BNC117-NDs. ACH-2 cell death and extracellular p24 were measured via flow cytometry and ELISA, respectively. Stable PLGA NDs co-encapsulated TNF-α and 3BNC117 with high efficiencies and released these agents in physiological conditions. NK92 phenotype remained similar in the presence of TNF-α-3BNC117-NDs. TNF-α released from NDs efficiently reactivated HIV in ACH-2 cells, as measured by a 3.0-fold increase in the frequency of intracellular p24 positive cells. Released 3BNC117 neutralized and bound reactivated virus, targeting 57.5% of total ACH-2 cells. Critically, TNF-α-3BNC117-NDs significantly enhanced NK92 cell-mediated killing of ACH-2 cells (1.9-fold) and reduced extracellular levels of p24 to baseline. These findings suggest the therapeutic potential of our novel ND-based tripartite strategy to reactivate HIV from latently infected cells, generate an HIV-specific site for bnAb binding, and enhance the killing of reactivated HIV-infected target cells by NK92 cells.