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病毒样颗粒表面嵌入膜的HIV-1包膜引发的免疫反应比可溶性包膜更广泛。

Membrane embedded HIV-1 envelope on the surface of a virus-like particle elicits broader immune responses than soluble envelopes.

作者信息

McBurney Sean P, Young Kelly R, Ross Ted M

机构信息

Center for Vaccine Research for Emerging Diseases and Biodefense, University of Pittsburgh School of Medicine, 9047 Biomedical Sciences Tower 3, 3501 Fifth Avenue, Pittsburgh, PA 15261, USA.

出版信息

Virology. 2007 Feb 20;358(2):334-46. doi: 10.1016/j.virol.2006.08.032. Epub 2006 Sep 28.

Abstract

Virally regulated HIV-1 particles were expressed from DNA plasmids encoding Gag, protease, reverse transcriptase, Vpu, Tat, Rev, and Env. The sequences for integrase, Vpr, Vif, Nef, and the long terminal repeats (LTRs) were deleted. Mutations were engineered into the VLP genome to produce particles deficient in activities associated with viral reverse transcriptase, RNase H, and RNA packaging. Each plasmid efficiently secreted particles from primate cells in vitro and particles were purified from the supernatants and used as immunogens. Mice (BALB/c) were vaccinated intranasally (day 1 and weeks 3 and 6) with purified VLPs and the elicited immunity was compared to particles without Env (Gag(p55)), to soluble monomeric Env(gp120), or to soluble trimerized Env(gp140). Only mice vaccinated with VLPs had robust anti-Env cellular immunity. In contrast, all mice had high titer anti-Env serum antibody (IgG). However, VLP-vaccinated mice had antisera that detected a broader number of linear Env peptides, had anti-Env mucosal IgA and IgG, as well as higher titers of serum neutralizing antibodies. VLPs elicited high titer antibodies that recognized linear regions in V4-C5 and the ectodomain of gp41, but did not recognize V3. These lentiviral VLPs are effective mucosal immunogens that elicit broader immunity against Env determinants in both the systemic and mucosal immune compartments than soluble forms of Env.

摘要

病毒调控的HIV-1颗粒由编码Gag、蛋白酶、逆转录酶、Vpu、Tat、Rev和Env的DNA质粒表达。整合酶、Vpr、Vif、Nef和长末端重复序列(LTR)的序列被删除。在病毒样颗粒(VLP)基因组中引入突变,以产生缺乏与病毒逆转录酶、核糖核酸酶H和RNA包装相关活性的颗粒。每个质粒在体外能有效地从灵长类细胞分泌颗粒,颗粒从培养上清中纯化出来并用作免疫原。用纯化的VLP对小鼠(BALB/c)进行鼻内接种(第1天以及第3周和第6周),并将诱导的免疫与不含Env的颗粒(Gag(p55))、可溶性单体Env(gp120)或可溶性三聚体Env(gp140)进行比较。只有接种VLP的小鼠具有强大的抗Env细胞免疫。相比之下,所有小鼠都有高滴度的抗Env血清抗体(IgG)。然而,接种VLP的小鼠的抗血清能检测到更多种类的线性Env肽,具有抗Env黏膜IgA和IgG以及更高滴度的血清中和抗体。VLP诱导产生的高滴度抗体能识别V4-C5区域的线性表位以及gp41的胞外结构域,但不能识别V3。这些慢病毒VLP是有效的黏膜免疫原,与可溶性形式的Env相比,能在全身和黏膜免疫区室中诱导针对Env决定簇更广泛的免疫。

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