Baylor Institute for Immunology Research, Dallas, TX, USA.
J Immunol Methods. 2011 Feb 28;365(1-2):27-37. doi: 10.1016/j.jim.2010.11.002. Epub 2010 Nov 18.
In the search for a therapeutic HIV-1 vaccine, we describe herein the development of a monocyte-derived dendritic cell (DC) vaccine loaded with a mixture of HIV-1-antigen lipopeptides (ANRS HIV-LIPO-5 Vaccine). LIPO-5 is comprised of five HIV-1-antigen peptides (Gag(17-35), Gag(253-284), Nef(66-97), Nef(116-145), and Pol(325-355)), each covalently linked to a palmitoyl-lysylamide moiety. Monocytes enriched from HIV-1-infected highly active antiretroviral therapy (HAART)-treated patients were cultured for three days with granulocyte-macrophage colony-stimulating factor and alpha-interferon. At day 2, the DCs were loaded with ANRS HIV-LIPO-5 vaccine, activated with lipopolysaccharide, harvested at day 3 and frozen. Flow cytometry analysis of thawed DC vaccines showed expression of DC differentiation markers: CD1b/c, CD14, HLA-DR, CD11c, co-stimulatory molecule CD80 and DC maturation marker CD83. DCs were capable of eliciting an HIV-1-antigen-specific response, as measured by expansion of autologous CD4(+) and CD8(+) T-cells. The expanded T-cells secreted gamma-IFN and interleukin (IL)-13, but not IL-10. The safety and immunogenicity of this DC vaccine are being evaluated in a Phase I/II clinical trial in chronically HIV-1-infected patients on HAART (clinicaltrials.gov identifier: NCT00796770).
在寻找治疗性 HIV-1 疫苗的过程中，我们在此描述了一种单核细胞衍生的树突状细胞（DC）疫苗的开发，该疫苗负载有 HIV-1 抗原脂肽混合物（ANRS HIV-LIPO-5 疫苗）。LIPO-5 由五个 HIV-1 抗原肽（Gag(17-35)、Gag(253-284)、Nef(66-97)、Nef(116-145)和 Pol(325-355)）组成，每个肽都与棕榈酰赖氨酸酰胺部分共价连接。从接受高效抗逆转录病毒治疗（HAART）的 HIV-1 感染的患者中富集单核细胞，用粒细胞-巨噬细胞集落刺激因子和 α-干扰素培养三天。在第 2 天，将 DC 用 ANRS HIV-LIPO-5 疫苗加载，用脂多糖激活，在第 3 天收获并冷冻。解冻后的 DC 疫苗的流式细胞术分析显示出 DC 分化标志物的表达：CD1b/c、CD14、HLA-DR、CD11c、共刺激分子 CD80 和 DC 成熟标志物 CD83。DC 能够引发 HIV-1 抗原特异性反应，这可以通过扩增自体 CD4(+)和 CD8(+)T 细胞来衡量。扩增的 T 细胞分泌 γ-IFN 和白细胞介素（IL）-13，但不分泌 IL-10。该 DC 疫苗的安全性和免疫原性正在接受接受 HAART 的慢性 HIV-1 感染患者的 I/II 期临床试验评估（clinicaltrials.gov 标识符：NCT00796770）。
