Strong HIV-Specific CD4+ and CD8+ T-Lymphocyte Proliferative Responses in Healthy Individuals Immunized with an HIV-1 DNA Vaccine and Boosted with Recombinant Modified Vaccinia Virus Ankara Expressing HIV-1 Genes
Strong HIV-Specific CD4+ and CD8+ T-Lymphocyte Proliferative Responses in Healthy Individuals Immunized with an HIV-1 DNA Vaccine and Boosted with Recombinant Modified Vaccinia Virus Ankara Expressing HIV-1 Genes
复制标题
DOI：
10.1128/cvi.00008-10
复制
发表时间：
2010-07-01
影响因子：
--
通讯作者：
Godoy-Ramirez, Karina
中科院分区：
文献类型：
--
作者：
Aboud, Said;Nilsson, Charlotta;Godoy-Ramirez, Karina
我们研究了HIV-1疫苗在健康志愿者中诱导的淋巴细胞增殖反应，（有或没有佐剂粒细胞-巨噬细胞集落刺激因子[GM-CSF]蛋白）与表达HIV-1 gag、env、rev和rt的DNA在0、1和3个月时使用Biojector进行，并在9个月时用表达异源HIV-1 gag、env、rev和rt的修饰的安卡拉牛痘病毒（MVA）加强，和pol（HIV-MVA）。在HIV-MVA加强后2周，通过[H-3]胸苷摄取试验和活化全血中特异性细胞介导免疫应答的流式细胞术测定（FASCIA-WB），检测对醛硫醇-2（AT-2）灭活的HIV-1抗原的增殖反应（n = 38）。还采用了使用外周血单核细胞的FASCIA（FASCIA-PBMC）（n = 14）。35/38例（92%）接种者通过[H-3]胸苷摄取试验呈反应性。38名疫苗接种者中有32名（84%）对CD 4（+）T细胞FASCIA-WB反应，38名疫苗接种者中有7名（18%）也表现出CD 8（+）T细胞反应。[H-3]胸腺嘧啶核苷摄取法测定的增殖反应与CD 4（+）T细胞FASCIA-WB法测定的增殖反应之间有较强的相关性（r = 0.68; P < 0.01）。使用所有三种测定法分析了14名疫苗接种者。FASCIA-WB和FASCIA-PBMC中分别有10/14例（71%）和11/14例（79%）表现出CD 4（+）T细胞应答。使用FASCIA-WB和FASCIA-PBMC分别在3/14（21%）和7/14（50%）中观察到CD 8（+）T细胞反应性。通过[H-3]胸苷摄取试验，所有14例均具有反应性。采用任何检测方法的疫苗接种者中的总体HIV特异性T细胞增殖应答为100%（38/38）。一个标准化的FASCIA-PBMC，它允许同时进行表型分析，可能是一个选项，[H-3]胸苷摄取试验评估疫苗诱导的T细胞增殖，特别是在同位素限制的设置。
We investigated HIV-1 vaccine-induced lymphoproliferative responses in healthy volunteers immunized intradermally or intramuscularly (with or without adjuvant granulocyte-macrophage colony-stimulating factor [GM-CSF] protein) with DNA expressing HIV-1 gag, env, rev, and rt at months 0, 1, and 3 using a Biojector and boosted at 9 months with modified vaccinia virus Ankara (MVA) expressing heterologous HIV-1 gag, env, and pol (HIV-MVA). Lymphoproliferative responses to aldrithiol-2 (AT-2)-inactivated-HIV-1 antigen were tested by a [H-3] thymidine uptake assay and a flow-cytometric assay of specific cell-mediated immune response in activated whole blood (FASCIA-WB) 2 weeks after the HIV-MVA boost (n = 38). A FASCIA using peripheral blood mononuclear cells (FASCIA-PBMC) was also employed (n = 14). Thirty-five of 38 (92%) vaccinees were reactive by the [H-3] thymidine uptake assay. Thirty-two of 38 (84%) vaccinees were reactive by the CD4(+) T-cell FASCIA-WB, and 7 of 38 (18%) also exhibited CD8(+) T-cell responses. There was strong correlation between the proliferative responses measured by the [H-3] thymidine uptake assay and CD4(+) T-cell FASCIA-WB (r = 0.68; P < 0.01). Fourteen vaccinees were analyzed using all three assays. Ten of 14 (71%) and 11/14 (79%) demonstrated CD4(+) T-cell responses in FASCIA-WB and FASCIA-PBMC, respectively. CD8(+) T-cell reactivity was observed in 3/14 (21%) and 7/14 (50%) using the FASCIA-WB and FASCIA-PBMC, respectively. All 14 were reactive by the [H-3] thymidine uptake assay. The overall HIV-specific T-cell proliferative response in the vaccinees employing any of the assays was 100% (38/38). A standardized FASCIA-PBMC, which allows simultaneous phenotyping, may be an option to the [H-3] thymidine uptake assay for assessment of vaccine-induced T-cell proliferation, especially in isotope-restricted settings.