Cytomegalovirus Vaccine Strain Towne-Derived Dense Bodies Induce Broad Cellular Immune Responses and Neutralizing Antibodies That Prevent Infection of Fibroblasts and Epithelial Cells
Cytomegalovirus Vaccine Strain Towne-Derived Dense Bodies Induce Broad Cellular Immune Responses and Neutralizing Antibodies That Prevent Infection of Fibroblasts and Epithelial Cells
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DOI：
10.1128/jvi.01554-13
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发表时间：
2013-10-01
影响因子：
5.4
通讯作者：
McCormick, A. Louise
中科院分区：
文献类型：
--
作者：
Cayatte, Corinne;Schneider-Ohrum, Kirsten;McCormick, A. Louise
人巨细胞病毒(HCMV)是一种β疱疹病毒，可导致免疫抑制患者和先天性感染后的严重疾病。预防先天性感染可能需要一种同时诱导体液免疫和细胞免疫的疫苗。致密小体(DB)是由感染HCMV的细胞产生的复杂的非传染性颗粒，可能代表一种疫苗选择。由于对DB的抗原性和免疫原性的了解尚不完全，我们探索了DB的表征方法，确定了DB的制备方法，并对DB材料在BALB/c小鼠体内的中和和细胞免疫应答进行了系统的评价。用病毒终止酶抑制剂2-溴-5，6-二氯-1-β-D-呋喃核糖基苯并咪唑核苷(BDCRB)处理Towne感染培养物中纯化的树突状细胞，用纳米颗粒跟踪分析(NTA)、二维荧光差示凝胶电泳法(2D-DGE)、免疫印迹、定量酶联免疫吸附试验等方法对其进行了表征。对DBS的体液和细胞免疫应答与加佐剂AddaVax的糖蛋白B(GB/AddaVax)的免疫原性进行了比较。DBS诱导中和抗体，可防止病毒感染培养的成纤维细胞和上皮细胞，并对多种病毒蛋白(包括pp65、GB和UL48)产生强大的细胞介导免疫反应。相比之下，GB/AddaVax未能诱导中和抗体来防止上皮细胞感染，这突显了这些候选疫苗诱导的体液反应的关键差异。我们的数据促进了DB疫苗方法的潜力，展示了重要的免疫原性，并有力地支持了DBs作为CMV候选疫苗的进一步评估。
Human cytomegalovirus (HCMV), a betaherpesvirus, can cause severe disease in immunosuppressed patients and following congenital infection. A vaccine that induces both humoral and cellular immunity may be required to prevent congenital infection. Dense bodies (DBs) are complex, noninfectious particles produced by HCMV-infected cells and may represent a vaccine option. As knowledge of the antigenicity and immunogenicity of DB is incomplete, we explored characterization methods and defined DB production methods, followed by systematic evaluation of neutralization and cell-mediated immune responses to the DB material in BALB/c mice. DBs purified from Towne-infected cultures treated with the viral terminase inhibitor 2-bromo-5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole riboside (BDCRB) were characterized by nanoparticle tracking analysis (NTA), two-dimensional fluorescence difference gel electrophoresis (2D-DIGE), immunoblotting, quantitative enzyme-linked immunosorbent assay, and other methods. The humoral and cellular immune responses to DBs were compared to the immunogenicity of glycoprotein B (gB) administered with the adjuvant AddaVax (gB/AddaVax). DBs induced neutralizing antibodies that prevented viral infection of cultured fibroblasts and epithelial cells and robust cell-mediated immune responses to multiple viral proteins, including pp65, gB, and UL48. In contrast, gB/AddaVax failed to induce neutralizing antibodies that prevented infection of epithelial cells, highlighting a critical difference in the humoral responses induced by these vaccine candidates. Our data advance the potential for the DB vaccine approach, demonstrate important immunogenicity properties, and strongly support the further evaluation of DBs as a CMV vaccine candidate.