Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Normalistas 800, Colinas de la Normal, Guadalajara, Jalisco 44270, México.
Virol J. 2012 Mar 27;9:75. doi: 10.1186/1743-422X-9-75.
The amino terminus of the tobacco etch virus (TEV) capsid protein is located on the external surface of infectious TEV particles, as proposed by previous studies and an in silico model. The epsilon amino groups on the exposed lysine residues are available for chemical conjugation to any given protein, and can thus act as antigen carriers. The availability of amino groups on the surfaces of TEV particles was determined and the immune response to TEV evaluated.
Using a biotin-tagged molecule that reacts specifically with amino groups, we found that the TEV capsid protein has amino groups on its surface available for coupling to other molecules via crosslinkers. Intraperitoneal TEV was administered to female BALB/c mice, and both their humoral and cellular responses measured. Different IgG isotypes, particularly IgG2a, directed against TEV were induced. In a cell proliferation assay, only spleen cells from vaccinated mice that were stimulated in vitro with TEV showed significant proliferation of CD3+/CD4+ and CD3+/CD8+ subpopulations and secreted significant amounts of interferon γ.
TEV has surface amino groups that are available for chemical coupling. TEV induces both humoral and cellular responses when administered alone intraperitoneally to mice. Therefore, TEV should be evaluated as a vaccine adjuvant when chemically coupled to antigens of choice.
烟草蚀纹病毒(TEV)衣壳蛋白的氨基末端位于传染性 TEV 粒子的外表面,这是以前的研究和计算模型提出的。暴露的赖氨酸残基上的ε氨基基团可用于与任何给定的蛋白质进行化学偶联,因此可以作为抗原载体。确定了 TEV 粒子表面上的氨基基团的可用性,并评估了对 TEV 的免疫反应。
使用与氨基基团特异性反应的生物素标记分子,我们发现 TEV 衣壳蛋白表面上有氨基基团可通过交联剂与其他分子偶联。将 TEV 腹腔内给予雌性 BALB/c 小鼠,并测量其体液和细胞反应。针对 TEV 诱导了不同的 IgG 同种型,特别是 IgG2a。在细胞增殖测定中,只有用 TEV 体外刺激的接种疫苗的小鼠的脾细胞显示出 CD3+/CD4+和 CD3+/CD8+亚群的显著增殖,并分泌大量的干扰素 γ。
TEV 具有可用于化学偶联的表面氨基基团。当单独腹腔内给予小鼠时,TEV 会引起体液和细胞反应。因此,当 TEV 与所选抗原化学偶联时,应将其评估为疫苗佐剂。