Construction and Immunological Evaluation of Dual Cell Surface Display of HIV-1 Gag and Salmonella enterica Serovar Typhimurium FliC in Lactobacillus acidophilus for Vaccine Delivery
Construction and Immunological Evaluation of Dual Cell Surface Display of HIV-1 Gag and Salmonella enterica Serovar Typhimurium FliC in Lactobacillus acidophilus for Vaccine Delivery
复制标题
DOI：
10.1128/cvi.00049-12
复制
发表时间：
2012-09-01
影响因子：
--
通讯作者：
Dean, Gregg
中科院分区：
文献类型：
--
作者：
Kajikawa, Akinobu;Zhang, Lin;Dean, Gregg
引起粘膜免疫应答的口服疫苗可能对人类免疫缺陷病毒1型（HIV-1）有效，因为其传播主要发生在粘膜。本研究的目的是构建重组乳酸杆菌，用于口服HIV-1口服疫苗，并评价其免疫原性。利用嗜酸乳杆菌（Lactobacillus acidophilus，L. acidophilus）粘液结合蛋白（Mub）的信号肽和锚基序融合，构建了HIV-1 Gag的重组嗜酸乳杆菌。在具有或不具有与不同的Mub信号肽和锚融合的肠道沙门氏菌血清型鼠伤寒沙门氏菌鞭毛蛋白（FliC）的共表达的情况下，使用经工程改造以表达Toll样受体5（TLR 5）的HEK 293细胞，测定FliC在细菌细胞表面上的生物活性。表面暴露的鞭毛蛋白保留了其TLR 5刺激活性，表明具有Gag和FliC双重展示的重组菌株可能提供与仅表达Gag的菌株不同的免疫效力。通过与人髓样树突状细胞（DCs）共培养来评估重组菌株的免疫学特性。细胞表面的异源抗原影响DC的成熟和细胞因子反应。还通过小鼠的胃内免疫来研究获得性免疫应答。酶联免疫吸附斑点试验表明，诱导γ-干扰素产生细胞在局部粘膜免疫小鼠后，与GAG生产菌株。同时，用L.同时显示Gag和FliC的嗜酸乳杆菌导致Gag特异性IgA分泌细胞的增加。这些结果表明，Gag展示L.嗜酸乳杆菌引起特异性免疫应答，并且FliC的共存赋予局部伊加产生的佐剂作用。
Oral vaccines that elicit a mucosal immune response may be effective against human immunodeficiency virus type 1 (HIV-1) because its transmission occurs mainly at the mucosa. The aim of this study was to construct recombinant Lactobacillus for oral delivery of oral vaccines against HIV-1 and to evaluate their immunogenicity. A recombinant Lactobacillus acidophilus strain expressing the HIV-1 Gag on the bacterial cell surface was established by fusion with the signal peptide and anchor motif of a mucus binding protein (Mub) from L. acidophilus with or without coexpression of Salmonella enterica serovar Typhimurium flagellin (FliC) fused to a different Mub signal peptide and anchor. Using HEK293 cells engineered to express Toll-like receptor 5 (TLR5), the biological activity of FliC on the bacterial cell surfaces was determined. The surface-exposed flagellin retained its TLR5-stimulating activity, suggesting that the recombinant strain with Gag and FliC dual display might provide a different immunopotency than the strain expressing only Gag. The immunological properties of the recombinant strains were assessed by coculture with human myeloid dendritic cells (DCs). The heterologous antigens on the cell surface affected maturation and cytokine responses of DCs. Acquired immune responses were also investigated by intragastric immunization of mice. The enzyme-linked immunosorbent spot assay showed induction of gamma interferon-producing cells at local mucosa after immunization of mice with the Gag-producing strain. Meanwhile, the immunization with L. acidophilus displaying both Gag and FliC resulted in an increase of Gag-specific IgA-secreting cells. These results suggested that the Gag-displaying L. acidophilus elicited specific immune responses and the coexistence of FliC conferred an adjuvant effect on local IgA production.