Pasetti Marcela F, Salerno-Gonçalves Rosangela, Sztein Marcelo B
Center for Vaccine Development, Department of Pediatrics, University of Maryland School of Medicine, Baltimore 21201, USA.
Infect Immun. 2002 Aug;70(8):4009-18. doi: 10.1128/IAI.70.8.4009-4018.2002.
We investigated the ability of live attenuated Salmonella enterica serovar Typhi strains delivered to mice intranasally to induce specific cytotoxic T-lymphocyte (CTL) responses at regional and systemic levels. Mice immunized with two doses (28 days apart) of Salmonella serovar Typhi strain Ty21a, the licensed oral typhoid vaccine, and genetically attenuated mutants CVD 908 (DeltaaroC DeltaaroD), CVD 915 (DeltaguaBA), and CVD 908-htrA (DeltaaroC DeltaaroD DeltahtrA) induced CTL specific for Salmonella serovar Typhi-infected cells in spleens and cervical lymph nodes. CTL were detected in effector T cells that had been expanded in vitro for 7 days in the presence of Salmonella-infected syngeneic splenocytes. A second round of stimulation further enhanced the levels of specific cytotoxicity. CTL activity was observed in sorted alphabeta+ CD8+ T cells, which were remarkably increased after expansion, but not in CD4+ T cells. CTL from both cervical lymph nodes and spleens failed to recognize Salmonella-infected major histocompatibility complex (MHC)-mismatched cells, indicating that the responses were MHC restricted. Studies in which MHC blocking antibodies were used showed that H-2L(d) was the restriction element. This is the first demonstration that Salmonella serovar Typhi vaccines delivered intranasally elicit CD8+ MHC class I-restricted CTL. The results further support the usefulness of the murine intranasal model for evaluating the immunogenicity of typhoid vaccine candidates at the preclinical level.
我们研究了经鼻内接种给小鼠的减毒活伤寒沙门氏菌菌株在局部和全身水平诱导特异性细胞毒性T淋巴细胞(CTL)反应的能力。用两剂(间隔28天)伤寒沙门氏菌Ty21a菌株(已获许可的口服伤寒疫苗)以及基因减毒突变体CVD 908(ΔaroC ΔaroD)、CVD 915(ΔguaBA)和CVD 908-htrA(ΔaroC ΔaroD ΔhtrA)免疫的小鼠,在脾脏和颈部淋巴结中诱导出了针对伤寒沙门氏菌感染细胞的CTL。在存在伤寒沙门氏菌感染的同基因脾细胞的情况下体外扩增7天的效应T细胞中检测到了CTL。第二轮刺激进一步提高了特异性细胞毒性水平。在分选的αβ+ CD8+ T细胞中观察到了CTL活性,这些细胞在扩增后显著增加,但在CD4+ T细胞中未观察到。来自颈部淋巴结和脾脏的CTL均未能识别伤寒沙门氏菌感染的主要组织相容性复合体(MHC)不匹配细胞,表明这些反应受MHC限制。使用MHC阻断抗体的研究表明,H-2L(d)是限制元件。这是首次证明经鼻内接种的伤寒沙门氏菌疫苗可引发CD8+ MHC I类限制性CTL。这些结果进一步支持了小鼠鼻内模型在临床前水平评估伤寒疫苗候选物免疫原性方面的实用性。