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ABSTRACT: Leptospirosis is an important zoonosis in humans. Immunity against leptospiral infection was thought to be primarily humoral, and limited studies have addressed the role of CD8(+)T cells. Leptospiral immunoglobulin-like protein A (LigA) is an important protective antigen of Leptospira and a potential target for Leptospira-specific cell-mediated immunity. In this study, twenty LigA-derived peptides were tested their binding affinity and stability for the HLA-A*0201 molecule. Peptides with high binding affinity and stability for HLA-A*0201 were then assessed their capacity to elicit specific cytotoxic T-lymphocyte (CTL) responses using cytotoxicity, ELISPOT assays for IFN-gamma and HLA-A*0201-peptide tetramer assays. We identified a HLA-A*0201-restricted epitope, LigA(305-313) KLIVTPAAL in Leptospira LigA. CTLs specific for LigA(305-313) were elicited both in HLA-A2.1/K(b) transgenic mice and in patients with a clinical and/or laboratory diagnosis of leptospirosis. Staining of the HLA-A*0201-LigA(305-313) tetramer revealed the presence of LigA(305-313)-specific CTLs in peripheral blood mononuclear cells (PBMCs) sourced from five patients infected with three different serovars of Leptospira. In conclusion, we report the existence of specific cytotoxic CD8(+)T cells in patients with leptospirosis and we suggest that the newly identified epitope, LigA(305-313), will be helpful in enhancing the understanding of the mechanism of immunity to leptospirosis.
Microbes and Infection 05/2010; 12(5):364-73. · 3.10 Impact Factor
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ABSTRACT: Postmitotic neurons in the developing cortex migrate along radial glial fibers to their proper location in the cortical plate and form the layered structure. Here we report that the radial migration of rat layer II/III cortical neurons requires guidance by the extracellular diffusible factor Semaphorin-3A (Sema3A). This factor is expressed in a descending gradient across the cortical layers, whereas its receptor neuropilin-1 (NP1) is highly expressed in migrating neurons. Downregulation or conditional knockout of NP1 in newborn cortical neurons impedes their radial migration by disrupting their radial orientation during migration without altering their cell fate. Studies in cultured cortical slices further show that the endogenous gradient of Sema3A is required for the proper migration of newborn neurons. In addition, transwell chemotaxis assays show that isolated newborn neurons are attracted by Sema3A. Thus, Sema3A may function as a chemoattractive guidance signal for the radial migration of newborn cortical neurons toward upper layers.
Nature Neuroscience 02/2008; 11(1):36-44. · 15.53 Impact Factor
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ABSTRACT: In order to investigate whether combined DNA vaccines are an ideal way to combine antigens in a single vaccine formulation, we immunized mice with three plasmids (pVSG, pVgD and pVE2), respectively, encoding the antigen of foot-and-mouth disease virus (FMDV), pseudorabies virus (PRV) and classic swine fever virus (CSFV), either alone or in a combined vaccine regimen. We also investigated the immune responses induced by a series of mixtures in which three plasmids were mixed in pairs. Then we further immunized mice with three different plasmids in separate sites and preformed an adoptive transfer experiment. While being given alone, each of the vaccine plasmids induced significant virus-specific antibody responses and splenocytes proliferative activity. But reduced immunogenicity of the pVSG plasmid was found in combined DNA vaccination, no matter whether it was injected in a single or a separate site. Removal single plasmid (pVgD or pVE2) from combined DNA vaccine led to significant increase in the immunogenicity of the pVSG plasmid (P<0.05). And the induction of immune suppression was not mediated by suppressor T cells, as demonstrated by an adoptive transfer experiment. Furthermore, by boosting with whole virus protein of FMDV, mice primed with either pVSG alone or combined DNA vaccine produced statistically significant increase in the FMDV-specific antibody titers (P<0.05). But after boosting, FMDV-specific splenocytes proliferative activity of mice primed with combined DNA vaccine was even lower than that of mice primed with pVSG alone (P<0.05). Taken together, this study reflected the immunogenicity of a single plasmid may be decreased in combined DNA immunization strategy, which still needs to be carefully evaluated before practical application.
Vaccine 06/2007; 25(22):4429-36. · 3.77 Impact Factor
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ABSTRACT: The development and widespread use of DNA-based vaccination against infectious pathogens have been a great triumph of medical science. Quality control of DNA vaccines as biopharmaceutical productions is a problem to solve. Residual genomic DNA of engineering bacteria has been identified as a potential risk factor, so whose level must be controlled under the regulatory standards. We report a dot-blot hybridization method to detect residual host cell DNA in purified DNA vaccines. The assay utilizes PCR amplified and digoxigenin-labeled Escherichia coli 16S rRNA gene as probe. The sensitivity of the dot-blot hybridization assay with E. coli 16S rRNA gene probe was evaluated in comparison with single copy UidR gene probe. The optimized dot-blot hybridization assay had both low background and a suitable sensitivity, detecting 10 pg of residual E. coli DNA. The method is suitable in the routine use of measuring the levels of residual E. coli DNA in the pharmaceutical-grade DNA vaccine.
Vaccine 04/2006; 24(14):2656-61. · 3.77 Impact Factor
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Ying-Jun Guo,
Shu-Han Sun,
Yi Zhang,
Zhu-Huan Chen, Kai-Yu Wang,
Li Huang,
Shu Zhang,
Hong-Ying Zhang,
Qing-Min Wang,
Dan Wu,
Wei-Jia Zhu
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ABSTRACT: In the present study, we investigated the duration of protection afforded to pigs immunized in two different prime-boost regimens: one is homologus priming and boosting with a protein vaccine, and the other is priming with a DNA vaccine and boosting with the protein vaccine. Groups of pigs that received the same vaccination regimen were then challenged with Taenia solium eggs at 6, 12 or 20 weeks post-immunization (wpi), respectively. The results showed that all vaccinated pigs challenged at 6 or 12 wpi showed significant (P < 0.05) reduction in the development of cysts. When challenged at 20 wpi, pigs primed with the DNA vaccine (pcDNA3-cC1) followed by two boosters of the protein vaccine (GST-cC1) showed significant (P < 0.05) protection against the challenge of T. solium eggs, whereas pigs receiving three injections of the protein vaccine showed no significant protection compared to non-vaccinated controls (P > 0.05). Antibody isotype assays showed that DNA prime-protein boost regimen induced a predominantly IgG2 response, compared to an IgG1 biased response for the protein prime-protein boost regimen. In addition, peripheral blood mononuclear cells (PBMC) obtained from the DNA prime-protein boost group proliferated strongly in response to GST-cC1 protein, and this responsiveness persisted until 20 wpi. Taken together, our data suggest that the use of a prime-boost strategy combining DNA and protein vaccines may be better than protein alone for the longevity of protection against the challenge of T. solium eggs.
Vaccine 09/2004; 22(29-30):3841-7. · 3.77 Impact Factor
