Oral Immunization with Attenuated Salmonella enterica Serovar Typhimurium Encoding Cryptosporidium parvum Cp23 and Cp40 Antigens Induces a Specific Immune Response in Mice
Oral Immunization with Attenuated Salmonella enterica Serovar Typhimurium Encoding Cryptosporidium parvum Cp23 and Cp40 Antigens Induces a Specific Immune Response in Mice
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DOI：
10.1128/cvi.00089-09
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发表时间：
2009-09-01
影响因子：
--
通讯作者：
Mead, Jan R.
中科院分区：
文献类型：
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作者：
Benitez, Alvaro J.;McNair, Nina;Mead, Jan R.
减毒沙门氏菌血清型鼠伤寒疫苗株SL 3261被用作针对两种隐孢子虫抗原Cp 23和Cp 40的小鼠口服免疫的抗原递送系统。将每种抗原亚克隆到pH7.1载体系统中，这允许它们在厌氧诱导型nirB启动子的控制下表达为与破伤风毒素的高免疫原性片段C的融合蛋白。将重组质粒导入鼠伤寒沙门氏菌疫苗株SL 3261中，用多克隆C.细小抗血清。小鼠分别经口接种单剂量的SL 3261/质粒-Cp 23或Cp 40，并在体外和体内证明质粒稳定性。免疫后35天，用酶联免疫吸附试验检测血清中抗Cp 23或Cp 40抗原的特异性免疫球蛋白G（IgG）抗体。此外，在用Cp 23免疫的30%的小鼠中检测到血清伊加和粘膜（粪便）伊加抗体。此外，用Cp 23和Cp 40 DNA疫苗载体进行初免-加强，随后进行沙门氏菌免疫，显著增加了对两种抗原的抗体应答。我们的数据表明，单次口服接种重组S。鼠伤寒沙门氏菌SL 3261可诱导抗C.表明重组沙门氏菌是一种可行的抗C.细小病毒感染
Attenuated Salmonella enterica serovar Typhimurium vaccine strain SL3261 was used as an antigen delivery system for the oral immunization of mice against two Cryptosporidium parvum antigens, Cp23 and Cp40. Each antigen was subcloned into the pTECH1 vector system, which allows them to be expressed as fusion proteins with highly immunogenic fragment C of tetanus toxin under the control of the anaerobically inducible nirB promoter. The recombinant vector was introduced into Salmonella Typhimurium vaccine strain SL3261, and the stable soluble expression of the chimeric protein was evaluated and confirmed by Western blotting with polyclonal C. parvum antisera. Mice were inoculated orally with a single dose of SL3261/pTECH-Cp23 or Cp40, respectively, and plasmid stability was demonstrated both in vitro and in vivo. Specific serum immunoglobulin G (IgG) antibodies against the Cp23 or Cp40 antigen were detected by enzyme-linked immunosorbent assay 35 days after immunization. Also, serum IgA and mucosal (feces) IgA antibodies were detected in 30% of the mice immunized with Cp23. In addition, prime-boosting with Cp23 and Cp40 DNA vaccine vectors followed by Salmonella immunization significantly increased antibody responses to both antigens. Our data show that a single oral inoculation with recombinant S. Typhimurium SL3261 can induce specific antibody responses to the Cp23 or Cp40 antigen from C. parvum in mice, suggesting that recombinant Salmonella is a feasible delivery system for a vaccine against C. parvum infection.