Gonzalez C, Hone D, Noriega F R, Tacket C O, Davis J R, Losonsky G, Nataro J P, Hoffman S, Malik A, Nardin E, Sztein M B, Heppner D G, Fouts T R, Isibasi A, Levine M M
Center for Vaccine Development, University of Maryland School of Medicine, Baltimore 21201.
J Infect Dis. 1994 Apr;169(4):927-31. doi: 10.1093/infdis/169.4.927.
rcsp, encoding amino acids 21-398 of Plasmodium falciparum circumsporozoite protein (CSP), under control of tacP was integrated into the chromosomal delta aroC locus of attenuated delta aroC, delta aroD Salmonella typhi CVD 908. By immunoblot and ELISA, rCSP expression was greater from a multicopy plasmid than from the single chromosomal gene. CVD 908 omega (delta aroC1019::tacP-rcsp) was well tolerated by 10 volunteers who were fed two doses of 5 x 10(7) organisms 8 days apart. Seven subjects excreted the vaccine strain for 1-3 days. All subjects developed serologic responses to O and H antigens of the live vector, whereas 3 vaccinees responded to the foreign antigen: 1 developed an 80-fold rise in serum anti-sporozoite antibody, another had a 4-fold rise in antibody to a recombinant portion of CSP (residues 309-345), while a third vaccinee developed CSP-specific CD8+ cytotoxic T lymphocyte activity. This is the first report of attenuated S. typhi eliciting a human serologic or a cytotoxic T lymphocyte response to a foreign protein. Improved foreign gene expression should enhance immunogenicity.
在 tacP 的控制下，编码恶性疟原虫环子孢子蛋白（CSP）第 21 - 398 位氨基酸的 rcsp 被整合到减毒的 aroC 缺失、aroD 缺失的伤寒沙门氏菌 CVD 908 的染色体 aroC 缺失位点。通过免疫印迹和酶联免疫吸附测定，多拷贝质粒中的 rCSP 表达量高于单染色体基因中的表达量。10 名志愿者间隔 8 天口服两剂 5×10⁷ 个生物体剂量的 CVD 908 omega（aroC1019 缺失::tacP - rcsp）后耐受性良好。7 名受试者排出疫苗菌株 1 - 3 天。所有受试者对活载体的 O 和 H 抗原产生了血清学反应，而 3 名接种者对外源抗原产生了反应：1 名接种者血清抗子孢子抗体升高了 80 倍，另一名接种者对 CSP 的重组部分（第 309 - 345 位氨基酸残基）的抗体升高了 4 倍，第三名接种者产生了 CSP 特异性 CD8⁺ 细胞毒性 T 淋巴细胞活性。这是减毒伤寒沙门氏菌引发人体对外源蛋白的血清学或细胞毒性 T 淋巴细胞反应的首次报道。改善的外源基因表达应能增强免疫原性。
