{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T21:41:54Z","timestamp":1791409314027,"version":"4.3.3"},"reference-count":50,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5376","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1998,7,24]]},"abstract":"<jats:p>\n            Avirulent strains of\n            <jats:italic>Salmonella typhimurium<\/jats:italic>\n            are being considered as antigen delivery vectors. During its intracellular stage in the host,\n            <jats:italic>S. typhimurium<\/jats:italic>\n            resides within a membrane-bound compartment and is not an efficient inducer of class I\u2013restricted immune responses. Viral epitopes were successfully delivered to the host-cell cytosol by using the type III protein secretion system of\n            <jats:italic>S. typhimurium<\/jats:italic>\n            . This resulted in class I\u2013restricted immune responses that protected vaccinated animals against lethal infection. This approach may allow the efficient use of\n            <jats:italic>S. typhimurium<\/jats:italic>\n            as an antigen delivery system to control infections by pathogens that require this type of immune response for protection.\n          <\/jats:p>","DOI":"10.1126\/science.281.5376.565","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:35:26Z","timestamp":1027762526000},"page":"565-568","source":"Crossref","is-referenced-by-count":247,"title":["Delivery of Epitopes by the\n            <i>Salmonella<\/i>\n            Type III Secretion System for Vaccine Development"],"prefix":"10.1126","volume":"281","author":[{"given":"Holger","family":"Ru\u0308ssmann","sequence":"first","affiliation":[{"name":"H. Ru\u0308ssmann, Y. Fu, J. E. Gala\u0301n, Department of Molecular Genetics and Microbiology, School of Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794\u20135222, USA. H. Shams, F. Poblete, R. O. Donis, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583\u20130905, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Homayoun","family":"Shams","sequence":"additional","affiliation":[{"name":"H. Ru\u0308ssmann, Y. Fu, J. E. Gala\u0301n, Department of Molecular Genetics and Microbiology, School of Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794\u20135222, USA. H. Shams, F. Poblete, R. O. Donis, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583\u20130905, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando","family":"Poblete","sequence":"additional","affiliation":[{"name":"H. Ru\u0308ssmann, Y. Fu, J. E. Gala\u0301n, Department of Molecular Genetics and Microbiology, School of Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794\u20135222, USA. H. Shams, F. Poblete, R. O. Donis, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583\u20130905, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yixin","family":"Fu","sequence":"additional","affiliation":[{"name":"H. Ru\u0308ssmann, Y. Fu, J. E. Gala\u0301n, Department of Molecular Genetics and Microbiology, School of Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794\u20135222, USA. H. Shams, F. Poblete, R. O. Donis, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583\u20130905, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jorge E.","family":"Gala\u0301n","sequence":"additional","affiliation":[{"name":"H. Ru\u0308ssmann, Y. Fu, J. E. Gala\u0301n, Department of Molecular Genetics and Microbiology, School of Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794\u20135222, USA. H. Shams, F. Poblete, R. O. Donis, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583\u20130905, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruben O.","family":"Donis","sequence":"additional","affiliation":[{"name":"H. Ru\u0308ssmann, Y. Fu, J. E. Gala\u0301n, Department of Molecular Genetics and Microbiology, School of Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794\u20135222, USA. H. Shams, F. Poblete, R. O. Donis, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583\u20130905, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","unstructured":"Anonymous Report WHO\/VRD\/GEN\/97.01 (World Health Organization Global Programme For Vaccines And Immunization Geneva 1997) ();"},{"key":"e_1_3_1_2_3","doi-asserted-by":"crossref","first-page":"1767","DOI":"10.1016\/S0140-6736(97)05358-0","volume":"350","author":"Katz S. L.","year":"1997","unstructured":"Katz S. L., Lancet 350, 1767 (1997);","journal-title":"Lancet"},{"key":"e_1_3_1_2_4","first-page":"45","volume":"87","author":"Lambert P. H.","year":"1996","unstructured":"Lambert P. H., Dev. Biol. Stand. 87, 45 (1996).","journal-title":"Dev. Biol. Stand."},{"key":"e_1_3_1_3_2","unstructured":"R. Curtiss III et al. Dev. Biol. 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Both plasmids are derived from the low-copy plasmid pWSK-II ["},{"key":"e_1_3_1_11_3","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/0378-1119(91)90366-J","volume":"100","author":"Wang R. F.","year":"1991","unstructured":"Wang R. F., Kushner S., Gene 100, 195 (1991);","journal-title":"Gene"},{"key":"e_1_3_1_11_4","unstructured":"]. Plasmid pSB776 which encodes a fusion between the first 179 amino acids of InvJ and residues 335 to 498 of the influenza virus nucleoprotein was constructed by polymerase chain reaction (PCR). This plasmid is derived from the low-copy plasmid pYA292 ["},{"key":"e_1_3_1_11_5","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/0378-1119(90)90464-3","volume":"94","author":"Gal\u00e1n J. E.","year":"1990","unstructured":"Gal\u00e1n J. E., Nakayama K., Curtiss R., Gene 94, 29 (1990);","journal-title":"Gene"},{"key":"e_1_3_1_11_6","unstructured":"]. Strain SB824 was constructed by introducing the sptP :: kan mutant allele from strain SB237 ( 8 ) into the \u0394 aroA strain SL3261 ["},{"key":"e_1_3_1_11_7","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1038\/291238a0","volume":"291","author":"Hoiseth S. K.","year":"1981","unstructured":"Hoiseth S. K., Stocker B. A., Nature 291, 238 (1981);","journal-title":"Nature"},{"key":"e_1_3_1_11_8","unstructured":"] by P22HT int transduction."},{"key":"e_1_3_1_12_2","doi-asserted-by":"crossref","first-page":"7078","DOI":"10.1128\/jb.177.24.7078-7085.1995","volume":"177","author":"Kaniga K.","year":"1995","unstructured":"Kaniga K., Trollinger D., Gal\u00e1n J. E., J. Bacteriol. 177, 7078 (1995).","journal-title":"J. Bacteriol."},{"key":"e_1_3_1_13_2","doi-asserted-by":"crossref","first-page":"2658","DOI":"10.4049\/jimmunol.151.5.2658","volume":"151","author":"Deckhut A.","year":"1993","unstructured":"Deckhut A., et al., J. Immunol. 151, 2658 (1993);","journal-title":"J. Immunol."},{"key":"e_1_3_1_13_3","doi-asserted-by":"crossref","first-page":"1745","DOI":"10.1084\/jem.162.6.1745","volume":"162","author":"Ljunggren H. G.","year":"1985","unstructured":"Ljunggren H. G., Karre K., J. Exp. Med. 162, 1745 (1985).","journal-title":"J. Exp. Med."},{"key":"e_1_3_1_14_2","unstructured":"RMA or RMA-S cells (C57BL\/6J mouse lymphoma) were used as antigen presenting cells (APCs) after infection with S. typhimurium strains grown as described in ( 12 15 ). About 3 \u00d7 10 8 bacterial CFU were used to infect 10 7 APCs in a 3-ml volume of Dulbecco's modified Eagle's medium (DMEM) at 37\u00b0C for 2 hours. Control APCs were prepared by infection with 10 7.5 plaque-forming units (PFU) of influenza A virus (strain A\/PR\/8\/34) or incubation in 1 \u03bcM IVNP 366\u2013374 synthetic peptide (ASNENMETM) for 2 hours. APCs subjected to the different procedures were washed incubated for 1 hour in DMEM containing gentamicin at 100 \u03bcg\/ml and distributed in quadruplicate 100-\u03bcl cultures in 96-well plates with 2 \u00d7 10 5 APCs each. Influenza NP-specific T cell hybridoma 12.164 (2 \u00d7 10 5 cells ) was added to the treated RMA or RMA-S cultures ( 12 ). Culture medium was collected after incubation for 44 hours at 37\u00b0C to determine the IL-2 concentration by a capture ELISA (Pharmingen)."},{"key":"e_1_3_1_15_2","unstructured":"SptP \u039435\u201362 -IVNP 366\u2013374 which carries a mutation within the binding site of SicP the SptP-specific chaperone [Y. Fu. and"},{"key":"e_1_3_1_15_3","doi-asserted-by":"crossref","first-page":"3393","DOI":"10.1128\/JB.180.13.3393-3399.1998","volume":"180","author":"Gal\u00e1n J. E.","year":"1998","unstructured":"Gal\u00e1n J. E., J. Bacteriol. 180, 3393 (1998);","journal-title":"J. Bacteriol."},{"key":"e_1_3_1_15_4","unstructured":"] was constructed by introducing a 17\u2013amino acid deletion in the SptP-IVNP 366\u2013374 coding sequence of pSB762 by PCR. The resulting mutant protein is secreted from the bacteria at levels indistinguishable from those of wild type but it is not translocated into host cells (Y. Fu and J. E. Gal\u00e1n unpublished results)."},{"key":"e_1_3_1_16_2","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1046\/j.1365-2958.1997.3781740.x","volume":"24","author":"Collazo C. M.","year":"1996","unstructured":"Collazo C. M., Gal\u00e1n J. E., Mol. Microbiol. 24, 747 (1996).","journal-title":"Mol. Microbiol."},{"key":"e_1_3_1_17_2","first-page":"25","volume":"15","author":"Collazo C. M.","year":"1995","unstructured":"Collazo C. M., Zierler M. K., Gal\u00e1n J. E., ibid. 15, 25 (1995).","journal-title":"ibid."},{"key":"e_1_3_1_18_2","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1038\/355647a0","volume":"355","author":"Attaya M.","year":"1992","unstructured":"Attaya M., et al., Nature 355, 647 (1992).","journal-title":"Nature"},{"key":"e_1_3_1_19_2","unstructured":"CTLs were quantitated as described in [J. Wunderlich and G. Shearer in Current Protocols in Immunology J. Coligan A. Kruisbeek D. Margulies E. Shevach W. Strober Eds. (Wiley New York (1991)]. Briefly spleen cells from two or three mice were pooled and restimulated in vitro by coculture with irradiated syngeneic splenocytes infected with A\/PR\/8\/34 influenza virus at a multiplicity of infection of 3 or treated with 1 \u03bcM synthetic peptide LCMVNP 118\u2013126 (RPQASGVYM) for 5 days. EL-4 ( H-2 b ) and P815 ( H-2 d ) target cells were incubated with 1 \u03bcM IVNP 366\u2013374 or LCMV NP 118\u2013126 peptides and labeled with 0.15 mCi of sodium [ 51 Cr]chromate. Appropriate numbers of effector cells were combined with constant numbers of target cells in quadruplicate cultures to assess target lysis by 51 Cr release after cocultivation for 4 hours. Target lysis was calculated as [(experimental release \u2212 spontaneous release)\/(maximum release \u2212 spontaneous release)] \u00d7 100. Maximum release was determined by Triton X-100 cell lysis. Spontaneous release was always <10% of maximum release. Data are presented as percent specific lysis with the percent lysis of unsensitized target cells subtracted from the percent lysis of cognate peptide-sensitized target cells."},{"key":"e_1_3_1_20_2","doi-asserted-by":"crossref","first-page":"2013","DOI":"10.4049\/jimmunol.143.6.2013","volume":"143","author":"Klavinskis L. S.","year":"1989","unstructured":"Klavinskis L. S., Tishon A., Oldstone M. B., J. Immunol. 143, 2013 (1989);","journal-title":"J. Immunol."},{"key":"e_1_3_1_20_3","doi-asserted-by":"crossref","first-page":"4303","DOI":"10.1128\/jvi.63.10.4303-4310.1989","volume":"63","author":"Whitton J. L.","year":"1989","unstructured":"Whitton J. L., et al., J. Virol. 63, 4303 (1989).","journal-title":"J. Virol."},{"key":"e_1_3_1_21_2","unstructured":"C57BL\/6J or BALB\/c mice were inoculated intragastrically with 1 to 2 \u00d7 10 8 CFU of appropriate live S. typhimurium strains grown as described in ( 15 ). Intragastric inoculation was repeated 4 and 6 weeks after the initial inoculation. Spleens were removed 2 weeks after the last bacterial inoculation to evaluate CTL activity in three mice from each group. To determine protective immunity we inoculated groups of six to eight vaccinated mice intracerebrally with 10 median lethal doses (LD 50 ) of LCMV strain Armstrong. For control purposes we inoculated mice intraperitoneally with 10 7.5 PFU of influenza virus A\/PR\/8\/34 10 6 PFU of vaccinia influenza NP ( 24 ) or 10 4 PFU of LCMV strain Armstrong 4 weeks before assessment of immunity."},{"key":"e_1_3_1_22_2","unstructured":"A group of five C57BL\/6J mice received live aroA sptP S. typhimurium mutant strain expressing SptP- LCMVNP 118\u2013126 intragastrically three times as described in ( 19 ). None of the mice in this group survived intracerebral inoculation with 10 LD 50 of LCMV strain Armstrong. In contrast another group of six C57BL\/6J mice that had been injected with 10 4 PFU of LCMV strain Armstrong 4 weeks before intracerebral inoculation with LCMV survived."},{"key":"e_1_3_1_22_3","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1016\/0958-1669(93)90085-B","volume":"4","author":"Hackett J.","year":"1993","unstructured":"Hackett J., Curr. Opin. Biotechnol. 4, 611 (1993);","journal-title":"Curr. Opin. 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Treat. 38 27 (1996).","DOI":"10.1007\/BF01803781"},{"key":"e_1_3_1_24_2","unstructured":"Abbreviations for amino acid residues are as follows: A Ala; C Cys; D Asp; E Glu; F Phe; G Gly; H His; I Ile; K Lys; L Leu; M Met; N Asn; P Pro; Q Gln; R Arg; S Ser; T Thr; V Val; W Trp Y Tyr."},{"key":"e_1_3_1_25_2","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1016\/0042-6822(87)90004-3","volume":"160","author":"Smith G. L.","year":"1987","unstructured":"Smith G. L., Levin J. Z., Palese P., Moss B., Virology 160, 336 (1987).","journal-title":"Virology"},{"key":"e_1_3_1_26_2","unstructured":"We thank P. Doherty for T cell hybridoma 12.164. Supported by grants from the NIH (J.E.G. and R.O.D.) and the American Heart Association (J.E.G.) and a fellowship from the German Bundesministerium f\u00fcr Forschung und Technologie-Aids Stipendienprogramm (H.R.)."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.281.5376.565","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T03:22:53Z","timestamp":1705116173000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.281.5376.565"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,7,24]]},"references-count":50,"journal-issue":{"issue":"5376","published-print":{"date-parts":[[1998,7,24]]}},"alternative-id":["10.1126\/science.281.5376.565"],"URL":"https:\/\/doi.org\/10.1126\/science.281.5376.565","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,7,24]]}}}