Abstract
We investigated the role of the 100-kilobase (kb) plasmid of Salmonella typhimurium in the virulence of this organism for mice. Three strains, LT2-Z, SR-11, and SL1344, which possessed 100-kb plasmids with identical restriction enzyme digestion profiles, were cured of their respective 100-kb plasmids after Tnmini-tet was used to label plasmids. Curing wild-type virulent strains SR-11 and SL1344 raised peroral 50% lethal doses from 3 x 10(5) and 6 x 10(4) CFU, respectively, to greater than 10(8) CFU. Both wild-type strains had intraperitoneal 50% lethal doses of less than 50 CFU, whereas the intraperitoneal 50% lethal doses for cured SR-11 and SL1344 were less than 50 and 400 CFU, respectively. Reintroduction of the Tnmini-tet-labeled, 100-kb plasmid restored wild-type virulence. Invasion from Peyer's patches to mesenteric lymph nodes and spleens after peroral inoculation was the stage of pathogenesis most affected by curing S. typhimurium of the 100-kb plasmid. Wild-type S. typhimurium replicated in spleens of mice inoculated intravenously to a greater extent than did plasmid-cured derivatives. Wild-type and cured strains equally adhered to and invaded Henle-407, HEp-2, and CHO cells; furthermore, the presence of the 100-kb plasmid was not necessary for replication of S. typhimurium within CHO cells. The 100-kb plasmid had no effect on phagocytosis and killing of S. typhimurium by murine peritoneal macrophages in vitro and in vivo. Similarly, wild-type and plasmid-cured strains were resistant to killing by 90% normal human, rabbit, and guinea pig sera. All wild-type and plasmid-cured S. typhimurium strains possessed complete lipopolysaccharide, as determined by silver staining solubilized cells in sodium dodecyl sulfate-polyacrylamide gels. We have confirmed the role of the 100-kb plasmid of S. typhimurium in virulence, primarily in invasion to mesenteric lymph nodes and spleens after peroral inoculation of mice. Involvement of the 100-kb plasmid in infection of mesenteric lymph nodes and spleens suggests a role for the plasmid in the complex interaction of S. typhimurium with cells of the reticuloendothelial system.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson E. S., Smith H. R. Fertility inhibition in strains of Salmonella typhimurium. Mol Gen Genet. 1972;118(1):79–84. [PubMed] [Google Scholar]
- Appleyard R K. Segregation of New Lysogenic Types during Growth of a Doubly Lysogenic Strain Derived from Escherichia Coli K12. Genetics. 1954 Jul;39(4):440–452. doi: 10.1093/genetics/39.4.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bagdasarian M., Hryniewicz M., Zdzienicka M. Integrative suppression of a dnaA mutation in Salmonella typhimurium. Mol Gen Genet. 1975 Aug 27;139(3):213–231. doi: 10.1007/BF00268973. [DOI] [PubMed] [Google Scholar]
- Birnboim H. C. A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol. 1983;100:243–255. doi: 10.1016/0076-6879(83)00059-2. [DOI] [PubMed] [Google Scholar]
- Blanden R. V., Mackaness G. B., Collins F. M. Mechanisms of acquired resistance in mouse typhoid. J Exp Med. 1966 Oct 1;124(4):585–600. doi: 10.1084/jem.124.4.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
- Briles D. E., Lehmeyer J., Forman C. Phagocytosis and killing of salmonella typhimurium by peritoneal exudate cells. Infect Immun. 1981 Aug;33(2):380–388. doi: 10.1128/iai.33.2.380-388.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carter P. B., Collins F. M. The route of enteric infection in normal mice. J Exp Med. 1974 May 1;139(5):1189–1203. doi: 10.1084/jem.139.5.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Centers for Disease Control (CDC) Human Salmonella isolates--United States, 1983. MMWR Morb Mortal Wkly Rep. 1984 Dec 14;33(49):693–695. [PubMed] [Google Scholar]
- Collins F. M. Immunity to enteric infection in mice. Infect Immun. 1970 Mar;1(3):243–250. doi: 10.1128/iai.1.3.243-250.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collins F. M. Salmonellosis in orally infected specific pathogen-free C57B1 mice. Infect Immun. 1972 Feb;5(2):191–198. doi: 10.1128/iai.5.2.191-198.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Colonna B., Nicoletti M., Visca P., Casalino M., Valenti P., Maimone F. Composite IS1 elements encoding hydroxamate-mediated iron uptake in FIme plasmids from epidemic Salmonella spp. J Bacteriol. 1985 Apr;162(1):307–316. doi: 10.1128/jb.162.1.307-316.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Craig S. W., Cebra J. J. Peyer's patches: an enriched source of precursors for IgA-producing immunocytes in the rabbit. J Exp Med. 1971 Jul 1;134(1):188–200. doi: 10.1084/jem.134.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Curtiss R., 3rd 1984 Kreshover lecture. Genetic analysis of Streptococcus mutans virulence and prospects for an anticaries vaccine. J Dent Res. 1986 Aug;65(8):1034–1045. doi: 10.1177/00220345860650080101. [DOI] [PubMed] [Google Scholar]
- Curtiss R., 3rd, Kelly S. M. Salmonella typhimurium deletion mutants lacking adenylate cyclase and cyclic AMP receptor protein are avirulent and immunogenic. Infect Immun. 1987 Dec;55(12):3035–3043. doi: 10.1128/iai.55.12.3035-3043.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Damiani G., Kiyotaki C., Soeller W., Sasada M., Peisach J., Bloom B. R. Macrophage variants in oxygen metabolism. J Exp Med. 1980 Oct 1;152(4):808–822. doi: 10.1084/jem.152.4.808. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Daskaleros P. A., Payne S. M. Cloning the gene for Congo red binding in Shigella flexneri. Infect Immun. 1985 Apr;48(1):165–168. doi: 10.1128/iai.48.1.165-168.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dowman J. E., Meynell G. G. Pleiotropic effects of de-repressed bacterial sex factors on colicinogeny and cell wall structure. Mol Gen Genet. 1970;109(1):57–68. doi: 10.1007/BF00334046. [DOI] [PubMed] [Google Scholar]
- Eisenstein T. K., Killar L. M., Stocker B. A., Sultzer B. M. Cellular immunity induced by avirulent Salmonella in LPS-defective C3H/HeJ mice. J Immunol. 1984 Aug;133(2):958–961. [PubMed] [Google Scholar]
- Fields P. I., Swanson R. V., Haidaris C. G., Heffron F. Mutants of Salmonella typhimurium that cannot survive within the macrophage are avirulent. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5189–5193. doi: 10.1073/pnas.83.14.5189. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hackett J., Kotlarski I., Mathan V., Francki K., Rowley D. The colonization of Peyer's patches by a strain of Salmonella typhimurium cured of the cryptic plasmid. J Infect Dis. 1986 Jun;153(6):1119–1125. doi: 10.1093/infdis/153.6.1119. [DOI] [PubMed] [Google Scholar]
- Hackett J., Wyk P., Reeves P., Mathan V. Mediation of serum resistance in Salmonella typhimurium by an 11-kilodalton polypeptide encoded by the cryptic plasmid. J Infect Dis. 1987 Mar;155(3):540–549. doi: 10.1093/infdis/155.3.540. [DOI] [PubMed] [Google Scholar]
- Hansen E. J., Frisch C. F., McDade R. L., Jr, Johnston K. H. Identification of immunogenic outer membrane proteins of Haemophilus influenzae type b in the infant rat model system. Infect Immun. 1981 Jun;32(3):1084–1092. doi: 10.1128/iai.32.3.1084-1092.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heffernan E. J., Fierer J., Chikami G., Guiney D. Natural history of oral Salmonella dublin infection in BALB/c mice: effect of an 80-kilobase-pair plasmid on virulence. J Infect Dis. 1987 Jun;155(6):1254–1259. doi: 10.1093/infdis/155.6.1254. [DOI] [PubMed] [Google Scholar]
- Helmuth R., Stephan R., Bunge C., Hoog B., Steinbeck A., Bulling E. Epidemiology of virulence-associated plasmids and outer membrane protein patterns within seven common Salmonella serotypes. Infect Immun. 1985 Apr;48(1):175–182. doi: 10.1128/iai.48.1.175-182.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hohmann A. W., Schmidt G., Rowley D. Intestinal colonization and virulence of Salmonella in mice. Infect Immun. 1978 Dec;22(3):763–770. doi: 10.1128/iai.22.3.763-770.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoiseth S. K., Stocker B. A. Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature. 1981 May 21;291(5812):238–239. doi: 10.1038/291238a0. [DOI] [PubMed] [Google Scholar]
- Husband A. J., Gowans J. L. The origin and antigen-dependent distribution of IgA-containing cells in the intestine. J Exp Med. 1978 Nov 1;148(5):1146–1160. doi: 10.1084/jem.148.5.1146. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones G. W., Rabert D. K., Svinarich D. M., Whitfield H. J. Association of adhesive, invasive, and virulent phenotypes of Salmonella typhimurium with autonomous 60-megadalton plasmids. Infect Immun. 1982 Nov;38(2):476–486. doi: 10.1128/iai.38.2.476-486.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koren H. S., Handwerger B. S., Wunderlich J. R. Identification of macrophage-like characteristics in a cultured murine tumor line. J Immunol. 1975 Feb;114(2 Pt 2):894–897. [PubMed] [Google Scholar]
- LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
- Lee J. C., Gibson C. W., Eisenstein T. K. Macrophage-mediated mitogenic suppression induced in mice of the C3H lineage by a vaccine strain of Salmonella typhimurium. Cell Immunol. 1985 Mar;91(1):75–91. doi: 10.1016/0008-8749(85)90033-4. [DOI] [PubMed] [Google Scholar]
- Liang-Takasaki C. J., Grossman N., Leive L. Salmonellae activate complement differentially via the alternative pathway depending on the structure of their lipopolysaccharide O-antigen. J Immunol. 1983 Apr;130(4):1867–1870. [PubMed] [Google Scholar]
- Liang-Takasaki C. J., Mäkelä P. H., Leive L. Phagocytosis of bacteria by macrophages: changing the carbohydrate of lipopolysaccharide alters interaction with complement and macrophages. J Immunol. 1982 Mar;128(3):1229–1235. [PubMed] [Google Scholar]
- Lissner C. R., Swanson R. N., O'Brien A. D. Genetic control of the innate resistance of mice to Salmonella typhimurium: expression of the Ity gene in peritoneal and splenic macrophages isolated in vitro. J Immunol. 1983 Dec;131(6):3006–3013. [PubMed] [Google Scholar]
- Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mauel J., Defendi V. Infection and transformation of mouse peritoneal macrophages by simian virus 40. J Exp Med. 1971 Aug 1;134(2):335–350. doi: 10.1084/jem.134.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maurelli A. T., Blackmon B., Curtiss R., 3rd Loss of pigmentation in Shigella flexneri 2a is correlated with loss of virulence and virulence-associated plasmid. Infect Immun. 1984 Jan;43(1):397–401. doi: 10.1128/iai.43.1.397-401.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakamura M., Sato S., Ohya T., Suzuki S., Ikeda S., Koeda T. Plasmid-cured Salmonella enteritidis AL1192 as a candidate for a live vaccine. Infect Immun. 1985 Nov;50(2):586–587. doi: 10.1128/iai.50.2.586-587.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pardon P., Popoff M. Y., Coynault C., Marly J., Miras I. Virulence-associated plasmids of Salmonella serotype Typhimurium in experimental murine infection. Ann Inst Pasteur Microbiol. 1986 Jul-Aug;137B(1):47–60. doi: 10.1016/s0769-2609(86)80093-x. [DOI] [PubMed] [Google Scholar]
- Ralph P., Prichard J., Cohn M. Reticulum cell sarcoma: an effector cell in antibody-dependent cell-mediated immunity. J Immunol. 1975 Feb;114(2 Pt 2):898–905. [PubMed] [Google Scholar]
- SCHNEIDER H. A., ZINDER N. D. Nutrition of the host and natural resistance to infection. V. An improved assay employing genetic markers in the double strain inoculation test. J Exp Med. 1956 Feb 1;103(2):207–223. doi: 10.1084/jem.103.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sakai T., Sasakawa C., Makino S., Kamata K., Yoshikawa M. Molecular cloning of a genetic determinant for Congo red binding ability which is essential for the virulence of Shigella flexneri. Infect Immun. 1986 Feb;51(2):476–482. doi: 10.1128/iai.51.2.476-482.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sato G., Asagi M., Oka C., Ishiguro N., Terakado N. Transmissible citrate-utilizing ability in Escherichia coli isolated from pigeons, pigs and cattle. Microbiol Immunol. 1978;22(6):357–360. doi: 10.1111/j.1348-0421.1978.tb00380.x. [DOI] [PubMed] [Google Scholar]
- Schmieger H. Phage P22-mutants with increased or decreased transduction abilities. Mol Gen Genet. 1972;119(1):75–88. doi: 10.1007/BF00270447. [DOI] [PubMed] [Google Scholar]
- Shannon K. P., Spratt B. G., Rowbury R. J. Cell division and the production of cells lacking nuclear bodies in a mutant of Salmonella typhimurium. Mol Gen Genet. 1972;118(2):185–197. doi: 10.1007/BF00267087. [DOI] [PubMed] [Google Scholar]
- Sheehy R. J., Allison D. P., Curtiss R., 3rd Cryptic plasmids in a minicell-producing strain of Salmonella typhimurium. J Bacteriol. 1973 Apr;114(1):439–442. doi: 10.1128/jb.114.1.439-442.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Small P. L., Isberg R. R., Falkow S. Comparison of the ability of enteroinvasive Escherichia coli, Salmonella typhimurium, Yersinia pseudotuberculosis, and Yersinia enterocolitica to enter and replicate within HEp-2 cells. Infect Immun. 1987 Jul;55(7):1674–1679. doi: 10.1128/iai.55.7.1674-1679.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith H. R., Humphreys G. O., Grindley N. D., Grindley J. N., Anderson E. S. Molecular studies of an fi+ plasmid from strains of Salmonella typhimurium. Mol Gen Genet. 1973 Nov 2;126(2):143–151. doi: 10.1007/BF00330989. [DOI] [PubMed] [Google Scholar]
- Smith H. W., Parsell Z., Green P. Thermosensitive H1 plasmids determining citrate utilization. J Gen Microbiol. 1978 Dec;109(2):305–311. doi: 10.1099/00221287-109-2-305. [DOI] [PubMed] [Google Scholar]
- Spratt B. G., Rowbury R. J. The plasmid of Salmonella typhimurium LT2. Mol Gen Genet. 1973 Mar 19;121(4):347–353. doi: 10.1007/BF00433233. [DOI] [PubMed] [Google Scholar]
- Tannock G. W., Blumershine R. V., Savage D. C. Association of Salmonella typhimurium with, and its invasion of, the ileal mucosa in mice. Infect Immun. 1975 Feb;11(2):365–370. doi: 10.1128/iai.11.2.365-370.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
- Vandenbosch J. L., Rabert D. K., Jones G. W. Plasmid-associated resistance of Salmonella typhimurium to complement activated by the classical pathway. Infect Immun. 1987 Nov;55(11):2645–2652. doi: 10.1128/iai.55.11.2645-2652.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
- ZINDER N. D., LEDERBERG J. Genetic exchange in Salmonella. J Bacteriol. 1952 Nov;64(5):679–699. doi: 10.1128/jb.64.5.679-699.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]