Isolation and Partial Characterization of an Immunogenic Moiety Obtained from<i>Salmonella typhimurium</i>

Venneman, Martin R.; Bigley, Nancy J.

doi:10.1128/jb.100.1.140-148.1969

Cited by 69 publications

(59 citation statements)

References 32 publications

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“…In more recent work (46), they have further suggested that the immunogenic activity of the fractions may be associated with one particular species of nucleic acid. 1 Preliminary results of this investigation were presented at the Annual Meeting of the American Society for Microbiology, Minneapolis, Minn., 2-7 May 1971. Immunization with bacterial ribosomal preparations has also been found to be an effective means of prophylaxis for infections with Staphylococcus aureus (41), Pseudomonas aeruginosa (40), Diplococcus pneumoniae (30), and Salmonella typhimurium (35). The immunogenic fractions of these organisms consist of approximately 60% RNA, 30% protein, and 10%7, "contaminant" material on a dry weight basis.…”

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confidence: 99%

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Purification of Immunogenically Active Ribonucleic Acid Preparations of Salmonella typhimurium : Molecular-Sieve and Anion-Exchange Chromatography

Venneman

1972

Infect Immun

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Immunogenic Salmonella typhimurium ribonucleic acid (RNA) preparations, prepared by differential centrifugation, phenol extraction at 65 C, and ethanol precipitation from 0.5% sodium dodecyl sulfate. solution, maintained their immunogenicity through lyophilization. As measured by survival, differential pathogen counts 5 days postchallenge, or clearance of the infecting organism from the tissues, immunization with 50 Ag (dry weight) of the lyophilized preparation proved as effective as immunization with 0.1 LD50 of attenuated S. typhimurium cells. Chromatography of the immunogenic fraction through Biogel P-6 (exclusion limit > 4,600) or through Biogel P-300 (exclusion limit > 300,000) resulted in only one immunogenically active protein of the eluate found in the void volume of the columns. Diethylaminoethyl (DEAE) cellulose anion-exchange chromatography of the RNA preparations showed that the immunogenic activity was eluted from the column at 0.8 to 1.0 M NaCl in a linear 0.1 to 2.0 M NaCl gradient. Nonimmunogenic, protein-containing minor peaks were eluted at 0.1 to 0.5 M NaCl. Serial fractionation of the crude RNA preparations over Biogel P-6 to DEAE cellulose to Biogel P-300 molecular-sieve or anion-exchange columns did not alter the immunogenicity of the RNA preparation. Incorporation of the column fractions into Freund's incomplete adjuvant did not increase their relative effectiveness in eliciting anti-salmonella resistance. Chemical analysis of the immunogenic preparations indicated that they were lacking in detectable protein, lipid, and deoxyribonucleic acid. These results suggest that the immunogenic moiety of the crude nucleic acid fraction is either RNA or an as yet undefined polysaccharide of greater than 300,000 molecular weight.Within the past 5 years, an increasing body of evidence has accumulated to indicate that bacterial subcellular preparations are effective in inducing resistance to several bacterial parasites. Youmans and Youmans have elegantly demonstrated that ribosomal and crude ethanol-precipitated ribonucleic acid (RNA) preparations of Mycobacterium tuberculosis induce an effective immune response in mice to challenge infection with the homologous organism (42)(43)(44)(45). In more recent work (46), they have further suggested that the immunogenic activity of the fractions may be associated with one particular species of nucleic acid.

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“…Preparation of crude RNA fractions. The procedure employed for the preparation of the immunogenic RNA fractions was that described previously with modification (35,36). An 8-liter broth culture of S. typhimurium SR-11 in midto late-log growth was harvested by continuous-flow centrifugation at 40,000 X g in a Sorvall refrigerated high-speed centrifuge.…”

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Purification of Immunogenically Active Ribonucleic Acid Preparations of Salmonella typhimurium : Molecular-Sieve and Anion-Exchange Chromatography

Venneman

1972

Infect Immun

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“…Preparation of bacterial cultures for nucleic acid extractions. The techniques for culturing, harvesting, and rupturing the virulent S. typhimurium were those of Venneman and Bigley (18) with the following exceptions. A 0.01 M phosphate buffer, pH 7.1, containing 4 X 10-4% polyvinyl sulfate (20) was used in the washing and harvesting of the bacteria.…”

Section: Methodsmentioning

confidence: 99%

“…Both RNA and protein moieties appeared to be necessary components of the effective immunogen. Venneman and Bigley (18) and Venneman et al (19) have shown that both a ribosomal subfraction from Salmonella typhimurium SR-1I and its subunits are effective immunogens in stimulat-ing immunity to lethal salmonellosis in mice. They also observed that an ethanol-precipitated, crude RNA fraction of the ribosomal preparations induced protective immunity comparable to that developed after immunization with the whole ribosomal fraction.…”

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Location and Description of Spiral-Shaped Microorganisms in the Normal Rat Cecum

1972

Infect Immun

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Mice were injected with virulent Salmonella typhimurium SR-11 subfractions containing varied amounts of ribonucleic acid (RNA) and protein or with living attenuated S. typhimurium RIA. In these mice, maximal resistance to lethal infection by 1,000 or 5,000 median lethal doses of S. typhimurium SR-11 was seen 2 to 3 weeks after immunization. The S. typhimurium RIA vaccine and a crude ethanol-precipitated RNA fraction (E-RNA) prepared from lysates of S. typhimurium SR-1 1 were the most efficient immunogens inducing protection against salmonellosis. The contribution of the components present in the E-RNA fractions to host protection against lethal salmonella infection was also examined. RNA-rich fractions (P-RNA) prepared from lysates of the virulent salmonellae contained several bands of protein when examined by disc electrophoresis. P-RNA fractions stimulated protective immunity in mice to infection with S. typhimurium SR-11 but to a much lesser degree than did the E-RNA fractions or strain RIA vaccine. Protein-rich fractions (NP), separated from E-RNA by salt precipitation, exhibited the same number and distribution of protein bands by disc electrophoresis as did the parent E-RNA fractions. Mixtures of either bovine liver soluble RNA or various synthetic polynucleotides and NP were examined, as was NP fraction alone, for the ability to confer protection in mice to challenge infections by the virulent strain of salmonella. Polyadenylic-uridylic acid plus NP conferred significant protective immunity to challenge infections in mice immunized with this mixture, being nearly as effective an immunogen as were the E-RNA fractions of S. typhimurium SR-11 or the attenuated S. typhimurium RIA. fection in mice.The immunogenic efficacy of RNA-protein fractions from lysates of S. typhimurium SR-1I and the corresponding RNA-rich and proteinrich subfractions were compared for the capacity to stimulate protective immunity to challenge in-377 on August 1, 2020 by guest http://iai.asm.org/ Downloaded from

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“…A number of studies on salmonella ribosomal vaccines have been reported since Venneman and Bigley [1] demonstrated the immunogenicity of a subcellular, ribonucleic acid preparation of Salmonella typhimurium. Many investigators indicated that the efficacy of salmonella ribosomal vaccines varies with mouse strains [2], and that contaminating lipopolysaccharide (LPS) exerted the major effect [3,4].…”

Section: Introductionmentioning

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Requirement of the conformational stability of a Salmonella ribosomal vaccine for its mouse protection

Kita

1991

FEMS Microbiology Letters

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Summary The 43‐kDa non‐O antigenic component isolated from the crude ribosomal fraction of Salmonella typhimurium [9] was further purified by affinity chromatography (43‐kDa protein: 43‐kDp). Immunization with 43‐kDp did not induce complete mouse protection in CF1 mice to 500 LD50 of S. typhimurium, although it elicited a substantial IgG antibody response. The 43‐kDp exhibited the mitogenicity to splenocytes (CF1 and C3H/HeJ) and B cell‐rich populations (CF1). Complexing 43‐kDp with the compact ribosomes of Streptococcus pyogenes by formaldehyde (complex vaccine: CV) elicited both IgM and IgG antibodies to 43‐kDp. CN induced a boosting effect to enhance IgG antibody response. Moreover, CV generated delayed‐type hypersensitivity to salmonella antigens and also conferred complete protection against 500 LD50 challenge of S. typhimurium to CF1 mice. These abilities of CV were reduced or impaired by RNase digestion. CV was able to induce partial or complete protection in inbred mouse strains (C3H/HeN, C3H/HeJ, DBA/2 and A/J). These data, in addition to other reports, suggest that conformational stability between ribosomes and contaminating substances such as 43‐kDp or O‐antigens might be required for the overall effects of the ribosomal vaccine.

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Isolation and Partial Characterization of an Immunogenic Moiety Obtained fromSalmonella typhimurium

Cited by 69 publications

References 32 publications

Purification of Immunogenically Active Ribonucleic Acid Preparations of Salmonella typhimurium : Molecular-Sieve and Anion-Exchange Chromatography

Purification of Immunogenically Active Ribonucleic Acid Preparations of Salmonella typhimurium : Molecular-Sieve and Anion-Exchange Chromatography

Location and Description of Spiral-Shaped Microorganisms in the Normal Rat Cecum

Requirement of the conformational stability of a Salmonella ribosomal vaccine for its mouse protection

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