Home LiteratureArticle Details
PMID: 2323818 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of genetically inactivated pertussis toxin mutants: candidates for a new vaccine against whooping cough.

Infection and immunity ·Vol. 58 ·No. 5 ·1990-05-00 ·Pages 1308-15

Nencioni L, Pizza M, Bugnoli M, De Magistris T, Di Tommaso A, Giovannoni F, Manetti R, Marsili I, Matteucci G, Nucci D

Abstract

the introduction of two amino acid substitutions within the enzymatically active subunit S1 of pertussis toxin (PT) abolishes its ADP-ribosyltransferase activity and toxicity on CHO cells (Pizza et al., Science 246:497-500, 1989). These genetically inactivated molecules are also devoid of other in vivo adverse reactions typical of PT, such as induction of leukocytosis, potentiation of anaphylaxis, stimulation of insulin secretion, and histamine sensitivity. However, the mutant PT molecules are indistinguishable from wild-type PT in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and maintain all the physical and chemical properties of PT, including affinity for toxin-neutralizing poly- and monoclonal antibodies. Either alone or stabilized with formaldehyde, PT mutants are able to induce high levels of neutralizing antibodies and to protect mice in a dose-dependent fashion against intracerebral challenge with virulent B. pertussis. These results clearly show that these genetically inactivated PT molecules are nontoxic but still immunogenic and justify their development as a component of a new, safer acellular vaccine against whooping cough.

MeSH Terms
Adjuvants, Immunologic Amino Acids/analysis Animals Antibodies, Bacterial/biosynthesis Antibodies, Monoclonal/immunology Bordetella pertussis/genetics,immunology Dose-Response Relationship, Immunologic Electrophoresis, Polyacrylamide Gel Formaldehyde Mice Pertussis Toxin Vaccines/immunology Vaccines, Synthetic/immunology Virulence Factors, Bordetella/genetics Whooping Cough/prevention & control
Chemicals
Adjuvants, Immunologic Amino Acids Antibodies, Bacterial Antibodies, Monoclonal Vaccines Vaccines, Synthetic Virulence Factors, Bordetella Formaldehyde Pertussis Toxin
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Nencioni L
Sclavo Research Center, Siena, Italy.
Pizza M
Bugnoli M
De Magistris T
Di Tommaso A
Giovannoni F
Manetti R
Marsili I
Matteucci G
Nucci D
References (46)
46 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Acellular pertussis vaccines: evaluation of reversion in a nude mouse model.
    J Biol Stand. 1988 Apr;16(2):99-108 PMID: 3259580
  3. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes.
    J Immunol. 1972 Jul;109(1):129-35 PMID: 4113792
  4. Distribution of labeled lymph node cells in mice during the lymphocytosis induced by Bordetella pertussis.
    J Exp Med. 1972 Dec 1;136(6):1581-93 PMID: 4345107
  5. ADP-ribosyltransferase mutations in the catalytic S-1 subunit of pertussis toxin.
    Infect Immun. 1988 Aug;56(8):1934-41 PMID: 3135265
  6. Mortality and morbidity from invasive bacterial infections during a clinical trial of acellular pertussis vaccines in Sweden.
    Pediatr Infect Dis J. 1988 Sep;7(9):637-45 PMID: 3050858
  7. Subunit S1 of pertussis toxin: mapping of the regions essential for ADP-ribosyltransferase activity.
    Proc Natl Acad Sci U S A. 1988 Oct;85(20):7521-5 PMID: 2902632
  8. Pertussis toxin S1 mutant with reduced enzyme activity and a conserved protective epitope.
    Science. 1988 Oct 7;242(4875):72-4 PMID: 2459776
  9. Clinical, metabolic, and antibody responses of adult volunteers to an investigational vaccine composed of pertussis toxin inactivated by hydrogen peroxide.
    J Pediatr. 1988 Nov;113(5):806-13 PMID: 3263485
  10. The efficacy of acellular pertussis vaccine.
    JAMA. 1989 Jan 27;261(4):560 PMID: 2909797
  11. Pertussis toxin effects on T lymphocytes are mediated through CD3 and not by pertussis toxin catalyzed modification of a G protein.
    J Immunol. 1989 Mar 1;142(5):1631-8 PMID: 2521885
  12. Human T cell clones define S1 subunit as the most immunogenic moiety of pertussis toxin and determine its epitope map.
    J Exp Med. 1989 May 1;169(5):1519-32 PMID: 2469760
  13. Role of pertussigen (pertussis toxin) on the mouse protective activity of vaccines made from Bordetella species.
    Microbiol Immunol. 1989;33(4):341-55 PMID: 2549344
  14. Mutants of pertussis toxin suitable for vaccine development.
    Science. 1989 Oct 27;246(4929):497-500 PMID: 2683073
  15. Characterization of the protective capacity and immunogenicity of the 69-kD outer membrane protein of Bordetella pertussis.
    J Exp Med. 1990 Jan 1;171(1):63-73 PMID: 2295882
  16. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  17. Binding of Cibacron blue F3GA to proteins containing the dinucleotide fold.
    Proc Natl Acad Sci U S A. 1976 Feb;73(2):361-5 PMID: 174104
  18. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  19. Immunogenic correlation between cross-reacting material (CRM197) produced by a mutant of Corynebacterium diphtheriae and diphtheria toxoid.
    J Infect Dis. 1980 Nov;142(5):716-24 PMID: 6780629
  20. Pertussis immunisation and serious acute neurological illness in children.
    Br Med J (Clin Res Ed). 1981 May 16;282(6276):1595-9 PMID: 6786580
  21. Biological activities of crystalline pertussigen from Bordetella pertussis.
    Infect Immun. 1981 Sep;33(3):820-6 PMID: 6269999
  22. Nature and rates of adverse reactions associated with DTP and DT immunizations in infants and children.
    Pediatrics. 1981 Nov;68(5):650-60 PMID: 7031583
  23. Direct modification of the membrane adenylate cyclase system by islet-activating protein due to ADP-ribosylation of a membrane protein.
    Proc Natl Acad Sci U S A. 1982 May;79(10):3129-33 PMID: 6954463
  24. Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin.
    Infect Immun. 1983 Jun;40(3):1198-203 PMID: 6682833
  25. A role of the B-oligomer moiety of islet-activating protein, pertussis toxin, in development of the biological effects on intact cells.
    J Biol Chem. 1983 Jun 10;258(11):6756-61 PMID: 6343381
  26. Separation and purification of the hemagglutinins from Bordetella pertussis.
    Infect Immun. 1983 Jul;41(1):313-20 PMID: 6305841
  27. Pertussis toxin. Affinity purification of a new ADP-ribosyltransferase.
    J Biol Chem. 1983 Dec 10;258(23):14647-51 PMID: 6315733
  28. Molecular mechanisms of lymphocyte extravasation. I. Studies of two selective inhibitors of lymphocyte recirculation.
    J Immunol. 1984 Jan;132(1):354-62 PMID: 6537815
  29. Proliferative response of immune mouse T-lymphocytes to the lymphocytosis-promoting factor of Bordetella pertussis.
    Infect Immun. 1984 Apr;44(1):1-6 PMID: 6608495
  30. A biphasic serum glucose response in mice to inoculation with pertussis vaccine.
    J Biol Stand. 1984;12(2):151-7 PMID: 6736046
  31. Dual mechanisms involved in development of diverse biological activities of islet-activating protein, pertussis toxin, as revealed by chemical modification of lysine residues in the toxin molecule.
    Biochim Biophys Acta. 1984 Sep 28;801(2):232-43 PMID: 6383483
  32. The concept of pertussis as a toxin-mediated disease.
    Pediatr Infect Dis. 1984 Sep-Oct;3(5):467-86 PMID: 6093069
  33. Bordetella pertussis infection in mice: correlation of specific antibodies against two antigens, pertussis toxin, and filamentous hemagglutinin with mouse protectivity in an intracerebral or aerosol challenge system.
    Infect Immun. 1984 Nov;46(2):415-21 PMID: 6542069
  34. Neutralizing antibodies to pertussis toxin in whooping cough.
    J Infect Dis. 1985 Apr;151(4):646-9 PMID: 2857752
  35. Whooping cough vaccine research revs up.
    Science. 1985 Mar 8;227(4691):1184-6 PMID: 2858128
  36. The standardization of an assay for pertussis toxin and antitoxin in microplate culture of Chinese hamster ovary cells.
    J Biol Stand. 1985 Jan;13(1):61-6 PMID: 4039324
  37. Pertussis toxin is required for pertussis vaccine encephalopathy.
    Proc Natl Acad Sci U S A. 1985 Dec;82(24):8733-6 PMID: 2867545
  38. Properties of dermonecrotic toxin prepared from sonic extracts Bordetella bronchiseptica.
    Infect Immun. 1986 May;52(2):370-7 PMID: 3699886
  39. Pertussis toxin gene: nucleotide sequence and genetic organization.
    Science. 1986 Jun 6;232(4755):1258-64 PMID: 3704651
  40. Cloning and sequencing of the pertussis toxin genes: operon structure and gene duplication.
    Proc Natl Acad Sci U S A. 1986 Jul;83(13):4631-5 PMID: 2873570
  41. Determination of antigen-specific immunoglobulin content in ascitic fluids and antisera.
    J Immunol Methods. 1986 Sep 27;92(2):189-93 PMID: 2428882
  42. Virulence factors of Bordetella pertussis.
    Annu Rev Microbiol. 1986;40:661-86 PMID: 2877614
  43. Bordetella parapertussis and Bordetella bronchiseptica contain transcriptionally silent pertussis toxin genes.
    J Bacteriol. 1987 Jun;169(6):2847-53 PMID: 3584073
  44. Acellular pertussis vaccine prepared by a simple extraction and toxoiding procedure.
    Vaccine. 1987 Sep;5(3):199-207 PMID: 2890244
  45. ADP-ribosyltransferase activity of pertussis toxin and immunomodulation by Bordetella pertussis.
    Science. 1988 Apr 29;240(4852):656-9 PMID: 2896387
  46. A simple chemically defined medium for the production of phase I Bordetella pertussis.
    J Gen Microbiol. 1970 Oct;63(2):211-20 PMID: 4324651
Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.58.5.1308-1315.1990
Published
1990-05-00
Pages
1308-15
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC258625
Subset
IM
External Links
PubMed source DOI: 10.1128/iai.58.5.1308-1315.1990 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2