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PMID: 9423849 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Immunodominant major outer membrane proteins of Ehrlichia chaffeensis are encoded by a polymorphic multigene family.

Infection and immunity ·Vol. 66 ·No. 1 ·1998-01-00 ·Pages 132-9

Ohashi N, Zhi N, Zhang Y, Rikihisa Y

Abstract

Several immunodominant major proteins ranging from 23 to 30 kDa were identified in the outer membrane fractions of Ehrlichia chaffeensis and Ehrlichia canis. The N-terminal amino acid sequence of a 28-kDa protein of E. chaffeensis (one of the major proteins) was determined. The gene (p28), almost full length, encoding the 28-kDa protein was cloned by PCR with primers designed based on the N-terminal sequence of the E. chaffeensis 28-kDa protein and the consensus sequence between the C termini of the Cowdria ruminantium MAP-1 and Anaplasma marginale MSP-4 proteins. The p28 gene was overexpressed, and antibody to the recombinant protein was raised in a rabbit. The antibody and serum from a patient infected with E. chaffeensis reacted with the recombinant protein, three proteins (29, 28, and 25 kDa) of E. chaffeensis, and a 30-kDa protein of E. canis. Immunoelectron microscopy with the rabbit antibody revealed that the antigenic epitope of the 28-kDa protein was exposed on the surface of E. chaffeensis. Southern blot analysis with a 32P-labeled p28 gene probe revealed multiple copies of genes homologous to p28 in the E. chaffeensis genome. Six copies of the p28 gene were cloned and sequenced from the genomic DNA by using the same probe. The open reading frames of these gene copies were tandemly arranged with intergenic spaces. They were nonidentical genes and contained a semivariable region and three hypervariable regions in the predicted protein molecules. One of the gene copies encoded a protein with an internal amino acid sequence identical to the chemically determined N-terminal amino acid sequence of a 23-kDa protein of E. chaffeensis. Immunization with the recombinant P28 protein protected mice from infection with E. chaffeensis. These findings suggest that the 30-kDa-range proteins of E. chaffeensis represent a family of antigenically related homologous proteins encoded by a single gene family.

MeSH Terms
Amino Acid Sequence Animals Antibodies, Bacterial/biosynthesis,immunology Antigens, Bacterial Bacterial Outer Membrane Proteins/genetics,immunology Bacterial Proteins/genetics Cloning, Molecular DNA, Bacterial/analysis,genetics Ehrlichia/genetics,immunology Ehrlichia chaffeensis/genetics,immunology,ultrastructure Gene Expression Genes, Bacterial Genome, Bacterial Humans Immunodominant Epitopes Male Membrane Proteins/genetics Mice Microscopy, Immunoelectron Molecular Sequence Data Neutralization Tests Open Reading Frames Phylogeny Polymerase Chain Reaction RNA, Ribosomal, 16S/genetics Rabbits Recombinant Proteins/immunology Restriction Mapping Sequence Alignment Sequence Analysis Sequence Analysis, DNA Sequence Homology, Amino Acid
Chemicals
Antibodies, Bacterial Antigens, Bacterial Bacterial Outer Membrane Proteins Bacterial Proteins DNA, Bacterial Immunodominant Epitopes Membrane Proteins Msp4 protein, Anaplasma marginale RNA, Ribosomal, 16S Recombinant Proteins major antigenic protein 1, Cowdria ruminantium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ohashi N
Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus 43210-1093, USA.
Zhi N
Zhang Y
Rikihisa Y
References (40)
40 references, click to expand
  1. Analysis and ultrastructural localization of Ehrlichia chaffeensis proteins with monoclonal antibodies.
    Am J Trop Med Hyg. 1996 Apr;54(4):405-12 PMID: 8615456
  2. Ehrlichia chaffeensis, a new species associated with human ehrlichiosis.
    J Clin Microbiol. 1991 Dec;29(12):2838-42 PMID: 1757557
  3. Anaplasma marginale major surface protein 3 is encoded by a polymorphic, multigene family.
    Infect Immun. 1997 Jan;65(1):156-63 PMID: 8975906
  4. Cloning and sequencing of the gene for a 120-kDa immunodominant protein of Ehrlichia chaffeensis.
    Gene. 1997 Jan 15;184(2):149-54 PMID: 9031621
  5. Genetic and antigenic diversity of Ehrlichia chaffeensis: comparative analysis of a novel human strain from Oklahoma and previously isolated strains.
    J Infect Dis. 1997 Apr;175(4):856-63 PMID: 9086141
  6. Antigenic variation in Lyme disease borreliae by promiscuous recombination of VMP-like sequence cassettes.
    Cell. 1997 Apr 18;89(2):275-85 PMID: 9108482
  7. Ehrlichia sennetsu groE operon and antigenic properties of the GroEL homolog.
    FEMS Immunol Med Microbiol. 1997 May;18(1):39-46 PMID: 9215585
  8. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate.
    J Bacteriol. 1973 Sep;115(3):717-22 PMID: 4580564
  9. Analyses of Ehrlichia canis and a canine granulocytic Ehrlichia infection.
    J Clin Microbiol. 1992 Jan;30(1):143-8 PMID: 1734046
  10. Detection of the etiologic agent of human ehrlichiosis by polymerase chain reaction.
    J Clin Microbiol. 1992 Apr;30(4):775-80 PMID: 1374076
  11. Phylogenetic relationship of Cowdria ruminantium, agent of heartwater, to Anaplasma marginale and other members of the order Rickettsiales determined on the basis of 16S rRNA sequence.
    Int J Syst Bacteriol. 1992 Apr;42(2):270-4 PMID: 1581187
  12. Diversity of immunodominant 56-kDa type-specific antigen (TSA) of Rickettsia tsutsugamushi. Sequence and comparative analyses of the genes encoding TSA homologues from four antigenic variants.
    J Biol Chem. 1992 Jun 25;267(18):12728-35 PMID: 1618776
  13. Ribonuclease S-peptide as a carrier in fusion proteins.
    Protein Sci. 1993 Mar;2(3):348-56 PMID: 8453373
  14. Human ehrlichiosis: hematopathology and immunohistologic detection of Ehrlichia chaffeensis.
    Hum Pathol. 1993 Apr;24(4):391-6 PMID: 8491479
  15. Ehrlichia chaffeensis expresses an immunoreactive protein homologous to the Escherichia coli GroEL protein.
    Infect Immun. 1993 Aug;61(8):3536-9 PMID: 8101510
  16. An immunoblotting diagnostic assay for heartwater based on the immunodominant 32-kilodalton protein of Cowdria ruminantium detects false positives in field sera.
    J Clin Microbiol. 1993 Oct;31(10):2729-37 PMID: 8253974
  17. Derivation of the complete msp4 gene sequence of Anaplasma marginale without cloning.
    Gene. 1993 Dec 22;136(1-2):291-4 PMID: 8294020
  18. Identification of the antigenic constituents of Ehrlichia chaffeensis.
    Am J Trop Med Hyg. 1994 Jan;50(1):52-8 PMID: 8304572
  19. Detection of Ehrlichia chaffeensis in human tissue by using a species-specific monoclonal antibody.
    J Clin Microbiol. 1993 Dec;31(12):3284-8 PMID: 7508458
  20. Molecular cloning, sequence analysis, and expression of the gene encoding the immunodominant 32-kilodalton protein of Cowdria ruminantium.
    Infect Immun. 1994 Apr;62(4):1451-6 PMID: 8132352
  21. Linear DNA of Borrelia species and antigenic variation.
    Trends Microbiol. 1993 Sep;1(6):236-9 PMID: 8137122
  22. The immunoprotective Anaplasma marginale major surface protein 2 is encoded by a polymorphic multigene family.
    Infect Immun. 1994 Sep;62(9):3808-16 PMID: 8063397
  23. Inhibition of infection of macrophages with Ehrlichia risticii by cytochalasins, monodansylcadaverine, and taxol.
    Infect Immun. 1994 Nov;62(11):5126-32 PMID: 7927796
  24. Western immunoblot analysis of Ehrlichia chaffeensis, E. canis, or E. ewingii infections in dogs and humans.
    J Clin Microbiol. 1994 Sep;32(9):2107-12 PMID: 7814533
  25. Ehrlichia canis-like agent isolated from a man in Venezuela: antigenic and genetic characterization.
    J Clin Microbiol. 1996 Sep;34(9):2133-9 PMID: 8862572
  26. Lysozyme-promoted association of protein I molecules in the outer membrane of Escherichia coli.
    J Bacteriol. 1977 Nov;132(2):411-8 PMID: 334740
  27. Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis.
    Infect Immun. 1981 Mar;31(3):1161-76 PMID: 7228399
  28. Outer membrane proteins of Brucella abortus: isolation and characterization.
    Infect Immun. 1982 Mar;35(3):979-89 PMID: 6802764
  29. Protein secretion in Escherichia coli.
    Annu Rev Microbiol. 1985;39:615-48 PMID: 3904614
  30. The repertoire of silent pilus genes in Neisseria gonorrhoeae: evidence for gene conversion.
    Cell. 1986 Jan 17;44(1):107-15 PMID: 2866848
  31. Enzyme-linked immunosorbent assay for Potomac horse fever disease.
    J Clin Microbiol. 1987 Jan;25(1):31-6 PMID: 3539995
  32. Human infection with Ehrlichia canis, a leukocytic rickettsia.
    N Engl J Med. 1987 Apr 2;316(14):853-6 PMID: 3029590
  33. Isolation and characterization of a new strain of Ehrlichia chaffeensis from a patient with nearly fatal monocytic ehrlichiosis.
    J Clin Microbiol. 1995 Jul;33(7):1704-11 PMID: 7665632
  34. Expression of major surface protein 2 antigenic variants during acute Anaplasma marginale rickettsemia.
    Infect Immun. 1996 Mar;64(3):836-41 PMID: 8641789
  35. Energy metabolism of monocytic Ehrlichia.
    Proc Natl Acad Sci U S A. 1989 Mar;86(5):1674-8 PMID: 2922404
  36. Purification and partial characterization of a type-specific antigen of Rickettsia tsutsugamushi.
    Infect Immun. 1989 May;57(5):1427-31 PMID: 2496028
  37. Chlamydia trachomatis-host cell interactions: role of the chlamydial major outer membrane protein as an adhesin.
    Infect Immun. 1990 Apr;58(4):1017-25 PMID: 2318528
  38. Epidemiologic, clinical, and laboratory findings of human ehrlichiosis in the United States, 1988.
    JAMA. 1990 Nov 7;264(17):2251-8 PMID: 2214103
  39. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  40. Diversity of 16S rRNA genes of new Ehrlichia strains isolated from horses with clinical signs of Potomac horse fever.
    Int J Syst Bacteriol. 1995 Apr;45(2):315-8 PMID: 7537065
Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.66.1.132-139.1998
Published
1998-01-00
Pages
132-9
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC107868
Subset
IM
Grants
NIAID NIH HHS · R01 AI33123 · United States
Databases
GENBANK
AF021338, U72291
External Links
PubMed source DOI: 10.1128/IAI.66.1.132-139.1998 PMC Full Text View journal details
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