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PMID: 16299305 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Identification of novel antigenic proteins in a complex Anaplasma marginale outer membrane immunogen by mass spectrometry and genomic mapping.

Infection and immunity ·Vol. 73 ·No. 12 ·2005-12-00 ·Pages 8109-18

Lopez JE, Siems WF, Palmer GH, Brayton KA, McGuire TC, Norimine J, Brown WC

Abstract

Immunization with purified Anaplasma marginale outer membranes induces complete protection against infection that is associated with CD4+ T-lymphocyte-mediated gamma interferon secretion and immunoglobulin G2 (IgG2) antibody titers. However, knowledge of the composition of the outer membrane immunogen is limited. Recent sequencing and annotation of the A. marginale genome predicts at least 62 outer membrane proteins (OMP), enabling a proteomic and genomic approach for identification of novel OMP by use of IgG serum antibody from outer membrane vaccinates. Outer membrane proteins were separated by two-dimensional electrophoresis, and proteins recognized by total IgG and IgG2 in immune sera of outer membrane-vaccinated cattle were detected by immunoblotting. Immunoreactive protein spots were excised and subjected to liquid chromatography-tandem mass spectrometry. A database search of the A. marginale genome identified 24 antigenic proteins that were predicted to be outer membrane, inner membrane, or membrane-associated proteins. These included the previously characterized surface-exposed outer membrane proteins MSP2, operon associated gene 2 (OpAG2), MSP3, and MSP5 as well as recently identified appendage-associated proteins. Among the 21 newly described antigenic proteins, 14 are annotated in the A. marginale genome and include type IV secretion system proteins, elongation factor Tu, and members of the MSP2 superfamily. The identification of these novel antigenic proteins markedly expands current understanding of the composition of the protective immunogen and provides new candidates for vaccine development.

MeSH Terms
Anaplasma marginale/genetics,immunology Animals Antigens, Bacterial/analysis,genetics,immunology Bacterial Outer Membrane Proteins/analysis,genetics,immunology Bacterial Vaccines/genetics,immunology Cattle Chromosome Mapping Genomics Immunization Mass Spectrometry Proteomics
Chemicals
Antigens, Bacterial Bacterial Outer Membrane Proteins Bacterial Vaccines
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Lopez Job E
Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USA.
Siems William F
Palmer Guy H
Brayton Kelly A
McGuire Travis C
Norimine Junzo
Brown Wendy C
References (63)
63 references, click to expand
  1. Emergence of Anaplasma marginale antigenic variants during persistent rickettsemia.
    Infect Immun. 1999 Nov;67(11):5834-40 PMID: 10531237
  2. Antibodies highly effective in SCID mice during infection by the intracellular bacterium Ehrlichia chaffeensis are of picomolar affinity and exhibit preferential epitope and isotype utilization.
    J Immunol. 2002 Aug 1;169(3):1419-25 PMID: 12133967
  3. Identification of Brucella suis genes affecting intracellular survival in an in vitro human macrophage infection model by signature-tagged transposon mutagenesis.
    Infect Immun. 2000 Mar;68(3):1297-303 PMID: 10678941
  4. Antibody-mediated elimination of the obligate intracellular bacterial pathogen Ehrlichia chaffeensis during active infection.
    Infect Immun. 2000 Apr;68(4):2187-95 PMID: 10722619
  5. Bacterial type IV secretion: conjugation systems adapted to deliver effector molecules to host cells.
    Trends Microbiol. 2000 Aug;8(8):354-60 PMID: 10920394
  6. Antigenic variation of Anaplasma marginale by expression of MSP2 mosaics.
    Infect Immun. 2000 Nov;68(11):6133-8 PMID: 11035716
  7. Highly conserved regions of the immunodominant major surface protein 2 of the genogroup II ehrlichial pathogen Anaplasma marginale are rich in naturally derived CD4+ T lymphocyte epitopes that elicit strong recall responses.
    J Immunol. 2001 Jan 15;166(2):1114-24 PMID: 11145692
  8. Outer membrane protein-specific monoclonal antibodies protect SCID mice from fatal infection by the obligate intracellular bacterial pathogen Ehrlichia chaffeensis.
    J Immunol. 2001 Feb 1;166(3):1855-62 PMID: 11160232
  9. Immunopathology and ehrlichial propagation are regulated by interferon-gamma and interleukin-10 in a murine model of human granulocytic ehrlichiosis.
    Am J Pathol. 2001 May;158(5):1881-8 PMID: 11337387
  10. CD4(+) T lymphocytes from calves immunized with Anaplasma marginale major surface protein 1 (MSP1), a heteromeric complex of MSP1a and MSP1b, preferentially recognize the MSP1a carboxyl terminus that is conserved among strains.
    Infect Immun. 2001 Nov;69(11):6853-62 PMID: 11598059
  11. Persistent Ehrlichia chaffeensis infection occurs in the absence of functional major histocompatibility complex class II genes.
    Infect Immun. 2002 Jan;70(1):380-8 PMID: 11748204
  12. Characterization and transcriptional analysis of gene clusters for a type IV secretion machinery in human granulocytic and monocytic ehrlichiosis agents.
    Infect Immun. 2002 Apr;70(4):2128-38 PMID: 11895979
  13. Expression of Anaplasma marginale major surface protein 2 operon-associated proteins during mammalian and arthropod infection.
    Infect Immun. 2002 Nov;70(11):6005-12 PMID: 12379676
  14. Elongation factor Tu and E1 beta subunit of pyruvate dehydrogenase complex act as fibronectin binding proteins in Mycoplasma pneumoniae.
    Mol Microbiol. 2002 Nov;46(4):1041-51 PMID: 12421310
  15. Identification of novel protective antigens from Anaplasma marginale.
    Vaccine. 2003 May 16;21(17-18):1874-83 PMID: 12706672
  16. A novel antigen from Anaplasma marginale: characterization, expression and preliminary evaluation of the recombinant protein.
    Vaccine. 2004 Jan 2;22(3-4):407-15 PMID: 14670322
  17. Delayed clearance of Ehrlichia chaffeensis infection in CD4+ T-cell knockout mice.
    Infect Immun. 2004 Jan;72(1):159-67 PMID: 14688093
  18. Characterization of lymphocyte subpopulations and major histocompatibility complex haplotypes of mastitis-resistant and susceptible cows.
    J Vet Sci. 2004 Mar;5(1):29-39 PMID: 15028883
  19. Cell surface-associated elongation factor Tu mediates the attachment of Lactobacillus johnsonii NCC533 (La1) to human intestinal cells and mucins.
    Infect Immun. 2004 Apr;72(4):2160-9 PMID: 15039339
  20. Glycosylation of Anaplasma marginale major surface protein 1a and its putative role in adhesion to tick cells.
    Infect Immun. 2004 May;72(5):3022-30 PMID: 15102815
  21. Production of IFN-gamma by CD4 T cells is essential for resolving ehrlichia infection.
    J Immunol. 2004 Jun 1;172(11):6894-901 PMID: 15153508
  22. CD4+ T lymphocytes from Anaplasma marginale major surface protein 2 (MSP2) vaccinees recognize naturally processed epitopes conserved in MSP3.
    Infect Immun. 2004 Jun;72(6):3688-92 PMID: 15155686
  23. Improved prediction of signal peptides: SignalP 3.0.
    J Mol Biol. 2004 Jul 16;340(4):783-95 PMID: 15223320
  24. Mechanisms of immunity in typhus infections. 3. Influence of human immune serum and complement on the fate of Rickettsia mooseri within the human macrophages.
    Infect Immun. 1973 Oct;8(4):631-40 PMID: 4795469
  25. Abundance and membrane association of elongation factor Tu in E. coli.
    Nature. 1976 May 6;261(5555):23-6 PMID: 775340
  26. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  27. Mechanisms of immunity in typhus infections. VI. Differential opsonizing and neutralizing action of human typhus rickettsia-specific cytophilic antibodies in cultures of human macrophages.
    Infect Immun. 1976 Oct;14(4):1071-6 PMID: 825465
  28. Functional properties of bovine IgG1 and IgG2: interaction with complement, macrophages, neutrophils and skin.
    Immunology. 1979 Oct;38(2):249-56 PMID: 511218
  29. Effect of immune serum on infectivity of Rickettsia tsutsugamushi.
    Infect Immun. 1983 Oct;42(1):341-9 PMID: 6137458
  30. Immune serum against Anaplasma marginale initial bodies neutralizes infectivity for cattle.
    J Immunol. 1984 Aug;133(2):1010-5 PMID: 6736645
  31. Immunization with an isolate-common surface protein protects cattle against anaplasmosis.
    Science. 1986 Mar 14;231(4743):1299-302 PMID: 3945825
  32. Detection of an Anaplasma marginale common surface protein present in all stages of infection.
    J Clin Microbiol. 1986 Jun;23(6):1078-83 PMID: 3711298
  33. Characterization of a neutralization-sensitive epitope on the Am 105 surface protein of Anaplasma marginale.
    Int J Parasitol. 1987 Oct;17(7):1279-85 PMID: 2448258
  34. Immunization of cattle with a 36-kilodalton surface protein induces protection against homologous and heterologous Anaplasma marginale challenge.
    Infect Immun. 1988 Jun;56(6):1526-31 PMID: 3372018
  35. Immunization of cattle with the MSP-1 surface protein complex induces protection against a structurally variant Anaplasma marginale isolate.
    Infect Immun. 1989 Nov;57(11):3666-9 PMID: 2807542
  36. Molecular basis for surface antigen size polymorphisms and conservation of a neutralization-sensitive epitope in Anaplasma marginale.
    Proc Natl Acad Sci U S A. 1990 Apr;87(8):3220-4 PMID: 1691504
  37. Passive transfer of antibody to Ehrlichia risticii protects mice from ehrlichiosis.
    Infect Immun. 1991 Jun;59(6):2058-62 PMID: 2037365
  38. Induction of protective immunity by using Anaplasma marginale initial body membranes.
    Infect Immun. 1991 Sep;59(9):3199-204 PMID: 1715323
  39. Identification of Anaplasma marginale long-term carrier cattle by detection of serum antibody to isolated MSP-3.
    J Clin Microbiol. 1991 Apr;29(4):788-93 PMID: 1890178
  40. The Anaplasma marginale msp5 gene encodes a 19-kilodalton protein conserved in all recognized Anaplasma species.
    Infect Immun. 1992 Dec;60(12):5139-44 PMID: 1280624
  41. Extensive polymorphism of the BoLA-DRB3 gene distinguished by PCR-RFLP.
    Anim Genet. 1992;23(6):483-96 PMID: 1362859
  42. IFN-gamma stimulates IgG2 production from bovine B cells costimulated with anti-mu and mitogen.
    Cell Immunol. 1994 Apr 1;154(1):287-95 PMID: 8131203
  43. 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
  44. Producer immunity towards the lantibiotic Pep5: identification of the immunity gene pepI and localization and functional analysis of its gene product.
    Appl Environ Microbiol. 1994 Aug;60(8):2876-83 PMID: 8085827
  45. Molecular and biological characterization of a newly isolated Anaplasma marginale strain.
    J Vet Diagn Invest. 1994 Oct;6(4):435-41 PMID: 7858023
  46. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels.
    Anal Chem. 1996 Mar 1;68(5):850-8 PMID: 8779443
  47. Evaluation of Anaplasma marginale major surface protein 3 (MSP3) as a diagnostic test antigen.
    J Clin Microbiol. 1996 Feb;34(2):270-76 PMID: 8788999
  48. Anaplasma marginale major surface protein 3 is encoded by a polymorphic, multigene family.
    Infect Immun. 1997 Jan;65(1):156-63 PMID: 8975906
  49. Mapping and identification of the major cell wall-associated components of Mycobacterium leprae.
    Infect Immun. 1998 Jun;66(6):2625-31 PMID: 9596726
  50. Cloning of the gene encoding the 44-kilodalton antigen of the agent of human granulocytic ehrlichiosis and characterization of the humoral response.
    Infect Immun. 1998 Jul;66(7):3264-9 PMID: 9632594
  51. A hidden Markov model for predicting transmembrane helices in protein sequences.
    Proc Int Conf Intell Syst Mol Biol. 1998;6:175-82 PMID: 9783223
  52. CD4(+) T-lymphocyte and immunoglobulin G2 responses in calves immunized with Anaplasma marginale outer membranes and protected against homologous challenge.
    Infect Immun. 1998 Nov;66(11):5406-13 PMID: 9784551
  53. The repertoire of Anaplasma marginale antigens recognized by CD4(+) T-lymphocyte clones from protectively immunized cattle is diverse and includes major surface protein 2 (MSP-2) and MSP-3.
    Infect Immun. 1998 Nov;66(11):5414-22 PMID: 9784552
  54. Multiple p44 genes encoding major outer membrane proteins are expressed in the human granulocytic ehrlichiosis agent.
    J Biol Chem. 1999 Jun 18;274(25):17828-36 PMID: 10364227
  55. Associations of the bovine major histocompatibility complex DRB3 (BoLA-DRB3) with production traits in Canadian dairy cattle.
    Anim Genet. 1999 Apr;30(2):157-60 PMID: 10376308
  56. A homologue of the Agrobacterium tumefaciens VirB and Bordetella pertussis Ptl type IV secretion systems is essential for intracellular survival of Brucella suis.
    Mol Microbiol. 1999 Sep;33(6):1210-20 PMID: 10510235
  57. Identification of a novel Anaplasma marginale appendage-associated protein that localizes with actin filaments during intraerythrocytic infection.
    Infect Immun. 2004 Dec;72(12):7257-64 PMID: 15557651
  58. Identification of IgG2-specific antigens in Mexican Anaplasma marginale strains.
    Ann N Y Acad Sci. 2004 Oct;1026:84-94 PMID: 15604474
  59. Complete genome sequencing of Anaplasma marginale reveals that the surface is skewed to two superfamilies of outer membrane proteins.
    Proc Natl Acad Sci U S A. 2005 Jan 18;102(3):844-9 PMID: 15618402
  60. Rapid and long-term disappearance of CD4+ T lymphocyte responses specific for Anaplasma marginale major surface protein-2 (MSP2) in MSP2 vaccinates following challenge with live A. marginale.
    J Immunol. 2005 Jun 1;174(11):6702-15 PMID: 15905510
  61. Antigenic variation of Anaplasma marginale msp2 occurs by combinatorial gene conversion.
    Mol Microbiol. 2002 Mar;43(5):1151-9 PMID: 11918803
  62. Bacterial secrets of secretion: EuroConference on the biology of type IV secretion processes.
    Mol Microbiol. 2002 Mar;43(5):1359-65 PMID: 11918819
  63. Molecular basis for vaccine development against the ehrlichial pathogen Anaplasma marginale.
    Parasitol Today. 1999 Jul;15(7):281-6 PMID: 10377531
Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.73.12.8109-8118.2005
Published
2005-12-00
Pages
8109-18
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC1307060
Subset
IM
Grants
NIAID NIH HHS · R01 AI053692 · United States
NIAID NIH HHS · E01-AI053692 · United States
External Links
PubMed source DOI: 10.1128/IAI.73.12.8109-8118.2005 PMC Full Text View journal details
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