title: Enterotoxigenic Escherichia coli elicits immune responses to multiple surface proteins.
abstract: Enterotoxigenic Escherichia coli (ETEC) causes considerable morbidity and mortality due to diarrheal illness in developing countries, particularly in young children. Despite the global importance of these heterogeneous pathogens, a broadly protective vaccine is not yet available. While much is known regarding the immunology of well-characterized virulence proteins, in particular the heat-labile toxin (LT) and colonization factors (CFs), to date, evaluation of the immune response to other antigens has been limited. However, the availability of genomic DNA sequences for ETEC strains coupled with proteomics technology affords opportunities to examine novel uncharacterized antigens that might also serve as targets for vaccine development. Analysis of whole or fractionated bacterial proteomes with convalescent-phase sera can potentially accelerate identification of secreted or surface-expressed targets that are recognized during the course of infection. Here we report results of an immunoproteomics approach to antigen discovery with ETEC strain H10407. Immunoblotting of proteins separated by two-dimensional electrophoresis (2DE) with sera from mice infected with strain H10407 or with convalescent human sera obtained following natural ETEC infections demonstrated multiple immunoreactive molecules in culture supernatant, outer membrane, and outer membrane vesicle preparations, suggesting that many antigens are recognized during the course of infection. Proteins identified by this approach included established virulence determinants, more recently identified putative virulence factors, as well as novel secreted and outer membrane proteins. Together, these studies suggest that existing and emerging proteomics technologies can provide a useful complement to ongoing approaches to ETEC vaccine development.
authors.affiliation: Department of Medicine, University of Tennessee Health Sciences Center, Memphis, Tennessee, USA.
authors.forename: Koushik
authors.initials: K
authors.lastname: Roy
authors.forename: Scott
authors.initials: S
authors.lastname: Bartels
authors.forename: Firdausi
authors.initials: F
authors.lastname: Qadri
authors.forename: James M
authors.initials: JM
authors.lastname: Fleckenstein
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D054307:Enterotoxigenic Escherichia coli / Q000276:immunology*; D004927:Escherichia coli Infections / Q000276:immunology* / Q000382:microbiology; D005434:Flow Cytometry; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D056724:Immunity, Humoral / Q000276:immunology; D015151:Immunoblotting; D007525:Isoelectric Focusing; D051379:Mice
chemical_list: D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins
references.citation: Clin Microbiol Rev. 1997 Oct;10(4):569-84
references.pmid: 9336662
references.citation: J Bacteriol. 2007 Apr;189(8):3228-36
references.pmid: 17293413
references.citation: Electrophoresis. 1999 Dec;20(18):3551-67
references.pmid: 10612281
references.citation: Proc Natl Acad Sci U S A. 2005 Jan 18;102(3):547-52
references.pmid: 15647345
references.citation: J Bacteriol. 2008 Oct;190(20):6881-93
references.pmid: 18676672
references.citation: Nature. 2001 Apr 26;410(6832):1099-103
references.pmid: 11323673
references.citation: Infect Immun. 2006 Apr;74(4):2245-58
references.pmid: 16552055
references.citation: J Bacteriol. 1970 Nov;104(2):882-9
references.pmid: 4923077
references.citation: Nat Protoc. 2009;4(7):1083-92
references.pmid: 19707189
references.citation: Infect Immun. 2006 Feb;74(2):869-75
references.pmid: 16428729
references.citation: Infect Immun. 1994 Aug;62(8):3463-71
references.pmid: 8039917
references.citation: Infect Immun. 1977 Nov;18(2):348-51
references.pmid: 336543
references.citation: Infect Immun. 2007 Aug;75(8):3941-9
references.pmid: 17517861
references.citation: Expert Rev Vaccines. 2009 Dec;8(12):1653-5
references.pmid: 19943760
references.citation: Expert Rev Vaccines. 2008 Aug;7(6):795-804
references.pmid: 18665777
references.citation: Nature. 2009 Jan 29;457(7229):594-8
references.pmid: 19060885
references.citation: Clin Microbiol Rev. 2005 Jul;18(3):465-83
references.pmid: 16020685
references.citation: J Infect Dis. 1977 Aug;136 Suppl:S118-23
references.pmid: 330772
references.citation: Proc Natl Acad Sci U S A. 2009 Jun 30;106(26):10793-8
references.pmid: 19515814
references.citation: FEMS Microbiol Immunol. 1988 Dec;1(3):117-25
references.pmid: 3078739
references.citation: Infect Immun. 2005 May;73(5):3137-46
references.pmid: 15845522
references.citation: J Clin Microbiol. 1980 Jan;11(1):35-40
references.pmid: 6986402
references.citation: Cell Microbiol. 2006 Sep;8(9):1516-27
references.pmid: 16922869
references.citation: Lancet. 2003 Jul 26;362(9380):286-91
references.pmid: 12892959
references.citation: Mol Microbiol. 1999 Nov;34(4):767-79
references.pmid: 10564516
references.citation: Infect Immun. 1999 Dec;67(12):6234-41
references.pmid: 10569732
references.citation: Trends Microbiol. 1996 Nov;4(11):444-52
references.pmid: 8950814
references.citation: Mol Cell Proteomics. 2008 Mar;7(3):473-85
references.pmid: 17982123
references.citation: Infect Immun. 2007 Jan;75(1):278-89
references.pmid: 17074858
references.citation: Proteomics. 2002 May;2(5):580-90
references.pmid: 11987132
references.citation: Vaccine. 2007 Mar 30;25(14):2545-66
references.pmid: 17224212
references.citation: J Bacteriol. 2005 Sep;187(18):6509-16
references.pmid: 16159784
references.citation: BMC Microbiol. 2009 Oct 16;9:220
references.pmid: 19835618
references.citation: BMC Genomics. 2008 Feb 08;9:75
references.pmid: 18261238
references.citation: Infect Immun. 1977 Jun;16(3):781-8
references.pmid: 330395
references.citation: Infect Immun. 1975 Sep;12(3):656-67
references.pmid: 1100526
references.citation: Infect Immun. 1999 Aug;67(8):4084-91
references.pmid: 10417177
references.citation: Infect Immun. 2007 Jan;75(1):252-9
references.pmid: 17074855
references.citation: J Clin Microbiol. 2009 Jan;47(1):189-97
references.pmid: 18971368
references.citation: Proteomics. 2002 Oct;2(10):1435-44
references.pmid: 12422360
references.citation: J Bacteriol. 2007 May;189(9):3532-46
references.pmid: 17351047
references.citation: J Infect Dis. 1995 Apr;171(4):1045-9
references.pmid: 7706787
references.citation: Curr Opin Gastroenterol. 2005 Jan;21(1):15-9
references.pmid: 15687879
references.citation: J Clin Microbiol. 1999 Sep;37(9):2974-8
references.pmid: 10449484
references.citation: J Biol Chem. 1972 Jul 10;247(13):4160-4
references.pmid: 4113817
references.citation: J Infect Dis. 2005 Feb 15;191(4):562-70
references.pmid: 15655780
references.citation: N Engl J Med. 1988 May 12;318(19):1240-3
references.pmid: 3283555
references.citation: Infect Immun. 1996 Jun;64(6):2256-65
references.pmid: 8675335
references.citation: Infect Immun. 1994 Dec;62(12):5624-31
references.pmid: 7960144
references.citation: Infect Immun. 1980 Aug;29(2):704-13
references.pmid: 7011982
references.citation: J Bacteriol. 2007 Jul;189(14):5060-7
references.pmid: 17496090
references.citation: Clin Chem. 1980 Aug;26(9):1255-60
references.pmid: 6772340
references.citation: Infect Immun. 2006 Jul;74(7):3864-73
references.pmid: 16790758
references.citation: Infect Immun. 1979 Mar;23(3):729-36
references.pmid: 378834
references.citation: Vaccine. 2009 Jul 23;27(34):4601-8
references.pmid: 19523914
references.citation: PLoS Pathog. 2009 Sep;5(9):e1000586
references.pmid: 19806177
references.citation: Infect Immun. 2007 Jun;75(6):2841-52
references.pmid: 17353282
references.citation: J Biol Chem. 2000 Apr 28;275(17):12489-96
references.pmid: 10777535
references.citation: Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):17020-4
references.pmid: 12471157
references.citation: Microbes Infect. 2010 Feb;12(2):89-98
references.pmid: 19883790
references.citation: Infect Immun. 2004 Mar;72(3):1786-94
references.pmid: 14977988
references.citation: Infect Immun. 2008 May;76(5):2106-12
references.pmid: 18285493
references.citation: Infect Immun. 1978 Aug;21(2):638-47
references.pmid: 80383
references.citation: Infect Immun. 2007 Aug;75(8):3961-8
references.pmid: 17548483