Evaluation of leptospiral recombinant antigens MPL17 and MPL21 for serological diagnosis of leptospirosis by enzyme-linked immunosorbent assays.
Leptospirosis is a zoonosis of multisystem involvement caused by pathogenic strains of the genus Leptospira. In the last few years, intensive studies aimed at the development of a vaccine have provided important knowledge about the nature of the immunological mechanisms of the host. The purpose of this study was to analyze the immune responses to two recombinant proteins, MPL17 and MPL21 (encoded by the genes LIC10765 and LIC13131, respectively) of Leptospira interrogans serovar Copenhageni in individuals during infection. The recombinant proteins were expressed in Escherichia coli as six-His tag fusion proteins and were purified from the soluble bacterial fraction by affinity chromatography with Ni(2+)-charged resin. The recombinant proteins were used to evaluate their ability to bind to immunoglobulin G (IgG) (and IgG subclass) or IgM antibodies in serum samples from patients in the early and convalescent phases of leptospirosis (n = 52) by enzyme-linked immunosorbent assays. The prevalences of total IgG antibodies against MPL17 and MPL21 were 38.5% and 21.2%, respectively. The titers achieved with MPL17 were statistically significantly higher than those obtained by the reference microscopic agglutination test. The specificity of the assay was estimated to be 95.5% for MPL17 and 80.6% for MPL21 when serum samples from individuals with unrelated febrile diseases and control healthy donors were tested. The proteins are conserved among Leptospira strains that cause human and animal diseases. MPL17 and MPL21 are most likely new surface proteins of leptospires, as revealed by liquid-phase immunofluorescence assays with living organisms. Our results demonstrate that these recombinant proteins are highly immunogenic and, when they are used together, might be useful as a means of diagnosing leptospirosis.
Oliveira
Tatiane R
TR
Centro de Biotecnologia, Instituto Butantan, São Paulo, SP, Brazil.
Longhi
Mariana T
MT
de Morais
Zenaide M
ZM
Romero
Eliete C
EC
Blanco
Roberta M
RM
Kirchgatter
Karin
K
Vasconcellos
Silvio A
SA
Nascimento
Ana L T O
AL
D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D007921:Leptospira interrogans / Q000235:genetics / Q000276:immunology*; D007922:Leptospirosis / Q000175:diagnosis*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011993:Recombinant Fusion Proteins
J Clin Microbiol. 2006 May;44(5):1853-5
16672421
J Clin Microbiol. 2001 Sep;39(9):3303-10
11526167
J Bacteriol. 1997 Jul;179(13):4403-6
9209061
Arch Microbiol. 2007 Nov;188(5):523-32
17609931
Nucleic Acids Res. 2001 Jul 15;29(14):2994-3005
11452024
Malar J. 2003 Nov 14;2(1):39
14617378
Microbiology (Reading). 2006 Jan;152(Pt 1):113-121
16385121
J Infect Dis. 2003 Jul 1;188(1):153-64
12825185
J Med Microbiol. 2004 Oct;53(Pt 10):975-984
15358819
Clin Vaccine Immunol. 2008 Mar;15(3):492-8
18184827
J Biomol NMR. 2004 Aug;29(4):553-4
15243195
Clin Diagn Lab Immunol. 2001 Jan;8(1):166-9
11139212
Nature. 2003 Apr 24;422(6934):888-93
12712204
Infect Immun. 2002 Sep;70(9):4936-45
12183539
J Clin Microbiol. 2007 May;45(5):1528-34
17360842
Braz J Med Biol Res. 2004 Apr;37(4):459-77
15064809
J Bacteriol. 2004 Apr;186(7):2164-72
15028702
J Mol Biol. 1999 Sep 17;292(2):195-202
10493868
Am J Trop Med Hyg. 2002 Jun;66(6):745-8
12224584
Infect Immun. 2006 Jul;74(7):4172-9
16790792
Anal Biochem. 1976 May 7;72:248-54
942051
Proteins. 1991;11(2):95-110
1946347
PLoS One. 2008 Feb 13;3(2):e1607
18270594
Vaccine. 2008 Jan 10;26(2):277-87
18055070
J Mol Biol. 2004 Jul 16;340(4):783-95
15223320
Infect Immun. 2005 Aug;73(8):4853-63
16040999
Infect Immun. 2007 Feb;75(2):766-73
17101664
J Microbiol Methods. 2008 Jan;72(1):73-81
18079011
J Clin Microbiol. 2003 Feb;41(2):803-9
12574287
FEMS Immunol Med Microbiol. 2001 Jul;31(1):73-81
11476985
Am J Trop Med Hyg. 1999 Nov;61(5):731-4
10586903
Malar J. 2006 Oct 06;5:83
17026752
Lancet Infect Dis. 2003 Dec;3(12):757-71
14652202
Methods Enzymol. 1990;185:60-89
2199796
J Med Microbiol. 2005 Jan;54(Pt 1):45-49
15591254
Proc Natl Acad Sci U S A. 2006 Sep 26;103(39):14560-5
16973745
Indian J Med Microbiol. 2008 Jan-Mar;26(1):45-9
18227597
Protein Eng. 1993 Jun;6(4):383-90
8332596
Infect Immun. 2001 Aug;69(8):4958-68
11447174
Protein Sci. 2003 Aug;12(8):1652-62
12876315
Comput Appl Biosci. 1993 Apr;9(2):197-9
8481823
Microbes Infect. 2000 Aug;2(10):1265-76
11008116
FEMS Microbiol Lett. 2005 Mar 15;244(2):305-13
15766783
Clin Infect Dis. 2003 Feb 15;36(4):447-52
12567302