title: Production of TNF-alpha, IL-12(p40) and IL-17 can discriminate between active TB disease and latent infection in a West African cohort.
abstract: BACKGROUND
  Mycobacterium tuberculosis (MTb) infects approximately 2 billion people world-wide resulting in almost 2 million deaths per year. Determining biomarkers that distinguish different stages of tuberculosis (TB) infection and disease will provide tools for more effective diagnosis and ultimately aid in the development of new vaccine candidates. The current diagnostic kits utilising production of IFN-gamma in response to TB antigens can detect MTb infection but are unable to distinguish between infection and disease. The aim of this study was to assess if the use of a longer term assay and the analysis of multiple cytokines would enhance diagnosis of active TB in a TB-endemic population.
  
  
  METHODS
  We compared production of multiple cytokines (TNF-alpha, IFN-gamma, IL-10, IL-12(p40), IL-13, IL-17 and IL-18) following long-term (7 days) stimulation of whole-blood with TB antigens (ESAT-6/CFP-10 (EC), PPD or TB10.4) from TB cases (n = 36) and their Mycobacterium-infected (TST+; n = 20) or uninfected (TST-; n = 19) household contacts (HHC).
  
  
  CONCLUSIONS
  We found that TNF-alpha production following EC stimulation and TNF-alpha and IL-12(p40) following TB10.4 stimulation were significantly higher from TB cases compared to TST+ HHC, while production of IFN-gamma and IL-13 were significantly higher from TST+ compared to TST- HHC following PPD or EC stimulation. Combined analysis of TNF-alpha, IL-12(p40) and IL-17 following TB10.4 stimulation resulted in 85% correct classification into TB cases or TST+ HHC. 74% correct classification into TST+ or TST- HHC was achieved with IFN-gamma alone following TB10.4 stimulation (69% following EC) and little enhancement was seen with additional cytokines. We also saw a tendency for TB cases infected with M. africanum to have increased TNF-alpha and IL-10 production compared to those infected with M. tuberculosis. Our results provide further insight into the pathogenesis of tuberculosis and may enhance the specificity of the currently available diagnostic tests, particularly for diagnosis of active TB.
authors.affiliation: Bacterial Diseases Programme, Medical Research Council Laboratories, Banjul, The Gambia. jsutherland@mrc.gm
authors.forename: Jayne S
authors.initials: JS
authors.lastname: Sutherland
authors.forename: Bouke C
authors.initials: BC
authors.lastname: de Jong
authors.forename: David J
authors.initials: DJ
authors.lastname: Jeffries
authors.forename: Ifedayo M
authors.initials: IM
authors.lastname: Adetifa
authors.forename: Martin O C
authors.initials: MO
authors.lastname: Ota
mesh_terms: D000293:Adolescent; D000354:Africa, Western; D000942:Antigens, Bacterial / Q000276:immunology; D015331:Cohort Studies; D017077:Culture Media, Conditioned / Q000378:metabolism; D003937:Diagnosis, Differential; D006801:Humans; D018664:Interleukin-12 / Q000096:biosynthesis*; D020381:Interleukin-17 / Q000096:biosynthesis*; D055985:Latent Tuberculosis / Q000175:diagnosis* / Q000276:immunology / Q000378:metabolism*; D008297:Male; D015999:Multivariate Analysis; D009169:Mycobacterium tuberculosis / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013997:Time Factors; D014373:Tuberculin / Q000276:immunology; D014374:Tuberculin Test; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*; D055815:Young Adult
chemical_list: D000942:Antigens, Bacterial; D017077:Culture Media, Conditioned; C505464:ESAT-6-CFP10 fusion protein; D020381:Interleukin-17; D011993:Recombinant Fusion Proteins; C501710:TB10.4 antigen, Mycobacterium tuberculosis; D014373:Tuberculin; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12
references.citation: PLoS One. 2010 Feb 04;5(2):e9051
references.pmid: 20140219
references.citation: J Immunol. 2006 Aug 1;177(3):1416-20
references.pmid: 16849446
references.citation: Clin Vaccine Immunol. 2009 Aug;16(8):1203-12
references.pmid: 19553548
references.citation: J Clin Microbiol. 2009 Apr;47(4):994-1001
references.pmid: 19193842
references.citation: Nature. 1999 Oct 14;401(6754):708-12
references.pmid: 10537110
references.citation: N Engl J Med. 1996 Dec 26;335(26):1941-9
references.pmid: 8960473
references.citation: Clin Diagn Lab Immunol. 2000 Mar;7(2):155-60
references.pmid: 10702486
references.citation: Respirology. 2005 Jun;10(3):295-9
references.pmid: 15955140
references.citation: Clin Infect Dis. 2002 Jul 15;35(2):210-2
references.pmid: 12087531
references.citation: Drug Discov Today. 2010 Feb;15(3-4):148-57
references.pmid: 19854295
references.citation: N Engl J Med. 2001 Oct 11;345(15):1098-104
references.pmid: 11596589
references.citation: J Infect Dis. 2008 Oct 1;198(7):1037-43
references.pmid: 18702608
references.citation: BMC Pulm Med. 2009 May 16;9:21
references.pmid: 19445695
references.citation: Tuberculosis (Edinb). 2007 Nov;87(6):465-73
references.pmid: 17719276
references.citation: FEMS Immunol Med Microbiol. 1997 Jul;18(3):203-7
references.pmid: 9271171
references.citation: Lancet Infect Dis. 2009 Mar;9(3):162-72
references.pmid: 19246020
references.citation: J Immunol. 2007 Sep 15;179(6):3973-81
references.pmid: 17785835
references.citation: Clin Exp Immunol. 2009 Apr;156(1):69-77
references.pmid: 19196252
references.citation: J Infect Dis. 2008 Apr 1;197(7):941-3
references.pmid: 18419531
references.citation: J Immunol. 2008 Feb 1;180(3):1962-70
references.pmid: 18209095
references.citation: Int J Tuberc Lung Dis. 2005 Jul;9(7):753-9
references.pmid: 16013770
references.citation: BMC Infect Dis. 2007 Jul 26;7:83
references.pmid: 17655752
references.citation: J Infect Dis. 2006 May 1;193(9):1279-86
references.pmid: 16586366
references.citation: Nat Immunol. 2007 Apr;8(4):369-77
references.pmid: 17351619
references.citation: Eur Respir J. 2010 Jul;36(1):135-42
references.pmid: 19926735
references.citation: Eur J Immunol. 2009 Mar;39(3):723-9
references.pmid: 19224636
references.citation: Clin Vaccine Immunol. 2006 Feb;13(2):179-86
references.pmid: 16467323
references.citation: BMC Infect Dis. 2009 Nov 21;9:182
references.pmid: 19930588