Evaluation of cytokine responses against novel Mtb antigens as diagnostic markers for TB disease.
We investigated the accuracy of host markers detected in Mtb antigen-stimulated whole blood culture supernatant in the diagnosis of TB.



Prospectively, blood from 322 individuals with presumed TB disease from six African sites was stimulated with four different Mtb antigens (Rv0081, Rv1284, ESAT-6/CFP-10, and Rv2034) in a 24 h whole blood stimulation assay (WBA). The concentrations of 42 host markers in the supernatants were measured using the Luminex multiplex platform. Diagnostic biosignatures were investigated through the use of multivariate analysis techniques.



17% of the participants were HIV infected, 106 had active TB disease and in 216 TB was excluded. Unstimulated concentrations of CRP, SAA, ferritin and IP-10 had better discriminating ability than markers from stimulated samples. Accuracy of marker combinations by general discriminant analysis (GDA) identified a six analyte model with 77% accuracy for TB cases and 84% for non TB cases, with a better performance in HIV uninfected patients.



A biosignature of 6 cytokines obtained after stimulation with four Mtb antigens has moderate potential as a diagnostic tool for pulmonary TB disease individuals and stimulated marker expression had no added value to unstimulated marker performance.
Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg 7505, Cape Town, South Africa.
Dolapo O
DO
Awoniyi
Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg 7505, Cape Town, South Africa.
Andrea
A
Teuchert
Medical Research Council Unit, Fajara, Gambia.
Jayne S
JS
Sutherland
Department of Medicine, Makerere University, Kampala, Uganda.
Harriet
H
Mayanja-Kizza
Armauer Hansen Research Institute, Addis Ababa, Ethiopia.
Rawleigh
R
Howe
Armauer Hansen Research Institute, Addis Ababa, Ethiopia.
Adane
A
Mihret
Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg 7505, Cape Town, South Africa.
Andre G
AG
Loxton
University of Namibia, Faculty of Health Sciences, School of Medicine, Namibia.
Jacob
J
Sheehama
Ethiopian Health and Nutrition Research Institute, Addis Ababa, Ethiopia.
Desta
D
Kassa
Karonga Prevention Study, Chilumba, Malawi; London School of Hygiene and Tropical Medicine, London, UK.
Amelia C
AC
Crampin
London School of Hygiene and Tropical Medicine, London, UK.
Hazel M
HM
Dockrell
Centre for Statistical Analysis, Stellenbosch University, South Africa.
Martin
M
Kidd
Statens Serum Institut, Copenhagen, Denmark.
Ida
I
Rosenkrands
Department of Infectious Diseases, Leiden University Medical Centre, Leiden, The Netherlands.
Annemieke
A
Geluk
Department of Infectious Diseases, Leiden University Medical Centre, Leiden, The Netherlands.
Tom H M
TH
Ottenhoff
Department of Molecular Cell Biology, Leiden University Medical Centre, Leiden, The Netherlands.
P L A M
PL
Corstjens
Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg 7505, Cape Town, South Africa.
Novel N
NN
Chegou
Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg 7505, Cape Town, South Africa. Electronic address: gwalzl@sun.ac.za.
Gerhard
G
Walzl
D000328:Adult; D000942:Antigens, Bacterial / Q000097:blood / Q000276:immunology* / Q000494:pharmacology; D015415:Biomarkers / Q000097:blood; D016207:Cytokines / Q000097:blood* / Q000276:immunology; D005260:Female; D015658:HIV Infections / Q000276:immunology; D006801:Humans; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000276:immunology; D011446:Prospective Studies; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000382:microbiology
D000942:Antigens, Bacterial; D015415:Biomarkers; D016207:Cytokines
Antigen; Biosignature; Host markers