Combinatorial use of antibodies to secreted mycobacterial proteins in a host immune system-independent test for tuberculosis

C. P. Landowski, H. P. Godfrey, S. I. Bentley-Hibbert, X. Liu, Z. Huang, R. Sepulveda, K. Huygen, M. L. Gennaro, F. H. Moy, S. A. Lesley, M. Haak-Frendscho

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Abstract

Laboratory diagnosis of tuberculosis is often difficult. Immunodetection of circulating Mycobacterium tuberculosis proteins shed during active infection would not depend on an intact host immune response and could take advantage of the speed and low costs afforded by antibody-based assays. We previously showed that patients with active tuberculosis had increased levels of circulating antigen 85 (Ag85) proteins independent of their tuberculin skin test status (S. I. Bentley-Hibbert, X. Quan, T. Newman, K. Huygen, and H. P. Godfrey, Infect. Immun. 67:581-588, 1999). To extend these observations to a Mycobacterium bovis BCG-vaccinated population and to another secreted mycobacterial protein, Ag85 and PstS-1 (protein antigen B, p38 antigen) were quantified in sera from 97 Chilean tuberculosis patients and healthy controls (many of whom had received BCG as children) using dot immunobinding, mouse monoclonal anti-BCG Ag85 complex antibody, and chicken antipeptide antibodies reactive with M. tuberculosis Ag85B and PstS-1. The latter antibodies had been raised to peptide-derived immunogens expressed on a novel proprietary protein carrier in Escherichia coli. Median serum Ag85 levels measured by using either anti-Ag85 antibody were significantly higher in patients with active tuberculosis than in healthy controls (P, <0.001 to 0.01); the median serum PstS-1 levels were similar in patients and controls. The sensitivity of significantly elevated circulating Ag85 levels in patients with pulmonary tuberculosis measured by anti-Ag85 complex or anti-Ag85B antibodies was 60 and 55%, respectively, but increased to 77% when results obtained with both anti-Ag85 antibodies were considered jointly (P < 0.02). The corresponding specificities for individual and joint consideration were 95, 85, and 80%, respectively. These results indicate that elevated Ag85 levels can be detected in patients with active tuberculosis even after BCG vaccination and suggest that combinatorial use of antibodies directed at different epitopes of this protein could provide a viable strategy for developing new host immune response-independent diagnostic tests for tuberculosis.

Original languageEnglish
Pages (from-to)2418-2424
Number of pages7
JournalJournal of Clinical Microbiology
Volume39
Issue number7
DOIs
Publication statusPublished - 14 Jul 2001
MoE publication typeA1 Journal article-refereed

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Immune System
Tuberculosis
Antigens
Antibodies
Mycobacterium bovis
Proteins
Mycobacterium tuberculosis
Serum
Tuberculin Test
Clinical Laboratory Techniques
Skin Tests
Antigen-Antibody Complex
Pulmonary Tuberculosis
Routine Diagnostic Tests
Epitopes
Anti-Idiotypic Antibodies
Chickens
Carrier Proteins
Vaccination
Joints

Cite this

Landowski, C. P. ; Godfrey, H. P. ; Bentley-Hibbert, S. I. ; Liu, X. ; Huang, Z. ; Sepulveda, R. ; Huygen, K. ; Gennaro, M. L. ; Moy, F. H. ; Lesley, S. A. ; Haak-Frendscho, M. / Combinatorial use of antibodies to secreted mycobacterial proteins in a host immune system-independent test for tuberculosis. In: Journal of Clinical Microbiology. 2001 ; Vol. 39, No. 7. pp. 2418-2424.
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abstract = "Laboratory diagnosis of tuberculosis is often difficult. Immunodetection of circulating Mycobacterium tuberculosis proteins shed during active infection would not depend on an intact host immune response and could take advantage of the speed and low costs afforded by antibody-based assays. We previously showed that patients with active tuberculosis had increased levels of circulating antigen 85 (Ag85) proteins independent of their tuberculin skin test status (S. I. Bentley-Hibbert, X. Quan, T. Newman, K. Huygen, and H. P. Godfrey, Infect. Immun. 67:581-588, 1999). To extend these observations to a Mycobacterium bovis BCG-vaccinated population and to another secreted mycobacterial protein, Ag85 and PstS-1 (protein antigen B, p38 antigen) were quantified in sera from 97 Chilean tuberculosis patients and healthy controls (many of whom had received BCG as children) using dot immunobinding, mouse monoclonal anti-BCG Ag85 complex antibody, and chicken antipeptide antibodies reactive with M. tuberculosis Ag85B and PstS-1. The latter antibodies had been raised to peptide-derived immunogens expressed on a novel proprietary protein carrier in Escherichia coli. Median serum Ag85 levels measured by using either anti-Ag85 antibody were significantly higher in patients with active tuberculosis than in healthy controls (P, <0.001 to 0.01); the median serum PstS-1 levels were similar in patients and controls. The sensitivity of significantly elevated circulating Ag85 levels in patients with pulmonary tuberculosis measured by anti-Ag85 complex or anti-Ag85B antibodies was 60 and 55{\%}, respectively, but increased to 77{\%} when results obtained with both anti-Ag85 antibodies were considered jointly (P < 0.02). The corresponding specificities for individual and joint consideration were 95, 85, and 80{\%}, respectively. These results indicate that elevated Ag85 levels can be detected in patients with active tuberculosis even after BCG vaccination and suggest that combinatorial use of antibodies directed at different epitopes of this protein could provide a viable strategy for developing new host immune response-independent diagnostic tests for tuberculosis.",
author = "Landowski, {C. P.} and Godfrey, {H. P.} and Bentley-Hibbert, {S. I.} and X. Liu and Z. Huang and R. Sepulveda and K. Huygen and Gennaro, {M. L.} and Moy, {F. H.} and Lesley, {S. A.} and M. Haak-Frendscho",
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Landowski, CP, Godfrey, HP, Bentley-Hibbert, SI, Liu, X, Huang, Z, Sepulveda, R, Huygen, K, Gennaro, ML, Moy, FH, Lesley, SA & Haak-Frendscho, M 2001, 'Combinatorial use of antibodies to secreted mycobacterial proteins in a host immune system-independent test for tuberculosis', Journal of Clinical Microbiology, vol. 39, no. 7, pp. 2418-2424. https://doi.org/10.1128/JCM.39.7.2418-2424.2001

Combinatorial use of antibodies to secreted mycobacterial proteins in a host immune system-independent test for tuberculosis. / Landowski, C. P.; Godfrey, H. P.; Bentley-Hibbert, S. I.; Liu, X.; Huang, Z.; Sepulveda, R.; Huygen, K.; Gennaro, M. L.; Moy, F. H.; Lesley, S. A.; Haak-Frendscho, M.

In: Journal of Clinical Microbiology, Vol. 39, No. 7, 14.07.2001, p. 2418-2424.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Liu, X.

AU - Huang, Z.

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AU - Huygen, K.

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