Synergistic antigen combinations for the development of interferon gamma release assays for paucibacillary leprosy
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- 作者：R. M. Oliveira (1)
E. M. Hungria (1)
A. de Araújo Freitas (1)
A. L. O. M. de Sousa (1)
M. B. Costa (1)
S. G. Reed (2)
M. S. Duthie (2)
M. M. A. Stefani (1) - 刊名：European Journal of Clinical Microbiology & Infectious Diseases
- 出版年：2014
- 出版时间：August 2014
- 年：2014
- 卷：33
- 期：8
- 页码：1415-1424
- 全文大小：554 KB
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E. M. Hungria (1)
A. de Araújo Freitas (1)
A. L. O. M. de Sousa (1)
M. B. Costa (1)
S. G. Reed (2)
M. S. Duthie (2)
M. M. A. Stefani (1)
1. Tropical Pathology and Public Health Institute, Federal University of Goiás, Goiania, GO, Brazil
2. Infectious Disease Research Institute, 1124 Columbia Street, Suite 400, Seattle, WA, 98104, USA - ISSN：1435-4373
文摘
The development of immunodiagnostic tests for paucibacillary leprosy (PB) is based on Mycobacterium leprae specific-cell mediated immunity (CMI)/IFN-γ production. Recently, novel M. leprae protein antigens that stimulate CMI have been described. This study evaluated different M. leprae antigen combinations in whole blood assay (WBA). Five study groups were tested (20 per group): newly diagnosed, untreated PB patients and multibacillary leprosy patients (MB); household contacts of MB patients (HHC); healthy endemic controls (EC); pulmonary tuberculosis patients (TB). WBA (heparinized, 24?h 37?°C 5?% CO2) were stimulated with: 10?μg/ml of each individual M. leprae recombinant protein (rML) and five combinations of rML (46f-?LID-1, ML0276-?LID-1, ML2055-?ML1632-?ML2044, ML0276--6f, ML2055-?LID-1)-em class="a-plus-plus">M. leprae cell sonicate (MLCS, 10?μg/ml), PHA (1?μg/ml), and PBS alone. Human IFN-γ ELISA (QuantiFERON?-TB Gold/QFT-G, Cellestis) was performed using stimulated plasma (arbitrary cut-off--0?pg/ml). Three out of five antigen combinations (46f-?LID-1, ML0276-?LID-1, ML2055-?ML1632-?ML2044) were able to increase the levels of IFN-γ production in WBA in a larger number of responders among both PB leprosy and contacts. However, the magnitude of IFN-γ responses was higher among contacts. The antigen combination (46f-?ML0276) stimulated IFN-γ only in symptomatic PB leprosy patients and not in asymptomatic contacts. Few controls (EC, TB) responded to combinations (0-5?%), indicating the specificity of the response in an endemic area with high BCG coverage. The synergistic effect of new combinations of M. leprae proteins upon IFN-γ production in WBA indicates their potential use for the development of an interferon gamma release assay/IGRA for the diagnosis of PB leprosy.