Multiple cytokines are released when blood from patients with tuberculosis is stimulated with Mycobacterium tuberculosis antigens

Kathryn L. Kellar, Jennifer Gehrke, Stephen Weis, Aida Mahmutovic-Mayhew, Blachy Davila, Margan J. Zajdowicz, Robin Scarborough, Philip A. LoBue, Alfred A. Lardizabal, Charles L. Daley, Randall R. Reves, John Bernardo, Brandon H. Campbell, William C. Whitworth, Gerald H. Mazurek

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Abstract

Background: Mycobacterium tuberculosis (Mtb) infection may cause overt disease or remain latent. Interferon gamma release assays (IGRAs) detect Mtb infection, both latent infection and infection manifesting as overt disease, by measuring whole-blood interferon gamma (IFN-γ) responses to Mtb antigens such as early secreted antigenic target-6 (ESAT-6), culture filtrate protein 10 (CFP-10), and TB7.7. Due to a lack of adequate diagnostic standards for confirming latent Mtb infection, IGRA sensitivity for detecting Mtb infection has been estimated using patients with culture-confirmed tuberculosis (CCTB) for whom recovery of Mtb confirms the infection. In this study, cytokines in addition to IFN-γ were assessed for potential to provide robust measures of Mtb infection. Methods: Cytokine responses to ESAT-6, CFP-10, TB7.7, or combinations of these Mtb antigens, for patients with CCTB were compared with responses for subjects at low risk for Mtb infection (controls). Three different multiplexed immunoassays were used to measure concentrations of 9 to 20 different cytokines. Responses were calculated by subtracting background cytokine concentrations from cytokine concentrations in plasma from blood stimulated with Mtb antigens. Results: Two assays demonstrated that ESAT-6, CFP-10, ESAT-6+CFP-10, and ESAT-6+CFP-10+TB7.7 stimulated the release of significantly greater amounts of IFN-γ, IL-2, IL-8, MCP-1 and MIP-1β for CCTB patients than for controls. Responses to combination antigens were, or tended to be, greater than responses to individual antigens. A third assay, using whole blood stimulation with ESAT-6+CFP-10+TB7.7, revealed significantly greater IFN-γ, IL-2, IL-6, IL-8, IP-10, MCP-1, MIP-1β, and TNF-α responses among patients compared with controls. One CCTB patient with a falsely negative IFN-γ response had elevated responses with other cytokines. Conclusions: Multiple cytokines are released when whole blood from patients with CCTB is stimulated with Mtb antigens. Measurement of multiple cytokine responses may improve diagnostic sensitivity for Mtb infection compared with assessment of IFN-γ alone.

Original languageEnglish
Article numbere26545
JournalPLoS ONE
Volume6
Issue number11
DOIs
StatePublished - 21 Nov 2011

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Mycobacterium tuberculosis
Mycobacterium Infections
tuberculosis
Tuberculosis
Blood
cytokines
Cytokines
antigens
blood
culture filtrates
Assays
Interferon-gamma Release Tests
infection
Interferon-gamma
Proteins
interferon-gamma
Interleukin-8
Interleukin-2
proteins
Infection

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Kellar, K. L., Gehrke, J., Weis, S., Mahmutovic-Mayhew, A., Davila, B., Zajdowicz, M. J., ... Mazurek, G. H. (2011). Multiple cytokines are released when blood from patients with tuberculosis is stimulated with Mycobacterium tuberculosis antigens. PLoS ONE, 6(11), [e26545]. https://doi.org/10.1371/journal.pone.0026545
Kellar, Kathryn L. ; Gehrke, Jennifer ; Weis, Stephen ; Mahmutovic-Mayhew, Aida ; Davila, Blachy ; Zajdowicz, Margan J. ; Scarborough, Robin ; LoBue, Philip A. ; Lardizabal, Alfred A. ; Daley, Charles L. ; Reves, Randall R. ; Bernardo, John ; Campbell, Brandon H. ; Whitworth, William C. ; Mazurek, Gerald H. / Multiple cytokines are released when blood from patients with tuberculosis is stimulated with Mycobacterium tuberculosis antigens. In: PLoS ONE. 2011 ; Vol. 6, No. 11.
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abstract = "Background: Mycobacterium tuberculosis (Mtb) infection may cause overt disease or remain latent. Interferon gamma release assays (IGRAs) detect Mtb infection, both latent infection and infection manifesting as overt disease, by measuring whole-blood interferon gamma (IFN-γ) responses to Mtb antigens such as early secreted antigenic target-6 (ESAT-6), culture filtrate protein 10 (CFP-10), and TB7.7. Due to a lack of adequate diagnostic standards for confirming latent Mtb infection, IGRA sensitivity for detecting Mtb infection has been estimated using patients with culture-confirmed tuberculosis (CCTB) for whom recovery of Mtb confirms the infection. In this study, cytokines in addition to IFN-γ were assessed for potential to provide robust measures of Mtb infection. Methods: Cytokine responses to ESAT-6, CFP-10, TB7.7, or combinations of these Mtb antigens, for patients with CCTB were compared with responses for subjects at low risk for Mtb infection (controls). Three different multiplexed immunoassays were used to measure concentrations of 9 to 20 different cytokines. Responses were calculated by subtracting background cytokine concentrations from cytokine concentrations in plasma from blood stimulated with Mtb antigens. Results: Two assays demonstrated that ESAT-6, CFP-10, ESAT-6+CFP-10, and ESAT-6+CFP-10+TB7.7 stimulated the release of significantly greater amounts of IFN-γ, IL-2, IL-8, MCP-1 and MIP-1β for CCTB patients than for controls. Responses to combination antigens were, or tended to be, greater than responses to individual antigens. A third assay, using whole blood stimulation with ESAT-6+CFP-10+TB7.7, revealed significantly greater IFN-γ, IL-2, IL-6, IL-8, IP-10, MCP-1, MIP-1β, and TNF-α responses among patients compared with controls. One CCTB patient with a falsely negative IFN-γ response had elevated responses with other cytokines. Conclusions: Multiple cytokines are released when whole blood from patients with CCTB is stimulated with Mtb antigens. Measurement of multiple cytokine responses may improve diagnostic sensitivity for Mtb infection compared with assessment of IFN-γ alone.",
author = "Kellar, {Kathryn L.} and Jennifer Gehrke and Stephen Weis and Aida Mahmutovic-Mayhew and Blachy Davila and Zajdowicz, {Margan J.} and Robin Scarborough and LoBue, {Philip A.} and Lardizabal, {Alfred A.} and Daley, {Charles L.} and Reves, {Randall R.} and John Bernardo and Campbell, {Brandon H.} and Whitworth, {William C.} and Mazurek, {Gerald H.}",
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Kellar, KL, Gehrke, J, Weis, S, Mahmutovic-Mayhew, A, Davila, B, Zajdowicz, MJ, Scarborough, R, LoBue, PA, Lardizabal, AA, Daley, CL, Reves, RR, Bernardo, J, Campbell, BH, Whitworth, WC & Mazurek, GH 2011, 'Multiple cytokines are released when blood from patients with tuberculosis is stimulated with Mycobacterium tuberculosis antigens', PLoS ONE, vol. 6, no. 11, e26545. https://doi.org/10.1371/journal.pone.0026545

Multiple cytokines are released when blood from patients with tuberculosis is stimulated with Mycobacterium tuberculosis antigens. / Kellar, Kathryn L.; Gehrke, Jennifer; Weis, Stephen; Mahmutovic-Mayhew, Aida; Davila, Blachy; Zajdowicz, Margan J.; Scarborough, Robin; LoBue, Philip A.; Lardizabal, Alfred A.; Daley, Charles L.; Reves, Randall R.; Bernardo, John; Campbell, Brandon H.; Whitworth, William C.; Mazurek, Gerald H.

In: PLoS ONE, Vol. 6, No. 11, e26545, 21.11.2011.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Multiple cytokines are released when blood from patients with tuberculosis is stimulated with Mycobacterium tuberculosis antigens

AU - Kellar, Kathryn L.

AU - Gehrke, Jennifer

AU - Weis, Stephen

AU - Mahmutovic-Mayhew, Aida

AU - Davila, Blachy

AU - Zajdowicz, Margan J.

AU - Scarborough, Robin

AU - LoBue, Philip A.

AU - Lardizabal, Alfred A.

AU - Daley, Charles L.

AU - Reves, Randall R.

AU - Bernardo, John

AU - Campbell, Brandon H.

AU - Whitworth, William C.

AU - Mazurek, Gerald H.

PY - 2011/11/21

Y1 - 2011/11/21

N2 - Background: Mycobacterium tuberculosis (Mtb) infection may cause overt disease or remain latent. Interferon gamma release assays (IGRAs) detect Mtb infection, both latent infection and infection manifesting as overt disease, by measuring whole-blood interferon gamma (IFN-γ) responses to Mtb antigens such as early secreted antigenic target-6 (ESAT-6), culture filtrate protein 10 (CFP-10), and TB7.7. Due to a lack of adequate diagnostic standards for confirming latent Mtb infection, IGRA sensitivity for detecting Mtb infection has been estimated using patients with culture-confirmed tuberculosis (CCTB) for whom recovery of Mtb confirms the infection. In this study, cytokines in addition to IFN-γ were assessed for potential to provide robust measures of Mtb infection. Methods: Cytokine responses to ESAT-6, CFP-10, TB7.7, or combinations of these Mtb antigens, for patients with CCTB were compared with responses for subjects at low risk for Mtb infection (controls). Three different multiplexed immunoassays were used to measure concentrations of 9 to 20 different cytokines. Responses were calculated by subtracting background cytokine concentrations from cytokine concentrations in plasma from blood stimulated with Mtb antigens. Results: Two assays demonstrated that ESAT-6, CFP-10, ESAT-6+CFP-10, and ESAT-6+CFP-10+TB7.7 stimulated the release of significantly greater amounts of IFN-γ, IL-2, IL-8, MCP-1 and MIP-1β for CCTB patients than for controls. Responses to combination antigens were, or tended to be, greater than responses to individual antigens. A third assay, using whole blood stimulation with ESAT-6+CFP-10+TB7.7, revealed significantly greater IFN-γ, IL-2, IL-6, IL-8, IP-10, MCP-1, MIP-1β, and TNF-α responses among patients compared with controls. One CCTB patient with a falsely negative IFN-γ response had elevated responses with other cytokines. Conclusions: Multiple cytokines are released when whole blood from patients with CCTB is stimulated with Mtb antigens. Measurement of multiple cytokine responses may improve diagnostic sensitivity for Mtb infection compared with assessment of IFN-γ alone.

AB - Background: Mycobacterium tuberculosis (Mtb) infection may cause overt disease or remain latent. Interferon gamma release assays (IGRAs) detect Mtb infection, both latent infection and infection manifesting as overt disease, by measuring whole-blood interferon gamma (IFN-γ) responses to Mtb antigens such as early secreted antigenic target-6 (ESAT-6), culture filtrate protein 10 (CFP-10), and TB7.7. Due to a lack of adequate diagnostic standards for confirming latent Mtb infection, IGRA sensitivity for detecting Mtb infection has been estimated using patients with culture-confirmed tuberculosis (CCTB) for whom recovery of Mtb confirms the infection. In this study, cytokines in addition to IFN-γ were assessed for potential to provide robust measures of Mtb infection. Methods: Cytokine responses to ESAT-6, CFP-10, TB7.7, or combinations of these Mtb antigens, for patients with CCTB were compared with responses for subjects at low risk for Mtb infection (controls). Three different multiplexed immunoassays were used to measure concentrations of 9 to 20 different cytokines. Responses were calculated by subtracting background cytokine concentrations from cytokine concentrations in plasma from blood stimulated with Mtb antigens. Results: Two assays demonstrated that ESAT-6, CFP-10, ESAT-6+CFP-10, and ESAT-6+CFP-10+TB7.7 stimulated the release of significantly greater amounts of IFN-γ, IL-2, IL-8, MCP-1 and MIP-1β for CCTB patients than for controls. Responses to combination antigens were, or tended to be, greater than responses to individual antigens. A third assay, using whole blood stimulation with ESAT-6+CFP-10+TB7.7, revealed significantly greater IFN-γ, IL-2, IL-6, IL-8, IP-10, MCP-1, MIP-1β, and TNF-α responses among patients compared with controls. One CCTB patient with a falsely negative IFN-γ response had elevated responses with other cytokines. Conclusions: Multiple cytokines are released when whole blood from patients with CCTB is stimulated with Mtb antigens. Measurement of multiple cytokine responses may improve diagnostic sensitivity for Mtb infection compared with assessment of IFN-γ alone.

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