Identification and evaluation of new Mycobacterium bovis antigens in the in vitro interferon gamma release assay for bovine tuberculosis diagnosis

Eirin, María Emilia; Macias, Analía; Magnano, Gabriel; Morsella, Claudia; Méndez, Laura; Blanco, Federico Carlos; Bianco, María Verónica; Severina, Walter; Alito, A.; Pando, María A.; Singh, Mahavir; Spallek, Ralf; Paolicchi, Fernando Alberto; Bigi, Fabiana; Cataldi, Angel

doi:10.1016/j.tube.2015.07.009

Cited by 12 publications

(9 citation statements)

References 29 publications

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“…The worldwide used diagnostic test for the control and surveillance of bTB is the TST, based on the detection of cell mediated immunity under the exposure to bPPD, the composition of which is highly complex and remains ill-defined [4, 12]. …”

Section: Discussionmentioning

confidence: 99%

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Proteomic analysis of protein purified derivative of Mycobacterium bovis

et al. 2017

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BackgroundTuberculin skin test based on in vivo intradermal inoculation of purified protein derivative from Mycobacterium bovis (bPPD) is the diagnostic test for the control and surveillance of bovine tuberculosis (bTB).MethodsProteomic analysis was performed on different bPPD preparations from M. bovis, strain AN5. Proteins were precipitated from bPPD solutions by TCA precipitation. The proteome of bPPD preparations was investigated by bottom-up proteomics, which consisted in protein digestion and nano-LC–MS/MS analysis. Mass spectrometry analysis was performed on a Q-exactive hybrid quadrupole-Orbitrap mass spectrometer coupled online to an Easy nano-LC1000 system.ResultsThree hundred and fifty-six proteins were identified and quantified by at least 2 peptides (99% confidence per peptide). One hundred and ninety-eight proteins, which had not been previously described, were detected; furthermore, the proteomic profile shared 80 proteins with previous proteomes from bPPDs from the United Kingdom and Brazil and 139 protein components from bPPD from Korea. Locus name of M. bovis (Mb) with orthologs from M. tuberculosis H37Rv, comparative gene and protein length, molecular mass, functional categories, gene name and function of each protein were reported. Ninety-two T cell mycobacterial antigens responsible for delayed-type hypersensitivity were detected, fifty-two of which were not previously reported in any bPPD proteome. Data are available via ProteomeXchange with identifier PXD005920.ConclusionsThis study represents the highest proteome coverage of bPPD preparations to date. Since proteins perform cellular functions essential to health and/or disease, obtaining knowledge of their presence and variance is of great importance in understanding disease states and for advancing translational studies. Therefore, to better understand Mycobacterium tuberculosis complex biology during infection, survival, and persistence, the reproducible evaluation of the proteins that catalyze and control these processes is critically important. More active and more specific tuberculins would be desirable. Indeed, many antigens contained within bPPD are currently responsible for the cross-reactivity resulting in false-positive results as they are shared between non-tuberculous and tuberculous mycobacteria.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-017-1172-1) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

“…Those animals that react to PPD are isolated and slaughtered [11, 12]. Despite intensive eradication efforts over decades, bTB persists as a costly disease with adverse impacts on animal health and welfare, trade of animals and animal products, and livelihoods of producers, and continues to be a problem with global perspectives [4, 13].…”

Section: Introductionmentioning

confidence: 99%

Proteomic analysis of protein purified derivative of Mycobacterium bovis

et al. 2017

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“…This may explain the finding where a truly infected animal was incorrectly classified as negative. Because of this, for future research it will be necessary the use of specific antigens that have already been shown to be able to differentiate animals infected with M. bovis, as in the case of ESAT-6; CFP-10 [18,39] and Mb2845c [20].…”

Section: Discussion Effect On the Sensitivity Of Sctt And Ifn-γ Test mentioning

confidence: 99%

“…This in-vitro test has some advantages, such as the greater specificity compared to the single tuberculin tests [13,16]. Also, the use of specific antigens, like ESAT-6, CFP-10 [17][18][19], and more recently Mb2845c [20], are capable of reduce the presentation of cross-reactions. On the other hand, the phenomenon of desensitization does not occur because it is an in-vitro test, what allows repetitions of the test without the need of waiting any period [12,17,21].…”

Section: Introductionmentioning

confidence: 99%

Effect of tuberculin tests and gamma interferon test repeatedly used in tuberculosis-infected and non-infected cattle of Uruguay

Crosi¹,

Fernández²,

Núñez³

et al. 2018

Anim Husb Dairy Vet Sci

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The first study evaluated the effect of repeated use of single cervical tuberculin test (SCTT) and gamma-interferon (IFN-γ) test at different time intervals in infected cattle. The hypothesis was that the IFN-γ test could be repeatedly used in desensitized cattle for bovine tuberculosis diagnosis without affecting its sensitivity. 32 dairy cattle positive to cervical comparative tuberculin test (CCTT) were selected from an infected herd and randomly divided into 4 groups (n=8). All animals were tested with the in-vitro test on day 0, then a second tuberculin test and IFN-γ test was performed at different time intervals according to the group, at 15, 30, 45 and 60 days post-CCTT. They all were submitted to post-mortem inspection, histopathological analysis and Mycobacterium bovis culture. In comparison with the first tuberculin test, the mean skin thickness decreased significantly when the SCTT was performed at 15 and 30 days, with presentation of false negative results even after 45 days. The sensitivity of the IFN-γ test applied repeatedly in these desensitized animals was not affected. The second investigation evaluated the specificity of the caudal fold test (CFT) and SCTT used repeatedly in non-infected cattle. The hypothesis was that this repeated used generate a sensitization to tuberculin, with the presentation of false positive results. 100 beef cattle were classified into two groups (n=50). One group received 6 rounds of CFT and the other 6 of SCTT, with 60-day intervals between tests. The IFN-γ test was performed in all the animals on day 0 obtaining a specificity of 97%. Regarding the SCTT, a significant increase (0.48mm, p=0.0034) in the mean skin thickness was observed after six rounds of the test compared to the first test, but no positive animal was observed. All the animals remained negative to CFT during the course of this experiment, without affecting its specificity.

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“…Prior work with H1N1 [ref. 12], Hepatitis B 13 , and Mycobacterial infections 14, 15 has demonstrated the utility of epitope recognition in the development of novel diagnostics and vaccines. Therefore, we propose that detection of a specific lingering cytotoxic T-lymphocyte (CTL) memory response against Zika and/or Dengue, unlike antibody responses, would allow these infections to be distinguished.…”

Section: Introductionmentioning

confidence: 99%

Identification of putative unique immunogenic ZIKV and DENV1-4 peptides for diagnostic cellular based tests

Oom

Smith

Akrami

2017

Sci Rep

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Since the re-emergence of Zika virus in 2014 and subsequent association with microcephaly, much work has focused on the development of a vaccine to halt its spread throughout the world. The mosquito vector that transmits this virus is widespread and responsible for the spread of other arboviridae including Dengue. Current diagnostic methods rely on serologic testing that are complicated by cross reactivity and therefore unable to distinguish Zika from Dengue infection in the absence of virus isolation. We performed an in silico analysis to identify potential epitopes that may stimulate a unique T-lymphocyte response to distinguish prior infection with Zika or Dengue. From this analysis, we not only identified epitopes unique to Zika and Dengue, but also identified epitopes unique to each Dengue serotype. These peptides contribute to a pool of peptides identified for vaccine development that can be tested in vitro to confirm immunogenicity, absence of homology and global population coverage. The current lack of accurate diagnostic testing hampers our ability to understand the scope of the epidemic, implications for vaccine implementation and complications related to monoinfection and co-infection with these two closely related viruses.

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Identification and evaluation of new Mycobacterium bovis antigens in the in vitro interferon gamma release assay for bovine tuberculosis diagnosis

Abstract: Identification and evaluation of new Mycobacterium bovis antigens in the in vitro interferon gamma release assay for bovine tuberculosis diagnosis, Tuberculosis (2015),

Cited by 12 publications

References 29 publications

Proteomic analysis of protein purified derivative of Mycobacterium bovis

Proteomic analysis of protein purified derivative of Mycobacterium bovis

Effect of tuberculin tests and gamma interferon test repeatedly used in tuberculosis-infected and non-infected cattle of Uruguay

Identification of putative unique immunogenic ZIKV and DENV1-4 peptides for diagnostic cellular based tests

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