Recombinant GroES in combination with CpG oligodeoxynucleotides protects mice against Mycobacterium avium infection

Fattorini, Lanfranco; Creti, Roberta; Nisini, Roberto; Pietrobono, Roberta; Fan, Yuming; Stringaro, Annarita; Arancia, Giuseppe; Serlupi-Crescenzi, Ottaviano; Iona, Elisabetta; Orefici, Graziella

doi:10.1099/0022-1317-51-12-1071

Cited by 8 publications

(7 citation statements)

References 33 publications

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“…Our results are correlating with earlier publications and many of the proteins identified in the PPD of M. tuberculosis , have already been analyzed for their potential as a diagnostic markers. Some of the hits we have found are GroES [52,53], GroEL [54,55], protein EsxB (EsxB) [56] heat shock protein HspX (HspX) [57], hypothetical protein (TB15.3), hypothetical protien (TB16.3) [58], 50S ribosomal protein L7/L12 (RplL) [59], hypothetical protein (EsxA) [60], immunogenic protein Mpt63 (Mpt63) [61], Mpt64 [62], ESAT-6 like protein EsxJ (EsxJ), and ESAT-6 like protein EsxO (EsxO) [63]. Based on these observations, many more M. tuberculosis PPD can be analyzed and the abundant antigens be evaluated for their potential as diagnostics biomarkers.…”

Section: Resultsmentioning

confidence: 99%

Proteomic analysis of purified protein derivative of Mycobacterium tuberculosis

et al. 2013

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BackgroundPurified protein derivative (PPD) has been used for more than half a century as an antigen for the diagnosis of tuberculosis infection based on delayed type hypersensitivity. Although designated as “purified,” in reality, the composition of PPD is highly complex and remains ill-defined. In this report, high resolution mass spectrometry was applied to understand the complexity of its constituent components. A comparative proteomic analysis of various PPD preparations and their functional characterization is likely to help in short-listing the relevant antigens required to prepare a less complex and more potent reagent for diagnostic purposes.ResultsProteomic analysis of Connaught Tuberculin 68 (PPD-CT68), a tuberculin preparation generated from M. tuberculosis, was carried out in this study. PPD-CT68 is the protein component of a commercially available tuberculin preparation, Tubersol, which is used for tuberculin skin testing. Using a high resolution LTQ-Orbitrap Velos mass spectrometer, we identified 265 different proteins. The identified proteins were compared with those identified from PPD M. bovis, PPD M. avium and PPD-S2 from previous mass spectrometry-based studies. In all, 142 proteins were found to be shared between PPD-CT68 and PPD-S2 preparations. Out of the 354 proteins from M. tuberculosis–derived PPDs (i.e. proteins in either PPD-CT68 or PPD-S2), 37 proteins were found to be shared with M. avium PPD and 80 were shared with M. bovis PPD. Alignment of PPD-CT68 proteins with proteins encoded by 24 lung infecting bacteria revealed a number of similar proteins (206 bacterial proteins shared epitopes with 47 PPD-CT68 proteins), which could potentially be involved in causing cross-reactivity. The data have been deposited to the ProteomeXchange with identifier PXD000377.ConclusionsProteomic and bioinformatics analysis of different PPD preparations revealed commonly and differentially represented proteins. This information could help in delineating the relevant antigens represented in various PPDs, which could further lead to development of a lesser complex and better defined skin test antigen with a higher specificity and sensitivity.

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Section: Resultsmentioning

confidence: 99%

Proteomic analysis of purified protein derivative of Mycobacterium tuberculosis

et al. 2013

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“…We compared nine recombinant M. tuberculosis vaccine candidates (Table 1) [27], [28], [29], [30], [31], [32], [33], [34], [35], [36], [37], [38], [39] for their ability to be recognized by T cells at 3 and 30 weeks following i.n. vaccination with BCG.…”

Section: Resultsmentioning

confidence: 99%

Cellular Immune Responses to Nine Mycobacterium tuberculosis Vaccine Candidates following Intranasal Vaccination

et al. 2011

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BackgroundThe identification of Mycobacterium tuberculosis vaccines that elicit a protective immune response in the lungs is important for the development of an effective vaccine against tuberculosis.Methods and Principal FindingsIn this study, a comparison of intranasal (i.n.) and subcutaneous (s.c.) vaccination with the BCG vaccine demonstrated that a single moderate dose delivered intranasally induced a stronger and sustained M. tuberculosis-specific T-cell response in lung parenchyma and cervical lymph nodes of BALB/c mice than vaccine delivered subcutaneously. Both BCG and a multicomponent subunit vaccine composed of nine M. tuberculosis recombinant proteins induced strong antigen-specific T-cell responses in various local and peripheral immune compartments. Among the nine recombinant proteins evaluated, the alanine proline rich antigen (Apa, Rv1860) was highly antigenic following i.n. BCG and immunogenic after vaccination with a combination of the nine recombinant antigens. The Apa-induced responses included induction of both type 1 and type 2 cytokines in the lungs as evaluated by ELISPOT and a multiplexed microsphere-based cytokine immunoassay. Of importance, i.n. subunit vaccination with Apa imparted significant protection in the lungs and spleen of mice against M. tuberculosis challenge. Despite observed differences in the frequencies and location of specific cytokine secreting T cells both BCG vaccination routes afforded comparable levels of protection in our study.Conclusion and SignificanceOverall, our findings support consideration and further evaluation of an intranasally targeted Apa-based vaccine to prevent tuberculosis.

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“…Spot CS12-5.13 was identified as a 10-kDa chaperonin M. paratuberculosis protein (also called GroES or HSP10) (15,23), and spot CS29-5.51 was confirmed to be fibronectin-binding antigen 85 complex B (also called Ag85B) (18). Spot CS29-5.37 was identified as an M. avium alpha antigen protein.…”

Section: Resultsmentioning

confidence: 99%

Possible Association of GroES and Antigen 85 Proteins with Heat Resistance ofMycobacterium paratuberculosis

Sung¹,

Takayama

Collins³

2004

Appl Environ Microbiol

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Conflicting reports on the heat resistance of Mycobacterium paratuberculosis prompted an examination of the effect of culture medium on this property of the organism. M. paratuberculosis was cultured in three types of media (fatty acid-containing medium 7H9-OADC (oleic acid-albumin-dextrose-catalase supplement) and glycerol-containing media WR-GD and 7H9-GD [glycerol-dextrose supplement]) at pH 6.0. M. paratuberculosis grown under these three culture conditions was then tested for heat resistance in distilled water at 65°C. Soluble proteins and mycolic acids of M. paratuberculosis were evaluated by two-dimensional electrophoresis (2-DE) and thin-layer chromatography (TLC), respectively. The type of culture medium used significantly affected the heat resistance of M. paratuberculosis. The decimal reduction times at 65°C (D 65°C values; times required to reduce the concentration of bacteria by a factor of 10 at 65°C) for M. paratuberculosis strains grown in 7H9-OADC were significantly higher than those for the organisms grown in WR-GD medium (P < 0.01). When the glycerol-dextrose supplement of WR was substituted for the fatty acid supplement (OADC) in 7H9 medium (resulting in 7H9-GD), the D 65°C value was significantly lower than that for the organism grown in 7H9-OADC medium (P ‫؍‬ 0.022) but higher than that when it was cultured in WR-GD medium (P ‫؍‬ 0.005). Proteomic analysis by 2-DE of soluble proteins extracted from M. paratuberculosis grown without heat stress in the three media (7H9-OADC, 7H9-GD, and WR-GD) revealed that seven proteins were more highly expressed in 7H9-OADC medium than in the other two media. When the seven proteins were subjected to matrix-assisted laser desorption ionization-mass spectrometric analysis, four of the seven protein spots were unidentifiable. The other three proteins were identified as GroES heat shock protein, alpha antigen, and antigen 85 complex B (Ag85B; fibronectin-binding protein). These proteins may be associated with the heat resistance of M. paratuberculosis. Alpha antigen and Ag85B are both trehalose mycolyltransferases involved in mycobacterial cell wall assembly. TLC revealed that 7H9-OADC medium supported production of more trehalose dimycolates and cell wall-bound mycolic acids than did WR-GD medium. The present study shows that in vitro culture conditions significantly affect heat resistance, cell wall synthesis, and protein expression of M. paratuberculosis and emphasize the importance of culture conditions on in vitro and ex vivo studies to estimate heat resistance.Mycobacterium paratuberculosis (also known as Mycobacterium avium subsp. paratuberculosis) is the cause of Johne's disease in animals, in particular dairy cattle (14). Numerous studies report a significant association of this pathogen with the chronic inflammatory bowel disease of humans called Crohn's disease (10,17,21,45,50). Consequently, M. paratuberculosis has come under scrutiny as a potential food-borne pathogen of dairy products (16). For this reason, studies of M. paratuberculosis heat re...

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Recombinant GroES in combination with CpG oligodeoxynucleotides protects mice against Mycobacterium avium infection

Cited by 8 publications

References 33 publications

Proteomic analysis of purified protein derivative of Mycobacterium tuberculosis

Proteomic analysis of purified protein derivative of Mycobacterium tuberculosis

Cellular Immune Responses to Nine Mycobacterium tuberculosis Vaccine Candidates following Intranasal Vaccination

Possible Association of GroES and Antigen 85 Proteins with Heat Resistance ofMycobacterium paratuberculosis

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