Diagnosis of tuberculosis infection based on synthetic peptides from Mycobacterium tuberculosis antigen 85 complex

Kashyap, Rajpal S.; Shekhawat, Seema D.; Nayak, Amit R.; Purohit, Hemant J.; Taori, Girdhar M.; Daginawala, Hatim F.

doi:10.1016/j.clineuro.2012.07.031

Cited by 28 publications

(26 citation statements)

References 29 publications

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“…T-Cell epitopes for CD4 regions from reference Ag85B sequence were identified from available literature [ 8 , 9 ]. Peptides P1 (P 25 -P 40 ) (LPGLVG LAGGAATAGA) and P2 (P 91 -P 108 ) (NG WDINTPAFEW YYQSG), of Ag85B were designed by protocol described elsewhere [ 10 ].…”

Section: Methodsmentioning

confidence: 99%

Comparative evaluation of booster efficacies of BCG, Ag85B, and Ag85B peptides based vaccines to boost BCG induced immunity in BALB/c mice: a pilot study

Husain

Warke

Daginawala

et al. 2015

Clin Exp Vaccine Res

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PurposeIn the present study booster efficacies of Ag85 B, Bacillus Calmette-Guerin (BCG), and Ag85B peptides were evaluated using prime boost regimes in BALB/c mice.Materials and MethodsMice were primed with BCG vaccine and subsequently boosted with Ag85B, BCG and cocktail of Ag85B peptides.ResultsBased on analysis of immune response it was observed mice boosted with Ag85B peptides showed significant (p < 0.001) cytokines levels (interferon γ, interleukin 12) and BCG specific antibodies (anti-BCG and anti-purified protein derivative titre) compared to booster dose of BCG, Ag85B and BCG alone.ConclusionOur pilot results suggest that prime boost regimes with Ag85B peptides can boost waning BCG induced immunity and may improve immunogenicity of BCG vaccine. However, lot of work is further needed using experimental model of tuberculosis infection to justify the result.

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Section: Methodsmentioning

confidence: 99%

Comparative evaluation of booster efficacies of BCG, Ag85B, and Ag85B peptides based vaccines to boost BCG induced immunity in BALB/c mice: a pilot study

Husain

Warke

Daginawala

et al. 2015

Clin Exp Vaccine Res

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“…ELISA assay was performed with the method of [26] with minor modifications. Microplates 96-wells high-binding capacity flat-bottom (Greiner-bio-one GmbH, Frickenhausen Germany) were coated with different concentrations (400, 200, 100 ng/well) of Ag85B, TB10.4, His- full 2 in 50 mM bicarbonate buffer (pH 9.6) and incubated at room temperature overnight.…”

Section: Methodsmentioning

confidence: 99%

Optimizing Escherichia coli as a protein expression platform to produce Mycobacterium tuberculosis immunogenic proteins

et al. 2013

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BackgroundA number of valuable candidates as tuberculosis vaccine have been reported, some of which have already entered clinical trials. The new vaccines, especially subunit vaccines, need multiple administrations in order to maintain adequate life-long immune memory: this demands for high production levels and degree of purity.ResultsIn this study, TB10.4, Ag85B and a TB10.4-Ag85B chimeric protein (here-after referred as full) - immunodominant antigens of Mycobacterium tuberculosis - were expressed in Escherichia coli and purified to homogeneity. The rational design of expression constructs and optimization of fermentation and purification conditions allowed a marked increase in solubility and yield of the recombinant antigens. Indeed, scaling up of the process guaranteed mass production of all these three antigens (2.5-25 mg of pure protein/L cultivation broth). Quality of produced soluble proteins was evaluated both by mass spectrometry to assess the purity of final preparations, and by circular dichroism spectroscopy to ascertain the protein conformation. Immunological tests of the different protein products demonstrated that when TB10.4 was fused to Ag85B, the chimeric protein was more immunoreactive than either of the immunogenic protein alone.ConclusionsWe reached the goal of purifying large quantities of soluble antigens effective in generating immunological response against M. tuberculosis by a robust, controlled, scalable and economically feasible production process.

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“…Active and latent TB are distinct phases of the disease that can be characterized by their antigen expression, and these antigens are effective at inducing an immune response [27] . Our group and others have demonstrated that patients with active pulmonary TB and latently infected individuals respond differently to several Mtb antigens, such as antigen 85 complex proteins, MPT51 and HspX [28] – [31] . In our previous work, we developed a fusion protein (CMX) composed of the immunodominant antigens from Mtb: Ag85C, MPT51 and the entire HspX protein, [32] which are expressed in different stages of TB (active and latent phases of the disease).…”

Section: Introductionmentioning

confidence: 83%

A New Recombinant BCG Vaccine Induces Specific Th17 and Th1 Effector Cells with Higher Protective Efficacy against Tuberculosis

et al. 2014

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Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb) that is a major public health problem. The vaccine used for TB prevention is Mycobacterium bovis bacillus Calmette-Guérin (BCG), which provides variable efficacy in protecting against pulmonary TB among adults. Consequently, several groups have pursued the development of a new vaccine with a superior protective capacity to that of BCG. Here we constructed a new recombinant BCG (rBCG) vaccine expressing a fusion protein (CMX) composed of immune dominant epitopes from Ag85C, MPT51, and HspX and evaluated its immunogenicity and protection in a murine model of infection. The stability of the vaccine in vivo was maintained for up to 20 days post-vaccination. rBCG-CMX was efficiently phagocytized by peritoneal macrophages and induced nitric oxide (NO) production. Following mouse immunization, this vaccine induced a specific immune response in cells from lungs and spleen to the fusion protein and to each of the component recombinant proteins by themselves. Vaccinated mice presented higher amounts of Th1, Th17, and polyfunctional specific T cells. rBCG-CMX vaccination reduced the extension of lung lesions caused by challenge with Mtb as well as the lung bacterial load. In addition, when this vaccine was used in a prime-boost strategy together with rCMX, the lung bacterial load was lower than the result observed by BCG vaccination. This study describes the creation of a new promising vaccine for TB that we hope will be used in further studies to address its safety before proceeding to clinical trials.

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Diagnosis of tuberculosis infection based on synthetic peptides from Mycobacterium tuberculosis antigen 85 complex

Cited by 28 publications

References 29 publications

Comparative evaluation of booster efficacies of BCG, Ag85B, and Ag85B peptides based vaccines to boost BCG induced immunity in BALB/c mice: a pilot study

Comparative evaluation of booster efficacies of BCG, Ag85B, and Ag85B peptides based vaccines to boost BCG induced immunity in BALB/c mice: a pilot study

Optimizing Escherichia coli as a protein expression platform to produce Mycobacterium tuberculosis immunogenic proteins

A New Recombinant BCG Vaccine Induces Specific Th17 and Th1 Effector Cells with Higher Protective Efficacy against Tuberculosis

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