Oral Immunogenicity of Plant-Made<i>Mycobacterium tuberculosis</i>ESAT6 and CFP10

Уварова, Е. А.; Belavin, P. A.; Permyakova, N. V.; Zagorskaya, A. A.; Nosareva, Olesya V.; Kakimzhanova, Almagul; Дейнеко, Е. В.

doi:10.1155/2013/316304

Cited by 40 publications

(38 citation statements)

References 25 publications

(41 reference statements)

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“…Previously, number of studies have reported ESAT‐6 expression in plants using nuclear and chloroplast transformation . In the current study, we expressed ESAT‐6 antigen in broccoli via Agrobacterium ‐mediated transformation.…”

Section: Discussionmentioning

confidence: 98%

“…It has been proven that stimulation of cell‐mediated immune mechanisms, including γδ T cells, CD4+ T cells, CD8+ T cells, and CD1‐restricted and HLA‐E‐restricted T cells, have the potential to kill the mycobacterium . Some of the most promising antigens of MTB that could be used for subunit vaccine development are: ESAT‐6, MTB72F, Ag85B, and LiPY . 6 kDa early secretory antigenic target (ESAT‐6) is one of the most important subunit vaccine antigen of MTB, which is encoded in the region of deletion (RD‐1) in all strains of MTB except in the BCG vaccine strains .…”

Section: Introductionmentioning

confidence: 99%

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Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Saba

Sameeullah

Asghar

et al. 2020

Biotech and App Biochem

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Tuberculosis (TB) is one of the major infectious diseases caused by Mycobacterium tuberculosis. The development of an effective and economical vaccine for controlling TB is essential especially for developing countries. Edible plants can serve as biofactories to produce vaccine antigens. In this study, 6 kDa early secretory antigenic target (ESAT‐6) of M. tuberculosis was expressed in Brassica oleracea var. italica via Agrobacterium‐mediated transformation to facilitate oral delivery of antigen. ESAT‐6 gene was cloned using Gateway® cloning strategy. Transformation and presence of transgene was confirmed through PCR. Expression level of transgene was calculated via quantitative real‐time PCR (qRT‐PCR) and the maximum integrated transgene number was two. Maximum amount of total soluble fraction of ESAT‐6 was evaluated by immunoblotting, estimated to accumulate up to 0.5% of total soluble protein. The recombinant ESAT‐6 protein was further purified and detected using silver staining and Western blotting. ESAT‐6 protein induced humoral immune response in mice immunized orally and subcutaneously. The expression of M. tuberculosis antigen in edible plants could aid in the development of cost‐effective and oral delivery of an antigen‐based subunit vaccine against TB. To the best our knowledge, it is the first report of expression of a vaccine antigen in broccoli.

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Saba

Sameeullah

Asghar

et al. 2020

Biotech and App Biochem

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“…Several in vitro assay systems based on the detection of immune reactivity against an M. tuberculosis-specific antigen have been established and widely accepted in recent years. Early secreted antigenic target 6-kDa protein (ESAT6) and culture filtrate protein of 10 kDa (CFP10), two major M. tuberculosis-specific antigens, have been reported to play key roles in the virulence of M. tuberculosis and elicit strong T-cell responses (3,4). Interferon gamma release assays (IGRAs) based on immune recognition against ESAT6 and CFP10 have been used to help diagnose activated and latent M. tuberculosis infection (5).…”

mentioning

confidence: 99%

Safety of Recombinant Fusion Protein ESAT6-CFP10 as a Skin Test Reagent for Tuberculosis Diagnosis: an Open-Label, Randomized, Single-Center Phase I Clinical Trial

Zhou

et al. 2016

Clin Vaccine Immunol

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cThis trial was conducted to explore the safety of recombinant fusion protein ESAT6-CFP10 as a skin test reagent for the diagnosis of Mycobacterium tuberculosis infection. Twenty-four healthy adult volunteers were recruited and randomized into four groups (groups A to D) to study four increasing doses of ESAT6-CFP10. All subjects in each dose group received an intradermal injection of reagent (0.1 ml) via the Mantoux technique. Then, the vital signs of all subjects were monitored, and skin reactions around injection sites and adverse events were recorded at different detection time points after the skin test. No serious adverse events were observed in this study. A total of 3 subjects had unexpected events. One subject in group A developed subcutaneous hemorrhage 24 h after the skin test, one subject in group B was found with red spots 15 min after the skin test, and another subject in group A showed abnormity during a chest X-ray after the skin test without affecting her health. One of three adverse events (red spots) was probably related to the recombinant ESAT6-CFP10 reagent. A single dose of 1, 5, 10, or 20 g/ml of recombinant ESAT6-CFP10 as a skin test reagent for M. tuberculosis infection diagnosis is well tolerated and safe in China. (This study has been registered at ClinicalTrials.gov under registration no. NCT01999231.)A lthough tuberculosis (TB) has been a curable disease, it still remains a major global problem that seriously threatens human health. According to the WHO Global TB report 2015, there were 9.6 million new cases and nearly 1.5 million TB-related deaths in 2014 (1). Thus, exploring diagnostic techniques with high sensitivity and specificity is crucial for controlling TB development and its drug resistance.TB is caused by infection with Mycobacterium tuberculosis, an intracellular pathogen transmitted by air (2). Several in vitro assay systems based on the detection of immune reactivity against an M. tuberculosis-specific antigen have been established and widely accepted in recent years. Early secreted antigenic target 6-kDa protein (ESAT6) and culture filtrate protein of 10 kDa (CFP10), two major M. tuberculosis-specific antigens, have been reported to play key roles in the virulence of M. tuberculosis and elicit strong T-cell responses (3, 4). Interferon gamma release assays (IGRAs) based on immune recognition against ESAT6 and CFP10 have been used to help diagnose activated and latent M. tuberculosis infection (5). IGRAs have an increased specificity compared to the conventional tuberculin skin test (TST) because the antigens included in IGRAs, namely, ESAT6 and CFP10, have been selected based on their absence in many environmental nontuberculous mycobacteria and the vaccine strain Mycobacterium bovis Bacillus Calmette-Guérin (BCG), which can only be detected in a number of pathogenic mycobacteria species, including M. tuberculosis, and a minority of nontuberculous mycobacteria (Mycobacteria kansasii, Mycobacteria szulgai, Mycobacteria marinum, and Mycobacteria riyadhense), altogether l...

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“…Uvarova et al [62] have reported in 2013 the individual expression of the M. tuberculosis ESAT-6 and CFP10 antigens in carrot roots, at levels above 0.056% of the total soluble protein (TSP) for ESAT-6 and 0.002% of TSP for CFP10. These levels of expression are in the range of expression obtained through nuclear transformation but still very low when compared with transplastomic or viral strategies.…”

Section: Carrot-derived Esat-6 and Cfp10 Antigensmentioning

confidence: 99%

“…In the case of the plant-based vaccines, the three candidates evaluated in oral immunization schemes obtaining promising findings in terms of immunogenicity [59,60,62] reflect the potential of inducing robust immunoprotective responses by this route. However, further research efforts are required to define this potential fully characterizing the involved mechanisms.…”

Section: Expert Commentarymentioning

confidence: 99%

An overview of tuberculosis plant-derived vaccines

Rosales‐Mendoza¹,

Ríos-Huerta²,

Angulo³

2015

Expert Review of Vaccines

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Tuberculosis (TB) is a leading fatal infectious disease to which the current BCG vaccine has a questionable efficacy in adults. Thus, the development of improved vaccines against TB is needed. In addition, decreasing the cost of vaccine formulations is required for broader vaccination coverage through global vaccination programs. In this regard, the use of plants as biofactories and delivery vehicles of TB vaccines has been researched over the last decade. These studies are systematically analyzed in the present review and placed in perspective. It is considered that substantial preclinical trials are still required to address improvements in expression levels as well as immunological data. Approaches for testing additional antigenic configurations with higher yields and improved immunogenic properties are also discussed.

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Oral Immunogenicity of Plant-MadeMycobacterium tuberculosisESAT6 and CFP10

Cited by 40 publications

References 25 publications

Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Expression of ESAT‐6 antigen from Mycobacterium tuberculosis in broccoli: An edible plant

Safety of Recombinant Fusion Protein ESAT6-CFP10 as a Skin Test Reagent for Tuberculosis Diagnosis: an Open-Label, Randomized, Single-Center Phase I Clinical Trial

An overview of tuberculosis plant-derived vaccines

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