Identification of the new T-cell-stimulating antigens from<i>Mycobacterium tuberculosis</i>culture filtrate

Lim, Jae-Hyun; Kim, Hwa Jung; Lee, Kil-Soo; Jo, Eun-Kyeong; Song, Chang‐Hwa; Jung, Sungyup; Kim, Su‐Young; Lee, Ji-Sook; Paik, Tae-Hyun; Park, Jeong‐Kyu

doi:10.1016/s0378-1097(04)00018-7

Cited by 21 publications

(15 citation statements)

References 38 publications

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“…MTSP14 (Rv1827), a mycobacterial protein described as a T-cell antigen in this study and in a previous study by Lim et al (30), should be considered the same protein as CFP17 identified by Weldingh et al (55). Evaluation of purified polypeptides for recognition by PFMCs of tuberculosis pleuritis patients.…”

Section: Isolation Of Culture Filtrate Proteins Of M Tuberculosis H supporting

confidence: 72%

Peripheral Blood and Pleural Fluid Mononuclear Cell Responses to Low-Molecular-Mass Secretory Polypeptides ofMycobacterium tuberculosisin Human Models of Immunity to Tuberculosis

et al. 2005

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A total of 104 polypeptides were purified from the low-molecular-mass secretory proteome of Mycobacterium tuberculosis H 37 Rv using a combination of anion exchange column chromatography and high resolution preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroelution. The goal of this study was to identify polypeptides from a low-molecular-mass secretory proteome recognized by human subjects infected with M. tuberculosis and to ascertain the differences in specificity of antigen recognition by the peripheral blood mononuclear cells (PBMCs) and pleural fluid mononuclear cells (PFMCs) of these individuals. The study identified CFP-8 (Rv0496), CFP-11 (Rv2433c), CFP-14.5 (Rv2445c), and CFP-31 (Rv0831c) as novel T-cell antigens apart from previously characterized ESAT-6, TB10.4, CFP10, GroES, MTSP14, MTSP17, CFP21, MPT64, Ag85A, and Ag85B on the basis of recognition by PBMCs of tuberculosis contacts and treated tuberculosis patients. Further, polypeptides prominently recognized by PFMCs of tuberculous pleurisy patients were the same as those recognized by PBMCs of healthy contacts and treated tuberculosis patients. The results of our study indicate the homogeneity of antigenic target recognition by lymphocytes at the site of infection and at the periphery in the human subjects studied and the need to evaluate these antigenic targets as components of future antituberculous vaccines.

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Section: Isolation Of Culture Filtrate Proteins Of M Tuberculosis H supporting

confidence: 72%

Peripheral Blood and Pleural Fluid Mononuclear Cell Responses to Low-Molecular-Mass Secretory Polypeptides ofMycobacterium tuberculosisin Human Models of Immunity to Tuberculosis

et al. 2005

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“…Multiple studies provide evidence that antigens recognized by the protected group, but not active TB patients, can be considered for vaccine development strategies by using IFN-␥ response as a protective correlate (29 -31). Although donor to donor variation exists in antigen recognition and magnitude of response, this approach is considered a highly viable method to identify the protective antigens (28). In agreement with a previous study (32), healthy contacts in our study displayed a strong IFN-␥ response to the 2D LPE fractions as compared with that of TB patients.…”

Section: Discussionsupporting

confidence: 91%

“…Like many previous T cell antigen discovery efforts (15,16,28), we focused on the secreted proteins of M. tuberculosis. However, our current efforts differ from earlier studies, which used experimental animals (15), whereas our study aims at immunological responses to human TB.…”

Section: Discussionmentioning

confidence: 99%

“…It is generally believed that low molecular mass proteins of M. tuberculosis are predominantly recognized by human T cells (61,62), and most of the studies on human T cell antigens have focused on low molecular mass or selective pooling of fractions (16,28) and reported that antigens Ͻ15 kDa were highly immunogenic. In the present study, only 14.8% of the immunodominant T cell antigens were represented by antigens with a predicted mass of less than 15 kDa.…”

Section: Biological and Immunological Roles Of Proteins Identifiedmentioning

confidence: 99%

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Immunoproteomic Identification of Human T Cell Antigens of Mycobacterium tuberculosis That Differentiate Healthy Contacts from Tuberculosis Patients

Deenadayalan

Heaslip

Rajendiran

et al. 2010

Molecular & Cellular Proteomics

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Identification of Mycobacterium tuberculosis antigens inducing cellular immune responses is required to improve the diagnosis of and vaccine development against tuberculosis. To identify the antigens of M. tuberculosis that differentiated between tuberculosis (TB) patients and healthy contacts based on T cell reactivity, the culture filtrate of in vitro grown M. tuberculosis was fractionated by two-dimensional liquid phase electrophoresis and tested for the ability to stimulate T cells in a whole blood assay. This approach separated the culture filtrate into 350 fractions with sufficient protein quantity (at least 200 g of protein) for mass spectrometry and immunological analyses. High levels of interferon-␥ (IFN-␥) secretion were induced by 105 fractions in healthy contacts compared with TB patients (p < 0.05). Most interesting was the identification of 10 fractions that specifically induced strong IFN-␥ production in the healthy contact population but not in TB patients. Other immunological measurements showed 42 fractions that induced significant lymphocyte proliferative responses in the healthy contact group compared with the TB patients. The tumor necrosis factor-␣ response for most of the fractions did not significantly differ in the tested groups, and the interleukin-4 response was below the detectable range for all fractions and both study groups. Proteomic characterization of the 105 fractions that induced a significant IFN-␥ response in the healthy contacts compared with the TB patients led to the identification of 59 proteins of which 24 represented potentially novel T cell antigens. Likewise, the protein identification in the 10 healthy "contact-specific fractions" revealed 16 proteins that are key candidates as vaccine or diagnostic targets. Molecular & Cellular Proteomics 9:538 -549, 2010. Tuberculosis (TB)1 is a major health problem throughout the world. A recent World Health Organization report shows that TB has been increasing at a rate of 1% per year, and an estimated 9.2 million new cases arise each year (1). Although TB is preventable, there has been an increase in its incidence in recent years. Re-emergence of TB is mainly due to its association with human immunodeficiency virus infection (2) and also due to the occurrence of multidrugresistant strains of the causative agent, Mycobacterium tuberculosis (3).Vaccination of general population is cost effective and represents one of the best biological measures for disease control. The current vaccine against tuberculosis, Bacille Calmette-Gué rin (BCG), has been administered to more people than any other vaccine. The side effects of BCG are tolerable, and it prevents miliary and meningeal tuberculosis in young children. In striking contrast, it affords limited and highly variable protection (0 -80%) against pulmonary TB (4). Thus, BCG does not seem to be a satisfactory vaccine (5, 6) and necessitates exploration of newer strategies to improve BCG or to develop a more effective vaccine.One of the potential strategies for the development of an im...

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“…Novel vaccines are required for a variety of infectious diseases, including tuberculosis, for which no truly efficacious vaccine has yet been designed (16). A number of antigens have been considered for developing new tuberculosis vaccines (3,19,33). Early secreted antigenic target 6-kDa protein (ESAT-6) is found in Mycobacterium bovis and Mycobacterium tuberculosis but not in the vaccine strain Mycobacterium bovis BCG (12).…”

mentioning

confidence: 99%

Bacterial Polyester Inclusions Engineered To Display Vaccine Candidate Antigens for Use as a Novel Class of Safe and Efficient Vaccine Delivery Agents

Parlane

Wedlock

Buddle

et al. 2009

Appl Environ Microbiol

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Bioengineered bacterial polyester inclusions have the potential to be used as a vaccine delivery system. The biopolyester beads were engineered to display a fusion protein of the polyester synthase PhaC and the two key antigens involved in immune response to the infectious agent that causes tuberculosis, Mycobacterium tuberculosis, notably antigen 85A (Ag85A) and the 6-kDa early secreted antigenic target (ESAT-6) from Mycobacterium tuberculosis. Polyester beads displaying the respective fusion protein at a high density were successfully produced (henceforth called Ag85A-ESAT-6 beads) by recombinant Escherichia coli. The ability of the Ag85A-ESAT-6 beads to enhance mouse immunity to the displayed antigens was investigated. The beads were not toxic to the animals, as determined by weight gain and absence of lesions at the inoculation site in immunized animals. In vivo injection of the Ag85A-ESAT-6 beads in mice induced significant humoral and cell-mediated immune responses to both Ag85A and ESAT-6. Vaccination with Ag85A-ESAT-6 beads was efficient at stimulating immunity on their own, and this ability was enhanced by administration of the beads in an oil-in-water emulsion. In addition, vaccination with the Ag85A-ESAT-6 beads induced significantly stronger humoral and cell-mediated immune responses than vaccination with an equivalent dose of the fusion protein Ag85A-ESAT-6 alone. The immune response induced by the beads was of a mixed Th1/Th2 nature, as assessed from the induction of the cytokine gamma interferon (Th1 immune response) and increased levels of immunoglobulin G1 (Th2 immune response). Hence, engineered biopolyester beads displaying foreign antigens represent a new class of versatile, safe, and biocompatible vaccines.Bioengineered nano-/microstructures manufactured by microorganisms are becoming increasingly attractive because of their functional properties suitable for applications in various fields, particularly the medical sciences (9, 25, 29). Biopolyester beads comprising polyhydroxyalkanoate (PHA) are produced as intracellular inclusions by a wide range of bacteria and archaea when a carbon source is available in excess (30). PHA synthesis requires the key enzyme, polyester synthase, to catalyze the stereoselective polymerization of (R)-3-hydroxyacyl-coenzyme A to PHA. Self-assembly of polyester chains results in the formation of polymer granules with a hydrophobic core, and the PHA synthase protein remains covalently attached at the surface (28). These spherical granules range in size from 50 to 300 nm and accumulate in the intracellular space (34).Such biopolyester beads can be engineered to display the PHA synthase protein and its fusion partners on the surface at a high density (24). There have been recent examples where biopolyester beads were specifically engineered, produced in bacteria, and then harvested for their potential applications

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Identification of the new T-cell-stimulating antigens fromMycobacterium tuberculosisculture filtrate

Cited by 21 publications

References 38 publications

Peripheral Blood and Pleural Fluid Mononuclear Cell Responses to Low-Molecular-Mass Secretory Polypeptides ofMycobacterium tuberculosisin Human Models of Immunity to Tuberculosis

Peripheral Blood and Pleural Fluid Mononuclear Cell Responses to Low-Molecular-Mass Secretory Polypeptides ofMycobacterium tuberculosisin Human Models of Immunity to Tuberculosis

Immunoproteomic Identification of Human T Cell Antigens of Mycobacterium tuberculosis That Differentiate Healthy Contacts from Tuberculosis Patients

Bacterial Polyester Inclusions Engineered To Display Vaccine Candidate Antigens for Use as a Novel Class of Safe and Efficient Vaccine Delivery Agents

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