Use of<i>Mycobacterium tuberculosis</i>Complex-Specific Antigen Cocktails for a Skin Test Specific for Tuberculosis

Lyashchenko, Konstantin P.; Manca, Claudia; Colangeli, Roberto; Heijbel, Anna; Williams, A. M.; Gennaro, Maria Laura

doi:10.1128/iai.66.8.3606-3610.1998

Cited by 47 publications

(12 citation statements)

References 34 publications

(23 reference statements)

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“…There are some differences in the time of maximum responses between the animal model and humans. The maximum response in the guinea pig model was between 24 and 48 h after antigen injection, while in humans the optimal time is 48-72 h following the injection; this is consistent with other studies of DTH in humans and animals [12,18,19].…”

Section: Discussionsupporting

confidence: 88%

Recombinant early secreted antigen target 6 protein as a skin test antigen for the specific detection of Mycobacterium tuberculosis infection

Zhang

et al. 2008

Clinical and Experimental Immunology

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SummaryAlthough the delayed-type hypersensitivity skin test reaction to tuberculin purified protein derivative (PPD) is used worldwide for tuberculosis (TB) detection, it is incapable of distinguishing Mycobacterium tuberculosis (MTB) infection from bacille Calmette-Guérin (BCG) vaccination or infection with non-tuberculous Mycobacteria. As a result, there is an urgent need for a more specific diagnostic tool for TB. This study reports the skin reactions of guinea pigs and human volunteers to recombinant early secreted antigen target 6 (rESAT6), a secretory protein found only in MTB, M. bovis and few other mycobacterial species. These volunteers had varying histories of BCG vaccination and exposure to MTB, allowing us to determine the specificity of their response to TB exposure. Our results show that 1·0 mg of the purified MTB rESAT6 antigen elicited a positive skin response in both animals and humans exposed to MTB, as well as in animals exposed to M. bovis and M. marinum, all species of Mycobacteria that contain the gene for early secreted antigen target 6 (ESAT6). ESAT6 appears to be more specific to MTB infection than PPD, as demonstrated by the fact that we saw no skin responses in the BCGvaccinated volunteers, nor in the guinea pigs sensitized with BCG vaccine, or with Mycobacteria that do not contain the gene encoding ESAT6. We believe that this is the first report of the use of a rESAT6 protein in a skin test in human volunteers, and that these data support its use in the specific detection of MTB infection.

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Section: Discussionsupporting

confidence: 88%

Recombinant early secreted antigen target 6 protein as a skin test antigen for the specific detection of Mycobacterium tuberculosis infection

Zhang

et al. 2008

Clinical and Experimental Immunology

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“…We have also demonstrated that stronger responses can be induced with reagent pools than with their individual components. Recently, protein pools composed of mycobacterial antigens which provide improved specificity over tuberculin in a guinea pig model have been defined (23). A further advantage in using diagnostic antigen cocktails is that it reduces the possibility of selecting "escape mutants", i.e., strains of tubercle bacilli lacking the diagnostic antigen.…”

Section: Discussionmentioning

confidence: 99%

Development of Diagnostic Reagents To Differentiate betweenMycobacterium bovisBCG Vaccination andM. bovisInfection in Cattle

Vordermeier¹,

Cockle²,

Whelan³

et al. 1999

Clin Diagn Lab Immunol

135

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In Great Britain a recent independent scientific review for the government has concluded that the development of a cattle vaccine against Mycobacterium bovis holds the best long-term prospect for tuberculosis control in British herds. A sine qua non for vaccination is the development of a complementary diagnostic test to differentiate between vaccinated animals and those infected withM. bovis so that test-and-slaughter-based control strategies can continue alongside vaccination. In order to assess the feasibility of developing a differential diagnostic test for a live vaccine, we chose M. bovis BCG Pasteur as a model system. Recombinant forms of antigens which are expressed in M. bovis but not, or only at low levels, in BCG Pasteur (ESAT-6, MPB64, MPB70, and MPB83) were produced. These reagents were tested either alone or in combination by using peripheral blood mononuclear cells from M. bovis-infected, BCG-vaccinated, andMycobacterium avium-sensitized calves. All four antigens induced in vitro proliferation and gamma interferon responses only inM. bovis-infected animals. A cocktail composed of ESAT-6, MPB64, and MPB83 identified infected animals but not those vaccinated with BCG. In addition, promiscuous T-cell epitopes of ESAT-6, MPB64, and MPB83 were formulated into a peptide cocktail. In T-cell assays with this peptide cocktail, infected animals were identified with frequencies similar to those obtained in assays with the protein cocktail, while BCG-vaccinated or M. avium-sensitized animals did not respond. In summary, our results suggest that peptide and protein cocktails can be designed to discriminate between M. bovis infection and BCG vaccination.

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“…The protocol usually yields protein preparations that are . 99% pure from protein contaminants and that have a content of lipopolysaccharide lower than 10 pg/mg of protein [11], a level that is below the sensitivity of guinea pig skin tests [12,13].…”

Section: Methodsmentioning

confidence: 95%

Immunological Characterization of Antigens Encoded by the RD1 Region of the Mycobacterium tuberculosis Genome

Brusasca¹,

Colangeli²,

Lyashchenko³

et al. 2001

Scand J Immunol

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Development of immunoassays specific for the diagnosis of tuberculosis requires antigens unique to Mycobacterium tuberculosis. In a search for such antigens we tested six proteins encoded by RD1, a region present in M. tuberculosis and virulent M. bovis genomes but missing from the DNA of all substrains of M. bovis Bacillus Calmette-Guerin (BCG). The six proteins (Rv3871, Rv3872, Rv3873, MTSA-10, ESAT-6 and Rv3878) were purified to near-homogeneity from recombinant Escherichia coli. When tested for the ability to elicit antibody responses and delayed type hypersensitivity in tuberculous guinea pigs, only two of six antigens, ESAT-6 and MTSA-10, elicited strong skin reactions, while vigorous antibody responses were observed to all six proteins. When antibody responses to RD1 antigens were evaluated in sera from patients having pulmonary tuberculosis and from control subjects (patients having mycobacterioses other than tuberculosis, and healthy persons), a sizeable proportion (25%) of tuberculosis patients but none of the control subjects, had antibodies against MTSA-10 and/or ESAT-6. We conclude that MTSA-10 and ESAT-6 are promising candidates for immunodiagnostic assays specific for tuberculosis.

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Use ofMycobacterium tuberculosisComplex-Specific Antigen Cocktails for a Skin Test Specific for Tuberculosis

Cited by 47 publications

References 34 publications

Recombinant early secreted antigen target 6 protein as a skin test antigen for the specific detection of Mycobacterium tuberculosis infection

Recombinant early secreted antigen target 6 protein as a skin test antigen for the specific detection of Mycobacterium tuberculosis infection

Development of Diagnostic Reagents To Differentiate betweenMycobacterium bovisBCG Vaccination andM. bovisInfection in Cattle

Immunological Characterization of Antigens Encoded by the RD1 Region of the Mycobacterium tuberculosis Genome

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