Molecular analysis of genetic differences between Mycobacterium bovis BCG and virulent M. bovis

Mahairas, Gregory G.; Sabo, Peterj .; Hickey, Mark J.; Singh, D C; Stover, C. Kendall

doi:10.1128/jb.178.5.1274-1282.1996

Cited by 960 publications

(783 citation statements)

References 42 publications

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“…Importantly, the gene encoding ESAT-6 is lacking in all strains of BCG tested whereas it is found in virulent M. bovis [176]. In agreement with this finding ESAT-6 was recently identified in the RD1 9.5 kb segment deleted in all BCG substrains believed to be the original mutation leading to attenuation of M. bovis [170]. No signal sequence was identified [145].…”

Section: Mpt51 (27 Kda) and Mpt64 (26 Kda)mentioning

confidence: 69%

Host Responses and Antigens Involved in Protective Immunity to Mycobacterium tuberculosis

1997

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Andersen P. Host Responses and Antigens Involved in Protective Immunity to Mycobacterium tuberculosis. Scand J Immunol 1997;45:115-131 Tuberculosis (TB) is the largest single infectious cause of human mortality. The incidence of TB has remained high in most of the developing world and the disease has recently re-emerged as a public health problem in industrialized countries. The development of a new improved TB vaccine is a highly prioritized international research area, which has been further stimulated by the appearance of multi-drug resistant strains of Mycobacterium tuberculosis. The present status of the attempts to characterize the protective immune response to TB will be reviewed with special emphasis on recent progress in the identification and characterization of target molecules recognized by protective cells. This paper will focus on proteins released from live bacteria and discuss their role in the host-pathogen interaction and the ongoing attempts to use these molecules in TB subunit vaccines.

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Section: Mpt51 (27 Kda) and Mpt64 (26 Kda)mentioning

confidence: 69%

Host Responses and Antigens Involved in Protective Immunity to Mycobacterium tuberculosis

1997

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“…Pure ORF-14 protein was then obtained by cleavage of the purified GST-ORF-14 fusion protein with thrombin protease. nucleotides 11731-12018 within the RD1 region [16,17]. The DNA sequences of forward (F) and reverse (R) primers for the amplification of ORF-10, ORF-14 and ORF-15 genes, based on the N-and C-terminal sequences of these ORF genes from M. bovis [16], are shown below.…”

Section: Methodsmentioning

confidence: 99%

“…Recent studies by Mahairas et al [16] have highlighted the genomic differences between avirulent BCG strains and virulent or clinical M. tuberculosis and M. bovis strains. One of these genomic segments of difference, designated RD1, that was present in all virulent and clinical strains of M. tuberculosis and M. bovis, was found to be deleted from the genomes of all BCG strains [16].…”

Section: Introductionmentioning

confidence: 99%

“…The RD1 region of M. bovis was shown potentially to encode eight proteins that are not found in BCG strains [16]. More rigorous analyses with Genetics Computer Group (GCG) software have indicated the presence of up to 20 open reading frames (ORFs), some of which were neither recognized in the original study [16] nor shown as encoding proteins by Cole et al [17]. A total of 14 ORFs (ORF-2 to ORF-15) are virulent M. tuberculosis-and M. bovisspecific and are deleted in BCG strains.…”

Section: Introductionmentioning

confidence: 99%

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Identification of a Novel Protein Antigen Encoded by a Mycobacterium tuberculosis‐Specific RD1 Region Gene

et al. 1999

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A genomic DNA region, designated RD1, that is present in virulent and clinical strains of Mycobacterium tuberculosis and M. bovis, has been shown to be deleted in bacillus Calmette Guérin (BCG). The DNA segments corresponding to three open reading frames (ORFs: ORF‐10, ORF‐14 and ORF‐15) of the RD1 region, that are deleted in BCG strains, were amplified from M. tuberculosis genomic DNA by polymerase chain reaction (PCR), subcloned into pGEX‐4T vector system and expressed in Escherichia coli as fusion proteins with glutathione‐S‐transferase (GST). The recombinant proteins appeared as major cellular proteins in SDS‐PAGE gels at the expected molecular mass. The identity of each fusion protein was confirmed by reactivity with anti‐GST antibodies in Western immunoblots. When pooled human sera from 11 tuberculosis (TB) patients were used as the source of antibodies, only GST‐ORF‐14 fusion protein reacted in Western immunoblots. The protein corresponding to ORF‐14 was then purified to near homogeneity and isolated free of its fusion partner (GST) by treating the purified GST‐ORF‐14 fusion protein with thrombin protease. In Western immunoblots, the purified ORF‐14 protein reacted with antibodies in 26 of 57 human sera (46%) from TB patients while no reactivity was seen with 11 sera from M. bovis BCG‐vaccinated healthy subjects. Interestingly, sera from nine of 15 (60%) long‐term contacts of TB patients also had antibodies reactive to the ORF‐14 protein. These results suggest that the ORF‐14 protein in combination with other immunodominant proteins could be useful in the serodiagnosis of individuals infected with M. tuberculosis.

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“…The recent completion of the genome sequences of both Mycobacterium tuberculosis (8) and Mycobacterium leprae (9) provides an opportunity to identify leprosy-specific antigens that might serve this purpose. A similar comparative approach has allowed identification of genes in the RD1 region that are deleted from Mycobacterium bovis BCG but present in M. tuberculosis and can therefore distinguish between infection with M. tuberculosis and vaccination with BCG (22). Among the deleted antigens are two low-molecular-weight proteins, culture filtrate protein 10 (CFP-10) (Rv3874) and ESAT-6 (Rv3875) (3,14).…”

mentioning

confidence: 99%

Comparative Analysis of B- and T-Cell Epitopes ofMycobacterium lepraeandMycobacterium tuberculosisCulture Filtrate Protein 10

et al. 2004

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Culture filtrate protein 10 (CFP-10) from Mycobacterium tuberculosis is a well-characterized immunodominant 10-kDa protein antigen known to elicit a very potent early gamma interferon response in T cells from M. tuberculosis-infected mice and humans. The sequence of the Mycobacterium leprae homologue of CFP-10 shows only 40% identity (60% homology) at the protein level with M. tuberculosis CFP-10 and thus has the potential for development as a T-or B-cell reactive antigen for specific diagnosis of leprosy. Antisera raised in mice or rabbits against recombinant M. leprae and M. tuberculosis CFP-10 proteins reacted only with homologous peptides from arrays of overlapping synthetic peptides, indicating that there was no detectable cross-reactivity at the antibody level. Sera from leprosy and tuberculosis patients were also specific for the homologous protein or peptides and showed distinct patterns of recognition for either M. leprae or M. tuberculosis CFP-10 peptides. At the cellular level, only 2 of 45 mouse T-cell hybridomas raised against either M. leprae or M. tuberculosis CFP-10 displayed a cross-reactive response against the N-terminal heterologous CFP-10 peptide, the region that exhibits the highest level of identity in the two proteins; however, the majority of peptide epitopes recognized by mouse T-cell hybridomas specific for each protein did not cross-react with heterologous peptides. Coupled with the human serology data, these results raise the possibility that peptides that could be used to differentiate infections caused by these two related microorganisms could be developed. Immunohistochemical staining of sections of M. leprae-infected nude mouse footpads resulted in strongly positive staining in macrophages and dendritic cells, as well as weaker staining in extracellular areas, suggesting that M. leprae CFP-10, like its homologue in M. tuberculosis, is a secreted protein.The number of cases of leprosy worldwide has been reduced dramatically through aggressive World Health Organizationsponsored multidrug therapy, from approximately 10 million cases in 1985 to a little fewer than 700,000 cases today (46-48). However, with the unabated emergence of more than 600,000 new cases per year, there is the possibility that the source of infection is not being addressed (16). The single greatest need in leprosy research is the development of definitive diagnostic tools that identify sources of leprosy and routes of transmission. The recent completion of the genome sequences of both Mycobacterium tuberculosis (8) and Mycobacterium leprae (9) provides an opportunity to identify leprosy-specific antigens that might serve this purpose. A similar comparative approach has allowed identification of genes in the RD1 region that are deleted from Mycobacterium bovis BCG but present in M. tuberculosis and can therefore distinguish between infection with M. tuberculosis and vaccination with BCG (22). Among the deleted antigens are two low-molecular-weight proteins, culture filtrate protein 10 (CFP-10) (Rv3874) and ESAT-6...

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Molecular analysis of genetic differences between Mycobacterium bovis BCG and virulent M. bovis

Cited by 960 publications

References 42 publications

Host Responses and Antigens Involved in Protective Immunity to Mycobacterium tuberculosis

Host Responses and Antigens Involved in Protective Immunity to Mycobacterium tuberculosis

Identification of a Novel Protein Antigen Encoded by a Mycobacterium tuberculosis‐Specific RD1 Region Gene

Comparative Analysis of B- and T-Cell Epitopes ofMycobacterium lepraeandMycobacterium tuberculosisCulture Filtrate Protein 10

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