Heterogenous Expression of the Related MBP70 and MPB83 Proteins Distinguish Various Substrains of <i>Mycobacterium bovis</i> BCG and <i>Mycobacterium tuberculosis</i> H37Rv

Wiker, Harald G.; Nagai, Shoichiro; Hewinson, R. Glyn; Russell, Wayne; Harboe, Morten

doi:10.1046/j.1365-3083.1996.d01-61.x

Cited by 69 publications

(68 citation statements)

References 25 publications

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“…Other mycobacterial proteins show a similar irregular migration (Prestidge et a[., 1995). (Harboe & Wiker, 1992) and MPB83 (Hewinson et al, 1996;Wiker et al, 1996;Matsuo et al, 1996) are examples of well-characterized mycobacterial lipoproteins. These differ from P27 in that they are also found in the culture supernatant (Fifis et al, 1990;Harboe & Wiker, 1992 strains gave rise to a faint amplification band, suggesting that a homologous gene exists in these species.…”

Section: Discussionmentioning

confidence: 92%

A novel 27 kDa lipoprotein antigen from Mycobacterium bovis

Bigi¹,

Espitia²,

Alito

et al. 1997

Microbiology

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A novel Mycobacterium bovis antigen was identified from an expression library using sera from naturally infected cattle. The Escherichia coli recombinant clone expressed a 27 kDa protein, named P27. A rabbit serum against the recombinant antigen recognized a protein of 27 kDa in cellular extracts from M. bovis and M. tuberculosis. No protein was recognized in the culture supernatant. Sequence analysis indicated that P27 has a molecular mass of 24 kDa, showing a characteristic signal sequence for lipoprotein modification (a signal peptidase type II site). The gene is identical t o a gene identified in the M. tuberculosis genome sequencing project. Cellular fractionation experiments suggested that P27 is an integral membrane protein. The antigen was recognized by individual sera and peripheral blood mononuclear cells (PBMC) from diseased cattle. PCR experiments with specific primers directed t o the P27 structural gene indicated that it is only present in the M. tuberculosis species complex. In conclusion, a novel immunogenic lipoprotein in M. bovis/M. tuberculosis has been identified. The results presented here and elsewhere suggest that mycobacterial lipoproteins should be considered in the design of new recombinant vaccines and diagnostic methods.

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Section: Discussionmentioning

confidence: 92%

A novel 27 kDa lipoprotein antigen from Mycobacterium bovis

Bigi¹,

Espitia²,

Alito

et al. 1997

Microbiology

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“…The gene for MPB70 is present in M. bovis BCG but expressed to greater or lesser extents in different strains (i.e., lower expression in Glaxo and Pasteur strains than in Tokyo or virulent M. bovis) (64,67). Evidence for a smaller gene of 4.6 kb in M. kansasii that hybridizes to a MPB83 gene probe has also been obtained using Southern blotting (65).…”

Section: Discussionmentioning

confidence: 94%

Gamma Interferon Responses Induced by a Panel of Recombinant and Purified Mycobacterial Antigens in Healthy, Non-Mycobacterium bovisBCG-Vaccinated Malawian Young Adults

Black

Weir

Chaguluka

et al. 2003

Clin Vaccine Immunol

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We have previously shown that young adults living in a rural area of northern Malawi showed greater gamma interferon (IFN-␥) responses to purified protein derivatives (PPD) prepared from environmental mycobacteria than to PPD from Mycobacterium tuberculosis. In order to define the mycobacterial species to which individuals living in a rural African population have been exposed and sensitized, we tested T-cell recognition of recombinant and purified antigens from M. tuberculosis (38 kDa, MPT64, and ESAT-6), M. bovis There is great variation in the protection that Mycobacterium bovis BCG vaccines can provide against tuberculosis in different locations (17). In two large randomized controlled studies of BCG vaccination in Malawi and the United Kingdom, we have shown that prior to the vaccination of previously unvaccinated young adults in Malawi, over 60% of individuals show gamma interferon (IFN-␥) responses to purified protein derivative (PPD) from M. tuberculosis and that BCG vaccination provides only a moderate increase in this T-cell response (8). When PPDs from a number of nontuberculous environmental (atypical) mycobacteria were used to stimulate whole-blood cultures from the same subjects, PPDs from members of the M. avium-M. intracellulare-M. scrofulaceum complex induced stronger IFN-␥ responses than PPD from M. tuberculosis (6). To further define the mycobacterial species to which these subjects had been exposed, we have now used a panel of recombinant and purified antigens, with known species distributions, to test IFN-␥ responses, as a measure of the type 1 T-cell response induced by mycobacterial antigens, in day 6 supernatants from diluted whole-blood cultures. We have also assessed the potential of these recombinant antigens for identifying infection with M. tuberculosis and M. leprae, by studying the association between the IFN-␥ response to these antigens and the skin test response to M. tuberculosis PPD.Most studies of T-cell recognition of individual mycobacterial antigens have used patients infected with pathogenic mycobacteria or the contacts of these patients. Our study is unusual in that non-BCG-vaccinated, human immunodeficiency virus-negative, healthy individuals living in a rural area of northern Malawi have been tested for their ability to make a type 1 cytokine response to a panel of seven recombinant mycobacterial antigens and one purified native mycobacterial antigen. These subjects were clinically well at the time of testing and were recruited in their villages and homes.

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“…Our results suggest that vaccine efficacy could now be monitored in the field by assessing this serological parameter, particularly in developing countries, where in vitro cytokine assays may be difficult to perform. MPB70 and MPB83 are both expressed constitutively in M. bovis yet are expressed only at very low levels in BCG strains that have lost the RD2 regions, such as BCG Pasteur, which was used in the present study (20,33). MPB70 is a secreted protein (21), whereas MPB83 is a cell wall-associated glycolipoprotein that is also found in culture filtrates (11).…”

Section: Fig 1 Boost Of Mpb70 and Mpb83mentioning

confidence: 99%