A comparison of the rate of oxygen uptake and the adenosine triphosphate content of routine and experimental BCG preparations

Borremans, Martine; Weckx, Maurice; Verhofstadt, Roger

doi:10.1016/s0092-1157(83)80007-9

Cited by 6 publications

(2 citation statements)

References 7 publications

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“…Gels were run overnight (see Fig. 2) or for 4 hours (see all other figures). After completion of SDS-PAGE, the proteins were electrophoretically transferred to nitrocellulose sheets by the method of Towbin et al (31).…”

Section: Methodsmentioning

confidence: 99%

Repertoires of antibodies to culture filtrate antigens in different mouse strains infected with Mycobacterium bovis BCG

Huygen¹,

Ljungqvist²,

Berg³

et al. 1990

Infect Immun

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Two susceptible (Bcg') mouse strains, BALB/c and C57BL/6, were compared by Western blot (immunoblot) analysis for their immunoglobulin G response to 14-day-old BCG culture filtrate (CF) following intravenous infection with live Mycobacterium bovis BCG. The two strains demonstrated a completely different antibody repertoire. BALB/c antibodies were directed against a wide range of CF antigens between 20 and about 100 kilodaltons (kDa), with a preferential recognition of the 65-kDa heat shock protein and the 32-kDa fibronectin-binding protein. C57BL/6 sera, on the other hand, showed a much more restricted antibody pattern, almost exclusively directed against three antigens with estimated molecular sizes of 37, 38, and 40 kDa. Whereas the 37-and 38-kDa antigens were also recognized by BALB/c mice, the 40-kDa antigen was very intensely stained by C57BL/6 sera only. F1 mice had the restricted antibody pattern of C57BL/6 after one injection of BCG and had a hybrid BALB/c-C57BL/6 phenotype following a boost injection of BCG 2 months after the initial infection. Analysis of seven recombinant inbred strains derived from the BALB/c x C57BL/6 cross and of congenic mice differing in major histocompatibility complex-coding chromosome 17 fragments suggests that a gene in the K-IA region of the H-2 locus is associated with the preferential recognition of certain CF antigens. Inoculation with the same dose of killed BCG failed to elicit an antibody response to these filtrate antigens.

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Section: Methodsmentioning

confidence: 99%

Repertoires of antibodies to culture filtrate antigens in different mouse strains infected with Mycobacterium bovis BCG

Huygen¹,

Ljungqvist²,

Berg³

et al. 1990

Infect Immun

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“…C. glutamicum was transformed by electroporation as described previously (7). The Mycobacterium strain used was M. bovis BCG GL2 (Pasteur Institute of Brussels), which was grown for 2 weeks as a surface pellicle on synthetic Sauton culture medium at 37.5°C (10).…”

Section: Methodsmentioning

confidence: 99%

Heterologous expression of the Mycobacterium tuberculosis gene encoding antigen 85A in Corynebacterium glutamicum

Salim¹,

Haedens²,

Content³

et al. 1997

Appl Environ Microbiol

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By using appropriate Corynebacterium glutamicum-Escherichia coli shuttle plasmids, the gene encoding the fibronectin-binding protein 85A (85A) from Mycobacterium tuberculosis was expressed in C. glutamicum, also an actinomycete and nonsporulating gram-positive rod bacterium, which is widely used in industrial amino acid production. The 85A gene was weakly expressed in C. glutamicum under the control of the ptac promoter from E. coli, but it was produced efficiently under the control of the promoter of the cspB gene encoding PS2, one of the two major secreted proteins from C. glutamicum. The 85A protein was produced in various forms, with or without its own signal sequence and with or without the signal sequence and the NH 2-terminal (18-amino-acid) mature sequence of PS2. Western blot analysis with monoclonal antibodies raised against the M. tuberculosis antigen 85 complex showed that recombinant 85A protein was present in the corynebacterial cell wall extract and also released in extracellular culture medium. NH 2-terminal microsequencing of recombinant 85A secreted by C. glutamicum showed that signal peptide was effectively cleaved off at the predicted site. The recombinant 85A protein was biologically active in vitro, inducing significant secretion of Th1 T-cell cytokines, particularly interleukin-2 and gamma interferon, in spleen cell cultures from mice vaccinated with live Mycobacterium bovis BCG. Heterologous expression of mycobacterial antigens in C. glutamicum now offers a potent tool for further immunological characterization and large scale preparation of these recombinant proteins.

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The production of mycobacterial antigens by homogeneous culture in a fermentor

Nyabenda¹,

Bautens²,

Borremans³

et al. 1988

Journal of Biological Standardization

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A comparison of the rate of oxygen uptake and the adenosine triphosphate content of routine and experimental BCG preparations

Cited by 6 publications

References 7 publications

Repertoires of antibodies to culture filtrate antigens in different mouse strains infected with Mycobacterium bovis BCG

Repertoires of antibodies to culture filtrate antigens in different mouse strains infected with Mycobacterium bovis BCG

Heterologous expression of the Mycobacterium tuberculosis gene encoding antigen 85A in Corynebacterium glutamicum

The production of mycobacterial antigens by homogeneous culture in a fermentor

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