The Mycobacterium tuberculosis 38-kDa antigen: overproduction in Escherichia coli, purification and characterization

Singh, Mandeep; Andersen, Åse Bengård; Mccarthy, J. E. G.; Rohde, Manfred; Schütte, H. R.; Sanders, Ernst A.; Timmis, K. N.

doi:10.1016/0378-1119(92)90489-c

Cited by 67 publications

(41 citation statements)

References 20 publications

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“…This modified vector expresses the 38-kDa antigen on the cell surface, allowing detection and tagging of transduced cells by an anti-Ag38 monoclonal antibody (MAb), HBT12. 9 The supernatant of the producer cell lines was tested for viral titer and antigen expression by FACScan analysis with MAb HBT12 and used for infection of B16-B78 murine melanoma cells. 11 Transduced cells expressed the Mycobacterium tuberculosis Ag38 transcript and produced the bacterial protein as detected by RT-PCR ( Figure 2a) and Western blot (Figure 2b), respectively.…”

Section: Resultsmentioning

confidence: 99%

“…24,25 G418-resistant individual colonies of producer cells were screened for cell surface Ag38 expression by FACScan analysis using antiAg38 MAb HBT12. 9 Virus produced by the best packaging cell line, in terms of viral titer and antigen expression, were used to infect murine melanoma cells.…”

Section: Injected Antigenmentioning

confidence: 99%

“…8 If the expression of this gene leads to enhanced Th1 cell maturation, a T cell response might also be induced against the tumor cell antigens and eventually result in the selective destruction of tumor cells through a specific antitumor immune response. 2 In this study, we transduced the murine melanoma cell line B16-B78 with a Mycobacterium tuberculosis (strain H37Rv) gene encoding a 38-kDa protein (Ag38), 9 which is one of the most immunogenic of the Mycobacterium tuberculosis antigens. 10 Transduced cells were injected into mice and the immune response against parental B16-B78 melanoma cells was evaluated.…”

Section: Introductionmentioning

confidence: 99%

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Anti-tumor immunity induced by murine melanoma cells transduced with the Mycobacterium tuberculosis gene encoding the 38-kDa antigen

et al. 1998

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Section: Resultsmentioning

confidence: 99%

Section: Injected Antigenmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Anti-tumor immunity induced by murine melanoma cells transduced with the Mycobacterium tuberculosis gene encoding the 38-kDa antigen

et al. 1998

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“…Recombinant Mycobacterium tuberculosis HSP70 was prepared from the Escherichia coli pop strain (20). HSP70 was purified by ion-exchange chromatography using Q-Sepharose, followed by ATP affinity chromatography.…”

Section: Preparation Of Microbial Hsp70mentioning

confidence: 99%

Stimulation of Cell Surface CCR5 and CD40 Molecules by Their Ligands or by HSP70 Up-Regulates APOBEC3G Expression in CD4+ T Cells and Dendritic Cells

Pido-Lopez

Whittall

Wang

et al. 2007

The Journal of Immunology

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Apolipoprotein B mRNA-editing, enzyme-catalytic, polypeptide-like-3G (A3G) is an intracellular innate antiviral factor that deaminates retroviral cytidine to uridine. In an attempt to harness the anti-HIV effect of A3G, we searched for an agent that would up-regulate A3G and identify the receptors involved. Stimulation of cell surface CCR5 with CCL3 and CD40 with CD40L or both molecules with microbial 70-kDa heat shock protein (HSP)70 up-regulated A3G mRNA and protein expression in human CD4+ T cells and monocyte-derived dendritic cells (DC), demonstrated by real-time PCR and Western blots, respectively. The specificity of CCR5 and CD40 stimulation was established by inhibition with TAK 779 and mAb to CD40, as well as using human embryonic kidney 293 cells transfected with CCR5 and CD40, respectively. A dose-dependent increase of A3G in CCL3- or HSP70-stimulated CD4+ T cells was associated with inhibition in HIV-1 infectivity. To differentiate between the inhibitory effect of HSP70-induced CCR5 binding and that of A3G, GFP-labeled pseudovirions were used to infect human embryonic kidney 293 cells, which showed inhibition of pseudovirion uptake, consistent with A3G being responsible for the inhibitory effect. Ligation of cell surface CCR5 receptors by CCL3 or CD40 by CD40L activated the ERK1/2 and p38 MAPK signaling pathways that induced A3G mRNA expression and production of the A3G protein. These in vitro results were corroborated by in vivo studies in rhesus macaques in which A3G was significantly up-regulated following immunization with SIVgp120 and p27 linked to HSP70. This novel preventive approach may in addition to adaptive immunity use the intracellular innate antiviral effect of A3G.

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“…A soluble extract from M. tuberculosis H37Rv (MTSE) was prepared as described earlier [8]. The recombinant 38-kD protein [13] was obtained through the WHO protein bank (Braunschweig, Germany); concanavalin A (Con A) from Sigma (Poole, UK). Synthetic peptides were produced by standard automated solid-phase Clin Exp Immunol 1996; 106:312-316 synthesis for Fmoc amino acid pentafluorophenyl esters as described earlier [14].…”

Section: Bacteria and Antigensmentioning

confidence: 99%

Increase of tuberculous infection in the organs of B cell-deficient mice

Vordermeier

Venkataprasad

Harris

et al. 1996

Clinical and Experimental Immunology

142

106

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SUMMARYProtective immunity against infection with Mycobacterium tuberculosis is imparted by T cells rather than antibodies, but B cells can play a role as antigen-presenting cells and in granuloma formation. We re-evaluated the role of B cells in the course of tuberculous infection in ¹ -chain knock-out (Ig ÿ ) mice. Surprisingly, the organs of M. tuberculosis-infected Ig ÿ mice were found to have three-to eight-fold elevated counts of viable bacilli compared with normal littermates at 3-6 weeks post-infection. Splenic interferon-gamma responses to whole antigen were unimpaired, whilst proliferation to certain mycobacterial peptides was found to be diminished. However, bacille Calmette-Guérin (BCG) vaccination significantly reduced the infection in Ig ÿ mice. The mechanisms by which B cells can influence primary tuberculous infection need further study.

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The Mycobacterium tuberculosis 38-kDa antigen: overproduction in Escherichia coli, purification and characterization

Cited by 67 publications

References 20 publications

Anti-tumor immunity induced by murine melanoma cells transduced with the Mycobacterium tuberculosis gene encoding the 38-kDa antigen

Anti-tumor immunity induced by murine melanoma cells transduced with the Mycobacterium tuberculosis gene encoding the 38-kDa antigen

Stimulation of Cell Surface CCR5 and CD40 Molecules by Their Ligands or by HSP70 Up-Regulates APOBEC3G Expression in CD4+ T Cells and Dendritic Cells

Increase of tuberculous infection in the organs of B cell-deficient mice

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