Differential T-Cell Recognition of Native and Recombinant Mycobacterium tuberculosis GroES
Autor: | Peter Højrup, Paolo Mascagni, Ida Rosenkrands, Anthony R.M. Coates, Mahavir Singh, Peter Birk Rasmussen, Karin Weldingh, Lise Ostergaard Brandt, Peter Andersen, Pernille Ravn |
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Rok vydání: | 1999 |
Předmět: |
Recombinant Fusion Proteins
T-Lymphocytes Immunology medicine.disease_cause Microbiology law.invention Mice Maltose-binding protein Affinity chromatography law Chaperonin 10 Escherichia coli medicine Animals Humans Gel electrophoresis Mice Inbred BALB C biology Mycobacterium tuberculosis Bacterial Infections GroES Fusion protein Molecular biology Mice Inbred C57BL Infectious Diseases Biochemistry Chaperone (protein) biology.protein Recombinant DNA Parasitology |
Zdroj: | Infection and Immunity. 67:5552-5558 |
ISSN: | 1098-5522 0019-9567 |
Popis: | Mycobacterium tuberculosis GroES was purified from culture filtrate, and its identity was confirmed by immunoblot analysis and N-terminal sequencing. Comparing the immunological recognition of native and recombinant GroES, we found that whereas native GroES elicited a strong proliferative response and release of gamma interferon-γ by peripheral blood mononuclear cells from healthy tuberculin reactors, the recombinant protein failed to do so. The same difference in immunological recognition was observed in a mouse model of TB infection. Both the native and recombinant preparations were recognized by mice immunized with the recombinant protein. Biochemical characterization including sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two-dimensional electrophoresis, and mass spectrometry analysis of both proteins demonstrated no differences between the native and recombinant forms of GroES except for the eight additional N-terminal amino acids derived from the fusion partner in recombinant GroES. The recombinant fusion protein, still tagged with the maltose binding protein, was recognized by T cells isolated from TB-infected mice if mixed with culture filtrate before affinity purification on an amylose column. The maltose binding protein treated in the same manner as a control preparation was not recognized. Based on the data presented, we suggest that the association of biologically active molecules from culture filtrate with the chaperone GroES may be responsible for the observed T-cell recognition of the native preparation. |
Databáze: | OpenAIRE |
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