RRC ID 42077
Author Higuchi K, Sekiya Y, Harada N.
Title Characterization of M. Tuberculosis-derived IL-12-inducing material by alveolar macrophages.
Journal Vaccine
Abstract We have investigated the substance derived from Mycobacterium tuberculosis (Mtb) that induces interleukin (IL)-12 production by alveolar macrophages (AMs) in vitro. The cytosol fraction of live Mtb H37Rv induced IL-12 production by AMs in a dose-dependent manner. The addition of interferon-gamma (IFN-gamma) augmented IL-12 production. IL-12-inducing activity by AMs (termed as surely active keeping rescue antigen, SAKRA) was purified by gel filtration and ion exchange column chromatography, and the molecular weight of SAKRA was estimated by gel filtration to be more than 700 kDa. SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotting of SAKRA using rabbit anti-SAKRA antibody suggested that SAKRA is composed with several low molecular weight proteins. Amino acids sequence analysis of several bands after SDS-PAGE suggested that SAKRA is a part of ribosomes. RT-PCR showed that SAKRA induced not only expression of IL-12 p40 mRNA, but expression of tumor necrosis factor (TNF)-alpha and inducible nitric oxide synthase (iNOS) mRNA at least 6 h after stimulation, suggesting that SAKRA activates the bactericidal activity of macrophages. To investigate the potential use of SAKRA as a vaccine against tuberculosis, SAKRA was administered to BALB/c mouse that had been immunized with BCG for 18 months, and mouse were infected with Mtb H37Rv via a respiratory route. Replication of Mtb in lungs and spleens was examined 6 weeks after infection. Administration of SAKRA to BCG-vaccinated mice significantly reduced the numbers of Mtb in lungs and spleens as compared with BCG-vaccinated control mice. Taken together, these results suggest that SAKRA is one of the Mtb-derived immunomodulatory substances which induce IL-12 production during infection and also increases mycobactericidal activities of macrophages, and that SAKRA may be a promising new vaccine candidate against tuberculosis.
Volume 22(5-6)
Pages 724-34
Published 2004-1-26
DOI 10.1016/j.vaccine.2003.08.018
PII S0264410X03006108
PMID 14741165
MeSH Adjuvants, Immunologic* Amino Acid Sequence Animals Antibodies, Bacterial / biosynthesis Blotting, Western Bronchoalveolar Lavage Fluid / cytology Bronchoalveolar Lavage Fluid / immunology Cell Line Cells, Cultured Chromatography, Gel Chromatography, Ion Exchange Cytosol / immunology Enzyme-Linked Immunosorbent Assay Female Humans Indicators and Reagents Interleukin-12 / biosynthesis* Macrophages, Alveolar / immunology* Mice Mice, Inbred BALB C Molecular Sequence Data Mycobacterium tuberculosis / chemistry Mycobacterium tuberculosis / immunology* Reverse Transcriptase Polymerase Chain Reaction Subcellular Fractions / immunology Tuberculosis Vaccines / immunology*
IF 3.285
Times Cited 2
Human and Animal Cells