title: Polyclonal antibody against conserved sequences of mce1A protein blocks MTB infection in macrophages.
abstract: The pathogenesis of Mycobacterium tuberculosis is largely due to its ability to enter and survive within human macrophages. It is suggested that a specific protein namely mammalian cell entry protein is involved in the pathogenesis and the specific gene for this protein mce1A has been identified in several pathogenic organisms such as Rickettsia, Shigella, Escherichia coli, Helicobacter, Streptomyces, Klebsiella, Vibrio, Neisseria, Rhodococcus, Nocardioides, Saccharopolyspora erthyrae, and Pseudomonas. Analysis of mce1 operons in the above mentioned organisms through bioinformatics tools has revealed the presence of unique sequences (conserved regions) suggesting that these sequences may be involved in the process of infection. Presently, the mce1A full-length (1,365 bp) region from Mycobacterium bovis and its conserved regions (303 bp) were cloned in to an expression vector and the purified expressed proteins of molecular weight ~47 and ~11 kDa, respectively, were injected to rabbits to raise the polyclonal antibodies. The purified polyclonal antibodies were checked for their ability to inhibit the Mycobacterium infection in cultured human macrophages. In macrophage invasion assay, when antibody added at high concentration, decrease in viable counts was observed in all cell cultures within the first 5 days after infection, where the intracellular bacterial CFU obtained from the infected MTB increased by the 3rd day at low concentration of antibody. The macrophage invasion assay has indicated that the purified antibodies of mce1A conserved region can inhibit the infection of Mycobacterium.
authors.affiliation: Department of Genome Medicine, Osho Biotech Research Institute (OBRI), Annanagar, Madurai, Tamil Nadu, India.
authors.forename: Sasikala
authors.initials: S
authors.lastname: Sivagnanam
authors.forename: Nalini
authors.initials: N
authors.lastname: Namasivayam
authors.forename: Rajamanickam
authors.initials: R
authors.lastname: Chellam
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000907:Antibodies, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D017124:Conserved Sequence; D006801:Humans; D008264:Macrophages / Q000276:immunology* / Q000382:microbiology; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000737:chemistry / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology*; D011817:Rabbits; D014376:Tuberculosis / Q000276:immunology* / Q000382:microbiology
chemical_list: D000906:Antibodies; D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; C473802:mammalian cell entry protein Mce1A, Mycobacterium tuberculosis