The role of Mycobacterium tuberculosis Rv3166c protein-derived high-activity binding peptides in inhibiting invasion of human cell lines

Given the urgent need for designing a new antituberculosis vaccine conferring total protection on patients of all ages, following the line of research adopted by our institute, this work has identified Mycobacterium tuberculosis (Mtb) Rv3166c protein high-activity binding peptides (HABPs) which are able to inhibit bacterial invasion of U937 (monocyte-derived macrophages) and A549 (type II alveolar epithelial cells) cell lines. The presence and transcription of the rv3166c gene in the Mtb species complex was confirmed by polymerase chain reaction (PCR) and reverse transcriptase-PCR; Rv3166c expression was evaluated by western blot and cellular localisation confirmed by immunoelectron microscopy. Its presence was mainly determined on cell surface. Sixteen peptides covering its entire length were chemically synthesised and tested for their ability to bind to U937 and A549 cells. Two U937 HABPs were identified and three for A549, one of them being shared by both cell lines. The four HABPs found inhibited Mtb entry by 15.0794.06. These results led us to including Rv3166c HABPs as candidates for further studies contributing towards the search for a multiepitope, chemically synthesised, subunit-based antituberculosis vaccine. © 2012 The Author.

• Fecha de publicación: 2012
• Institución: Universidad del Rosario
• Repositorio: Repositorio EdocUR - U. Rosario
• Idioma: eng
• OAI Identifier: oai:repository.urosario.edu.co:10336/22890
• Acceso en línea: https://doi.org/10.1093/protein/gzs011; https://repository.urosario.edu.co/handle/10336/22890
• Palabra clave: A549 cells; Alveolar epithelial cells; Bacterial invasion; Binding peptide; Cell lines; Cell surfaces; Human cell lines; Immunoelectron microscopy; Localisation; Monocyte-derived macrophages; Mycobacterium tuberculosis; Synthetic peptide; Type II; Western blots; Cell membranes; Genes; Peptides; Polymerase chain reaction; Transcription; Tubes (components); Vaccines; Cell culture; Bacterial protein; Carrier protein; High activity binding peptide; Protein rv 3166c; Unclassified drug; Amino acid sequence; Article; Cell invasion; Cell line; Cell strain U937; Cell surface; Cellular distribution; Controlled study; Epithelium cell; Gene amplification; Genetic transcription; Immunoelectron microscopy; Lung alveolus epithelium; Macrophage; Mycobacterium tuberculosis; Nonhuman; Priority journal; Protein analysis; Protein binding; Protein synthesis; Reverse transcription polymerase chain reaction; Western blotting; Animals; Bacterial Proteins; Humans; Macrophages; Membrane Proteins; Mycobacterium tuberculosis; Pulmonary Alveoli; Rabbits; Tuberculosis Vaccines; U937 Cells; Bacteria (microorganisms); Mycobacterium tuberculosis; Antituberculosis vaccine; Human cell line; Invasion inhibition; Sub-cellular localisation; Synthetic peptide; Immunoelectron; Bacterial; Antibodies; Microscopy
• License: Abierto (Texto Completo)