ABSTRACT
Mycobacterium tuberculosis infection continues to be a major cause of morbidity and mortality throughout the world. The vast complexity of the intracellular pathogen M. tuberculosis and the diverse mechanisms by which it can invade host cells highlight the importance of developing a fully protective vaccine. Our vaccine development strategy consists of including fragments from multiple mycobacterial proteins involved in cell invasion. The aim of this study was to identify high activity binding peptides (HABPs) in the immunogenic protein Rv1980c from M. tuberculosis H37Rv with the ability to inhibit mycobacterial invasion into U937 monocyte-derived macrophages and A549 cells. The presence and transcription of the Rv1980c gene was assessed in members belonging to the M. tuberculosis complex and other nontuberculous mycobacteria by PCR and RT-PCR, respectively. Cell surface localization was confirmed by immuno-electron microscopy. Three peptides binding with high activity to U937 cells and one to A549 cells were identified. HABPs 31100, 31101, and 31107 inhibited invasion of M. tuberculosis into A549 and U937 cells and therefore could be promising candidates for the design of a subunit-based antituberculous vaccine.
Subject(s)
Bacterial Proteins/immunology , Bacterial Proteins/metabolism , Bacterial Vaccines/immunology , Mycobacterium tuberculosis/physiology , Peptide Fragments/metabolism , Vaccines, Synthetic/immunology , Alveolar Epithelial Cells/cytology , Alveolar Epithelial Cells/immunology , Alveolar Epithelial Cells/microbiology , Bacterial Proteins/analysis , Bacterial Proteins/chemistry , Bacterial Vaccines/chemistry , Binding Sites , Blotting, Western , Cells, Cultured , Humans , Models, Molecular , Mycobacterium tuberculosis/genetics , Peptide Fragments/analysis , Peptide Fragments/chemistry , Peptide Fragments/immunology , Transcription, Genetic , Vaccines, Synthetic/chemistryABSTRACT
A population of cells exhibiting bona fide dendritic cell (DC) morphological and functional characteristics was obtained by treating Aotus spp. monocytes with human IL-4 and GM-CSF. Although the purity of mature DCs was relatively low IL-4/GM-CSF-treated monocytes (hereafter called Aotus spp. DCs) down-regulated CD14 and up-regulated discrete levels of CD80, MHC-Class II and CD1b molecules in response to different maturation stimuli. Aotus spp. DCs generated a potent allogeneic in vitro response evidenced in mixed lymphocyte reaction (MLR) where DCs were 2- to 10-fold more efficient than peripheral blood mononuclear cells (PBMCs). Aotus spp. DC ability to boost T-cells or priming naive T-cells in vivo was proved by vaccinating Aotus spp. with autologous DCs pulsed with tetanus toxoid (TT). A single dose of TT-pulsed DCs was sufficient to increase cellular response to TT in these experiments as assessed by lymphoproliferation and cytokine production. Since Aotus spp. represents a suitable animal model for evaluating anti-Plasmodium falciparum malaria vaccine, the results shown here suggest that using antigen-pulsed Aotus spp. DCs as vaccines might lead to identifying new prospects for malarial vaccines unidentified to date because they are being formulated in less efficient adjuvants.
