ABSTRACT
The Bacille Calmette-Guérin (BCG) vaccine comprises a family of genetically different strains derived by the loss of genomic regions (RDs) and other mutations. In BCG Moreau, loss of RD16 inactivates rv3405c * , encoding a transcriptional repressor that negatively regulates the expression of Rv3406, an alkyl sulfatase. To evaluate the impact of this loss on the BCG and host cell viability and the cytokine profile, THP-1 cells were infected with BCG Moreau (harbouring the empty vector) and a complemented strain carrying a functional copy of rv3405c. Viability of the host cells and bacteria as well as the pattern of cytokine secretion were evaluated. Our results show that the viability of BCG Moreau is higher than that of the complemented strain in an axenic medium, suggesting a possible functional gain associated with the constitutive expression of Rv3406. Viability of the host cells did not vary significantly between recombinant strains, but differences in the profiles of the cytokine secretion (IL-1ß and IL-6) were observed. Our results suggest an example of a functional gain due to gene loss contributing to the elucidation of the impact of RD16 on the physiology of BCG Moreau.
Subject(s)
BCG Vaccine/pharmacology , Cell Survival/genetics , Cytokines/drug effects , Gain of Function Mutation/genetics , Macrophages/microbiology , Mycobacterium bovis/genetics , Transcription, Genetic/genetics , BCG Vaccine/genetics , Cell Survival/drug effects , Cytokines/genetics , Gain of Function Mutation/drug effects , Humans , Mycobacterium bovis/physiology , Time Factors , Transcription, Genetic/drug effects , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/microbiologyABSTRACT
Tuberculosis has great public health impact with high rates of mortality and the only prophylactic measure for it is the Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccine. The present study evaluated the release of cytokines [interleukin (IL)-1, tumour necrosis factor and IL-6] and chemokines [macrophage inflammatory protein (MIP)-1α and MIP-1ß] by THP-1 derived macrophages infected with BCG vaccine obtained by growing mycobacteria in Viscondessa de Moraes Institute medium medium (oral) or Sauton medium (intradermic) to compare the effects of live and heat-killed (HK) mycobacteria. Because BCG has been reported to lose viability during the lyophilisation process and during storage, we examined whether exposing BCG to different temperatures also triggers differences in the expression of some important cytokines and chemokines of the immune response. Interestingly, we observed that HK mycobacteria stimulated cytokine and chemokine production in a different pattern from that observed with live mycobacteria.
Subject(s)
Chemokines/metabolism , Macrophages/immunology , Microbial Viability/immunology , Mycobacterium bovis/classification , Cell Line , Chemokine CCL3/metabolism , Chemokine CCL4/metabolism , Cytokines/metabolism , Humans , Interleukin-1/metabolism , Interleukin-6/metabolism , Macrophages/classification , Macrophages/drug effects , Mycobacterium bovis/immunology , Tumor Necrosis Factor-alpha/metabolism , Vaccines, InactivatedABSTRACT
Tuberculosis has great public health impact with high rates of mortality and the only prophylactic measure for it is the Mycobacterium bovisbacillus Calmette-Guérin (BCG) vaccine. The present study evaluated the release of cytokines [interleukin (IL)-1, tumour necrosis factor and IL-6] and chemokines [macrophage inflammatory protein (MIP)-1α and MIP-1β] by THP-1 derived macrophages infected with BCG vaccine obtained by growing mycobacteria in Viscondessa de Moraes Institute medium medium (oral) or Sauton medium (intradermic) to compare the effects of live and heat-killed (HK) mycobacteria. Because BCG has been reported to lose viability during the lyophilisation process and during storage, we examined whether exposing BCG to different temperatures also triggers differences in the expression of some important cytokines and chemokines of the immune response. Interestingly, we observed that HK mycobacteria stimulated cytokine and chemokine production in a different pattern from that observed with live mycobacteria.
Subject(s)
Humans , Chemokines , Macrophages/immunology , Microbial Viability/immunology , Mycobacterium bovis/classification , Cell Line , Cytokines , Interleukin-1 , Macrophages/classification , Macrophages/drug effects , Mycobacterium bovis/immunology , Tumor Necrosis Factor-alpha , Vaccines, InactivatedABSTRACT
The bacillus Calmette-Guérin (BCG) vaccine is the only licensed vaccine for human use against tuberculosis (TB). Although controversy exists about its efficacy, the BCG vaccine is able to protect newborns and children against disseminated forms of TB, but fails to protect adults against active forms of TB. In the last few years, interest in the mucosal delivery route for the vaccine has been increasing owing to its increased capacity to induce protective immune responses both in the mucosal and the systemic immune compartments. Here, we show the importance of this route of vaccination in newly developed vaccines, especially for vaccines against TB.
Subject(s)
BCG Vaccine/administration & dosage , Tuberculosis, Pulmonary/prevention & control , Vaccination/methods , Administration, Oral , Adolescent , Adult , BCG Vaccine/history , BCG Vaccine/immunology , Child , History, 20th Century , History, 21st Century , Humans , Immunity, Mucosal , Immunologic Memory , Infant, Newborn , Lymphoid Tissue/immunologyABSTRACT
The bacillus Calmette-Guérin (BCG) vaccine is the only licensed vaccine for human use against tuberculosis (TB). Although controversy exists about its efficacy, the BCG vaccine is able to protect newborns and children against disseminated forms of TB, but fails to protect adults against active forms of TB. In the last few years, interest in the mucosal delivery route for the vaccine has been increasing owing to its increased capacity to induce protective immune responses both in the mucosal and the systemic immune compartments. Here, we show the importance of this route of vaccination in newly developed vaccines, especially for vaccines against TB.
