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B cells expressing IL-10 mRNA modulate memory T cells after DNA-Hsp65 immunization.
- Source :
-
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas [Braz J Med Biol Res] 2015 Dec; Vol. 48 (12), pp. 1095-100. Date of Electronic Publication: 2015 Sep 18. - Publication Year :
- 2015
-
Abstract
- In DNA vaccines, the gene of interest is cloned into a bacterial plasmid that is engineered to induce protein production for long periods in eukaryotic cells. Previous research has shown that the intramuscular immunization of BALB/c mice with a naked plasmid DNA fragment encoding the Mycobacterium leprae 65-kDa heat-shock protein (pcDNA3-Hsp65) induces protection against M. tuberculosis challenge. A key stage in the protective immune response after immunization is the generation of memory T cells. Previously, we have shown that B cells capture plasmid DNA-Hsp65 and thereby modulate the formation of CD8+ memory T cells after M. tuberculosis challenge in mice. Therefore, clarifying how B cells act as part of the protective immune response after DNA immunization is important for the development of more-effective vaccines. The aim of this study was to investigate the mechanisms by which B cells modulate memory T cells after DNA-Hsp65 immunization. C57BL/6 and BKO mice were injected three times, at 15-day intervals, with 100 µg naked pcDNA-Hsp65 per mouse. Thirty days after immunization, the percentages of effector memory T (TEM) cells (CD4+ and CD8+/CD44high/CD62Llow) and memory CD8+ T cells (CD8+/CD44high/CD62Llow/CD127+) were measured with flow cytometry. Interferon γ, interleukin 12 (IL-12), and IL-10 mRNAs were also quantified in whole spleen cells and purified B cells (CD43-) with real-time qPCR. Our data suggest that a B-cell subpopulation expressing IL-10 downregulated proinflammatory cytokine expression in the spleen, increasing the survival of CD4+ TEM cells and CD8+ TEM/CD127+ cells.
- Subjects :
- Animals
B-Lymphocytes metabolism
Flow Cytometry
Gene Expression genetics
Heat-Shock Proteins therapeutic use
Immunologic Memory physiology
Immunophenotyping classification
Inflammation Mediators analysis
Interferon-gamma analysis
Interleukin-10 immunology
Interleukin-12 analysis
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
RNA, Messenger genetics
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Spleen cytology
Spleen immunology
T-Lymphocyte Subsets classification
Vaccines, DNA immunology
Vaccines, DNA therapeutic use
B-Lymphocytes immunology
Heat-Shock Proteins immunology
Immunomodulation genetics
Interleukin-10 genetics
RNA, Messenger immunology
T-Lymphocyte Subsets immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1414-431X
- Volume :
- 48
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas
- Publication Type :
- Academic Journal
- Accession number :
- 26397973
- Full Text :
- https://doi.org/10.1590/1414-431X20154409