The effect of adjuvants and delivery systems on Th1, Th2, Th17 and Treg cytokine responses in mice immunized with Mycobacterium tuberculosis-specific proteins

PLoS One. 2020 Feb 6;15(2):e0228381. doi: 10.1371/journal.pone.0228381. eCollection 2020.

Abstract

Tuberculosis (TB) is a major health problem of global concern. The control of this disease requires appropriate preventive measures, including vaccines. In TB, T helper (Th)1 cytokines provide protection whereas Th2 and T regulatory (Treg) cytokines contribute to the pathogenesis and Th17 cytokines play a role in both protection and pathogenesis. Previous studies with Mycobacterium tuberculosis-specific proteins have identified seven low molecular weight proteins, PE35, ESXA, ESXB, Rv2346c, Rv2347c, Rv3619c, and Rv3620c, as immunodominant antigens inducing Th1-cell responses in humans following natural infection with M. tuberculosis. The aim of this study was to characterize the cytokine responses induced in mice immunized with these proteins, using various adjuvants and delivery systems, i.e. chemical adjuvants (Alum and IFA), non-pathogenic mycobacteria (M. smegmatis and M. vaccae) and a DNA vaccine plasmid (pUMVC6). The immune responses were monitored by quantifying the marker cytokines secreted by Th1 (IFN-ɣ), Th2 (IL-5), Treg (IL-10), and Th17 (IL-17A) cells. DNA corresponding to pe35, esxa, esxb, rv2346c, rv2347c, rv3619c, and rv3620c genes were cloned into the expression vectors pGES-TH-1, pDE22 and pUMVC6 for expression in Escherichia coli, mycobacteria and eukaryotic cells, respectively. Mice were immunized with the recombinants using different adjuvants and delivery systems, and spleen cells were stimulated in vitro with peptides of immunizing proteins to investigate antigen-specific secretion of Th1 (IFN-ɣ), Th2 (IL-5), Treg (IL-10), and Th17 (IL-17A) cytokines. The results showed that spleen cells, from mice immunized with all antigens, secreted the protective Th1 cytokine IFN-ɣ, except ESXB, with one or more adjuvants and delivery systems. However, only Rv3619c consistently induced Th1-biased responses, without the secretion of significant concentrations of Th2, Th17 and Treg cytokines, with all adjuvants and delivery systems. Rv3619c also induced antigen-specific IgG antibodies in immunized mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / administration & dosage*
  • Adjuvants, Pharmaceutic / administration & dosage
  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / immunology*
  • Cells, Cultured
  • Cytokines / metabolism*
  • Drug Delivery Systems / methods*
  • Female
  • Immunization / methods
  • Mice
  • Mice, Inbred BALB C
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / immunology*
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / metabolism
  • Th1 Cells / immunology
  • Th1 Cells / metabolism
  • Th17 Cells / immunology
  • Th17 Cells / metabolism
  • Th2 Cells / immunology
  • Th2 Cells / metabolism
  • Tuberculosis / immunology
  • Tuberculosis / metabolism
  • Tuberculosis / prevention & control*

Substances

  • Adjuvants, Immunologic
  • Adjuvants, Pharmaceutic
  • Bacterial Proteins
  • Cytokines

Grants and funding

ASM HAA AE HAS: A graduate research grant from the College of Graduate Studies, Kuwait University (kuniv.edu/COGS/index.htm) ASM HAA AE HAS: grant numbers YM06/15 and SRU02/13 from the Research Sector, Kuwait University (www.ovpr.kuniv.edu/) ASM HAS: grant number CB17-63MM-03 from Kuwait Foundation for the Advancement of Sciences (kfas.org.kw/) The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.