J Vet Sci.  2018 Jul;19(4):536-542. 10.4142/jvs.2018.19.4.536.

A Salmonella Typhi ghost induced by the E gene of phage φX174 stimulates dendritic cells and efficiently activates the adaptive immune response

• Won G
• Eo SK
• Park SY
• Hur J
• Lee JH

• KMID: 2417569
• DOI: http://doi.org/10.4142/jvs.2018.19.4.536

Abstract

Previously, we genetically engineered a Salmonella Typhi bacterial ghost (STG) as a novel inactivated vaccine candidate against typhoid fever. The underlying mechanism employed by the ghost in stimulating the adaptive immune response remains to be investigated. In this study, we aimed to evaluate the immunostimulatory effect of STG on mouse bone marrow-derived dendritic cells (BMDCs) and its activation of the adaptive immune response in vitro. Immature BMDCs were stimulated with STG, which efficiently stimulated maturation events in BMDCs, as indicated by upregulated expressions of CD40, CD80, and major histocompatibility complex class II molecules on CD11⁺ BMDCs. Immature BMDCs responded to STG stimulation by significantly increasing the expression of interleukin (IL)-6, which might indicate the induction of dendritic cell maturation in vivo (p < 0.05). In addition, ghost-stimulated murine BMDCs showed significant expressions of interferon gamma and IL-4, which can drive the development of Th1 and Th2 cells, respectively, in co-cultured CD4⁺ T cells in vitro. These results suggest that STG can effectively stimulate maturation of BMDCs and facilitate subsequent immune responses via potent immunomodulatory cytokine responses.

Keyword

Salmonella Typhi; dendritic cells; innate immunity

MeSH Terms

Adaptive Immunity*
Animals
Bacteriophages*
Dendritic Cells*
Immunity, Innate
In Vitro Techniques
Interferons
Interleukin-4
Interleukins
Major Histocompatibility Complex
Mice
Salmonella typhi*
Salmonella*
T-Lymphocytes
Th2 Cells
Typhoid Fever
Interferons
Interleukin-4
Interleukins

Figure

• 

  Fig. 1 Morphological observation of bone marrow-derived dendritic cells (BMDCs) during the differentiation procedures. The pictures were captured by inverted microscope equipped with a digital imaging system (Leica DMi1; Leica Microsystems, Germany). (A) BMDCs observed at day 2 of culture. (B) Morphology of the non-stimulated BMDCs (arrows) at day 7 of culture. (C) Morphology of DCs (arrows) pulsed with lipopolysaccharide at day 9 of culture. (D) Morphology of DCs (arrows) stimulated with Salmonella Typhi ghost at day 9 of culture. Scale bars = 50 µm (A–D).

• 

  Fig. 2 Co-stimulatory molecule expressions on dendritic cells (DC) surfaces was assessed by gating on CD11c+ cell populations using fluorescence-activated cell sorting (FACS). (A) Representative FACS histogram of the surface marker-positive cell population. (B) Percentages of co-stimulatory molecule-positive DCs. NS, non-stimulated; LPS, lipopolysaccharide. *p < 0.05 (vs. non-stimulated DCs).

• 

  Fig. 3 Cytokine mRNA upregulated in dendritic cells co-cultured with lipopolysaccharide (LPS) or Salmonella Typhi ghost (JOL1502 ghost). Relative fold changes were calculated based on 2−ΔΔCt method. IL, interleukin; TNF-α, tumor necrosis factor alpha. *p < 0.05.

• 

  Fig. 4 Expression of T-cell-derived cytokines induced by indirect activation of naive CD4+ T cells by dendritic cells primed with the JOL1502 ghost (Salmonella Typhi ghost) or lipopolysaccharide (LPS). Relative fold changes were assessed by using the 2−ΔΔCt method. IL, interleukin; INF-β, interferon gamma. *p < 0.05.

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