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Interferon-α curbs production of interleukin-22 by human peripheral blood mononuclear cells exposed to live Borrelia burgdorferi.

Berner A, Bachmann M, Pfeilschifter J, Kraiczy P, Mühl H - J. Cell. Mol. Med. (2015)

Bottom Line: Cytokine networks initiated by means of innate immunity are regarded as a major determinant of host defence in response to acute infection by bacteria including Borrelia burgdorferi.Herein, we demonstrate that interferon (IFN)-α, either endogenously produced after exposure of cells to toll-like receptor-9-activating CpG oligonucleotides or provided as recombinant cytokine, weakens activation of the anti-bacterial interleukin (IL)-1/IL-22 axis in human peripheral blood mononuclear cells exposed to viable B. burgdorferi.As IFN-α has been related to pathological dissemination of the spirochaete, data suggest an immunoregulatory role of type I IFN in this context that is able to significantly modify cytokine profiles thereby possibly determining early course of B. burgdorferi infection.

View Article: PubMed Central - PubMed

Affiliation: Pharmazentrum Frankfurt/ZAFES, University Hospital Goethe-University Frankfurt, Frankfurt am Main, Germany.

No MeSH data available.


Related in: MedlinePlus

(A) PBMC were kept as an unstimulated control or exposed to Borrelia burgdorferi (MOI = 0.1) in presence or absence of IFN-α (5 ng/ml) or with IFN-α (5 ng/ml) alone. After 24 hrs, total RNA was isolated and IL-22 mRNA expression was determined by realtime PCR. IL-22 mRNA was normalized to that of GAPDH and is shown as mean fold-induction compared to unstimulated control ± SEM (n = 4); **P < 0.01 compared with unstimulated control, #P < 0.05. (B) PBMC were kept as unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 8); ***P < 0.001 compared with unstimulated control, ###P < 0.001. Inset: PBMC were kept as unstimulated control, were exposed to B. burgdorferi (MOI = 0.1) in the presence or absence of the indicated concentrations of IFN-α (10 or 50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data (% of B. burgdorferi alone) are expressed as means ± SEM (n = 8); ##P < 0.05, ###P < 0.001 compared with B. burgdorferi alone. (C) PBMC were kept as an unstimulated control or were activated by anti-CD3 (5 μg/ml) in the presence or absence of IFN-α (50 ng/ml) or by IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 4); ***P < 0.001 compared with unstimulated control. n.s. denotes, not significant. (D and E) PBMC were kept as an unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-1β (D) or IL-8 (E) was determined by ELISA. Data are expressed as means ± SEM (n = 8); **P < 0.01; ***P < 0.001 compared with unstimulated control; ###P < 0.001. C or B, denote cultivation of unstimulated PBMC or exposure of PBMC to B. burgdorferi, respectively.
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fig02: (A) PBMC were kept as an unstimulated control or exposed to Borrelia burgdorferi (MOI = 0.1) in presence or absence of IFN-α (5 ng/ml) or with IFN-α (5 ng/ml) alone. After 24 hrs, total RNA was isolated and IL-22 mRNA expression was determined by realtime PCR. IL-22 mRNA was normalized to that of GAPDH and is shown as mean fold-induction compared to unstimulated control ± SEM (n = 4); **P < 0.01 compared with unstimulated control, #P < 0.05. (B) PBMC were kept as unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 8); ***P < 0.001 compared with unstimulated control, ###P < 0.001. Inset: PBMC were kept as unstimulated control, were exposed to B. burgdorferi (MOI = 0.1) in the presence or absence of the indicated concentrations of IFN-α (10 or 50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data (% of B. burgdorferi alone) are expressed as means ± SEM (n = 8); ##P < 0.05, ###P < 0.001 compared with B. burgdorferi alone. (C) PBMC were kept as an unstimulated control or were activated by anti-CD3 (5 μg/ml) in the presence or absence of IFN-α (50 ng/ml) or by IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 4); ***P < 0.001 compared with unstimulated control. n.s. denotes, not significant. (D and E) PBMC were kept as an unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-1β (D) or IL-8 (E) was determined by ELISA. Data are expressed as means ± SEM (n = 8); **P < 0.01; ***P < 0.001 compared with unstimulated control; ###P < 0.001. C or B, denote cultivation of unstimulated PBMC or exposure of PBMC to B. burgdorferi, respectively.

Mentions: To further evaluate a potential regulatory function of IFN-α on B. burgdorferi-stimulated IL-22, PBMC activation was analysed under the influence of this type I IFN. In fact, Figure 2A demonstrates significant suppression of B. burgdorferi-induced IL-22 mRNA expression by IFN-α. Modulation of IL-22 mRNA likewise translated to the level of cytokine secretion (Fig. 2B). Dose-response analysis revealed that effects of IFN-α were optimal at 50 ng/ml (Fig. 2B, inset), which was used in further experiments. Modulation of IL-22 secretion by IFN-α essentially connected to the induction of innate immunity, as this regulatory path was not detectable in case of IL-22 production in response to polyclonal T cell activation using an agonistic anti-CD3 antibody (Fig. 2C). Notably, type I IFN, namely IFN-β, has previously been reported to inhibit spontaneous release of IL-22 from otherwise unstimulated CD4+ T cells obtained from multiple sclerosis patients 17. In this context it is noteworthy that memory helper CD4+ and cytotoxic CD8+ T cells but also naïve CD4+ T cells exposed to an inflammatory environment can produce ample amounts of cytokines in an antigen-/T cell receptor-independent manner 18,19. In accord with this notion, we recently identified T cells, mainly CD4+ but also CD8+ T cells, as chief cellular IL-22 source in PBMC exposed to B. burgdorferi6.


Interferon-α curbs production of interleukin-22 by human peripheral blood mononuclear cells exposed to live Borrelia burgdorferi.

Berner A, Bachmann M, Pfeilschifter J, Kraiczy P, Mühl H - J. Cell. Mol. Med. (2015)

(A) PBMC were kept as an unstimulated control or exposed to Borrelia burgdorferi (MOI = 0.1) in presence or absence of IFN-α (5 ng/ml) or with IFN-α (5 ng/ml) alone. After 24 hrs, total RNA was isolated and IL-22 mRNA expression was determined by realtime PCR. IL-22 mRNA was normalized to that of GAPDH and is shown as mean fold-induction compared to unstimulated control ± SEM (n = 4); **P < 0.01 compared with unstimulated control, #P < 0.05. (B) PBMC were kept as unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 8); ***P < 0.001 compared with unstimulated control, ###P < 0.001. Inset: PBMC were kept as unstimulated control, were exposed to B. burgdorferi (MOI = 0.1) in the presence or absence of the indicated concentrations of IFN-α (10 or 50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data (% of B. burgdorferi alone) are expressed as means ± SEM (n = 8); ##P < 0.05, ###P < 0.001 compared with B. burgdorferi alone. (C) PBMC were kept as an unstimulated control or were activated by anti-CD3 (5 μg/ml) in the presence or absence of IFN-α (50 ng/ml) or by IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 4); ***P < 0.001 compared with unstimulated control. n.s. denotes, not significant. (D and E) PBMC were kept as an unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-1β (D) or IL-8 (E) was determined by ELISA. Data are expressed as means ± SEM (n = 8); **P < 0.01; ***P < 0.001 compared with unstimulated control; ###P < 0.001. C or B, denote cultivation of unstimulated PBMC or exposure of PBMC to B. burgdorferi, respectively.
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Related In: Results  -  Collection

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fig02: (A) PBMC were kept as an unstimulated control or exposed to Borrelia burgdorferi (MOI = 0.1) in presence or absence of IFN-α (5 ng/ml) or with IFN-α (5 ng/ml) alone. After 24 hrs, total RNA was isolated and IL-22 mRNA expression was determined by realtime PCR. IL-22 mRNA was normalized to that of GAPDH and is shown as mean fold-induction compared to unstimulated control ± SEM (n = 4); **P < 0.01 compared with unstimulated control, #P < 0.05. (B) PBMC were kept as unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 8); ***P < 0.001 compared with unstimulated control, ###P < 0.001. Inset: PBMC were kept as unstimulated control, were exposed to B. burgdorferi (MOI = 0.1) in the presence or absence of the indicated concentrations of IFN-α (10 or 50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data (% of B. burgdorferi alone) are expressed as means ± SEM (n = 8); ##P < 0.05, ###P < 0.001 compared with B. burgdorferi alone. (C) PBMC were kept as an unstimulated control or were activated by anti-CD3 (5 μg/ml) in the presence or absence of IFN-α (50 ng/ml) or by IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-22 was determined by ELISA. Data are expressed as means ± SEM (n = 4); ***P < 0.001 compared with unstimulated control. n.s. denotes, not significant. (D and E) PBMC were kept as an unstimulated control or exposed to B. burgdorferi (MOI = 0.1) in presence or absence of IFN-α (50 ng/ml) or with IFN-α (50 ng/ml) alone. After 65 hrs, release of IL-1β (D) or IL-8 (E) was determined by ELISA. Data are expressed as means ± SEM (n = 8); **P < 0.01; ***P < 0.001 compared with unstimulated control; ###P < 0.001. C or B, denote cultivation of unstimulated PBMC or exposure of PBMC to B. burgdorferi, respectively.
Mentions: To further evaluate a potential regulatory function of IFN-α on B. burgdorferi-stimulated IL-22, PBMC activation was analysed under the influence of this type I IFN. In fact, Figure 2A demonstrates significant suppression of B. burgdorferi-induced IL-22 mRNA expression by IFN-α. Modulation of IL-22 mRNA likewise translated to the level of cytokine secretion (Fig. 2B). Dose-response analysis revealed that effects of IFN-α were optimal at 50 ng/ml (Fig. 2B, inset), which was used in further experiments. Modulation of IL-22 secretion by IFN-α essentially connected to the induction of innate immunity, as this regulatory path was not detectable in case of IL-22 production in response to polyclonal T cell activation using an agonistic anti-CD3 antibody (Fig. 2C). Notably, type I IFN, namely IFN-β, has previously been reported to inhibit spontaneous release of IL-22 from otherwise unstimulated CD4+ T cells obtained from multiple sclerosis patients 17. In this context it is noteworthy that memory helper CD4+ and cytotoxic CD8+ T cells but also naïve CD4+ T cells exposed to an inflammatory environment can produce ample amounts of cytokines in an antigen-/T cell receptor-independent manner 18,19. In accord with this notion, we recently identified T cells, mainly CD4+ but also CD8+ T cells, as chief cellular IL-22 source in PBMC exposed to B. burgdorferi6.

Bottom Line: Cytokine networks initiated by means of innate immunity are regarded as a major determinant of host defence in response to acute infection by bacteria including Borrelia burgdorferi.Herein, we demonstrate that interferon (IFN)-α, either endogenously produced after exposure of cells to toll-like receptor-9-activating CpG oligonucleotides or provided as recombinant cytokine, weakens activation of the anti-bacterial interleukin (IL)-1/IL-22 axis in human peripheral blood mononuclear cells exposed to viable B. burgdorferi.As IFN-α has been related to pathological dissemination of the spirochaete, data suggest an immunoregulatory role of type I IFN in this context that is able to significantly modify cytokine profiles thereby possibly determining early course of B. burgdorferi infection.

View Article: PubMed Central - PubMed

Affiliation: Pharmazentrum Frankfurt/ZAFES, University Hospital Goethe-University Frankfurt, Frankfurt am Main, Germany.

No MeSH data available.


Related in: MedlinePlus