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Distinct Upstream Role of Type I IFN Signaling in Hematopoietic Stem Cell-Derived and Epithelial Resident Cells for Concerted Recruitment of Ly-6Chi Monocytes and NK Cells via CCL2-CCL3 Cascade.

Uyangaa E, Kim JH, Patil AM, Choi JY, Kim SB, Eo SK - PLoS Pathog. (2015)

Bottom Line: However, the crucial regulators and cell populations that are affected by IFN-I to establish the early environment of innate cells in HSV-infected mucosal tissues are largely unknown.Also, IFN-I signaling in HSC-derived leukocytes appeared to suppress Ly-6Ghi neutrophil recruitment to ameliorate immunopathology.Therefore, this cascade response of resident-to-hematopoietic-to-resident cells that drives cytokine-to-chemokine-to-cytokine production to recruit orchestrated innate cells is critical for attenuation of HSV replication in inflamed tissues.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University, Iksan, Republic of Korea.

ABSTRACT
Type I interferon (IFN-I)-dependent orchestrated mobilization of innate cells in inflamed tissues is believed to play a critical role in controlling replication and CNS-invasion of herpes simplex virus (HSV). However, the crucial regulators and cell populations that are affected by IFN-I to establish the early environment of innate cells in HSV-infected mucosal tissues are largely unknown. Here, we found that IFN-I signaling promoted the differentiation of CCL2-producing Ly-6Chi monocytes and IFN-γ/granzyme B-producing NK cells, whereas deficiency of IFN-I signaling induced Ly-6Clo monocytes producing CXCL1 and CXCL2. More interestingly, recruitment of Ly-6Chi monocytes preceded that of NK cells with the levels peaked at 24 h post-infection in IFN-I-dependent manner, which was kinetically associated with the CCL2-CCL3 cascade response. Early Ly-6Chi monocyte recruitment was governed by CCL2 produced from hematopoietic stem cell (HSC)-derived leukocytes, whereas NK cell recruitment predominantly depended on CC chemokines produced by resident epithelial cells. Also, IFN-I signaling in HSC-derived leukocytes appeared to suppress Ly-6Ghi neutrophil recruitment to ameliorate immunopathology. Finally, tissue resident CD11bhiF4/80hi macrophages and CD11chiEpCAM+ dendritic cells appeared to produce initial CCL2 for migration-based self-amplification of early infiltrated Ly-6Chi monocytes upon stimulation by IFN-I produced from infected epithelial cells. Ultimately, these results decipher a detailed IFN-I-dependent pathway that establishes orchestrated mobilization of Ly-6Chi monocytes and NK cells through CCL2-CCL3 cascade response of HSC-derived leukocytes and epithelium-resident cells. Therefore, this cascade response of resident-to-hematopoietic-to-resident cells that drives cytokine-to-chemokine-to-cytokine production to recruit orchestrated innate cells is critical for attenuation of HSV replication in inflamed tissues.

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Differentiation levels and cytokine expression profile of infiltrated CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN.(A,B) Differentiation levels of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN. The expression of differentiation markers was determined by flow cytometric analysis using cells prepared from vaginal tract (A) and iliac LN (B) by collagenase digestion at 24 h pi. (C,D) Cytokine expression profile of sorted CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes. The levels of cytokine and chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes sorted from vaginal tract (C) and iliac LN (D) of infected BL/6 or IFNAR KO mice at 24 h pi. Data denote the average ± SD of values derived from three individual experiments (n = 5–6). *, p<0.05; **, p<0.01; ***, p<0.001 for comparison between the indicated groups.
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ppat.1005256.g002: Differentiation levels and cytokine expression profile of infiltrated CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN.(A,B) Differentiation levels of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN. The expression of differentiation markers was determined by flow cytometric analysis using cells prepared from vaginal tract (A) and iliac LN (B) by collagenase digestion at 24 h pi. (C,D) Cytokine expression profile of sorted CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes. The levels of cytokine and chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes sorted from vaginal tract (C) and iliac LN (D) of infected BL/6 or IFNAR KO mice at 24 h pi. Data denote the average ± SD of values derived from three individual experiments (n = 5–6). *, p<0.05; **, p<0.01; ***, p<0.001 for comparison between the indicated groups.

Mentions: In the context of inflammation, CD11b+Ly-6Chi monocytes differentiate into inflammatory DCs that could exert tailored protective immunity [24–28]. Therefore, we were interested in testing whether CD11b+Ly-6Chi monocytes that infiltrated into vaginal and iliac LN tissues also have the ability to present Ag as professional antigen-presenting cells similar to DCs during mucosal inflammation caused by HSV-1 infection. To this end, we further characterized CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes for antigen-presenting molecules and activation markers. CD11b+Ly-6Chi monocytes from the vaginal tract of both WT BL/6 and IFNAR KO mice expressed higher levels of Ag presenting-related molecules (CD40, CD80, CD86, and MHC II) than CD11b+Ly-6Clo monocytes, and these molecules showed significantly higher expression levels in CD11b+Ly-6Chi monocytes derived from WT BL/6 mice compared to those from IFNAR KO mice (Fig 2A). Also, CD11b+Ly-6Chi monocytes derived from vaginal tract of BL/6 mice displayed upregulated expression of CCR2, the receptor of CCL2. Similarly, CD11b+Ly-6Chi monocytes in iliac LN of BL/6 mice also showed upregulated expression levels of Ag-presenting–related molecules and activation markers compared to those of IFNAR KO mice (Fig 2B). Since CD11b+Ly-6Chi monocytes also exert antimicrobial activity by secreting antimicrobial factors (TNF-α, NO), we next examined the expression of cytokines and chemokines in CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes derived from vaginal and iliac LN tissues. Intriguingly, CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes showed contrasting expression patterns of cytokines and chemokines in WT BL/6 and IFNAR KO mice (Fig 2C). CD11b+Ly-6Chi monocytes derived from vaginal tract of WT BL/6 mice showed markedly higher expression of TNF-α and iNOS cytokines (the main factors of TipDC) and the CCL2 chemokine (the ligand of CCR2 as a major monocyte chemotactic factor) than those of IFNAR KO mice. In contrast, CD11b+Ly-6Clo monocytes from vaginal tract of IFNAR KO mice expressed high mRNA levels of cytokines IL-23, IL-10, and TGF-β and CXC chemokines (CXCL1, CXCL2). Similarly, iliac CD11b+Ly-6Chi monocytes derived from WT BL/6 mice showed upregulated expression of CCL2, TNF-α, and iNOS, whereas the expression of CXC chemokines (CXCL1 and CXCL2) and cytokines (IL-23, IL-10, and TGF-β) was increased in iliac CD11b+Ly-6Clo monocytes of IFNAR KO mice (Fig 2D). Collectively, these results indicate that IFN-I signal is crucial for determining the characteristics of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes. Also, somewhat surprisingly, IFN-I signaling differentially affected the functional maturation of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes during mucosal HSV-1-induced inflammation.


Distinct Upstream Role of Type I IFN Signaling in Hematopoietic Stem Cell-Derived and Epithelial Resident Cells for Concerted Recruitment of Ly-6Chi Monocytes and NK Cells via CCL2-CCL3 Cascade.

Uyangaa E, Kim JH, Patil AM, Choi JY, Kim SB, Eo SK - PLoS Pathog. (2015)

Differentiation levels and cytokine expression profile of infiltrated CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN.(A,B) Differentiation levels of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN. The expression of differentiation markers was determined by flow cytometric analysis using cells prepared from vaginal tract (A) and iliac LN (B) by collagenase digestion at 24 h pi. (C,D) Cytokine expression profile of sorted CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes. The levels of cytokine and chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes sorted from vaginal tract (C) and iliac LN (D) of infected BL/6 or IFNAR KO mice at 24 h pi. Data denote the average ± SD of values derived from three individual experiments (n = 5–6). *, p<0.05; **, p<0.01; ***, p<0.001 for comparison between the indicated groups.
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Related In: Results  -  Collection

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ppat.1005256.g002: Differentiation levels and cytokine expression profile of infiltrated CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN.(A,B) Differentiation levels of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes in vaginal tract and iliac LN. The expression of differentiation markers was determined by flow cytometric analysis using cells prepared from vaginal tract (A) and iliac LN (B) by collagenase digestion at 24 h pi. (C,D) Cytokine expression profile of sorted CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes. The levels of cytokine and chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes sorted from vaginal tract (C) and iliac LN (D) of infected BL/6 or IFNAR KO mice at 24 h pi. Data denote the average ± SD of values derived from three individual experiments (n = 5–6). *, p<0.05; **, p<0.01; ***, p<0.001 for comparison between the indicated groups.
Mentions: In the context of inflammation, CD11b+Ly-6Chi monocytes differentiate into inflammatory DCs that could exert tailored protective immunity [24–28]. Therefore, we were interested in testing whether CD11b+Ly-6Chi monocytes that infiltrated into vaginal and iliac LN tissues also have the ability to present Ag as professional antigen-presenting cells similar to DCs during mucosal inflammation caused by HSV-1 infection. To this end, we further characterized CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes for antigen-presenting molecules and activation markers. CD11b+Ly-6Chi monocytes from the vaginal tract of both WT BL/6 and IFNAR KO mice expressed higher levels of Ag presenting-related molecules (CD40, CD80, CD86, and MHC II) than CD11b+Ly-6Clo monocytes, and these molecules showed significantly higher expression levels in CD11b+Ly-6Chi monocytes derived from WT BL/6 mice compared to those from IFNAR KO mice (Fig 2A). Also, CD11b+Ly-6Chi monocytes derived from vaginal tract of BL/6 mice displayed upregulated expression of CCR2, the receptor of CCL2. Similarly, CD11b+Ly-6Chi monocytes in iliac LN of BL/6 mice also showed upregulated expression levels of Ag-presenting–related molecules and activation markers compared to those of IFNAR KO mice (Fig 2B). Since CD11b+Ly-6Chi monocytes also exert antimicrobial activity by secreting antimicrobial factors (TNF-α, NO), we next examined the expression of cytokines and chemokines in CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes derived from vaginal and iliac LN tissues. Intriguingly, CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes showed contrasting expression patterns of cytokines and chemokines in WT BL/6 and IFNAR KO mice (Fig 2C). CD11b+Ly-6Chi monocytes derived from vaginal tract of WT BL/6 mice showed markedly higher expression of TNF-α and iNOS cytokines (the main factors of TipDC) and the CCL2 chemokine (the ligand of CCR2 as a major monocyte chemotactic factor) than those of IFNAR KO mice. In contrast, CD11b+Ly-6Clo monocytes from vaginal tract of IFNAR KO mice expressed high mRNA levels of cytokines IL-23, IL-10, and TGF-β and CXC chemokines (CXCL1, CXCL2). Similarly, iliac CD11b+Ly-6Chi monocytes derived from WT BL/6 mice showed upregulated expression of CCL2, TNF-α, and iNOS, whereas the expression of CXC chemokines (CXCL1 and CXCL2) and cytokines (IL-23, IL-10, and TGF-β) was increased in iliac CD11b+Ly-6Clo monocytes of IFNAR KO mice (Fig 2D). Collectively, these results indicate that IFN-I signal is crucial for determining the characteristics of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes. Also, somewhat surprisingly, IFN-I signaling differentially affected the functional maturation of CD11b+Ly-6Chi and CD11b+Ly-6Clo monocytes during mucosal HSV-1-induced inflammation.

Bottom Line: However, the crucial regulators and cell populations that are affected by IFN-I to establish the early environment of innate cells in HSV-infected mucosal tissues are largely unknown.Also, IFN-I signaling in HSC-derived leukocytes appeared to suppress Ly-6Ghi neutrophil recruitment to ameliorate immunopathology.Therefore, this cascade response of resident-to-hematopoietic-to-resident cells that drives cytokine-to-chemokine-to-cytokine production to recruit orchestrated innate cells is critical for attenuation of HSV replication in inflamed tissues.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University, Iksan, Republic of Korea.

ABSTRACT
Type I interferon (IFN-I)-dependent orchestrated mobilization of innate cells in inflamed tissues is believed to play a critical role in controlling replication and CNS-invasion of herpes simplex virus (HSV). However, the crucial regulators and cell populations that are affected by IFN-I to establish the early environment of innate cells in HSV-infected mucosal tissues are largely unknown. Here, we found that IFN-I signaling promoted the differentiation of CCL2-producing Ly-6Chi monocytes and IFN-γ/granzyme B-producing NK cells, whereas deficiency of IFN-I signaling induced Ly-6Clo monocytes producing CXCL1 and CXCL2. More interestingly, recruitment of Ly-6Chi monocytes preceded that of NK cells with the levels peaked at 24 h post-infection in IFN-I-dependent manner, which was kinetically associated with the CCL2-CCL3 cascade response. Early Ly-6Chi monocyte recruitment was governed by CCL2 produced from hematopoietic stem cell (HSC)-derived leukocytes, whereas NK cell recruitment predominantly depended on CC chemokines produced by resident epithelial cells. Also, IFN-I signaling in HSC-derived leukocytes appeared to suppress Ly-6Ghi neutrophil recruitment to ameliorate immunopathology. Finally, tissue resident CD11bhiF4/80hi macrophages and CD11chiEpCAM+ dendritic cells appeared to produce initial CCL2 for migration-based self-amplification of early infiltrated Ly-6Chi monocytes upon stimulation by IFN-I produced from infected epithelial cells. Ultimately, these results decipher a detailed IFN-I-dependent pathway that establishes orchestrated mobilization of Ly-6Chi monocytes and NK cells through CCL2-CCL3 cascade response of HSC-derived leukocytes and epithelium-resident cells. Therefore, this cascade response of resident-to-hematopoietic-to-resident cells that drives cytokine-to-chemokine-to-cytokine production to recruit orchestrated innate cells is critical for attenuation of HSV replication in inflamed tissues.

Show MeSH
Related in: MedlinePlus