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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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Resident CD11bhiF4/80hi macrophages and CD11chi DCs play a key role in initiating migration-based self-amplification of CD11b+Ly-6Chi monocytes through very early production of CCL2 protein.(A) Crucial role of resident CD11bhiF4/80hi macrophages in inducing early recruitment of CD11b+Ly-6Chi monocytes. BL/6 mice were treated with clodronate liposomes to deplete CD11b+ macrophages and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (B) Regulation of NK cell infiltration by CD11b+F4/80+ macrophages. CD3−NK1.1+DX5+ NK cells in vaginal tract were analyzed in clodronate-treated mice at 48 h pi. (C) Regulation of initial CD11b+Ly-6Chi monocyte infiltration by CD11chi DCs. CD11c-DTR Tg mice were treated with DT to deplete CD11chi DCs and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (D) Regulation of NK cell infiltration by CD11chi DCs. CD3−NK1.1+DX5+ NK cells in the vaginal tract were analyzed in DT-treated CD11c-DTR mice at 48 h pi. (E,F) Resident CD11bhiF4/80hi macrophages and CD11chi DCs provide very early CCL2 production. The levels of CCL2 and CCL3 proteins in vaginal lavages were determined in clodronate- and DT-treated mice at 6, 12, and 24 h pi. (G,H) Viral replication in vaginal tract of mice depleted for CD11bhiF4/80hi macrophages and CD11chi DCs. Viral titers in vaginal tract of clodronate- and DT-treated mice were determined by plaque assay at 6, 12, and 24 h pi. Data in the bar chart represent the average ± SD of values derived from three individual experiments (n = 4–5). *, p<0.05; **, p<0.01; ***, p<0.001 compared with the level of the indicated group.
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ppat.1005256.g008: Resident CD11bhiF4/80hi macrophages and CD11chi DCs play a key role in initiating migration-based self-amplification of CD11b+Ly-6Chi monocytes through very early production of CCL2 protein.(A) Crucial role of resident CD11bhiF4/80hi macrophages in inducing early recruitment of CD11b+Ly-6Chi monocytes. BL/6 mice were treated with clodronate liposomes to deplete CD11b+ macrophages and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (B) Regulation of NK cell infiltration by CD11b+F4/80+ macrophages. CD3−NK1.1+DX5+ NK cells in vaginal tract were analyzed in clodronate-treated mice at 48 h pi. (C) Regulation of initial CD11b+Ly-6Chi monocyte infiltration by CD11chi DCs. CD11c-DTR Tg mice were treated with DT to deplete CD11chi DCs and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (D) Regulation of NK cell infiltration by CD11chi DCs. CD3−NK1.1+DX5+ NK cells in the vaginal tract were analyzed in DT-treated CD11c-DTR mice at 48 h pi. (E,F) Resident CD11bhiF4/80hi macrophages and CD11chi DCs provide very early CCL2 production. The levels of CCL2 and CCL3 proteins in vaginal lavages were determined in clodronate- and DT-treated mice at 6, 12, and 24 h pi. (G,H) Viral replication in vaginal tract of mice depleted for CD11bhiF4/80hi macrophages and CD11chi DCs. Viral titers in vaginal tract of clodronate- and DT-treated mice were determined by plaque assay at 6, 12, and 24 h pi. Data in the bar chart represent the average ± SD of values derived from three individual experiments (n = 4–5). *, p<0.05; **, p<0.01; ***, p<0.001 compared with the level of the indicated group.

Mentions: Since tissue resident CD11bhiF4/80hi macrophages and CD11chiEpCAM+ DCs were proposed to be the initial key cell populations for recruiting early CD11b+Ly-6Chi monocytes for migration-based self-amplification, we investigated the contribution of these cell populations in establishing early orchestrated infiltration of CD11b+Ly-6Chi monocytes and NK cells in vaginal tract. To examine the role of CD11bhiF4/80hi macrophages we used the depletion model of CD11bhiF4/80hi cells using clodronate liposome, as WT BL/6 mice showed an approximately 60% reduction in CD11bhiF4/80hi macrophages after administration of clodronate liposomes (S6A and S6B Fig). Ablation of tissue resident CD11bhiF4/80hi macrophages reduced the accumulated number of infiltrated CD11b+Ly-6Chi monocytes, but the frequency of CD11b+Ly-6Chi monocytes was not obviously changed after gating on CD11b+ cells because of the effect of clodronate liposomes on total CD11b+ myeloid cells (Fig 8A). Thus, the accumulated number of total CD11b+ and CD11b+Ly-6Ghi granulocytes was also reduced. Since clodronate liposomes affect total CD11b+ myeloid cells, including tissue resident CD11bhiF4/80hi macrophages, we also used GdCl3, which has been known as macrophage-selective inhibitor [36]. Our data revealed that GdCl3 administration selectively reduced the frequency and accumulated number of CD11b+Ly-6Chi monocytes in vaginal tract (S7A Fig). Also, the recruitment of NK cells in vaginal tract was apparently decreased in both frequency and accumulated number by the inhibition of tissue resident CD11bhiF4/80hi macrophages using cloronate liposomes (Fig 8B) and GdCl3 (S7B Fig). In addition, we used CD11c-DTR mice that allow conditional DC depletion upon DT injection (S6C and S6D Fig) to examine the role of CD11chiEpCAM+ DCs in the recruitment of early CD11b+Ly-6Chi monocytes. Consistently, ablation of CD11chiEpCAM+ DCs reduced the frequency and accumulated number of CD11b+Ly-6Chi monocytes and NK cells in vaginal tract (Fig 8C and 8D). These results suggest that tissue resident CD11bhiF4/80hi macrophages and CD11chiEpCAM+ DCs play an important role in recruiting the early infiltration of CD11b+Ly-6Chi monocytes to foster the orchestrated infiltration of subsequent leukocytes.


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)

Resident CD11bhiF4/80hi macrophages and CD11chi DCs play a key role in initiating migration-based self-amplification of CD11b+Ly-6Chi monocytes through very early production of CCL2 protein.(A) Crucial role of resident CD11bhiF4/80hi macrophages in inducing early recruitment of CD11b+Ly-6Chi monocytes. BL/6 mice were treated with clodronate liposomes to deplete CD11b+ macrophages and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (B) Regulation of NK cell infiltration by CD11b+F4/80+ macrophages. CD3−NK1.1+DX5+ NK cells in vaginal tract were analyzed in clodronate-treated mice at 48 h pi. (C) Regulation of initial CD11b+Ly-6Chi monocyte infiltration by CD11chi DCs. CD11c-DTR Tg mice were treated with DT to deplete CD11chi DCs and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (D) Regulation of NK cell infiltration by CD11chi DCs. CD3−NK1.1+DX5+ NK cells in the vaginal tract were analyzed in DT-treated CD11c-DTR mice at 48 h pi. (E,F) Resident CD11bhiF4/80hi macrophages and CD11chi DCs provide very early CCL2 production. The levels of CCL2 and CCL3 proteins in vaginal lavages were determined in clodronate- and DT-treated mice at 6, 12, and 24 h pi. (G,H) Viral replication in vaginal tract of mice depleted for CD11bhiF4/80hi macrophages and CD11chi DCs. Viral titers in vaginal tract of clodronate- and DT-treated mice were determined by plaque assay at 6, 12, and 24 h pi. Data in the bar chart represent the average ± SD of values derived from three individual experiments (n = 4–5). *, p<0.05; **, p<0.01; ***, p<0.001 compared with the level of the indicated group.
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Show All Figures
getmorefigures.php?uid=PMC4664252&req=5

ppat.1005256.g008: Resident CD11bhiF4/80hi macrophages and CD11chi DCs play a key role in initiating migration-based self-amplification of CD11b+Ly-6Chi monocytes through very early production of CCL2 protein.(A) Crucial role of resident CD11bhiF4/80hi macrophages in inducing early recruitment of CD11b+Ly-6Chi monocytes. BL/6 mice were treated with clodronate liposomes to deplete CD11b+ macrophages and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (B) Regulation of NK cell infiltration by CD11b+F4/80+ macrophages. CD3−NK1.1+DX5+ NK cells in vaginal tract were analyzed in clodronate-treated mice at 48 h pi. (C) Regulation of initial CD11b+Ly-6Chi monocyte infiltration by CD11chi DCs. CD11c-DTR Tg mice were treated with DT to deplete CD11chi DCs and infected i.vag. with HSV-1. The infiltration of CD11b+Ly-6Chi monocytes was determined by flow cytometric analysis at 24 h pi. (D) Regulation of NK cell infiltration by CD11chi DCs. CD3−NK1.1+DX5+ NK cells in the vaginal tract were analyzed in DT-treated CD11c-DTR mice at 48 h pi. (E,F) Resident CD11bhiF4/80hi macrophages and CD11chi DCs provide very early CCL2 production. The levels of CCL2 and CCL3 proteins in vaginal lavages were determined in clodronate- and DT-treated mice at 6, 12, and 24 h pi. (G,H) Viral replication in vaginal tract of mice depleted for CD11bhiF4/80hi macrophages and CD11chi DCs. Viral titers in vaginal tract of clodronate- and DT-treated mice were determined by plaque assay at 6, 12, and 24 h pi. Data in the bar chart represent the average ± SD of values derived from three individual experiments (n = 4–5). *, p<0.05; **, p<0.01; ***, p<0.001 compared with the level of the indicated group.
Mentions: Since tissue resident CD11bhiF4/80hi macrophages and CD11chiEpCAM+ DCs were proposed to be the initial key cell populations for recruiting early CD11b+Ly-6Chi monocytes for migration-based self-amplification, we investigated the contribution of these cell populations in establishing early orchestrated infiltration of CD11b+Ly-6Chi monocytes and NK cells in vaginal tract. To examine the role of CD11bhiF4/80hi macrophages we used the depletion model of CD11bhiF4/80hi cells using clodronate liposome, as WT BL/6 mice showed an approximately 60% reduction in CD11bhiF4/80hi macrophages after administration of clodronate liposomes (S6A and S6B Fig). Ablation of tissue resident CD11bhiF4/80hi macrophages reduced the accumulated number of infiltrated CD11b+Ly-6Chi monocytes, but the frequency of CD11b+Ly-6Chi monocytes was not obviously changed after gating on CD11b+ cells because of the effect of clodronate liposomes on total CD11b+ myeloid cells (Fig 8A). Thus, the accumulated number of total CD11b+ and CD11b+Ly-6Ghi granulocytes was also reduced. Since clodronate liposomes affect total CD11b+ myeloid cells, including tissue resident CD11bhiF4/80hi macrophages, we also used GdCl3, which has been known as macrophage-selective inhibitor [36]. Our data revealed that GdCl3 administration selectively reduced the frequency and accumulated number of CD11b+Ly-6Chi monocytes in vaginal tract (S7A Fig). Also, the recruitment of NK cells in vaginal tract was apparently decreased in both frequency and accumulated number by the inhibition of tissue resident CD11bhiF4/80hi macrophages using cloronate liposomes (Fig 8B) and GdCl3 (S7B Fig). In addition, we used CD11c-DTR mice that allow conditional DC depletion upon DT injection (S6C and S6D Fig) to examine the role of CD11chiEpCAM+ DCs in the recruitment of early CD11b+Ly-6Chi monocytes. Consistently, ablation of CD11chiEpCAM+ DCs reduced the frequency and accumulated number of CD11b+Ly-6Chi monocytes and NK cells in vaginal tract (Fig 8C and 8D). These results suggest that tissue resident CD11bhiF4/80hi macrophages and CD11chiEpCAM+ DCs play an important role in recruiting the early infiltration of CD11b+Ly-6Chi monocytes to foster the orchestrated infiltration of subsequent leukocytes.

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