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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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IFN-I signaling on infiltrated leukocytes derived from HSC lineage is required for normal recruitment of CD11b+Ly-6Chi monocytes, but not NK cells.BM cells from BL/6 (WT) or IFNAR KO (KO) were grafted into lethally irradiated BL/6 or IFNAR KO recipient mice, which were infected i.vag. with HSV-1. (A) Leukocyte infiltration in vaginal tract and iliac LN of infected recipients. (B,C) Accumulated number of infiltrated leukocyte subsets. Cells were prepared from vaginal tract (B; VT) and iliac LN (C; ILN) by collagenase digestion at 24 h pi and subcellular proportions of CD11b+Ly-6Chi and CD11b+Ly-6Ghi leukocytes were determined using flow cytometric analysis. (D,E) Frequency and absolute number of infiltrated NK cells in VT and ILN of the recipients. Cells prepared from VT and ILN of the recipients were used to analyze the frequency (D) and total number (E) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (F) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (G) Chemokine secretion in vaginal lavages of the recipients. The levels of chemokine proteins were determined by CBA using vaginal lavages at 6, 12, 24, and 48 h pi. Values in dot-plots represent the average percentages of each population derived from four independent samples, and data in graphs 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.g005: IFN-I signaling on infiltrated leukocytes derived from HSC lineage is required for normal recruitment of CD11b+Ly-6Chi monocytes, but not NK cells.BM cells from BL/6 (WT) or IFNAR KO (KO) were grafted into lethally irradiated BL/6 or IFNAR KO recipient mice, which were infected i.vag. with HSV-1. (A) Leukocyte infiltration in vaginal tract and iliac LN of infected recipients. (B,C) Accumulated number of infiltrated leukocyte subsets. Cells were prepared from vaginal tract (B; VT) and iliac LN (C; ILN) by collagenase digestion at 24 h pi and subcellular proportions of CD11b+Ly-6Chi and CD11b+Ly-6Ghi leukocytes were determined using flow cytometric analysis. (D,E) Frequency and absolute number of infiltrated NK cells in VT and ILN of the recipients. Cells prepared from VT and ILN of the recipients were used to analyze the frequency (D) and total number (E) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (F) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (G) Chemokine secretion in vaginal lavages of the recipients. The levels of chemokine proteins were determined by CBA using vaginal lavages at 6, 12, 24, and 48 h pi. Values in dot-plots represent the average percentages of each population derived from four independent samples, and data in graphs 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 CD11b+Ly-6Chi monocytes and NK cells showed concerted recruitment with levels peaking at different time points associated with the cascade response of CCL2-CCL3 in the vaginal tract, we were interested in dissecting the contribution of resident (stromal and epithelial cells) and HSC-derived leukocytes in this process. To achieve this, we used a BM chimeric model of WT BL/6 and IFNAR KO mice. As expected, IFNAR KO recipients of IFNAR KO BM donor cells (KO-KO chimera) showed a diminished frequency of CD11b+Ly-6Chi monocytes in vaginal and iliac LN, compared to WT recipients of WT BM donor cells (WT-WT chimera) (Fig 5A). Also, somewhat intriguingly, IFNAR KO recipients of WT BM donor cells (WT-KO chimeras) showed comparable recruitment of CD11b+Ly-6Chi monocytes in the vaginal tract and iliac LN to WT-WT chimeras, but WT BL/6 recipients of IFNAR KO BM donor cells (KO-WT chimeras) contained a lower frequency of CD11b+Ly-6Chi monocytes with levels comparable to those of KO-KO chimeras, indicating that IFN-I receptors of some leukocytes derived from HSC lineage are essential for the recruitment of CD11b+Ly-6Chi monocytes in vaginal and iliac LN tissues. When the absolute number of recruited CD11b+Ly-6Chi monocytes was determined, the essential role of IFN-I receptor on leukocytes derived from the HSC lineage in vaginal CD11b+Ly-6Chi monocyte recruitment was revealed again, even though KO-WT chimeras showed slightly higher levels of total recruited CD11b+ cells compared to the other chimeras (Fig 5B). In contrast, IFN-I receptors in HSC-derived leukocytes appeared to be dispensable for the recruitment of CD11b+Ly-6Ghi neutrophils in the vaginal tract, because WT-KO chimeras showed a reduced number of neutrophils compared to KO-KO chimeras. Also, considerable recruitment of iliac CD11b+Ly-6Chi monocytes in both WT-KO and KO-WT chimeras highlighted the importance of IFN-I receptors in both HSC-derived and resident cells for lymphoid tissues (Fig 5C). Therefore, these results indicate that IFN-I signal in some leukocytes derived from the HSC lineage plays a crucial role in early recruitment of CD11b+Ly-6Chi monocytes in the vaginal tract with relatively less contribution to recruitment of CD11b+Ly-6Chi monocytes in draining LN, even though specific leukocytes derived from the HSC lineage to involve in the recruitment of Ly-6Chi monocytes were not defined. Another intriguing result is that, unlike vaginal CD11b+Ly-6Chi monocyte recruitment, the recruitment of NK cells in the vaginal tract and iliac LN obviously depended dominantly on IFN-I receptor in resident cells; hence, KO-WT chimera showed completely recovered recruitment of CD3−NK1.1+DX5+ NK cells in vaginal tract and iliac LN, compared to diminished recruitment of NK cells in KO-KO and WT-KO chimeras (Fig 5D and 5E). In addition, it was likely that recovered recruitment of CD11b+Ly-6Chi monocytes and NK cells in WT-KO and KO-WT chimeras contributed to the control of HSV-1 replication in the vaginal tract (Fig 5F). Collectively, these results suggest that IFN-I signaling in resident cells is dominantly responsible for recruiting NK cells in inflammatory mucosal tissues, in contrast to CD11b+Ly-6Chi monocytes that depend on the IFN-I signal in BM-derived HSCs.


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)

IFN-I signaling on infiltrated leukocytes derived from HSC lineage is required for normal recruitment of CD11b+Ly-6Chi monocytes, but not NK cells.BM cells from BL/6 (WT) or IFNAR KO (KO) were grafted into lethally irradiated BL/6 or IFNAR KO recipient mice, which were infected i.vag. with HSV-1. (A) Leukocyte infiltration in vaginal tract and iliac LN of infected recipients. (B,C) Accumulated number of infiltrated leukocyte subsets. Cells were prepared from vaginal tract (B; VT) and iliac LN (C; ILN) by collagenase digestion at 24 h pi and subcellular proportions of CD11b+Ly-6Chi and CD11b+Ly-6Ghi leukocytes were determined using flow cytometric analysis. (D,E) Frequency and absolute number of infiltrated NK cells in VT and ILN of the recipients. Cells prepared from VT and ILN of the recipients were used to analyze the frequency (D) and total number (E) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (F) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (G) Chemokine secretion in vaginal lavages of the recipients. The levels of chemokine proteins were determined by CBA using vaginal lavages at 6, 12, 24, and 48 h pi. Values in dot-plots represent the average percentages of each population derived from four independent samples, and data in graphs 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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Related In: Results  -  Collection

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ppat.1005256.g005: IFN-I signaling on infiltrated leukocytes derived from HSC lineage is required for normal recruitment of CD11b+Ly-6Chi monocytes, but not NK cells.BM cells from BL/6 (WT) or IFNAR KO (KO) were grafted into lethally irradiated BL/6 or IFNAR KO recipient mice, which were infected i.vag. with HSV-1. (A) Leukocyte infiltration in vaginal tract and iliac LN of infected recipients. (B,C) Accumulated number of infiltrated leukocyte subsets. Cells were prepared from vaginal tract (B; VT) and iliac LN (C; ILN) by collagenase digestion at 24 h pi and subcellular proportions of CD11b+Ly-6Chi and CD11b+Ly-6Ghi leukocytes were determined using flow cytometric analysis. (D,E) Frequency and absolute number of infiltrated NK cells in VT and ILN of the recipients. Cells prepared from VT and ILN of the recipients were used to analyze the frequency (D) and total number (E) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (F) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (G) Chemokine secretion in vaginal lavages of the recipients. The levels of chemokine proteins were determined by CBA using vaginal lavages at 6, 12, 24, and 48 h pi. Values in dot-plots represent the average percentages of each population derived from four independent samples, and data in graphs 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 CD11b+Ly-6Chi monocytes and NK cells showed concerted recruitment with levels peaking at different time points associated with the cascade response of CCL2-CCL3 in the vaginal tract, we were interested in dissecting the contribution of resident (stromal and epithelial cells) and HSC-derived leukocytes in this process. To achieve this, we used a BM chimeric model of WT BL/6 and IFNAR KO mice. As expected, IFNAR KO recipients of IFNAR KO BM donor cells (KO-KO chimera) showed a diminished frequency of CD11b+Ly-6Chi monocytes in vaginal and iliac LN, compared to WT recipients of WT BM donor cells (WT-WT chimera) (Fig 5A). Also, somewhat intriguingly, IFNAR KO recipients of WT BM donor cells (WT-KO chimeras) showed comparable recruitment of CD11b+Ly-6Chi monocytes in the vaginal tract and iliac LN to WT-WT chimeras, but WT BL/6 recipients of IFNAR KO BM donor cells (KO-WT chimeras) contained a lower frequency of CD11b+Ly-6Chi monocytes with levels comparable to those of KO-KO chimeras, indicating that IFN-I receptors of some leukocytes derived from HSC lineage are essential for the recruitment of CD11b+Ly-6Chi monocytes in vaginal and iliac LN tissues. When the absolute number of recruited CD11b+Ly-6Chi monocytes was determined, the essential role of IFN-I receptor on leukocytes derived from the HSC lineage in vaginal CD11b+Ly-6Chi monocyte recruitment was revealed again, even though KO-WT chimeras showed slightly higher levels of total recruited CD11b+ cells compared to the other chimeras (Fig 5B). In contrast, IFN-I receptors in HSC-derived leukocytes appeared to be dispensable for the recruitment of CD11b+Ly-6Ghi neutrophils in the vaginal tract, because WT-KO chimeras showed a reduced number of neutrophils compared to KO-KO chimeras. Also, considerable recruitment of iliac CD11b+Ly-6Chi monocytes in both WT-KO and KO-WT chimeras highlighted the importance of IFN-I receptors in both HSC-derived and resident cells for lymphoid tissues (Fig 5C). Therefore, these results indicate that IFN-I signal in some leukocytes derived from the HSC lineage plays a crucial role in early recruitment of CD11b+Ly-6Chi monocytes in the vaginal tract with relatively less contribution to recruitment of CD11b+Ly-6Chi monocytes in draining LN, even though specific leukocytes derived from the HSC lineage to involve in the recruitment of Ly-6Chi monocytes were not defined. Another intriguing result is that, unlike vaginal CD11b+Ly-6Chi monocyte recruitment, the recruitment of NK cells in the vaginal tract and iliac LN obviously depended dominantly on IFN-I receptor in resident cells; hence, KO-WT chimera showed completely recovered recruitment of CD3−NK1.1+DX5+ NK cells in vaginal tract and iliac LN, compared to diminished recruitment of NK cells in KO-KO and WT-KO chimeras (Fig 5D and 5E). In addition, it was likely that recovered recruitment of CD11b+Ly-6Chi monocytes and NK cells in WT-KO and KO-WT chimeras contributed to the control of HSV-1 replication in the vaginal tract (Fig 5F). Collectively, these results suggest that IFN-I signaling in resident cells is dominantly responsible for recruiting NK cells in inflammatory mucosal tissues, in contrast to CD11b+Ly-6Chi monocytes that depend on the IFN-I signal in BM-derived HSCs.

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