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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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Sequential responses of chemokines in response to mucosal infection with HSV-1.(A) Expression of chemokine mRNA in vaginal tract of BL/6 or IFNAR KO mice following mucosal infection with HSV-1. The levels of chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from vaginal tract (VT) tissues at 12, 24, 48, and 96 h pi. (B) Secreted levels of chemokine proteins in vaginal tract. (C) Serum chemokine protein levels. The levels of chemokine proteins were determined by CBA using vaginal lavages and sera at 12, 24, 48, and 96 h pi. Data show the average ± SD of values derived from three individual experiments (n = 5–6). Areas highlighted by gray color denote interesting points. *, p<0.05; **, p<0.01; ***, p<0.001 compared with the levels in IFNAR KO mice at the indicated time point.
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ppat.1005256.g004: Sequential responses of chemokines in response to mucosal infection with HSV-1.(A) Expression of chemokine mRNA in vaginal tract of BL/6 or IFNAR KO mice following mucosal infection with HSV-1. The levels of chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from vaginal tract (VT) tissues at 12, 24, 48, and 96 h pi. (B) Secreted levels of chemokine proteins in vaginal tract. (C) Serum chemokine protein levels. The levels of chemokine proteins were determined by CBA using vaginal lavages and sera at 12, 24, 48, and 96 h pi. Data show the average ± SD of values derived from three individual experiments (n = 5–6). Areas highlighted by gray color denote interesting points. *, p<0.05; **, p<0.01; ***, p<0.001 compared with the levels in IFNAR KO mice at the indicated time point.

Mentions: Sequential responses of chemokines appear to be necessary for the selective and tailored environment of infiltrated leukocytes that provides effective protection against mucosal HSV-1 infection, even though the chemokine responses can be redundant [32,33]. Moreover, our data demonstrated that infiltration of CD11b+Ly-6Chi monocytes preceded that of NK cells during mucosal inflammation; CD11b+Ly-6Chi monocyte infiltration peaked at around 24 h pi, whereas NK cell infiltration peaked at 48 h after mucosal HSV-1 infection. Therefore, we were interested in the regulatory molecules that are involved in establishing this tailored environment of sequential CD11b+Ly-6Chi monocyte and NK cell infiltration in vaginal tissues. To this end, we performed a kinetic examination of the expression and secretion of several CC and CXC chemokines in vaginal tissues and lavages at different time points up to 96 h pi. Somewhat intriguingly, our data revealed that CCL2 and CCL3 expression was evident early, with cascade responses peaking at subsequent time points. Hence, expression of CCL2, a ligand of CCR2, began as early as 12 h pi in vaginal tract of WT BL/6 mice and increased sharply in a time-dependent manner (Fig 4A). Maximal levels of vaginal CCL2 induction in WT mice were observed at around 24 h pi and had declined 80% by 96 h pi, whereas IFNAR KO mice showed a delayed and lower expression of vaginal CCL2 with levels gradually increasing after 48 h pi. Vaginal expression of CCL3, a ligand of CCR1 and CCR5, peaked at 48 h pi in WT mice, whereas IFNAR KO mice showed a drastically decreased expression level of CCL3. Other CXC chemokines (CXCL1, CXCL2) that are involved in recruitment of neutrophils via CXCR2 showed higher expression levels in the vaginal tract of IFNAR KO mice compared to WT BL/6 mice. Notably, CXCL2 expression gradually increased in IFNAR KO mice after 48 h pi, whereas WT BL/6 mice showed decreased levels at 96 h pi. Regarding secreted chemokine proteins in vaginal lavages, ligands of CCR2 (CCL2, CCL7) were secreted at a higher level in WT BL/6 mice with reaching a maximum level at 24 h pi, compared to secretion in IFNAR KO mice (Fig 4B). In contrast, ligands of CCR5 (CCL3, CCL4, CCL5) showed delayed peaks at around 48 h pi and such levels were markedly higher in WT mice than in IFNAR KO mice. These results indicate that cascade responses of CC chemokines, especially CCL2 and CCL3, appear to be closely associated with consecutive recruitment of CD11b+Ly-6Chi monocytes and NK cells in the vaginal tract of WT BL/6 mice, whereas enhanced induction of CXC chemokine (CXCL1, CXCL2) in IFNAR KO mice appears to be involved in neutrophil recruitment in the vaginal tract. However, systemic levels of most chemokine proteins were markedly higher in the sera of IFNAR KO mice compared to WT BL/6 mice (Fig 4C). Although cellular sources for huge amount of chemokine secretion in sera of IFNAR KO mice were not defined, it is likely that various cell types derived from IFNAR KO mice, including epithelial cells, fibroblast, and endothelial cells, are highly susceptible to HSV-1 replication, thereby secreting chemokines in sera by severe inflammation in whole body. In addition, this result suggests that enhanced chemokine levels in the serum may cause disturbed trafficking of various leukocytes in IFNAR KO mice.


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)

Sequential responses of chemokines in response to mucosal infection with HSV-1.(A) Expression of chemokine mRNA in vaginal tract of BL/6 or IFNAR KO mice following mucosal infection with HSV-1. The levels of chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from vaginal tract (VT) tissues at 12, 24, 48, and 96 h pi. (B) Secreted levels of chemokine proteins in vaginal tract. (C) Serum chemokine protein levels. The levels of chemokine proteins were determined by CBA using vaginal lavages and sera at 12, 24, 48, and 96 h pi. Data show the average ± SD of values derived from three individual experiments (n = 5–6). Areas highlighted by gray color denote interesting points. *, p<0.05; **, p<0.01; ***, p<0.001 compared with the levels in IFNAR KO mice at the indicated time point.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4664252&req=5

ppat.1005256.g004: Sequential responses of chemokines in response to mucosal infection with HSV-1.(A) Expression of chemokine mRNA in vaginal tract of BL/6 or IFNAR KO mice following mucosal infection with HSV-1. The levels of chemokine mRNAs were determined by real-time qRT-PCR using total RNA extracted from vaginal tract (VT) tissues at 12, 24, 48, and 96 h pi. (B) Secreted levels of chemokine proteins in vaginal tract. (C) Serum chemokine protein levels. The levels of chemokine proteins were determined by CBA using vaginal lavages and sera at 12, 24, 48, and 96 h pi. Data show the average ± SD of values derived from three individual experiments (n = 5–6). Areas highlighted by gray color denote interesting points. *, p<0.05; **, p<0.01; ***, p<0.001 compared with the levels in IFNAR KO mice at the indicated time point.
Mentions: Sequential responses of chemokines appear to be necessary for the selective and tailored environment of infiltrated leukocytes that provides effective protection against mucosal HSV-1 infection, even though the chemokine responses can be redundant [32,33]. Moreover, our data demonstrated that infiltration of CD11b+Ly-6Chi monocytes preceded that of NK cells during mucosal inflammation; CD11b+Ly-6Chi monocyte infiltration peaked at around 24 h pi, whereas NK cell infiltration peaked at 48 h after mucosal HSV-1 infection. Therefore, we were interested in the regulatory molecules that are involved in establishing this tailored environment of sequential CD11b+Ly-6Chi monocyte and NK cell infiltration in vaginal tissues. To this end, we performed a kinetic examination of the expression and secretion of several CC and CXC chemokines in vaginal tissues and lavages at different time points up to 96 h pi. Somewhat intriguingly, our data revealed that CCL2 and CCL3 expression was evident early, with cascade responses peaking at subsequent time points. Hence, expression of CCL2, a ligand of CCR2, began as early as 12 h pi in vaginal tract of WT BL/6 mice and increased sharply in a time-dependent manner (Fig 4A). Maximal levels of vaginal CCL2 induction in WT mice were observed at around 24 h pi and had declined 80% by 96 h pi, whereas IFNAR KO mice showed a delayed and lower expression of vaginal CCL2 with levels gradually increasing after 48 h pi. Vaginal expression of CCL3, a ligand of CCR1 and CCR5, peaked at 48 h pi in WT mice, whereas IFNAR KO mice showed a drastically decreased expression level of CCL3. Other CXC chemokines (CXCL1, CXCL2) that are involved in recruitment of neutrophils via CXCR2 showed higher expression levels in the vaginal tract of IFNAR KO mice compared to WT BL/6 mice. Notably, CXCL2 expression gradually increased in IFNAR KO mice after 48 h pi, whereas WT BL/6 mice showed decreased levels at 96 h pi. Regarding secreted chemokine proteins in vaginal lavages, ligands of CCR2 (CCL2, CCL7) were secreted at a higher level in WT BL/6 mice with reaching a maximum level at 24 h pi, compared to secretion in IFNAR KO mice (Fig 4B). In contrast, ligands of CCR5 (CCL3, CCL4, CCL5) showed delayed peaks at around 48 h pi and such levels were markedly higher in WT mice than in IFNAR KO mice. These results indicate that cascade responses of CC chemokines, especially CCL2 and CCL3, appear to be closely associated with consecutive recruitment of CD11b+Ly-6Chi monocytes and NK cells in the vaginal tract of WT BL/6 mice, whereas enhanced induction of CXC chemokine (CXCL1, CXCL2) in IFNAR KO mice appears to be involved in neutrophil recruitment in the vaginal tract. However, systemic levels of most chemokine proteins were markedly higher in the sera of IFNAR KO mice compared to WT BL/6 mice (Fig 4C). Although cellular sources for huge amount of chemokine secretion in sera of IFNAR KO mice were not defined, it is likely that various cell types derived from IFNAR KO mice, including epithelial cells, fibroblast, and endothelial cells, are highly susceptible to HSV-1 replication, thereby secreting chemokines in sera by severe inflammation in whole body. In addition, this result suggests that enhanced chemokine levels in the serum may cause disturbed trafficking of various leukocytes in IFNAR KO mice.

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