Limits...
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.

Show MeSH

Related in: MedlinePlus

CCL2 produced from infiltrated leukocytes derived from HSC lineage plays a dominant role in mucosal recruitment of CD11b+Ly-6Chi monocytes.BM cells from BL/6 (WT) or CCL2 KO (KO) mice were grafted to lethally irradiated BL/6 or CCL2 KO recipient mice, which were infected i.vag. with HSV-1. (A,B) CD11b+Ly-6Chi infiltration of infected recipients in vaginal tract. Cells were prepared from the vaginal tract 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. (C,D) Frequency and absolute number of infiltrated NK cells in vaginal tract of the recipients. Cells prepared from the vaginal tract of the recipients were used to analyze the frequency (C) and total number (D) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (E) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (F) 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.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4664252&req=5

ppat.1005256.g006: CCL2 produced from infiltrated leukocytes derived from HSC lineage plays a dominant role in mucosal recruitment of CD11b+Ly-6Chi monocytes.BM cells from BL/6 (WT) or CCL2 KO (KO) mice were grafted to lethally irradiated BL/6 or CCL2 KO recipient mice, which were infected i.vag. with HSV-1. (A,B) CD11b+Ly-6Chi infiltration of infected recipients in vaginal tract. Cells were prepared from the vaginal tract 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. (C,D) Frequency and absolute number of infiltrated NK cells in vaginal tract of the recipients. Cells prepared from the vaginal tract of the recipients were used to analyze the frequency (C) and total number (D) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (E) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (F) 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: To further confirm the differential dependence of CD11b+Ly-6Chi monocyte and NK cell infiltration on CC chemokines produced by HSC-derived leukocytes and mucosal resident cells, we used BM chimeric model of WT BL/6 and CCL2 KO mice. Since CCL2 KO recipients of WT BM donor cells (WT-KO) displayed a significantly higher frequency of vaginal CD11b+Ly-6Chi monocytes compared to CCL2 KO recipients of CCL2 KO BM donor cells (KO-KO) and WT recipients of CCL2 KO BM donor cells (KO-WT), CCL2 produced by HSC-derived leukocytes appeared to play a dominant role in the early recruitment of CD11b+Ly-6Chi monocytes in vaginal tract (Fig 6A). However, CC chemokines produced by vaginal resident cells were also considered to have a residual role in the recruitment of CD11b+Ly-6Chi monocytes because WT-KO chimeras still showed a lower frequency of vaginal CD11b+Ly-6Chi monocytes than WT recipients of WT BM donor cells (WT-WT). Supporting these findings, when the absolute number of CD11b+Ly-6Chi monocytes in the vaginal tract was determined, a significant role of CCL2 produced by HSC-derived leukocytes was evident in the early recruitment of CD11b+Ly-6Chi monocytes in the vaginal tract (Fig 6B). Moreover, CCL2 produced from HSC-derived leukocytes and resident cells was shown to partially contribute to vaginal recruitment of CD11b+Ly-6Ghi neutrophils, because KO-KO chimeras showed a decreased number of vaginal CD11b+Ly-6Ghi neutrophils compared to WT-KO and KO-WT chimeric models (Fig 6B). In addition, our data revealed that CCL2 played a certain role in recruiting NK cells, because KO-KO chimeras showed diminished NK cell frequency and number compared to WT-WT chimeras (Fig 6C and 6D). However, recruitment of CD3−NK1.1+DX5+ NK cells in vaginal tract appeared to dominantly depend on CC chemokines produced by resident cells together with CCL2, because KO-WT chimeras displayed a comparable frequency of NK cells to WT-WT chimeras. Also, WT-KO chimeras showed a higher frequency of NK cells in the vaginal tract than KO-KO chimeras, which indicates that CCL2 produced by HSC-derived leukocytes contributes in part to NK cell recruitment. Viral burden in vaginal lavages reflected the differential dependence of CD11b+Ly-6Chi monocytes and NK cells on CCL2 produced from HSC-derived and resident cells (Fig 6E). We also examined the levels of secreted CC and CXC chemokines in vaginal lavages of four CCL2 KO BM chimeric models. As expected, WT-WT chimeras showed the highest CCL2 levels, which peaked at 24 h pi and declined by 48 h pi, whereas a considerable amount of secreted CCL2 protein was detected in WT-KO chimeras with a delayed pattern that started at around 24 h pi and gradually increased until 48 h pi (Fig 6F), indicating that early infiltrated HSC-derived leukocytes producing CCL2 may be evident from around 24 h pi. In contrast with IFNAR KO BM chimera experiment, secretion of CCL2 protein was evident at higher levels in KO-WT chimeras at 24 h pi compared to WT-KO chimeras. This implies that vaginal resident cells could compensate for the CCL2 supply at a certain point in the absence of CCL2 production by infiltrated leukocytes. Other CC chemokines (CCL3, CCL4, CCL5) that are involved in NK and Th CD4+ T cell recruitment were detected at higher levels in WT-WT and KO-WT chimeras compared to WT-KO and KO-KO chimeras, suggesting that the supply of those CC chemokines is mostly accomplished by vaginal resident cells after 24 h pi. Moreover, maximum secretion of CCR5 ligands (CCL3, CCL4, CCL5) was observed in vaginal tract of KO-WT chimera at around 24 h pi, which implies that partial CCL2 ablation could change sequential production of other chemokines, due to compensation roles. Collectively, these results indicate that CCL2 produced by HSC-derived leukocytes is predominantly responsible for the accumulation of CD11b+Ly-6Chi monocytes within 24 pi, whereas recruitment of NK cells in the vaginal tract appears to be mediated by the CC chemokine pool achieved by vaginal resident cells from 24 to 48 h pi, together with CCL2.


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)

CCL2 produced from infiltrated leukocytes derived from HSC lineage plays a dominant role in mucosal recruitment of CD11b+Ly-6Chi monocytes.BM cells from BL/6 (WT) or CCL2 KO (KO) mice were grafted to lethally irradiated BL/6 or CCL2 KO recipient mice, which were infected i.vag. with HSV-1. (A,B) CD11b+Ly-6Chi infiltration of infected recipients in vaginal tract. Cells were prepared from the vaginal tract 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. (C,D) Frequency and absolute number of infiltrated NK cells in vaginal tract of the recipients. Cells prepared from the vaginal tract of the recipients were used to analyze the frequency (C) and total number (D) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (E) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (F) 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.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664252&req=5

ppat.1005256.g006: CCL2 produced from infiltrated leukocytes derived from HSC lineage plays a dominant role in mucosal recruitment of CD11b+Ly-6Chi monocytes.BM cells from BL/6 (WT) or CCL2 KO (KO) mice were grafted to lethally irradiated BL/6 or CCL2 KO recipient mice, which were infected i.vag. with HSV-1. (A,B) CD11b+Ly-6Chi infiltration of infected recipients in vaginal tract. Cells were prepared from the vaginal tract 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. (C,D) Frequency and absolute number of infiltrated NK cells in vaginal tract of the recipients. Cells prepared from the vaginal tract of the recipients were used to analyze the frequency (C) and total number (D) of CD3−NK1.1+DX5+ NK cells at 48 h pi. (E) Viral titers in vaginal lavages. Viral titers in vaginal lavages collected at 48 h pi were determined by plaque assay. (F) 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: To further confirm the differential dependence of CD11b+Ly-6Chi monocyte and NK cell infiltration on CC chemokines produced by HSC-derived leukocytes and mucosal resident cells, we used BM chimeric model of WT BL/6 and CCL2 KO mice. Since CCL2 KO recipients of WT BM donor cells (WT-KO) displayed a significantly higher frequency of vaginal CD11b+Ly-6Chi monocytes compared to CCL2 KO recipients of CCL2 KO BM donor cells (KO-KO) and WT recipients of CCL2 KO BM donor cells (KO-WT), CCL2 produced by HSC-derived leukocytes appeared to play a dominant role in the early recruitment of CD11b+Ly-6Chi monocytes in vaginal tract (Fig 6A). However, CC chemokines produced by vaginal resident cells were also considered to have a residual role in the recruitment of CD11b+Ly-6Chi monocytes because WT-KO chimeras still showed a lower frequency of vaginal CD11b+Ly-6Chi monocytes than WT recipients of WT BM donor cells (WT-WT). Supporting these findings, when the absolute number of CD11b+Ly-6Chi monocytes in the vaginal tract was determined, a significant role of CCL2 produced by HSC-derived leukocytes was evident in the early recruitment of CD11b+Ly-6Chi monocytes in the vaginal tract (Fig 6B). Moreover, CCL2 produced from HSC-derived leukocytes and resident cells was shown to partially contribute to vaginal recruitment of CD11b+Ly-6Ghi neutrophils, because KO-KO chimeras showed a decreased number of vaginal CD11b+Ly-6Ghi neutrophils compared to WT-KO and KO-WT chimeric models (Fig 6B). In addition, our data revealed that CCL2 played a certain role in recruiting NK cells, because KO-KO chimeras showed diminished NK cell frequency and number compared to WT-WT chimeras (Fig 6C and 6D). However, recruitment of CD3−NK1.1+DX5+ NK cells in vaginal tract appeared to dominantly depend on CC chemokines produced by resident cells together with CCL2, because KO-WT chimeras displayed a comparable frequency of NK cells to WT-WT chimeras. Also, WT-KO chimeras showed a higher frequency of NK cells in the vaginal tract than KO-KO chimeras, which indicates that CCL2 produced by HSC-derived leukocytes contributes in part to NK cell recruitment. Viral burden in vaginal lavages reflected the differential dependence of CD11b+Ly-6Chi monocytes and NK cells on CCL2 produced from HSC-derived and resident cells (Fig 6E). We also examined the levels of secreted CC and CXC chemokines in vaginal lavages of four CCL2 KO BM chimeric models. As expected, WT-WT chimeras showed the highest CCL2 levels, which peaked at 24 h pi and declined by 48 h pi, whereas a considerable amount of secreted CCL2 protein was detected in WT-KO chimeras with a delayed pattern that started at around 24 h pi and gradually increased until 48 h pi (Fig 6F), indicating that early infiltrated HSC-derived leukocytes producing CCL2 may be evident from around 24 h pi. In contrast with IFNAR KO BM chimera experiment, secretion of CCL2 protein was evident at higher levels in KO-WT chimeras at 24 h pi compared to WT-KO chimeras. This implies that vaginal resident cells could compensate for the CCL2 supply at a certain point in the absence of CCL2 production by infiltrated leukocytes. Other CC chemokines (CCL3, CCL4, CCL5) that are involved in NK and Th CD4+ T cell recruitment were detected at higher levels in WT-WT and KO-WT chimeras compared to WT-KO and KO-KO chimeras, suggesting that the supply of those CC chemokines is mostly accomplished by vaginal resident cells after 24 h pi. Moreover, maximum secretion of CCR5 ligands (CCL3, CCL4, CCL5) was observed in vaginal tract of KO-WT chimera at around 24 h pi, which implies that partial CCL2 ablation could change sequential production of other chemokines, due to compensation roles. Collectively, these results indicate that CCL2 produced by HSC-derived leukocytes is predominantly responsible for the accumulation of CD11b+Ly-6Chi monocytes within 24 pi, whereas recruitment of NK cells in the vaginal tract appears to be mediated by the CC chemokine pool achieved by vaginal resident cells from 24 to 48 h pi, together with CCL2.

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