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Persistent and compartmentalised disruption of dendritic cell subpopulations in the lung following influenza A virus infection.

Strickland DH, Fear V, Shenton S, Wikstrom ME, Zosky G, Larcombe AN, Holt PG, Berry C, von Garnier C, Stumbles PA - PLoS ONE (2014)

Bottom Line: A significant depletion in the percentage of AMDC was observed at day 4 post-infection that was associated with a change in steady-state CD11b+ and CD11b- AMDC subset frequencies and significantly elevated CD40 and CD80 expression and that returned to baseline by day 14 post-infection.In contrast, percentages and total numbers of PLDC were significantly elevated at day 14 and remained so until day 21 post-infection.Accompanying this was a change in CD11b+and CD11b- PLDC subset frequencies and significant increase in CD40 and CD80 expression at these time points.

View Article: PubMed Central - PubMed

Affiliation: Telethon Institute for Child Health Research and Centre for Child Health Research, University of Western Australia, Perth, W.A., Australia.

ABSTRACT
Immunological homeostasis in the respiratory tract is thought to require balanced interactions between networks of dendritic cell (DC) subsets in lung microenvironments in order to regulate tolerance or immunity to inhaled antigens and pathogens. Influenza A virus (IAV) poses a serious threat of long-term disruption to this balance through its potent pro-inflammatory activities. In this study, we have used a BALB/c mouse model of A/PR8/34 H1N1 Influenza Type A Virus infection to examine the effects of IAV on respiratory tissue DC subsets during the recovery phase following clearance of the virus. In adult mice, we found differences in the kinetics and activation states of DC residing in the airway mucosa (AMDC) compared to those in the parenchymal lung (PLDC) compartments. A significant depletion in the percentage of AMDC was observed at day 4 post-infection that was associated with a change in steady-state CD11b+ and CD11b- AMDC subset frequencies and significantly elevated CD40 and CD80 expression and that returned to baseline by day 14 post-infection. In contrast, percentages and total numbers of PLDC were significantly elevated at day 14 and remained so until day 21 post-infection. Accompanying this was a change in CD11b+and CD11b- PLDC subset frequencies and significant increase in CD40 and CD80 expression at these time points. Furthermore, mice infected with IAV at 4 weeks of age showed a significant increase in total numbers of PLDC, and increased CD40 expression on both AMDC and PLDC, when analysed as adults 35 days later. These data suggest that the rate of recovery of DC populations following IAV infection differs in the mucosal and parenchymal compartments of the lung and that DC populations can remain disrupted and activated for a prolonged period following viral clearance, into adulthood if infection occurred early in life.

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Kinetics of airway mucosal and parenchymal lung DC changes following IAV infection.(A) Representative FACS profiles showing gating for AMDC at the indicated time points p.i. (B and C) AMDC percentage frequencies (B) and total numbers (C) following IAV infection for control (open circles) and IAV infected mice (closed circles). (D) Representative FACS profiles showing gating for PLDC at the indicated time points p.i. (E and F) PLDC percentage frequencies (E) and total numbers (F) following IAV infection for control (open circles) and IAV infected mice (closed circles). (G) Representative FACS profiles showing gating for PLMac at the indicated time points p.i. (H and I) PLMac percentage frequencies (H) and total numbers (I) following IAV infection for control (open circles) and IAV infected mice (closed circles). Data are means +/− SEM for 3 independent infection experiments using pools of tissue from 3 to 4 mice for each experiment. *  =  p<0.05; **  =  p<0.01.
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pone-0111520-g004: Kinetics of airway mucosal and parenchymal lung DC changes following IAV infection.(A) Representative FACS profiles showing gating for AMDC at the indicated time points p.i. (B and C) AMDC percentage frequencies (B) and total numbers (C) following IAV infection for control (open circles) and IAV infected mice (closed circles). (D) Representative FACS profiles showing gating for PLDC at the indicated time points p.i. (E and F) PLDC percentage frequencies (E) and total numbers (F) following IAV infection for control (open circles) and IAV infected mice (closed circles). (G) Representative FACS profiles showing gating for PLMac at the indicated time points p.i. (H and I) PLMac percentage frequencies (H) and total numbers (I) following IAV infection for control (open circles) and IAV infected mice (closed circles). Data are means +/− SEM for 3 independent infection experiments using pools of tissue from 3 to 4 mice for each experiment. *  =  p<0.05; **  =  p<0.01.

Mentions: Our previous mouse studies have identified functionally distinct populations of DC in the airway mucosa (AMDC) and parenchymal lung (PLDC), with AMDC displaying more rapid turnover rates (<12 h) compared to PLDC (>7 days) and more rapid activation in response to aeroallergen challenge [15], [16]. Given that acute IAV infection is characterised by early infection and replication of the virus in epithelial cells of the airway mucosa, we initially examined the population dynamics of AMDC compared to their more peripheral PLDC counterparts following IAV infection. AMDC and PLDC were identified by flow cytometry using co-staining of tracheal and parenchymal lung tissue respectively for CD11c and MHC class II (I-A/E) as previously described [15], allowing gating of CD11c+ I-A/Ehi AMDC and PLDC following IAV infection (Figs. 4A and 4D). This combination of markers also allowed identification of CD11c+ I-A/Elow parenchymal lung macrophages (PLMac) [15], which were also tracked over the same time course post-IAV infection (Fig. 4G). In addition, as expression of I-A/E may possibly have modulated after IAV infection, we confirmed that these phenotypes remained stable by substituting the mouse DC marker CD205 for I-A/E (Fig. S1). A time course analysis of tracheal tissue showed a significant depletion of AMDC as a percentage of total cells (p<0.01) (Fig. 4A and 4B), but not total cell numbers (Fig. 4C), at day 4 p.i. with the percentages of AMDC returning to baseline levels by d7-14 p.i. In contrast, percentages (Fig. 4D and 4E) and total numbers (Fig. 4F) of PLDC in peripheral lung tissue remained unchanged from controls at d4 p.i., but were then significantly increased above control levels from d14 p.i (p <0.05). Over the same time-course, a decrease in percentages (Fig. 4G and 4H) and total numbers (Fig. 4I) of PLMac was observed from d4 p.i., with a significant decrease in the percentage of PLMac at d4 and d14 p.i. (p<0.01), returning to near-baseline levels at d21 p.i.


Persistent and compartmentalised disruption of dendritic cell subpopulations in the lung following influenza A virus infection.

Strickland DH, Fear V, Shenton S, Wikstrom ME, Zosky G, Larcombe AN, Holt PG, Berry C, von Garnier C, Stumbles PA - PLoS ONE (2014)

Kinetics of airway mucosal and parenchymal lung DC changes following IAV infection.(A) Representative FACS profiles showing gating for AMDC at the indicated time points p.i. (B and C) AMDC percentage frequencies (B) and total numbers (C) following IAV infection for control (open circles) and IAV infected mice (closed circles). (D) Representative FACS profiles showing gating for PLDC at the indicated time points p.i. (E and F) PLDC percentage frequencies (E) and total numbers (F) following IAV infection for control (open circles) and IAV infected mice (closed circles). (G) Representative FACS profiles showing gating for PLMac at the indicated time points p.i. (H and I) PLMac percentage frequencies (H) and total numbers (I) following IAV infection for control (open circles) and IAV infected mice (closed circles). Data are means +/− SEM for 3 independent infection experiments using pools of tissue from 3 to 4 mice for each experiment. *  =  p<0.05; **  =  p<0.01.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4232261&req=5

pone-0111520-g004: Kinetics of airway mucosal and parenchymal lung DC changes following IAV infection.(A) Representative FACS profiles showing gating for AMDC at the indicated time points p.i. (B and C) AMDC percentage frequencies (B) and total numbers (C) following IAV infection for control (open circles) and IAV infected mice (closed circles). (D) Representative FACS profiles showing gating for PLDC at the indicated time points p.i. (E and F) PLDC percentage frequencies (E) and total numbers (F) following IAV infection for control (open circles) and IAV infected mice (closed circles). (G) Representative FACS profiles showing gating for PLMac at the indicated time points p.i. (H and I) PLMac percentage frequencies (H) and total numbers (I) following IAV infection for control (open circles) and IAV infected mice (closed circles). Data are means +/− SEM for 3 independent infection experiments using pools of tissue from 3 to 4 mice for each experiment. *  =  p<0.05; **  =  p<0.01.
Mentions: Our previous mouse studies have identified functionally distinct populations of DC in the airway mucosa (AMDC) and parenchymal lung (PLDC), with AMDC displaying more rapid turnover rates (<12 h) compared to PLDC (>7 days) and more rapid activation in response to aeroallergen challenge [15], [16]. Given that acute IAV infection is characterised by early infection and replication of the virus in epithelial cells of the airway mucosa, we initially examined the population dynamics of AMDC compared to their more peripheral PLDC counterparts following IAV infection. AMDC and PLDC were identified by flow cytometry using co-staining of tracheal and parenchymal lung tissue respectively for CD11c and MHC class II (I-A/E) as previously described [15], allowing gating of CD11c+ I-A/Ehi AMDC and PLDC following IAV infection (Figs. 4A and 4D). This combination of markers also allowed identification of CD11c+ I-A/Elow parenchymal lung macrophages (PLMac) [15], which were also tracked over the same time course post-IAV infection (Fig. 4G). In addition, as expression of I-A/E may possibly have modulated after IAV infection, we confirmed that these phenotypes remained stable by substituting the mouse DC marker CD205 for I-A/E (Fig. S1). A time course analysis of tracheal tissue showed a significant depletion of AMDC as a percentage of total cells (p<0.01) (Fig. 4A and 4B), but not total cell numbers (Fig. 4C), at day 4 p.i. with the percentages of AMDC returning to baseline levels by d7-14 p.i. In contrast, percentages (Fig. 4D and 4E) and total numbers (Fig. 4F) of PLDC in peripheral lung tissue remained unchanged from controls at d4 p.i., but were then significantly increased above control levels from d14 p.i (p <0.05). Over the same time-course, a decrease in percentages (Fig. 4G and 4H) and total numbers (Fig. 4I) of PLMac was observed from d4 p.i., with a significant decrease in the percentage of PLMac at d4 and d14 p.i. (p<0.01), returning to near-baseline levels at d21 p.i.

Bottom Line: A significant depletion in the percentage of AMDC was observed at day 4 post-infection that was associated with a change in steady-state CD11b+ and CD11b- AMDC subset frequencies and significantly elevated CD40 and CD80 expression and that returned to baseline by day 14 post-infection.In contrast, percentages and total numbers of PLDC were significantly elevated at day 14 and remained so until day 21 post-infection.Accompanying this was a change in CD11b+and CD11b- PLDC subset frequencies and significant increase in CD40 and CD80 expression at these time points.

View Article: PubMed Central - PubMed

Affiliation: Telethon Institute for Child Health Research and Centre for Child Health Research, University of Western Australia, Perth, W.A., Australia.

ABSTRACT
Immunological homeostasis in the respiratory tract is thought to require balanced interactions between networks of dendritic cell (DC) subsets in lung microenvironments in order to regulate tolerance or immunity to inhaled antigens and pathogens. Influenza A virus (IAV) poses a serious threat of long-term disruption to this balance through its potent pro-inflammatory activities. In this study, we have used a BALB/c mouse model of A/PR8/34 H1N1 Influenza Type A Virus infection to examine the effects of IAV on respiratory tissue DC subsets during the recovery phase following clearance of the virus. In adult mice, we found differences in the kinetics and activation states of DC residing in the airway mucosa (AMDC) compared to those in the parenchymal lung (PLDC) compartments. A significant depletion in the percentage of AMDC was observed at day 4 post-infection that was associated with a change in steady-state CD11b+ and CD11b- AMDC subset frequencies and significantly elevated CD40 and CD80 expression and that returned to baseline by day 14 post-infection. In contrast, percentages and total numbers of PLDC were significantly elevated at day 14 and remained so until day 21 post-infection. Accompanying this was a change in CD11b+and CD11b- PLDC subset frequencies and significant increase in CD40 and CD80 expression at these time points. Furthermore, mice infected with IAV at 4 weeks of age showed a significant increase in total numbers of PLDC, and increased CD40 expression on both AMDC and PLDC, when analysed as adults 35 days later. These data suggest that the rate of recovery of DC populations following IAV infection differs in the mucosal and parenchymal compartments of the lung and that DC populations can remain disrupted and activated for a prolonged period following viral clearance, into adulthood if infection occurred early in life.

Show MeSH
Related in: MedlinePlus