Limits...
Dynamic imaging of experimental Leishmania donovani-induced hepatic granulomas detects Kupffer cell-restricted antigen presentation to antigen-specific CD8 T cells.

Beattie L, Peltan A, Maroof A, Kirby A, Brown N, Coles M, Smith DF, Kaye PM - PLoS Pathog. (2010)

Bottom Line: Kupffer cells (KCs) represent the major phagocytic population within the liver and provide an intracellular niche for the survival of a number of important human pathogens.Although KCs have been extensively studied in vitro, little is known of their in vivo response to infection and their capacity to directly interact with antigen-specific CD8(+) T cells.This restriction of antigen presentation to KCs within the Leishmania granuloma has important implications for the identification of new candidate vaccine antigens and for the design of novel immuno-therapeutic interventions.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Centre for Immunology and Infection, Hull York Medical School, University of York, York, United Kingdom.

ABSTRACT
Kupffer cells (KCs) represent the major phagocytic population within the liver and provide an intracellular niche for the survival of a number of important human pathogens. Although KCs have been extensively studied in vitro, little is known of their in vivo response to infection and their capacity to directly interact with antigen-specific CD8(+) T cells. Here, using a combination of approaches including whole mount and thin section confocal microscopy, adoptive cell transfer and intra-vital 2-photon microscopy, we demonstrate that KCs represent the only detectable population of mononuclear phagocytes within granulomas induced by Leishmania donovani infection that are capable of presenting parasite-derived peptide to effector CD8(+) T cells. This restriction of antigen presentation to KCs within the Leishmania granuloma has important implications for the identification of new candidate vaccine antigens and for the design of novel immuno-therapeutic interventions.

Show MeSH

Related in: MedlinePlus

CD8+ T cell dynamics in the liver following L. donovani infection.A) The entrance and B) exit rate of CFSE labelled memory-like OT-I T cells 5–14 h post-transfer into d14–21 infected hCD2.GFP mice calculated by dividing the number of OT-I cells entering or leaving each granuloma for each imaging session and dividing by the time of each imaging session to give a rate/min (n = 60 imaging sessions for WT and 71 for PINK infected mice, ** < 0.001). C) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of CFSE labelled memory-like OT-I T cells transferred into d14-21 WT L. donovani or D) PINK-infected mice. Comparison of the E) cell velocities, F) meandering index and G) track length of memory-like OT-I T cells transferred into d14-21 WT L. donovani- or PINK-infected mice (n = 266 for WT and 311 for PINK, *** P<0.0001, ** P<0.001). H) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of Hoechst labelled memory-like OT-I T cells (blue tracks) and CFSE labelled memory-like F5 cells (green) transferred into d14-21 PINK infected mice. I) Comparison of the cell velocities of memory-like F5 and OT-I T cells transferred into d14-21 PINK-infected mice (n = 105 for F5 and 87 for OT-I T cells). Data represents mean ± SEM, ** P<0.001.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2837408&req=5

ppat-1000805-g007: CD8+ T cell dynamics in the liver following L. donovani infection.A) The entrance and B) exit rate of CFSE labelled memory-like OT-I T cells 5–14 h post-transfer into d14–21 infected hCD2.GFP mice calculated by dividing the number of OT-I cells entering or leaving each granuloma for each imaging session and dividing by the time of each imaging session to give a rate/min (n = 60 imaging sessions for WT and 71 for PINK infected mice, ** < 0.001). C) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of CFSE labelled memory-like OT-I T cells transferred into d14-21 WT L. donovani or D) PINK-infected mice. Comparison of the E) cell velocities, F) meandering index and G) track length of memory-like OT-I T cells transferred into d14-21 WT L. donovani- or PINK-infected mice (n = 266 for WT and 311 for PINK, *** P<0.0001, ** P<0.001). H) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of Hoechst labelled memory-like OT-I T cells (blue tracks) and CFSE labelled memory-like F5 cells (green) transferred into d14-21 PINK infected mice. I) Comparison of the cell velocities of memory-like F5 and OT-I T cells transferred into d14-21 PINK-infected mice (n = 105 for F5 and 87 for OT-I T cells). Data represents mean ± SEM, ** P<0.001.

Mentions: Altered accumulation of CD8+ T cells within granulomas could be the result of altered rates of immigration or emigration. To distinguish between these possibilities, we examined the dynamics of OT-I T cell movement within individual granulomas in WT L. donovani and PINK-infected mice 5–14 h post-transfer of OT-I T cells. We calculated the rate at which OT-I T cells entered granulomas by dividing the number of cells entering or exiting the granuloma in each imaging period by the length of the imaging period in minutes. No significant differences were seen in the rate at which OT-I T cells entered granulomas in WT L. donovani- and PINK-infected mice (Figure 7A). In contrast, the rate at which OT-I T cells left granulomas in PINK-infected mice was slower than in WT L. donovani-infected mice (Figure 7B). The finding that exit rate, but not entrance rate, was influenced by the presence or absence of cognate antigen suggested that OT-I T cells behaved differently if antigen was available. To determine whether this was reflected in altered velocity, we calculated the average velocity of OT-I cells (n = 311 cells from 43 imaging fields) in PINK-infected and OT-I cells (n = 266 cells from 48 imaging fields) in WT L. donovani-infected hCD2.VaDs Red mice (here used to identify the border of the granuloma by endogenous labelling of all other T cells). The results of this analysis demonstrated that OT-I T cells moved significantly more slowly in the presence of cognate antigen (Figure 7C, D, E and Video S5). The meandering index (calculated by diving the displacement of the cell from its original starting point by the total track length of that cell) was significantly higher for OT-I T cells transferred into PINK-infected mice than those transferred into WT L. donovani- infected mice (Figure 7E). This was reflected by significantly lower track lengths for OT-I cells transferred into PINK infected mice and therefore in the presence of cognate antigen (Figure 7F).


Dynamic imaging of experimental Leishmania donovani-induced hepatic granulomas detects Kupffer cell-restricted antigen presentation to antigen-specific CD8 T cells.

Beattie L, Peltan A, Maroof A, Kirby A, Brown N, Coles M, Smith DF, Kaye PM - PLoS Pathog. (2010)

CD8+ T cell dynamics in the liver following L. donovani infection.A) The entrance and B) exit rate of CFSE labelled memory-like OT-I T cells 5–14 h post-transfer into d14–21 infected hCD2.GFP mice calculated by dividing the number of OT-I cells entering or leaving each granuloma for each imaging session and dividing by the time of each imaging session to give a rate/min (n = 60 imaging sessions for WT and 71 for PINK infected mice, ** < 0.001). C) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of CFSE labelled memory-like OT-I T cells transferred into d14-21 WT L. donovani or D) PINK-infected mice. Comparison of the E) cell velocities, F) meandering index and G) track length of memory-like OT-I T cells transferred into d14-21 WT L. donovani- or PINK-infected mice (n = 266 for WT and 311 for PINK, *** P<0.0001, ** P<0.001). H) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of Hoechst labelled memory-like OT-I T cells (blue tracks) and CFSE labelled memory-like F5 cells (green) transferred into d14-21 PINK infected mice. I) Comparison of the cell velocities of memory-like F5 and OT-I T cells transferred into d14-21 PINK-infected mice (n = 105 for F5 and 87 for OT-I T cells). Data represents mean ± SEM, ** P<0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2837408&req=5

ppat-1000805-g007: CD8+ T cell dynamics in the liver following L. donovani infection.A) The entrance and B) exit rate of CFSE labelled memory-like OT-I T cells 5–14 h post-transfer into d14–21 infected hCD2.GFP mice calculated by dividing the number of OT-I cells entering or leaving each granuloma for each imaging session and dividing by the time of each imaging session to give a rate/min (n = 60 imaging sessions for WT and 71 for PINK infected mice, ** < 0.001). C) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of CFSE labelled memory-like OT-I T cells transferred into d14-21 WT L. donovani or D) PINK-infected mice. Comparison of the E) cell velocities, F) meandering index and G) track length of memory-like OT-I T cells transferred into d14-21 WT L. donovani- or PINK-infected mice (n = 266 for WT and 311 for PINK, *** P<0.0001, ** P<0.001). H) Snapshot of the extended focus view of a time-lapse imaging sequence showing the cell tracks of Hoechst labelled memory-like OT-I T cells (blue tracks) and CFSE labelled memory-like F5 cells (green) transferred into d14-21 PINK infected mice. I) Comparison of the cell velocities of memory-like F5 and OT-I T cells transferred into d14-21 PINK-infected mice (n = 105 for F5 and 87 for OT-I T cells). Data represents mean ± SEM, ** P<0.001.
Mentions: Altered accumulation of CD8+ T cells within granulomas could be the result of altered rates of immigration or emigration. To distinguish between these possibilities, we examined the dynamics of OT-I T cell movement within individual granulomas in WT L. donovani and PINK-infected mice 5–14 h post-transfer of OT-I T cells. We calculated the rate at which OT-I T cells entered granulomas by dividing the number of cells entering or exiting the granuloma in each imaging period by the length of the imaging period in minutes. No significant differences were seen in the rate at which OT-I T cells entered granulomas in WT L. donovani- and PINK-infected mice (Figure 7A). In contrast, the rate at which OT-I T cells left granulomas in PINK-infected mice was slower than in WT L. donovani-infected mice (Figure 7B). The finding that exit rate, but not entrance rate, was influenced by the presence or absence of cognate antigen suggested that OT-I T cells behaved differently if antigen was available. To determine whether this was reflected in altered velocity, we calculated the average velocity of OT-I cells (n = 311 cells from 43 imaging fields) in PINK-infected and OT-I cells (n = 266 cells from 48 imaging fields) in WT L. donovani-infected hCD2.VaDs Red mice (here used to identify the border of the granuloma by endogenous labelling of all other T cells). The results of this analysis demonstrated that OT-I T cells moved significantly more slowly in the presence of cognate antigen (Figure 7C, D, E and Video S5). The meandering index (calculated by diving the displacement of the cell from its original starting point by the total track length of that cell) was significantly higher for OT-I T cells transferred into PINK-infected mice than those transferred into WT L. donovani- infected mice (Figure 7E). This was reflected by significantly lower track lengths for OT-I cells transferred into PINK infected mice and therefore in the presence of cognate antigen (Figure 7F).

Bottom Line: Kupffer cells (KCs) represent the major phagocytic population within the liver and provide an intracellular niche for the survival of a number of important human pathogens.Although KCs have been extensively studied in vitro, little is known of their in vivo response to infection and their capacity to directly interact with antigen-specific CD8(+) T cells.This restriction of antigen presentation to KCs within the Leishmania granuloma has important implications for the identification of new candidate vaccine antigens and for the design of novel immuno-therapeutic interventions.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Centre for Immunology and Infection, Hull York Medical School, University of York, York, United Kingdom.

ABSTRACT
Kupffer cells (KCs) represent the major phagocytic population within the liver and provide an intracellular niche for the survival of a number of important human pathogens. Although KCs have been extensively studied in vitro, little is known of their in vivo response to infection and their capacity to directly interact with antigen-specific CD8(+) T cells. Here, using a combination of approaches including whole mount and thin section confocal microscopy, adoptive cell transfer and intra-vital 2-photon microscopy, we demonstrate that KCs represent the only detectable population of mononuclear phagocytes within granulomas induced by Leishmania donovani infection that are capable of presenting parasite-derived peptide to effector CD8(+) T cells. This restriction of antigen presentation to KCs within the Leishmania granuloma has important implications for the identification of new candidate vaccine antigens and for the design of novel immuno-therapeutic interventions.

Show MeSH
Related in: MedlinePlus