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Influence of Trichobilharzia regenti (Digenea: Schistosomatidae) on the defence activity of Radix lagotis (Lymnaeidae) Haemocytes.

Skála V, Černíková A, Jindrová Z, Kašný M, Vostrý M, Walker AJ, Horák P - PLoS ONE (2014)

Bottom Line: When the infection was in the patent phase, haemocyte phagocytic activity and hydrogen peroxide production were significantly reduced in infected R. lagotis when compared to uninfected counterparts, whereas haemocyte abundance increased in infected snails.At a molecular level, protein kinase C (PKC) and extracellular-signal regulated kinase (ERK) were found to play an important role in regulating these defence reactions in R. lagotis.Moreover, haemocytes from snails with patent infection displayed lower PKC and ERK activity in cell adhesion assays when compared to those from uninfected snails, which may therefore be related to the reduced defence activities of these cells.

View Article: PubMed Central - PubMed

Affiliation: Charles University in Prague, Faculty of Science, Department of Parasitology, Prague, Czech Republic.

ABSTRACT
Radix lagotis is an intermediate snail host of the nasal bird schistosome Trichobilharzia regenti. Changes in defence responses in infected snails that might be related to host-parasite compatibility are not known. This study therefore aimed to characterize R. lagotis haemocyte defence mechanisms and determine the extent to which they are modulated by T. regenti. Histological observations of R. lagotis infected with T. regenti revealed that early phases of infection were accompanied by haemocyte accumulation around the developing larvae 2-36 h post exposure (p.e.) to the parasite. At later time points, 44-92 h p.e., no haemocytes were observed around T. regenti. Additionally, microtubular aggregates likely corresponding to phagocytosed ciliary plates of T. regenti miracidia were observed within haemocytes by use of transmission electron microscopy. When the infection was in the patent phase, haemocyte phagocytic activity and hydrogen peroxide production were significantly reduced in infected R. lagotis when compared to uninfected counterparts, whereas haemocyte abundance increased in infected snails. At a molecular level, protein kinase C (PKC) and extracellular-signal regulated kinase (ERK) were found to play an important role in regulating these defence reactions in R. lagotis. Moreover, haemocytes from snails with patent infection displayed lower PKC and ERK activity in cell adhesion assays when compared to those from uninfected snails, which may therefore be related to the reduced defence activities of these cells. These data provide the first integrated insight into the immunobiology of R. lagotis and demonstrate modulation of haemocyte-mediated responses in patent T. regenti infected snails. Given that immunomodulation occurs during patency, interference of snail-host defence by T. regenti might be important for the sustained production and/or release of infective cercariae.

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PMA-stimulated H2O2 production in haemocytes from uninfected Radix lagotis, and the effect of PKC inhibition on H2O2 production.H2O2 output by haemocyte monolayers in the presence of PMA (5 µM), GF109203X (5 µM) and PMA, DMSO (vehicle) and PMA, or SSS+ alone was detected by Amplex red and the intensity of fluorescence was measured by microplate reader over 60 min. The mean (± SEM; n = 7) relative fluorescence values shown represent the increase in H2O2 production over time in the various treatments. *p<0.05, **p<0.01, ***p<0.001, for PMA values compared to basal production, and **p<0.01, ***p<0.001 for GF109203X+PMA compared to DMSO+PMA; paired t-test or paired-samples Wilcoxon test combined with Fishers's combination test.
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pone-0111696-g009: PMA-stimulated H2O2 production in haemocytes from uninfected Radix lagotis, and the effect of PKC inhibition on H2O2 production.H2O2 output by haemocyte monolayers in the presence of PMA (5 µM), GF109203X (5 µM) and PMA, DMSO (vehicle) and PMA, or SSS+ alone was detected by Amplex red and the intensity of fluorescence was measured by microplate reader over 60 min. The mean (± SEM; n = 7) relative fluorescence values shown represent the increase in H2O2 production over time in the various treatments. *p<0.05, **p<0.01, ***p<0.001, for PMA values compared to basal production, and **p<0.01, ***p<0.001 for GF109203X+PMA compared to DMSO+PMA; paired t-test or paired-samples Wilcoxon test combined with Fishers's combination test.

Mentions: Treatment of haemocytes from uninfected R. lagotis with 5 µM PMA resulted in a 212% increase in H2O2 production after 60 min in contrast to an 80% increase in the absence of PMA; thus at this time point PMA stimulated H2O2 output approximately 2.6-fold when compared to controls (p<0.001; Figure 9). Next, the ability of PKC (GF109203X; 5 µM) and MEK (U0126; 5 µM) inhibitors to affect haemocyte H2O2 production was tested. GF109203X substantially attenuated H2O2 release by PMA-stimulated haemocytes when compared to haemocytes treated with DMSO (vehicle) and PMA at all time points (p<0.01, p<0.001; Figure 9), reducing H2O2 output to levels similar to those seen under basal conditions. In addition, DMSO did not significantly affect PMA-stimulated H2O2 production when compared to that of cells treated with PMA only. U0126 also significantly reduced PMA-stimulated H2O2 production by R. lagotis haemocytes (Figure 10). After 60 min, the increase in H2O2 production as a result of PMA exposure was reduced by 37% (p<0.001; Figure 10).


Influence of Trichobilharzia regenti (Digenea: Schistosomatidae) on the defence activity of Radix lagotis (Lymnaeidae) Haemocytes.

Skála V, Černíková A, Jindrová Z, Kašný M, Vostrý M, Walker AJ, Horák P - PLoS ONE (2014)

PMA-stimulated H2O2 production in haemocytes from uninfected Radix lagotis, and the effect of PKC inhibition on H2O2 production.H2O2 output by haemocyte monolayers in the presence of PMA (5 µM), GF109203X (5 µM) and PMA, DMSO (vehicle) and PMA, or SSS+ alone was detected by Amplex red and the intensity of fluorescence was measured by microplate reader over 60 min. The mean (± SEM; n = 7) relative fluorescence values shown represent the increase in H2O2 production over time in the various treatments. *p<0.05, **p<0.01, ***p<0.001, for PMA values compared to basal production, and **p<0.01, ***p<0.001 for GF109203X+PMA compared to DMSO+PMA; paired t-test or paired-samples Wilcoxon test combined with Fishers's combination test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4221104&req=5

pone-0111696-g009: PMA-stimulated H2O2 production in haemocytes from uninfected Radix lagotis, and the effect of PKC inhibition on H2O2 production.H2O2 output by haemocyte monolayers in the presence of PMA (5 µM), GF109203X (5 µM) and PMA, DMSO (vehicle) and PMA, or SSS+ alone was detected by Amplex red and the intensity of fluorescence was measured by microplate reader over 60 min. The mean (± SEM; n = 7) relative fluorescence values shown represent the increase in H2O2 production over time in the various treatments. *p<0.05, **p<0.01, ***p<0.001, for PMA values compared to basal production, and **p<0.01, ***p<0.001 for GF109203X+PMA compared to DMSO+PMA; paired t-test or paired-samples Wilcoxon test combined with Fishers's combination test.
Mentions: Treatment of haemocytes from uninfected R. lagotis with 5 µM PMA resulted in a 212% increase in H2O2 production after 60 min in contrast to an 80% increase in the absence of PMA; thus at this time point PMA stimulated H2O2 output approximately 2.6-fold when compared to controls (p<0.001; Figure 9). Next, the ability of PKC (GF109203X; 5 µM) and MEK (U0126; 5 µM) inhibitors to affect haemocyte H2O2 production was tested. GF109203X substantially attenuated H2O2 release by PMA-stimulated haemocytes when compared to haemocytes treated with DMSO (vehicle) and PMA at all time points (p<0.01, p<0.001; Figure 9), reducing H2O2 output to levels similar to those seen under basal conditions. In addition, DMSO did not significantly affect PMA-stimulated H2O2 production when compared to that of cells treated with PMA only. U0126 also significantly reduced PMA-stimulated H2O2 production by R. lagotis haemocytes (Figure 10). After 60 min, the increase in H2O2 production as a result of PMA exposure was reduced by 37% (p<0.001; Figure 10).

Bottom Line: When the infection was in the patent phase, haemocyte phagocytic activity and hydrogen peroxide production were significantly reduced in infected R. lagotis when compared to uninfected counterparts, whereas haemocyte abundance increased in infected snails.At a molecular level, protein kinase C (PKC) and extracellular-signal regulated kinase (ERK) were found to play an important role in regulating these defence reactions in R. lagotis.Moreover, haemocytes from snails with patent infection displayed lower PKC and ERK activity in cell adhesion assays when compared to those from uninfected snails, which may therefore be related to the reduced defence activities of these cells.

View Article: PubMed Central - PubMed

Affiliation: Charles University in Prague, Faculty of Science, Department of Parasitology, Prague, Czech Republic.

ABSTRACT
Radix lagotis is an intermediate snail host of the nasal bird schistosome Trichobilharzia regenti. Changes in defence responses in infected snails that might be related to host-parasite compatibility are not known. This study therefore aimed to characterize R. lagotis haemocyte defence mechanisms and determine the extent to which they are modulated by T. regenti. Histological observations of R. lagotis infected with T. regenti revealed that early phases of infection were accompanied by haemocyte accumulation around the developing larvae 2-36 h post exposure (p.e.) to the parasite. At later time points, 44-92 h p.e., no haemocytes were observed around T. regenti. Additionally, microtubular aggregates likely corresponding to phagocytosed ciliary plates of T. regenti miracidia were observed within haemocytes by use of transmission electron microscopy. When the infection was in the patent phase, haemocyte phagocytic activity and hydrogen peroxide production were significantly reduced in infected R. lagotis when compared to uninfected counterparts, whereas haemocyte abundance increased in infected snails. At a molecular level, protein kinase C (PKC) and extracellular-signal regulated kinase (ERK) were found to play an important role in regulating these defence reactions in R. lagotis. Moreover, haemocytes from snails with patent infection displayed lower PKC and ERK activity in cell adhesion assays when compared to those from uninfected snails, which may therefore be related to the reduced defence activities of these cells. These data provide the first integrated insight into the immunobiology of R. lagotis and demonstrate modulation of haemocyte-mediated responses in patent T. regenti infected snails. Given that immunomodulation occurs during patency, interference of snail-host defence by T. regenti might be important for the sustained production and/or release of infective cercariae.

Show MeSH
Related in: MedlinePlus