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Cyclooxygenase-2 and Prostaglandin E2 Signaling through Prostaglandin Receptor EP-2 Favor the Development of Myocarditis during Acute Trypanosoma cruzi Infection.

Guerrero NA, Camacho M, Vila L, Íñiguez MA, Chillón-Marinas C, Cuervo H, Poveda C, Fresno M, Gironès N - PLoS Negl Trop Dis (2015)

Bottom Line: Our results show an increase in the expression of several of these enzymes in acute T. cruzi infected heart.T. cruzi infections in COX-2 or PGE2-dependent prostaglandin receptor EP-2 deficient mice indicate that both, COX-2 and EP-2 signaling contribute significantly to the heart leukocyte infiltration and to the release of chemokines and inflammatory cytokines in the heart of T. cruzi infected mice.In conclusion, COX-2 plays a detrimental role in acute Chagas disease myocarditis and points to COX-2 as a potential target for immune intervention.

View Article: PubMed Central - PubMed

Affiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Madrid, Spain.

ABSTRACT
Inflammation plays an important role in the pathophysiology of Chagas disease, caused by Trypanosoma cruzi. Prostanoids are regulators of homeostasis and inflammation and are produced mainly by myeloid cells, being cyclooxygenases, COX-1 and COX-2, the key enzymes in their biosynthesis from arachidonic acid (AA). Here, we have investigated the expression of enzymes involved in AA metabolism during T. cruzi infection. Our results show an increase in the expression of several of these enzymes in acute T. cruzi infected heart. Interestingly, COX-2 was expressed by CD68+ myeloid heart-infiltrating cells. In addition, infiltrating myeloid CD11b+Ly6G- cells purified from infected heart tissue express COX-2 and produce prostaglandin E2 (PGE2) ex vivo. T. cruzi infections in COX-2 or PGE2-dependent prostaglandin receptor EP-2 deficient mice indicate that both, COX-2 and EP-2 signaling contribute significantly to the heart leukocyte infiltration and to the release of chemokines and inflammatory cytokines in the heart of T. cruzi infected mice. In conclusion, COX-2 plays a detrimental role in acute Chagas disease myocarditis and points to COX-2 as a potential target for immune intervention.

No MeSH data available.


Related in: MedlinePlus

COX-2 expression and activity in heart and infiltrating myeloid cells during T. cruzi infection.(A) Purified Ly6G+, Ly6G-CD11b+and Ly6G-CD11b- cells were obtained from infected C57BL/6 (14 d.p.i.) and BALB/c (21 d.p.i.) mice hearts, by magnetic cell separation. Arg-1, iNOS and COX-2 levels were analyzed by Western blot. Protein levels of Actin are shown as loading control. A representative experiment of the two performed is shown. (B) CD11b+ cells were obtained from pooled infected BALB/c mouse hearts or blood at 21 d.p.i (n = 15). Ptgs2 (COX-2) expression in blood and heart tissue was determined by real time qRT-PCR. Mean ± SEM of two independent experiments is shown. (C) PGE2 and 6-oxo-PGF1α levels in total heart extracts of BALB/c from non-infected (0 d.p.i.) and 21 d.p.i., were determined as described in Methods and represented as scaled imputed data (ScaledImpData) after normalizing raw data values respect to median values of each day run (*p<0.05). Prostanoid production in purified Ly6G+ (D) and Ly6G- CD11b+ (E) cells from BALB/c mice hearts at 21 d.p.i. was determined by incubation with labeled 25 μM [C14] AA and analysis by HPLC (black line and text). Prostanoid standards (red line and text) were run in parallel as described in Methods. A representative experiment out of two performed is shown.
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pntd.0004025.g002: COX-2 expression and activity in heart and infiltrating myeloid cells during T. cruzi infection.(A) Purified Ly6G+, Ly6G-CD11b+and Ly6G-CD11b- cells were obtained from infected C57BL/6 (14 d.p.i.) and BALB/c (21 d.p.i.) mice hearts, by magnetic cell separation. Arg-1, iNOS and COX-2 levels were analyzed by Western blot. Protein levels of Actin are shown as loading control. A representative experiment of the two performed is shown. (B) CD11b+ cells were obtained from pooled infected BALB/c mouse hearts or blood at 21 d.p.i (n = 15). Ptgs2 (COX-2) expression in blood and heart tissue was determined by real time qRT-PCR. Mean ± SEM of two independent experiments is shown. (C) PGE2 and 6-oxo-PGF1α levels in total heart extracts of BALB/c from non-infected (0 d.p.i.) and 21 d.p.i., were determined as described in Methods and represented as scaled imputed data (ScaledImpData) after normalizing raw data values respect to median values of each day run (*p<0.05). Prostanoid production in purified Ly6G+ (D) and Ly6G- CD11b+ (E) cells from BALB/c mice hearts at 21 d.p.i. was determined by incubation with labeled 25 μM [C14] AA and analysis by HPLC (black line and text). Prostanoid standards (red line and text) were run in parallel as described in Methods. A representative experiment out of two performed is shown.

Mentions: We next isolated myeloid cells from hearts of T. cruzi infected C57BL/6 and BALB/c mice at the times,14 and 21 d.p.i. respectively, when maximum Arg-1 and inducible nitric oxide synthase (iNOS) expression is observed [7]. Using anti-Ly6G antibody labeled magnetic microbeads we obtained the Ly6G+ population. CD11b+ cells were selected from the remaining Ly6G- population, (Fig 2A). As previously described [7], the CD11b+Ly6G- cell population expressed iNOS and Arg-1, and here we show that they also expressed COX-2 (Fig 2A). Interestingly, COX-2 gene expression was much higher in CD11b+ cells obtained from infected cardiac tissue than those from the blood (Fig 2B), pointing to infiltrating myeloid cells in inflamed tissue as the source of COX-2.


Cyclooxygenase-2 and Prostaglandin E2 Signaling through Prostaglandin Receptor EP-2 Favor the Development of Myocarditis during Acute Trypanosoma cruzi Infection.

Guerrero NA, Camacho M, Vila L, Íñiguez MA, Chillón-Marinas C, Cuervo H, Poveda C, Fresno M, Gironès N - PLoS Negl Trop Dis (2015)

COX-2 expression and activity in heart and infiltrating myeloid cells during T. cruzi infection.(A) Purified Ly6G+, Ly6G-CD11b+and Ly6G-CD11b- cells were obtained from infected C57BL/6 (14 d.p.i.) and BALB/c (21 d.p.i.) mice hearts, by magnetic cell separation. Arg-1, iNOS and COX-2 levels were analyzed by Western blot. Protein levels of Actin are shown as loading control. A representative experiment of the two performed is shown. (B) CD11b+ cells were obtained from pooled infected BALB/c mouse hearts or blood at 21 d.p.i (n = 15). Ptgs2 (COX-2) expression in blood and heart tissue was determined by real time qRT-PCR. Mean ± SEM of two independent experiments is shown. (C) PGE2 and 6-oxo-PGF1α levels in total heart extracts of BALB/c from non-infected (0 d.p.i.) and 21 d.p.i., were determined as described in Methods and represented as scaled imputed data (ScaledImpData) after normalizing raw data values respect to median values of each day run (*p<0.05). Prostanoid production in purified Ly6G+ (D) and Ly6G- CD11b+ (E) cells from BALB/c mice hearts at 21 d.p.i. was determined by incubation with labeled 25 μM [C14] AA and analysis by HPLC (black line and text). Prostanoid standards (red line and text) were run in parallel as described in Methods. A representative experiment out of two performed is shown.
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pntd.0004025.g002: COX-2 expression and activity in heart and infiltrating myeloid cells during T. cruzi infection.(A) Purified Ly6G+, Ly6G-CD11b+and Ly6G-CD11b- cells were obtained from infected C57BL/6 (14 d.p.i.) and BALB/c (21 d.p.i.) mice hearts, by magnetic cell separation. Arg-1, iNOS and COX-2 levels were analyzed by Western blot. Protein levels of Actin are shown as loading control. A representative experiment of the two performed is shown. (B) CD11b+ cells were obtained from pooled infected BALB/c mouse hearts or blood at 21 d.p.i (n = 15). Ptgs2 (COX-2) expression in blood and heart tissue was determined by real time qRT-PCR. Mean ± SEM of two independent experiments is shown. (C) PGE2 and 6-oxo-PGF1α levels in total heart extracts of BALB/c from non-infected (0 d.p.i.) and 21 d.p.i., were determined as described in Methods and represented as scaled imputed data (ScaledImpData) after normalizing raw data values respect to median values of each day run (*p<0.05). Prostanoid production in purified Ly6G+ (D) and Ly6G- CD11b+ (E) cells from BALB/c mice hearts at 21 d.p.i. was determined by incubation with labeled 25 μM [C14] AA and analysis by HPLC (black line and text). Prostanoid standards (red line and text) were run in parallel as described in Methods. A representative experiment out of two performed is shown.
Mentions: We next isolated myeloid cells from hearts of T. cruzi infected C57BL/6 and BALB/c mice at the times,14 and 21 d.p.i. respectively, when maximum Arg-1 and inducible nitric oxide synthase (iNOS) expression is observed [7]. Using anti-Ly6G antibody labeled magnetic microbeads we obtained the Ly6G+ population. CD11b+ cells were selected from the remaining Ly6G- population, (Fig 2A). As previously described [7], the CD11b+Ly6G- cell population expressed iNOS and Arg-1, and here we show that they also expressed COX-2 (Fig 2A). Interestingly, COX-2 gene expression was much higher in CD11b+ cells obtained from infected cardiac tissue than those from the blood (Fig 2B), pointing to infiltrating myeloid cells in inflamed tissue as the source of COX-2.

Bottom Line: Our results show an increase in the expression of several of these enzymes in acute T. cruzi infected heart.T. cruzi infections in COX-2 or PGE2-dependent prostaglandin receptor EP-2 deficient mice indicate that both, COX-2 and EP-2 signaling contribute significantly to the heart leukocyte infiltration and to the release of chemokines and inflammatory cytokines in the heart of T. cruzi infected mice.In conclusion, COX-2 plays a detrimental role in acute Chagas disease myocarditis and points to COX-2 as a potential target for immune intervention.

View Article: PubMed Central - PubMed

Affiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Madrid, Spain.

ABSTRACT
Inflammation plays an important role in the pathophysiology of Chagas disease, caused by Trypanosoma cruzi. Prostanoids are regulators of homeostasis and inflammation and are produced mainly by myeloid cells, being cyclooxygenases, COX-1 and COX-2, the key enzymes in their biosynthesis from arachidonic acid (AA). Here, we have investigated the expression of enzymes involved in AA metabolism during T. cruzi infection. Our results show an increase in the expression of several of these enzymes in acute T. cruzi infected heart. Interestingly, COX-2 was expressed by CD68+ myeloid heart-infiltrating cells. In addition, infiltrating myeloid CD11b+Ly6G- cells purified from infected heart tissue express COX-2 and produce prostaglandin E2 (PGE2) ex vivo. T. cruzi infections in COX-2 or PGE2-dependent prostaglandin receptor EP-2 deficient mice indicate that both, COX-2 and EP-2 signaling contribute significantly to the heart leukocyte infiltration and to the release of chemokines and inflammatory cytokines in the heart of T. cruzi infected mice. In conclusion, COX-2 plays a detrimental role in acute Chagas disease myocarditis and points to COX-2 as a potential target for immune intervention.

No MeSH data available.


Related in: MedlinePlus