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Inhibition of chemokine-glycosaminoglycan interactions in donor tissue reduces mouse allograft vasculopathy and transplant rejection.

Dai E, Liu LY, Wang H, McIvor D, Sun YM, Macaulay C, King E, Munuswamy-Ramanujam G, Bartee MY, Williams J, Davids J, Charo I, McFadden G, Esko JD, Lucas AR - PLoS ONE (2010)

Bottom Line: Donor tissue GAG or CCR2 deficiency markedly reduced inflammation and vasculopathy, whereas recipient deficiencies did not.M-T1 and M3 inhibited WT (Ccr2(+/+) and Ndst1(+/+), p< or =0.006) allograft vasculopathy, but did not block vasculopathy in Ccr2(-/-) (p = 0.61).Interruption of chemokine-GAG interactions, even in the absence of chemokine-receptor blockade, is a highly effective approach to reduction of allograft rejection, reducing vascular inflammation and prolonging allograft survival.

View Article: PubMed Central - PubMed

Affiliation: Vascular Biology Research Group, Robarts Research Institute, The University of Western Ontario, London, Ontario, Canada.

ABSTRACT

Background: Binding of chemokines to glycosaminoglycans (GAGs) is classically described as initiating inflammatory cell migration and creating tissue chemokine gradients that direct local leukocyte chemotaxis into damaged or transplanted tissues. While chemokine-receptor binding has been extensively studied during allograft transplantation, effects of glycosaminoglycan (GAG) interactions with chemokines on transplant longevity are less well known. Here we examine the impact of interrupting chemokine-GAG interactions and chemokine-receptor interactions, both locally and systemically, on vascular disease in allografts.

Methodology/principal findings: Analysis of GAG or CC chemokine receptor 2 (CCR2) deficiency were coupled with the infusion of viral chemokine modulating proteins (CMPs) in mouse aortic allograft transplants (n = 239 mice). Inflammatory cell invasion and neointimal hyperplasia were significantly reduced in N-deacetylase-N-sulfotransferase-1 (Ndst1(f/f)TekCre(+)) heparan sulfate (GAG)-deficient (Ndst1(-/-), p<0.044) and CCR2-deficient (Ccr2(-/-), p<0.04) donor transplants. Donor tissue GAG or CCR2 deficiency markedly reduced inflammation and vasculopathy, whereas recipient deficiencies did not. Treatment with three CMPs was also investigated; Poxviral M-T1 blocks CC chemokine receptor binding, M-T7 blocks C, CC, and CXC GAG binding, and herpesviral M3 binds receptor and GAG binding for all classes. M-T7 reduced intimal hyperplasia in wild type (WT) (Ccr2(+/+), p< or =0.003 and Ccr2(-/-), p

Conclusions/significance: Interruption of chemokine-GAG interactions, even in the absence of chemokine-receptor blockade, is a highly effective approach to reduction of allograft rejection, reducing vascular inflammation and prolonging allograft survival. Although chemokines direct both local and systemic cell migration, interruption of inherent chemokine responses in the donor tissue unexpectedly had a greater therapeutic impact on allograft vasculopathy.

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Related in: MedlinePlus

Local, but not systemic, GAG or receptor deficiency reduces neointimal hyperplasia in mouse aortic allograft transplants.Histology cross sections of aortic allograft transplants stained with Masson's trichrome demonstrating reduced inflammatory cell invasion with Ndst1-deficient (Ndst1−/−) (B) and Ccr2-deficient (Ccr2−/−) (D) donor aortic allografts when compared to wild type (WT)(A, B). Bar graphs demonstrate similar, significant reductions in mean neointimal area for Ccr2−/− (D) and Ndst1−/− (E) transplants. Reduced intimal area is detected with donor aortic deficiency, but not with recipient deficiency (F, reverse transplant WT into Ccr2−/− or Ndst1−/− deficient recipient mice). Measurements reported as mean ± S.E. Arrows bracket intimal plaque, arrow heads indicates mononuclear cell infiltrates. Mag 400X.
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pone-0010510-g001: Local, but not systemic, GAG or receptor deficiency reduces neointimal hyperplasia in mouse aortic allograft transplants.Histology cross sections of aortic allograft transplants stained with Masson's trichrome demonstrating reduced inflammatory cell invasion with Ndst1-deficient (Ndst1−/−) (B) and Ccr2-deficient (Ccr2−/−) (D) donor aortic allografts when compared to wild type (WT)(A, B). Bar graphs demonstrate similar, significant reductions in mean neointimal area for Ccr2−/− (D) and Ndst1−/− (E) transplants. Reduced intimal area is detected with donor aortic deficiency, but not with recipient deficiency (F, reverse transplant WT into Ccr2−/− or Ndst1−/− deficient recipient mice). Measurements reported as mean ± S.E. Arrows bracket intimal plaque, arrow heads indicates mononuclear cell infiltrates. Mag 400X.

Mentions: The effects of either GAG or chemokine receptor deficiency on vascular neointimal hyperplasia (plaque) growth, was examined in mouse aortic allograft transplant models with donor aortic genetic deficiency of Ndst1 or Ccr2 (Table 1, n = 79) and wild type recipients. Excess inflammatory cell infiltration and neointimal hyperplasia (accelerated plaque growth) were detected in aortic allograft transplants at 4 weeks follow up in control, saline treated, wild type (WT) mice, using either C57Bl/6 donor to Balb/c recipient mice (Fig. 1A,E) or Balb/c donor to C57Bl/6 recipient mice (Fig. 1 C, E, n = 19). Transplantation of conditionally HS deficient Ndst1−/− donor aorta (C57Bl/6 background) into WT recipient (Balb/c background, Ndst1+/+) mice significantly reduced plaque area (Fig. 1 B, E, n = 12), whether measured as the ratio of intimal to medial thickness (72.9% reduction, P<0.044) or by morphometric analysis of neointimal plaque area (72.4%, P<0.05), when compared to WT donor aortic transplant (Balb/c recipient) using littermate controls (Fig. 1 A, E). Similarly, transplant of Ccr2−/− donor aorta (Balb/c background) into WT (C57Bl/6 background, Ccr2+/+) recipient mice significantly reduced neointimal hyperplasia (Fig. 1 D, E, n = 23) measured as ratios of intimal to medial thickness (55.6%, P<0.040) or as total neointimal cross-section area (43.5%, P<0.021) when compared to WT (Ccr2+/+) littermate controls (C57Bl/6 recipient, Fig. 1C, E). Although Ndst1−/− are on a C57Bl/6 background and Ccr2−/− mice are on a Balb/c background, reductions in intimal plaque area were detected in each analysis using comparison to matched controls, e.g. WT C57Bl/6 to Balb/c and WT Balb/c to C57Bl/6 (ANOVA p<0.027). A significant difference in total intimal plaque area was detectable for saline treated WT C57Bl/6 to Balb/c when compared to the C57Bl/6 to Balb/c controls (n = 19, mean plaque area 0.15±0.037 mm2 for WT Balb/c versus 0.052±0.013 mm2 for WT C57Bl/6 donor allografts, P<0.034). However, no statistically significant increase was seen on analysis of intimal to medial thickness (Fig. 1E, n = 19, p = 0.521). Measurement of the intimal to medial thickness ratios normalizes intimal plaque size to arterial medial thickness. All analyses were performed using both intimal to medial thickness ratios together with morphometric analysis of plaque area.


Inhibition of chemokine-glycosaminoglycan interactions in donor tissue reduces mouse allograft vasculopathy and transplant rejection.

Dai E, Liu LY, Wang H, McIvor D, Sun YM, Macaulay C, King E, Munuswamy-Ramanujam G, Bartee MY, Williams J, Davids J, Charo I, McFadden G, Esko JD, Lucas AR - PLoS ONE (2010)

Local, but not systemic, GAG or receptor deficiency reduces neointimal hyperplasia in mouse aortic allograft transplants.Histology cross sections of aortic allograft transplants stained with Masson's trichrome demonstrating reduced inflammatory cell invasion with Ndst1-deficient (Ndst1−/−) (B) and Ccr2-deficient (Ccr2−/−) (D) donor aortic allografts when compared to wild type (WT)(A, B). Bar graphs demonstrate similar, significant reductions in mean neointimal area for Ccr2−/− (D) and Ndst1−/− (E) transplants. Reduced intimal area is detected with donor aortic deficiency, but not with recipient deficiency (F, reverse transplant WT into Ccr2−/− or Ndst1−/− deficient recipient mice). Measurements reported as mean ± S.E. Arrows bracket intimal plaque, arrow heads indicates mononuclear cell infiltrates. Mag 400X.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2865544&req=5

pone-0010510-g001: Local, but not systemic, GAG or receptor deficiency reduces neointimal hyperplasia in mouse aortic allograft transplants.Histology cross sections of aortic allograft transplants stained with Masson's trichrome demonstrating reduced inflammatory cell invasion with Ndst1-deficient (Ndst1−/−) (B) and Ccr2-deficient (Ccr2−/−) (D) donor aortic allografts when compared to wild type (WT)(A, B). Bar graphs demonstrate similar, significant reductions in mean neointimal area for Ccr2−/− (D) and Ndst1−/− (E) transplants. Reduced intimal area is detected with donor aortic deficiency, but not with recipient deficiency (F, reverse transplant WT into Ccr2−/− or Ndst1−/− deficient recipient mice). Measurements reported as mean ± S.E. Arrows bracket intimal plaque, arrow heads indicates mononuclear cell infiltrates. Mag 400X.
Mentions: The effects of either GAG or chemokine receptor deficiency on vascular neointimal hyperplasia (plaque) growth, was examined in mouse aortic allograft transplant models with donor aortic genetic deficiency of Ndst1 or Ccr2 (Table 1, n = 79) and wild type recipients. Excess inflammatory cell infiltration and neointimal hyperplasia (accelerated plaque growth) were detected in aortic allograft transplants at 4 weeks follow up in control, saline treated, wild type (WT) mice, using either C57Bl/6 donor to Balb/c recipient mice (Fig. 1A,E) or Balb/c donor to C57Bl/6 recipient mice (Fig. 1 C, E, n = 19). Transplantation of conditionally HS deficient Ndst1−/− donor aorta (C57Bl/6 background) into WT recipient (Balb/c background, Ndst1+/+) mice significantly reduced plaque area (Fig. 1 B, E, n = 12), whether measured as the ratio of intimal to medial thickness (72.9% reduction, P<0.044) or by morphometric analysis of neointimal plaque area (72.4%, P<0.05), when compared to WT donor aortic transplant (Balb/c recipient) using littermate controls (Fig. 1 A, E). Similarly, transplant of Ccr2−/− donor aorta (Balb/c background) into WT (C57Bl/6 background, Ccr2+/+) recipient mice significantly reduced neointimal hyperplasia (Fig. 1 D, E, n = 23) measured as ratios of intimal to medial thickness (55.6%, P<0.040) or as total neointimal cross-section area (43.5%, P<0.021) when compared to WT (Ccr2+/+) littermate controls (C57Bl/6 recipient, Fig. 1C, E). Although Ndst1−/− are on a C57Bl/6 background and Ccr2−/− mice are on a Balb/c background, reductions in intimal plaque area were detected in each analysis using comparison to matched controls, e.g. WT C57Bl/6 to Balb/c and WT Balb/c to C57Bl/6 (ANOVA p<0.027). A significant difference in total intimal plaque area was detectable for saline treated WT C57Bl/6 to Balb/c when compared to the C57Bl/6 to Balb/c controls (n = 19, mean plaque area 0.15±0.037 mm2 for WT Balb/c versus 0.052±0.013 mm2 for WT C57Bl/6 donor allografts, P<0.034). However, no statistically significant increase was seen on analysis of intimal to medial thickness (Fig. 1E, n = 19, p = 0.521). Measurement of the intimal to medial thickness ratios normalizes intimal plaque size to arterial medial thickness. All analyses were performed using both intimal to medial thickness ratios together with morphometric analysis of plaque area.

Bottom Line: Donor tissue GAG or CCR2 deficiency markedly reduced inflammation and vasculopathy, whereas recipient deficiencies did not.M-T1 and M3 inhibited WT (Ccr2(+/+) and Ndst1(+/+), p< or =0.006) allograft vasculopathy, but did not block vasculopathy in Ccr2(-/-) (p = 0.61).Interruption of chemokine-GAG interactions, even in the absence of chemokine-receptor blockade, is a highly effective approach to reduction of allograft rejection, reducing vascular inflammation and prolonging allograft survival.

View Article: PubMed Central - PubMed

Affiliation: Vascular Biology Research Group, Robarts Research Institute, The University of Western Ontario, London, Ontario, Canada.

ABSTRACT

Background: Binding of chemokines to glycosaminoglycans (GAGs) is classically described as initiating inflammatory cell migration and creating tissue chemokine gradients that direct local leukocyte chemotaxis into damaged or transplanted tissues. While chemokine-receptor binding has been extensively studied during allograft transplantation, effects of glycosaminoglycan (GAG) interactions with chemokines on transplant longevity are less well known. Here we examine the impact of interrupting chemokine-GAG interactions and chemokine-receptor interactions, both locally and systemically, on vascular disease in allografts.

Methodology/principal findings: Analysis of GAG or CC chemokine receptor 2 (CCR2) deficiency were coupled with the infusion of viral chemokine modulating proteins (CMPs) in mouse aortic allograft transplants (n = 239 mice). Inflammatory cell invasion and neointimal hyperplasia were significantly reduced in N-deacetylase-N-sulfotransferase-1 (Ndst1(f/f)TekCre(+)) heparan sulfate (GAG)-deficient (Ndst1(-/-), p<0.044) and CCR2-deficient (Ccr2(-/-), p<0.04) donor transplants. Donor tissue GAG or CCR2 deficiency markedly reduced inflammation and vasculopathy, whereas recipient deficiencies did not. Treatment with three CMPs was also investigated; Poxviral M-T1 blocks CC chemokine receptor binding, M-T7 blocks C, CC, and CXC GAG binding, and herpesviral M3 binds receptor and GAG binding for all classes. M-T7 reduced intimal hyperplasia in wild type (WT) (Ccr2(+/+), p< or =0.003 and Ccr2(-/-), p

Conclusions/significance: Interruption of chemokine-GAG interactions, even in the absence of chemokine-receptor blockade, is a highly effective approach to reduction of allograft rejection, reducing vascular inflammation and prolonging allograft survival. Although chemokines direct both local and systemic cell migration, interruption of inherent chemokine responses in the donor tissue unexpectedly had a greater therapeutic impact on allograft vasculopathy.

Show MeSH
Related in: MedlinePlus