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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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Interference with chemokine-CCR2 interactions reduces inflammatory cell invasion in aortic allografts.Immunohistochemical staining of aortic allograft transplant cross sections demonstrated increased lymphocyte (A) invasion in WT (Balb/c, Ccr2+/+) donor aortic transplants at 4 weeks. M-T7 reduced T cell invasion in the WT Balb/c donor transplants (B). In the Ccr2−/− transplants T cell invasion was reduced at 4 weeks in saline treated mice (C), without further significant reduction with M-T7 treatment (D). Bar graphs demonstrate reduced T cell invasion (P<0.0001), but not macrophage (P = 0.106) in Ccr2−/− saline treated allografts when compared to saline treated Ccr2+/+ allografts. M-T1, M-T7, and M3 treatment significantly reduced T cell (P<0.001)(E) and macrophage (P<0.029) (F) invasion in the WT (Balb/c, Ccr2+/+) donor aortic allograft transplants, but not in Ccr2−/− donor allografts (P = NS)(E,F). Measurements reported as mean ± S.E. Arrow heads point to brown, positively stained CD3+ T cells. Arrow points to blue stained suture seen in the top left corner of panel D. Mag 1000X.
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pone-0010510-g005: Interference with chemokine-CCR2 interactions reduces inflammatory cell invasion in aortic allografts.Immunohistochemical staining of aortic allograft transplant cross sections demonstrated increased lymphocyte (A) invasion in WT (Balb/c, Ccr2+/+) donor aortic transplants at 4 weeks. M-T7 reduced T cell invasion in the WT Balb/c donor transplants (B). In the Ccr2−/− transplants T cell invasion was reduced at 4 weeks in saline treated mice (C), without further significant reduction with M-T7 treatment (D). Bar graphs demonstrate reduced T cell invasion (P<0.0001), but not macrophage (P = 0.106) in Ccr2−/− saline treated allografts when compared to saline treated Ccr2+/+ allografts. M-T1, M-T7, and M3 treatment significantly reduced T cell (P<0.001)(E) and macrophage (P<0.029) (F) invasion in the WT (Balb/c, Ccr2+/+) donor aortic allograft transplants, but not in Ccr2−/− donor allografts (P = NS)(E,F). Measurements reported as mean ± S.E. Arrow heads point to brown, positively stained CD3+ T cells. Arrow points to blue stained suture seen in the top left corner of panel D. Mag 1000X.

Mentions: Comparisons of invasion of inflammatory mononuclear cells in Ccr2−/− donor allograft (to C57Bl/6 recipient) (Fig. 5) and in Ndst1−/− donor (to Ndst1+/+ Balb/c recipient) (Fig. 6) with WT aortic allograft transplants was assessed by immunohistochemical cell staining of aortic cross sections. Cell invasion was reduced at 4 weeks follow up for both Ccr2−/− (Fig. 5) and Ndst1−/− (Fig. 6) aortic donor allografts with saline control treatment, when compared to WT donor controls (Ccr2+/+and Ndst1+/+, respectively). Selective staining for CD3-positive T cells (CD3+) demonstrated significant reductions in CD3+ T cells in saline treated Ccr2−/− donor aortic allografts (Fig. 5C, E; p<0.0001) when compared to Ccr2+/+ controls (Fig. 5A, E). Compared to controls, macrophage counts were not significantly reduced in Ccr2−/− aortic allografts (Fig. 5F, p = 0.106).


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)

Interference with chemokine-CCR2 interactions reduces inflammatory cell invasion in aortic allografts.Immunohistochemical staining of aortic allograft transplant cross sections demonstrated increased lymphocyte (A) invasion in WT (Balb/c, Ccr2+/+) donor aortic transplants at 4 weeks. M-T7 reduced T cell invasion in the WT Balb/c donor transplants (B). In the Ccr2−/− transplants T cell invasion was reduced at 4 weeks in saline treated mice (C), without further significant reduction with M-T7 treatment (D). Bar graphs demonstrate reduced T cell invasion (P<0.0001), but not macrophage (P = 0.106) in Ccr2−/− saline treated allografts when compared to saline treated Ccr2+/+ allografts. M-T1, M-T7, and M3 treatment significantly reduced T cell (P<0.001)(E) and macrophage (P<0.029) (F) invasion in the WT (Balb/c, Ccr2+/+) donor aortic allograft transplants, but not in Ccr2−/− donor allografts (P = NS)(E,F). Measurements reported as mean ± S.E. Arrow heads point to brown, positively stained CD3+ T cells. Arrow points to blue stained suture seen in the top left corner of panel D. Mag 1000X.
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pone-0010510-g005: Interference with chemokine-CCR2 interactions reduces inflammatory cell invasion in aortic allografts.Immunohistochemical staining of aortic allograft transplant cross sections demonstrated increased lymphocyte (A) invasion in WT (Balb/c, Ccr2+/+) donor aortic transplants at 4 weeks. M-T7 reduced T cell invasion in the WT Balb/c donor transplants (B). In the Ccr2−/− transplants T cell invasion was reduced at 4 weeks in saline treated mice (C), without further significant reduction with M-T7 treatment (D). Bar graphs demonstrate reduced T cell invasion (P<0.0001), but not macrophage (P = 0.106) in Ccr2−/− saline treated allografts when compared to saline treated Ccr2+/+ allografts. M-T1, M-T7, and M3 treatment significantly reduced T cell (P<0.001)(E) and macrophage (P<0.029) (F) invasion in the WT (Balb/c, Ccr2+/+) donor aortic allograft transplants, but not in Ccr2−/− donor allografts (P = NS)(E,F). Measurements reported as mean ± S.E. Arrow heads point to brown, positively stained CD3+ T cells. Arrow points to blue stained suture seen in the top left corner of panel D. Mag 1000X.
Mentions: Comparisons of invasion of inflammatory mononuclear cells in Ccr2−/− donor allograft (to C57Bl/6 recipient) (Fig. 5) and in Ndst1−/− donor (to Ndst1+/+ Balb/c recipient) (Fig. 6) with WT aortic allograft transplants was assessed by immunohistochemical cell staining of aortic cross sections. Cell invasion was reduced at 4 weeks follow up for both Ccr2−/− (Fig. 5) and Ndst1−/− (Fig. 6) aortic donor allografts with saline control treatment, when compared to WT donor controls (Ccr2+/+and Ndst1+/+, respectively). Selective staining for CD3-positive T cells (CD3+) demonstrated significant reductions in CD3+ T cells in saline treated Ccr2−/− donor aortic allografts (Fig. 5C, E; p<0.0001) when compared to Ccr2+/+ controls (Fig. 5A, E). Compared to controls, macrophage counts were not significantly reduced in Ccr2−/− aortic allografts (Fig. 5F, p = 0.106).

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