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A novel clinically relevant strategy to abrogate autoimmunity and regulate alloimmunity in NOD mice.

Vergani A, D'Addio F, Jurewicz M, Petrelli A, Watanabe T, Liu K, Law K, Schuetz C, Carvello M, Orsenigo E, Deng S, Rodig SJ, Ansari JM, Staudacher C, Abdi R, Williams J, Markmann J, Atkinson M, Sayegh MH, Fiorina P - Diabetes (2010)

Bottom Line: Using allogeneic islet transplantation models as well as NOD mice with recent onset type 1 diabetes, we addressed the therapeutic efficacy and immunomodulatory mechanisms associated with a new immunoregulatory protocol based on prolonged low-dose mATG plus CTLA4-Ig.Immunologic analysis of mice receiving transplants revealed a complete abrogation of autoimmune responses and severe downregulation of alloimmunity in response to treatment.The capacity to regulate alloimmunity and to abrogate the autoimmune response in NOD mice in different settings confirmed that prolonged mATG+CTLA4-Ig treatment is a clinically relevant strategy to translate to humans with type 1 diabetes.

View Article: PubMed Central - PubMed

Affiliation: Transplantation Research Center, Children's Hospital and Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. paolo.fiorina@childrens.harvard.edu

ABSTRACT

Objective: To investigate a new clinically relevant immunoregulatory strategy based on treatment with murine Thymoglobulin mATG Genzyme and CTLA4-Ig in NOD mice to prevent allo- and autoimmune activation using a stringent model of islet transplantation and diabetes reversal.

Research design and methods: Using allogeneic islet transplantation models as well as NOD mice with recent onset type 1 diabetes, we addressed the therapeutic efficacy and immunomodulatory mechanisms associated with a new immunoregulatory protocol based on prolonged low-dose mATG plus CTLA4-Ig.

Results: BALB/c islets transplanted into hyperglycemic NOD mice under prolonged mATG+CTLA4-Ig treatment showed a pronounced delay in allograft rejection compared with untreated mice (mean survival time: 54 vs. 8 days, P < 0.0001). Immunologic analysis of mice receiving transplants revealed a complete abrogation of autoimmune responses and severe downregulation of alloimmunity in response to treatment. The striking effect on autoimmunity was confirmed by 100% diabetes reversal in newly hyperglycemic NOD mice and 100% indefinite survival of syngeneic islet transplantation (NOD.SCID into NOD mice).

Conclusions: The capacity to regulate alloimmunity and to abrogate the autoimmune response in NOD mice in different settings confirmed that prolonged mATG+CTLA4-Ig treatment is a clinically relevant strategy to translate to humans with type 1 diabetes.

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Related in: MedlinePlus

Analysis of Peripheral Blood Mononuclear Cells (PBMC) was performed at various time points after transplantation in diabetic NOD mice. CD4+ and CD8+ T-cell depletion is detected on day 7 after mATG treatment (untreated vs. all treatment, P < 0.05) (A–C). Addition of CTLA4-Ig slowed reconstitution after mATG depletion on day 14 (CD3+, CD4+, and CD8+ cells: prolonged mATG+CTLA4-Ig vs. prolonged mATG, P < 0.05). On day 28, peripheral T-cell percentage in animals treated with prolonged mATG+CTLA4-Ig was similar to that of animals treated with induction mATG+CTLA4-Ig (P = NS). On day 7, FoxP3+CD25+CD4+ (Tregs) was increased by both induction and prolonged mATG treatment (P < 0.05 vs. day 7 untreated), and on day 14 by prolonged mATG treatment (P < 0.05 vs. day 7 untreated) (D). Addition of CTLA4-Ig abrogated this increase (day 7: induction mATG+CTLA4-Ig vs. induction mATG, P = 0.04; day 7 and day 14 prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.002 and P = 0.008, respectively) (D). Peripheral Treg frequency at day 28 was increased with prolonged mATG + CTLA4-Ig treatment compared with induction mATG+CTLA4-Ig treatment (P < 0.05) (D). On day 14, a relative increase in CD44highCD62lowCD4+ cells (Teffs) was evident with prolonged mATG treatment (P < 0.05 vs. day 7 untreated); this increase was prevented by the addition of CTLA4-Ig (prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.0001) (E). On day 28, Teffs were increased in the prolonged mATG+CTLA4-Ig group compared with the induction group (P < 0.05) (E). *P < 0.05.
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Figure 1: Analysis of Peripheral Blood Mononuclear Cells (PBMC) was performed at various time points after transplantation in diabetic NOD mice. CD4+ and CD8+ T-cell depletion is detected on day 7 after mATG treatment (untreated vs. all treatment, P < 0.05) (A–C). Addition of CTLA4-Ig slowed reconstitution after mATG depletion on day 14 (CD3+, CD4+, and CD8+ cells: prolonged mATG+CTLA4-Ig vs. prolonged mATG, P < 0.05). On day 28, peripheral T-cell percentage in animals treated with prolonged mATG+CTLA4-Ig was similar to that of animals treated with induction mATG+CTLA4-Ig (P = NS). On day 7, FoxP3+CD25+CD4+ (Tregs) was increased by both induction and prolonged mATG treatment (P < 0.05 vs. day 7 untreated), and on day 14 by prolonged mATG treatment (P < 0.05 vs. day 7 untreated) (D). Addition of CTLA4-Ig abrogated this increase (day 7: induction mATG+CTLA4-Ig vs. induction mATG, P = 0.04; day 7 and day 14 prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.002 and P = 0.008, respectively) (D). Peripheral Treg frequency at day 28 was increased with prolonged mATG + CTLA4-Ig treatment compared with induction mATG+CTLA4-Ig treatment (P < 0.05) (D). On day 14, a relative increase in CD44highCD62lowCD4+ cells (Teffs) was evident with prolonged mATG treatment (P < 0.05 vs. day 7 untreated); this increase was prevented by the addition of CTLA4-Ig (prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.0001) (E). On day 28, Teffs were increased in the prolonged mATG+CTLA4-Ig group compared with the induction group (P < 0.05) (E). *P < 0.05.

Mentions: We tested the depleting potential of mATG on peripheral blood in an allogeneic fully mismatched model of islet transplantation (i.e., BALB/c islets transplanted into NOD mice). Mice were followed until rejection, as after rejection mice became lymphopenic and immunocompromised. In untreated mice, the numbers of peripheral CD3+ cells dropped soon after islet transplantation (Fig. 1A). An induction dose of mATG (500 μg at days 0 and 4) further depleted CD3+ cells in the peripheral blood of NOD transplanted mice (Fig. 1A), and CD4+ and CD8+ cells appeared to be equally affected (Fig. 1B and C). Prolonged mATG treatment (mATG 500 μg at days 0 and 4, then 100 μg twice weekly) did not appear to result in further depletion.


A novel clinically relevant strategy to abrogate autoimmunity and regulate alloimmunity in NOD mice.

Vergani A, D'Addio F, Jurewicz M, Petrelli A, Watanabe T, Liu K, Law K, Schuetz C, Carvello M, Orsenigo E, Deng S, Rodig SJ, Ansari JM, Staudacher C, Abdi R, Williams J, Markmann J, Atkinson M, Sayegh MH, Fiorina P - Diabetes (2010)

Analysis of Peripheral Blood Mononuclear Cells (PBMC) was performed at various time points after transplantation in diabetic NOD mice. CD4+ and CD8+ T-cell depletion is detected on day 7 after mATG treatment (untreated vs. all treatment, P < 0.05) (A–C). Addition of CTLA4-Ig slowed reconstitution after mATG depletion on day 14 (CD3+, CD4+, and CD8+ cells: prolonged mATG+CTLA4-Ig vs. prolonged mATG, P < 0.05). On day 28, peripheral T-cell percentage in animals treated with prolonged mATG+CTLA4-Ig was similar to that of animals treated with induction mATG+CTLA4-Ig (P = NS). On day 7, FoxP3+CD25+CD4+ (Tregs) was increased by both induction and prolonged mATG treatment (P < 0.05 vs. day 7 untreated), and on day 14 by prolonged mATG treatment (P < 0.05 vs. day 7 untreated) (D). Addition of CTLA4-Ig abrogated this increase (day 7: induction mATG+CTLA4-Ig vs. induction mATG, P = 0.04; day 7 and day 14 prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.002 and P = 0.008, respectively) (D). Peripheral Treg frequency at day 28 was increased with prolonged mATG + CTLA4-Ig treatment compared with induction mATG+CTLA4-Ig treatment (P < 0.05) (D). On day 14, a relative increase in CD44highCD62lowCD4+ cells (Teffs) was evident with prolonged mATG treatment (P < 0.05 vs. day 7 untreated); this increase was prevented by the addition of CTLA4-Ig (prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.0001) (E). On day 28, Teffs were increased in the prolonged mATG+CTLA4-Ig group compared with the induction group (P < 0.05) (E). *P < 0.05.
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Figure 1: Analysis of Peripheral Blood Mononuclear Cells (PBMC) was performed at various time points after transplantation in diabetic NOD mice. CD4+ and CD8+ T-cell depletion is detected on day 7 after mATG treatment (untreated vs. all treatment, P < 0.05) (A–C). Addition of CTLA4-Ig slowed reconstitution after mATG depletion on day 14 (CD3+, CD4+, and CD8+ cells: prolonged mATG+CTLA4-Ig vs. prolonged mATG, P < 0.05). On day 28, peripheral T-cell percentage in animals treated with prolonged mATG+CTLA4-Ig was similar to that of animals treated with induction mATG+CTLA4-Ig (P = NS). On day 7, FoxP3+CD25+CD4+ (Tregs) was increased by both induction and prolonged mATG treatment (P < 0.05 vs. day 7 untreated), and on day 14 by prolonged mATG treatment (P < 0.05 vs. day 7 untreated) (D). Addition of CTLA4-Ig abrogated this increase (day 7: induction mATG+CTLA4-Ig vs. induction mATG, P = 0.04; day 7 and day 14 prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.002 and P = 0.008, respectively) (D). Peripheral Treg frequency at day 28 was increased with prolonged mATG + CTLA4-Ig treatment compared with induction mATG+CTLA4-Ig treatment (P < 0.05) (D). On day 14, a relative increase in CD44highCD62lowCD4+ cells (Teffs) was evident with prolonged mATG treatment (P < 0.05 vs. day 7 untreated); this increase was prevented by the addition of CTLA4-Ig (prolonged mATG+CTLA4-Ig vs. prolonged mATG, P = 0.0001) (E). On day 28, Teffs were increased in the prolonged mATG+CTLA4-Ig group compared with the induction group (P < 0.05) (E). *P < 0.05.
Mentions: We tested the depleting potential of mATG on peripheral blood in an allogeneic fully mismatched model of islet transplantation (i.e., BALB/c islets transplanted into NOD mice). Mice were followed until rejection, as after rejection mice became lymphopenic and immunocompromised. In untreated mice, the numbers of peripheral CD3+ cells dropped soon after islet transplantation (Fig. 1A). An induction dose of mATG (500 μg at days 0 and 4) further depleted CD3+ cells in the peripheral blood of NOD transplanted mice (Fig. 1A), and CD4+ and CD8+ cells appeared to be equally affected (Fig. 1B and C). Prolonged mATG treatment (mATG 500 μg at days 0 and 4, then 100 μg twice weekly) did not appear to result in further depletion.

Bottom Line: Using allogeneic islet transplantation models as well as NOD mice with recent onset type 1 diabetes, we addressed the therapeutic efficacy and immunomodulatory mechanisms associated with a new immunoregulatory protocol based on prolonged low-dose mATG plus CTLA4-Ig.Immunologic analysis of mice receiving transplants revealed a complete abrogation of autoimmune responses and severe downregulation of alloimmunity in response to treatment.The capacity to regulate alloimmunity and to abrogate the autoimmune response in NOD mice in different settings confirmed that prolonged mATG+CTLA4-Ig treatment is a clinically relevant strategy to translate to humans with type 1 diabetes.

View Article: PubMed Central - PubMed

Affiliation: Transplantation Research Center, Children's Hospital and Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. paolo.fiorina@childrens.harvard.edu

ABSTRACT

Objective: To investigate a new clinically relevant immunoregulatory strategy based on treatment with murine Thymoglobulin mATG Genzyme and CTLA4-Ig in NOD mice to prevent allo- and autoimmune activation using a stringent model of islet transplantation and diabetes reversal.

Research design and methods: Using allogeneic islet transplantation models as well as NOD mice with recent onset type 1 diabetes, we addressed the therapeutic efficacy and immunomodulatory mechanisms associated with a new immunoregulatory protocol based on prolonged low-dose mATG plus CTLA4-Ig.

Results: BALB/c islets transplanted into hyperglycemic NOD mice under prolonged mATG+CTLA4-Ig treatment showed a pronounced delay in allograft rejection compared with untreated mice (mean survival time: 54 vs. 8 days, P < 0.0001). Immunologic analysis of mice receiving transplants revealed a complete abrogation of autoimmune responses and severe downregulation of alloimmunity in response to treatment. The striking effect on autoimmunity was confirmed by 100% diabetes reversal in newly hyperglycemic NOD mice and 100% indefinite survival of syngeneic islet transplantation (NOD.SCID into NOD mice).

Conclusions: The capacity to regulate alloimmunity and to abrogate the autoimmune response in NOD mice in different settings confirmed that prolonged mATG+CTLA4-Ig treatment is a clinically relevant strategy to translate to humans with type 1 diabetes.

Show MeSH
Related in: MedlinePlus