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IL-2 pathway blocking in combination with anti-CD154 synergistically establishes mixed macrochimerism with limited dose of bone marrow cells and prolongs skin graft survival in mice.

Lee JH, Ha J, Kim SH, Kim SJ - J. Korean Med. Sci. (2006)

Bottom Line: In graft versus host disease model using CFSE staining, CD4+ T cell and CD8+ T cell proliferation were reduced in groups treated with CTLA4-Ig or anti-IL-2R or both in combination with anti-CD154.However, anti-IL-2R was not so strong as CTLA4-Ig in terms of inhibition of T cell proliferation.In conclusion, IL-2 pathway blocking combined with anti-CD154 can establish macrochimerism with limited dose of BM transplantation and induce specific tolerance to allograft.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Hallym University College of Medicine, Chuncheon, Korea.

ABSTRACT
To facilitate the establishment of mixed chimerism with limited dose of bone marrow (BM) cells, and to achieve tolerance in skin graft model, combined blocking of costimulatory pathway and IL-2 pathway was used in minimally myeloablative model using busulfan. BM cells (2.5x10(7)) of BALB/c were injected into C57BL/6 mice at day 0 with full thickness skin graft after single dose injection of busulfan (25 mg/kg) on day-1. Recipients were grouped and injected the anti-CD154, CTLA4-Ig, anti-IL-2R at days 0, 2, 4, and 6 according to protocol. Mixed macrochimerism were induced in groups treated with anti-CD154+anti-CTLA4-Ig, anti-CD154+anti-IL-2R, and anti-CD154+anti-CTLA4 Ig+anti-IL-2R. Three groups having chimerism enjoyed prolonged graft survival more than 6 months. Superantigen deletion study revealed deletion of alloreactive T cells in combined blockade treated groups. In graft versus host disease model using CFSE staining, CD4+ T cell and CD8+ T cell proliferation were reduced in groups treated with CTLA4-Ig or anti-IL-2R or both in combination with anti-CD154. However, anti-IL-2R was not so strong as CTLA4-Ig in terms of inhibition of T cell proliferation. In conclusion, IL-2 pathway blocking combined with anti-CD154 can establish macrochimerism with limited dose of BM transplantation and induce specific tolerance to allograft.

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The utilization of Vβ5, Vβ8 and Vβ11 by CD4+ T cells in peripheral blood of control (C57BL/6, BALB/c) and treated mice. Allogeneic bone marrow transplant with busulfan. C57BL/6 recipients were treated with anti-CD154 (n=6), anti-CD154 and anti-IL-2R Ab (n=6), anti-CD154 and CTLA4-Ig (n=8), anti-CD154, anti-IL-2R and CTLA4-Ig (n=8).
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Figure 3: The utilization of Vβ5, Vβ8 and Vβ11 by CD4+ T cells in peripheral blood of control (C57BL/6, BALB/c) and treated mice. Allogeneic bone marrow transplant with busulfan. C57BL/6 recipients were treated with anti-CD154 (n=6), anti-CD154 and anti-IL-2R Ab (n=6), anti-CD154 and CTLA4-Ig (n=8), anti-CD154, anti-IL-2R and CTLA4-Ig (n=8).

Mentions: In normal condition, BALB/c mice delete Vβ11 and Vβ5.1/2 positive CD4+ T cells in the thymus due to their high affinity for endogenous retroviral superantigens (mouse mammary tumor virus [MMTV]) presented by I-E MHC class II molecules, whereas C57BL/6 mice do not express I-E and utilize 4-5% of Vβ11 positive CD4+ T cells and 2-3% of Vβ5.1/5.2 positive CD4+ T cells. Both BALB/c and C57BL/ 6 mice have 15-20% of CD4+ T cells which express Vβ8.1/8.2 (37, 38). Six months after the skin graft, we checked alloreactive T cell deletion of each group using flow cytometric analysis (Fig. 3). The group II, which had been administered anti-CD154 only, has both CD4+ Vβ11+ T cells (3.80±1.50%) and CD4+ Vβ5+ T cells (1.14±0.73%). In group III, IV, and V which had additional anti-IL-2R or CTLA4 Ig or both, CD4+ T cells which express Vβ11+ and Vβ5+ were deleted almost completely compared to normal level (Vβ11; 1.05±0.35% vs. 0.79±0.21% vs. 0.94±0.34%, Vβ5; 0.39±0.16% vs. 0.25±0.13% vs. 0.54±0.23%). However, CD4+ Vβ8+ T cells in all groups showed persistently. This means that chimeric BALB/c cells deleted donor reactive recipient T cells in the thymus.


IL-2 pathway blocking in combination with anti-CD154 synergistically establishes mixed macrochimerism with limited dose of bone marrow cells and prolongs skin graft survival in mice.

Lee JH, Ha J, Kim SH, Kim SJ - J. Korean Med. Sci. (2006)

The utilization of Vβ5, Vβ8 and Vβ11 by CD4+ T cells in peripheral blood of control (C57BL/6, BALB/c) and treated mice. Allogeneic bone marrow transplant with busulfan. C57BL/6 recipients were treated with anti-CD154 (n=6), anti-CD154 and anti-IL-2R Ab (n=6), anti-CD154 and CTLA4-Ig (n=8), anti-CD154, anti-IL-2R and CTLA4-Ig (n=8).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2721919&req=5

Figure 3: The utilization of Vβ5, Vβ8 and Vβ11 by CD4+ T cells in peripheral blood of control (C57BL/6, BALB/c) and treated mice. Allogeneic bone marrow transplant with busulfan. C57BL/6 recipients were treated with anti-CD154 (n=6), anti-CD154 and anti-IL-2R Ab (n=6), anti-CD154 and CTLA4-Ig (n=8), anti-CD154, anti-IL-2R and CTLA4-Ig (n=8).
Mentions: In normal condition, BALB/c mice delete Vβ11 and Vβ5.1/2 positive CD4+ T cells in the thymus due to their high affinity for endogenous retroviral superantigens (mouse mammary tumor virus [MMTV]) presented by I-E MHC class II molecules, whereas C57BL/6 mice do not express I-E and utilize 4-5% of Vβ11 positive CD4+ T cells and 2-3% of Vβ5.1/5.2 positive CD4+ T cells. Both BALB/c and C57BL/ 6 mice have 15-20% of CD4+ T cells which express Vβ8.1/8.2 (37, 38). Six months after the skin graft, we checked alloreactive T cell deletion of each group using flow cytometric analysis (Fig. 3). The group II, which had been administered anti-CD154 only, has both CD4+ Vβ11+ T cells (3.80±1.50%) and CD4+ Vβ5+ T cells (1.14±0.73%). In group III, IV, and V which had additional anti-IL-2R or CTLA4 Ig or both, CD4+ T cells which express Vβ11+ and Vβ5+ were deleted almost completely compared to normal level (Vβ11; 1.05±0.35% vs. 0.79±0.21% vs. 0.94±0.34%, Vβ5; 0.39±0.16% vs. 0.25±0.13% vs. 0.54±0.23%). However, CD4+ Vβ8+ T cells in all groups showed persistently. This means that chimeric BALB/c cells deleted donor reactive recipient T cells in the thymus.

Bottom Line: In graft versus host disease model using CFSE staining, CD4+ T cell and CD8+ T cell proliferation were reduced in groups treated with CTLA4-Ig or anti-IL-2R or both in combination with anti-CD154.However, anti-IL-2R was not so strong as CTLA4-Ig in terms of inhibition of T cell proliferation.In conclusion, IL-2 pathway blocking combined with anti-CD154 can establish macrochimerism with limited dose of BM transplantation and induce specific tolerance to allograft.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Hallym University College of Medicine, Chuncheon, Korea.

ABSTRACT
To facilitate the establishment of mixed chimerism with limited dose of bone marrow (BM) cells, and to achieve tolerance in skin graft model, combined blocking of costimulatory pathway and IL-2 pathway was used in minimally myeloablative model using busulfan. BM cells (2.5x10(7)) of BALB/c were injected into C57BL/6 mice at day 0 with full thickness skin graft after single dose injection of busulfan (25 mg/kg) on day-1. Recipients were grouped and injected the anti-CD154, CTLA4-Ig, anti-IL-2R at days 0, 2, 4, and 6 according to protocol. Mixed macrochimerism were induced in groups treated with anti-CD154+anti-CTLA4-Ig, anti-CD154+anti-IL-2R, and anti-CD154+anti-CTLA4 Ig+anti-IL-2R. Three groups having chimerism enjoyed prolonged graft survival more than 6 months. Superantigen deletion study revealed deletion of alloreactive T cells in combined blockade treated groups. In graft versus host disease model using CFSE staining, CD4+ T cell and CD8+ T cell proliferation were reduced in groups treated with CTLA4-Ig or anti-IL-2R or both in combination with anti-CD154. However, anti-IL-2R was not so strong as CTLA4-Ig in terms of inhibition of T cell proliferation. In conclusion, IL-2 pathway blocking combined with anti-CD154 can establish macrochimerism with limited dose of BM transplantation and induce specific tolerance to allograft.

Show MeSH
Related in: MedlinePlus