J Korean Med Sci.  2006 Dec;21(6):1005-1011. 10.3346/jkms.2006.21.6.1005.

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

• KMID: 1713109
• DOI: http://doi.org/10.3346/jkms.2006.21.6.1005

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 (2.5 x 10(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.

Keyword

ABI1793 monoclonal antibody; Asnti-CD154; abatacept; CTLA4-Ig; Anti-IL-2R; Mixed Chimerism; Transplantation Chimera; Bone Marrow Transplantation; Skin Transplantation; Busulfan

MeSH Terms

Skin Transplantation/*immunology/methods
Mice, Inbred BALB C
Mice
Male
Interleukin-2/*immunology
Immunoconjugates/*administration & dosage
Graft Survival/*immunology
Drug Combinations
CD40 Ligand/*immunology
Bone Marrow Transplantation/*immunology/methods
Antibodies/*administration & dosage/immunology
Animals

Figure

• 

  Fig. 1 Mixed chimerism was measured from peripheral blood of C57BL/6 recipients having BALB/c BM cells. Percentages of donor-derived T cells to the total number of T cells were expressed. Groups are allogeneinc bone marrow transplantation with busulfan only (△; n=6), treated with anti-CD154 (□; n=4), anti-CD154 and anti-IL-2R (○; n=5), anti-CD154 and CTLA4-Ig (■; n=8), anti-CD154, anti-IL-2R and CTLA-4 Ig (●; n=8).

• 

  Fig. 2 Survival of skin allografts. Combined use of costimulation blockade achieved indefinite graft survival. Groups are allogeneinc bone marrow transplantation with busulfan only (△; n=6), treated with anti-CD154 (□; n=4), anti-CD154 and anti-IL-2R (○; n=5), anti-CD154 and CTLA4-Ig (■; n=8), anti-CD154, anti-IL-2R and CTLA-4 Ig (●; n=8).

• 

  Fig. 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).

• 

  Fig. 4 (A) Representative samples from a flow cytometric assay for T cell proliferation in vivo. Supralethally irradiated BALB/c mice received CFSE-labeled T cells from naïve and treated mice. Cells were harvested at 66 hr after infusion. (B) Percentage of precursor A cells which divided 4 or more times.

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