Immune Netw.  2010 Apr;10(2):46-54. 10.4110/in.2010.10.2.46.

Roles of Host Nonhematopoietic Cells in Autoimmunity and Donor Cell Engraftment in Graft-versus-host Disease

• Kim J
• Park S
• Kim HA
• Jung D
• Kim HJ
• Choi HJ
• Cho HR
• Kwon B

• KMID: 2150659
• DOI: http://doi.org/10.4110/in.2010.10.2.46

Abstract

BACKGROUND: Graft-versus-host disease (GVHD) is initiated when alloreactive donor T cells are primed by host APCs to undergo clonal expansion and maturation. Since there is a controversy regarding the role of nonhematopoietic cells in GVHD, we wanted to investigate the influence of MHC disparity on nonhematopoietic cells on the pathogenesis of GVHD in the MHC-haplomismatched C57BL/6 (H-2(b)) or DBA/2 (H-2(d))-->unirradiated (C57BL/6xDBA/2) F(1)(BDF(1); H-2(b/d)) murine model of acute GVHD (aGVHD) or chronic GVHD (cGVHD).
METHODS
We generated (BDF(1)-->C57BL/6), (BDF(1)-->DBA/2), and (BDF(1)-->BDF(1)) chimeras and examined GVHD-related parameters and donor cell engraftment in those chimeras.
RESULTS
Using this experimental system, we found that 1) severe aGVHD across MHC Ag barrier depends on the expression of nonhematopoietically rather than hematopoietically derived alloAgs for maximal GVHD manifestations; 2) host APCs were sufficient to break B cell tolerance to self molecules in cGVHD, whereas host APCs were insufficient to induce autoimmunity in aGVHD; 3) donor cell engraftment was greatly enhanced in the host with MHC-matched nonhematopoietic cells.
CONCLUSION
Taken together, our results provide an insight into how MHC disparity on GVHD target organs contribute to the pathogenesis of GVHD.

Keyword

Autoimmunity; Chimera; Graft-versus-host disease; Immune tolerance

MeSH Terms

Autoimmunity
Chimera
Graft vs Host Disease
Humans
Immune Tolerance
T-Lymphocytes
Tissue Donors

Figure

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  Figure 1 Confirmation of BM reconstitution. B6, DBA/2 and BDF1 mice received 12 Gy irradiation and were reconstituted with 5×106 T cell-depleted BDF1 BM cells. PBMCs were analyzed by flow cytometry at days 45 and 55 after cell transfer. Percent of donor cells were counted by staining PBMCs with anti-H-2Kb or H-2Kd mAbs (n=10~20 per group).

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  Figure 2 MHC disparity on nonhematopoietic cells exacerbates aGVHD. Eighty-four days after BM reconstitution, aGVHD was induced by transferring 8×107 B6 spleen/lymph node cells into three sets of mice (n=20 per group). (A) Loss of body weight. (B) Percent of survival. (C) Pathological scores for livers and colons. Organs were harvested at day 54 after disease induction (n=5~8 per group). †p<0.001, **p<0.01 and *p<0.05 between the indicated groups.

• 

  Figure 3 MHC match on nonhematopoietic cells increases donor cell engraftment in aGVHD. Eighty-four days after BM reconstitution, aGVHD was induced by transferring 8×107 B6 spleen/lymph node cells into three sets of mice (n=20 per group). Splenocytes were harvested at day 54 after disease induction and stained with anti-H-2Kd plus anti-CD4, anti-CD8 or B220. (A) Percent of total donor cells. (B) Percent of donor B cells. (C) Percent of donor CD4+ T cells. (D) Percent of donor CD8+ T cells (n=5~8 per group). †p<0.001, **p<0.01 and *p<0.05 between the indicated groups.

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  Figure 4 MHC match on nonhematopoietic cells decreases activation of donor T cells in aGVHD. Eighty-four days after BM reconstitution, aGVHD was induced by transferring 8×107 B6 spleen/lymph node cells into three sets of mice (n=20 per group). Splenocytes were harvested at day 54 after disease induction and stained with anti-H-2Kd plus anti-CD62L and anti-CD4 or anti-CD8. (A) Percent of donor CD4+CD62Llow T cells. (B) Percent of donor CD8+CD62Llow T cells (n=8 per group). †p<0.001, **p<0.01 and *p<0.05 between the indication groups.

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  Figure 5 MHC disparity on nonhematopoietic cells does not affect the development of cGVHD. B6, DBA/2 and BDF1 mice received 12 Gy irradiation and were reconstituted with 5×106 T cell-depleted BDF1 BM cells. Eighty-four days after BM reconstitution, cGVHD was induced by transferring 8×107 DBA/2 spleen/lymph node cells into three sets of mice (n=10 per group). (A) Serum samples were collected every 2 wk and assayed in duplicate by ELISA for IgG1 anti-DNA autoAb. The OD of duplicate samples for each mouse was measured at 450 nm, using serially diluted serum samples. (B) Percent of survival (n=10 per group). (C) Histology of kidneys harvested at day 54 after disease induction. Representative kidney sections are shown for H&E staining. *p<0.05 between the indicated groups.

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  Figure 6 MHC match on nonhematopoietic cells increases donor lymphocyte engraftment in cGVHD. Eighty-four days after BM reconstitution, cGVHD was induced by transferring 5×106 DBA/2 spleen/lymph node cells into three sets of mice (n=20 per group). Splenocytes were harvested at day 54 after disease induction and stained with anti-H-2Kb plus anti-CD4, anti-CD8 or B220. (A) Percent of total donor cells. (B) Percent of donor B cells. (C) Percent of donor CD4+ T cells. (D) Percent of donor CD8+ T cells (n=5~8 per group). **p<0.01 and *p<0.05 between the indicated groups.

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  Figure 7 Donor lymphocyte engraftment correlates with donor CD8+ T cell activation in cGVHD. Eighty-four days after BM reconstitution, aGVHD was induced by transferring 5×106 DBA/2 spleen/lymph node cells into three sets of mice (n=10 per group). Splenocytes were harvested at day 54 after disease induction and stained with anti-H-2Kb plus anti-CD62L and anti-CD4 or anti-CD8. (A) Percent of donor CD4+ CD62Llow T cells. (B) Percent of donor CD8+CD62Llow T cells (n=8 per group). *p<0.05 between the indicated groups.

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