Immune Netw.  2011 Dec;11(6):368-375. 10.4110/in.2011.11.6.368.

Blockade of Vascular Endothelial Growth Factor (VEGF) Aggravates the Severity of Acute Graft-versus-host Disease (GVHD) after Experimental Allogeneic Hematopoietic Stem Cell Transplantation (allo-HSCT)

• Kim AR
• Lim JY
• Jeong DC
• Park G
• Lee BC
• Min CK

• KMID: 2150722
• DOI: http://doi.org/10.4110/in.2011.11.6.368

Abstract

BACKGROUND
Recent clinical observation reported that there was a significant correlation between change in circulating vascular endothelial growth factor (VEGF) levels and the occurrence of severe acute graft-versus-host disease (GVHD) following allogeneic hematopoietic stem cell transplantation (allo-HSCT), but the action mechanisms of VEGF in GVHD have not been demonstrated.
METHODS
This study investigated whether or not blockade of VEGF has an effect on acute GVHD in a lethally irradiated murine allo-HSCT model of B6 (H-2b)-->B6D2F1 (H-2b/d). Syngeneic or allogeneic recipient mice were injected subcutaneously with anti-VEGF peptides, dRK6 (50 microg/dose) or control diluent every other day for 2 weeks (total 7 doses).
RESULTS
Administration of the dRK6 peptide after allo-HSCT significantly reduced survival with greaterclinical GVHD scores and body weight loss. Allogeneic recipients injected with the dRK6 peptide exhibited significantly increased circulating levels of VEGF and expansion of donor CD3+ T cells on day +7 compared to control treated animals. The donor CD4+ and CD8+ T-cell subsets have differential expansion caused by the dRK6 injection. The circulating VEGF levels were reduced on day +14 regardless of blockade of VEGF.
CONCLUSION
Together these findings demonstrate that the allo-reactive responses after allo-HSCT are exaggerated by the blockade of VEGF. VEGF seems to be consumed during the progression of acute GVHD in this murine allo-HSCT model.

Keyword

Vascular endothelial growth factors (VEGF); Allogeneic hematopoietic stem cell transplantation; Acute graft-versus-host disease; VEGF blockade; dRK6

MeSH Terms

Animals
Body Weight
Graft vs Host Disease
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells
Humans
Mice
Oligopeptides
Peptides
T-Lymphocyte Subsets
T-Lymphocytes
Tissue Donors
Vascular Endothelial Growth Factor A
Oligopeptides
Peptides
Vascular Endothelial Growth Factor A

Figure

• 

  Figure 1 Treatment with anti-VEGF dRK6 peptide aggravates acute GVHD mortality. Effect of dRK6 on survival in experimental acute GVHD induced across MHC class I and II. B6D2F1 mice received 1100 cGy TBI and underwent transplantation with 10×106 BM cells and 20×106 splenocytes from allogeneic B6 (Allo, ▪ and •, n= 12, respectively) or syngeneic F1 donors (Syn, ▴ and ▾, n=6, respectively). Recipient animals were injected subcutaneously with either 50 µg dRK6 per mouse (▪ and ▴) or the diluent (• and ▾) every other day from day 0 to day +14 (total 7 doses). Survival was evaluated as detailed in "Materials and Methods". Data from 2 similar experiments are combined. ▪ vs •, p=0.012.

• 

  Figure 2 Treatment with anti-VEGF dRK6 peptide aggravates acute GVHD-specific morbidity. Effect of dRK6 on acute GVHD score (A) and change of body weight (B) in experimental acute GVHD induced across MHC class I and II. B6D2F1 mice received 1100 cGy TBI and underwent transplantation with 10×106 BM cells and 20×106 splenocytes from allogeneic B6 (Allo, ▪ and •, n=12, respectively) or syngeneic F1 donors (Syn, ▴ and ▾, n=6, respectively). Recipient animals were injected subcutaneously with either 50 µg dRK6 per mouse (▪ and ▴) or the diluent (• and ▾) every other day from day 0 to day +14 (total 7 doses). GVHD score was evaluated as detailed in "Materials and Methods". Mean±SE is shown. Data from 2 similar experiments are combined. *p<0.05.

• 

  Figure 3 (A) A representative of histopathology of liver, intestine and skin. (B) Histologic scoring. Animals that underwent transplantation (n=4/group) were killed; liver, intestine and skin were obtained for analysis on day +28 after HSCT. Coded slides were semi-quantitative to assess pathologic damage, as described in "Materials and Methods". Total GVHD score; mean±SE of the sum of scores for tissues from individual animals in each group.

• 

  Figure 4 Effect of dRK6 on donor T-cell expansion after allo-HSCT. B6D2F1 (Allo) or B6 (Syn) mice received 1100 cGy TBI and underwent transplantation with 10×106 BM cells and 20×106 splenocytes from allogeneic B6.Ly-5a (CD45.1+) donors as in Fig. 1. Allogeneic recipient were injected subcutaneously with either 50 µg dRK6 per mouse or the diluent as described in "Materials and Methods". (A-C) Splenocytes were harvested on day +7 from the recipints (n=4/group) and labeled with anti-CD45.1+ APC and anti-CD3+ PE (A), anti-CD4+ FITC (B) or anti-CD8+ FITC (C). The percentage of donor CD3+, CD4+ and CD8+ cells was determined by flow cytometry. dRK6 significantly increased donor CD3+ and CD4+ expansion but not CD8+ expansion (*p<0.05).

• 

  Figure 5 Effect of dRK6 on the kinetics of serum VEGF concentrations and VEGF protein expressions in mouse liver after allo-HSCT. B6D2F1 (Allo) or B6 (Syn) mice received 1100 cGy TBI and underwent transplantation with 10×106 BM cells and 20×106 splenocytes from allogeneic B6. Ly-5a (CD45.1+) donors as in Fig. 1. Allogeneic recipient were injected subcutaneously with either 50 µg dRK6 per mouse or the diluent as described in "Materials and Methods". Sera from the recipient animals were obtained on days +7 and +14 after transplant and were analyzed for VEGF as describe in "Materials and Methods" (*p=0.005). analyzed for VEGF as describe in "Materials and Methods" (*p=0.005).

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