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Conditioning with Fludarabine-Busulfan versus Busulfan-Cyclophosphamide Is Associated with Lower aGVHD and Higher Survival but More Extensive and Long Standing Bone Marrow Damage

He, Xin, Ye, YongBin, Xu, XiaoJun, Wang, Jing, Huang, YuXian, Weng, GuangYang, Zhang, MingWan, Guo, KunYuan·2016·BioMed Research International
DOI 10.1155/2016/3071214PMC PMC5098055
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Abstract

Acute graft-versus-host disease (aGVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT) and a major cause of nonrelapse mortality after allo-HSCT. A conditioning regimen plays a pivotal role in the development of aGVHD. To provide a platform for studying aGVHD and evaluating the impact of different conditioning regimens, we established a murine aGVHD model that simulates the clinical situation and can be conditioned with Busulfan-Cyclophosphamide (Bu-Cy) and Fludarabine-Busulfan (Flu-Bu). In our study, BALB/c mice were conditioned with Bu-Cy or Flu-Bu and transplanted with 2×107 bone marrow cells and 2×107 splenocytes from either allogeneic (C57BL/6) or syngeneic (BALB/c) donors. The allogeneic recipients conditioned with Bu-Cy had shorter survivals (P<0.05), more severe clinical manifestations, and higher hepatic and intestinal pathology scores, associated with increased INF-γ expression and diminished IL-4 expression in serum, compared to allogeneic recipients conditioned with Flu-Bu. Moreover, higher donor-derived T-cell infiltration and severely impaired B-cell development were seen in the bone marrow of mice, exhibiting aGVHD and conditioned with Flu-Bu. Our study showed that the conditioning regimen with Bu-Cy resulted in more severe aGVHD while the Flu-Bu regimen was associated with more extensive and long standing bone marrow damage.

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Figure 1 Survival analysis of different conditioning doses. Male BALB/c were conditioned with different doses of Bu-Cy or Flu-Bu and transplanted with 2 × 107 bone marrow cells and 2 × 107 splenocytes from C57BL/6. (a) Survival analysis of mice conditioned with Bu-Cy. (b) Survival analysis of mice conditioned with Flu-Bu.
Figure 1 Survival analysis of different conditioning doses. Male BALB/c were conditioned with different doses of Bu-Cy or Flu-Bu and transplanted with 2 × 107 bone marrow cells and 2 × 107 splenocytes from C57BL/6. (a) Survival analysis of mice conditioned with Bu-Cy. (b) Survival analysis of mice conditioned with Flu-Bu.CC BY · PubMed Central
Figure 2 aGVHD manifestations, survival, and aGVHD clinical scores. (a) Weight loss started after conditioning and bone marrow transplantation in allogeneic and syngeneic groups conditioned with Bu-Cy. (b) Weight loss started after conditioning and bone marrow transplantation in allogeneic and syngeneic groups conditioned with Flu-Bu. (c) Survival analysis of different treatment groups. (d) aGVHD clinical scores of allogeneic recipients. (e) Appearance of mice in Group A. (f–h). Appearance of allogeneically transplanted mice (conditioned with Bu-Cy). (i-j) Appearance of allogeneically transplanted mice (conditioned with Flu-Bu). Group A: Normal BALB/c in the same condition; Group B: BALB/c injected intraperitoneally with 0.0625 mL DMSO; Group C: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg); Group D: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from BALB/c; Group E: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6; Group F: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg); Group G: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from BALB/c; Group H: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. All data are presented as mean ± s.e. for 3–7 animals in each group per time point. ∗ P < 0.05.
Figure 2 aGVHD manifestations, survival, and aGVHD clinical scores. (a) Weight loss started after conditioning and bone marrow transplantation in allogeneic and syngeneic groups conditioned with Bu-Cy. (b) Weight loss started after conditioning and bone marrow transplantation in allogeneic and syngeneic groups conditioned with Flu-Bu. (c) Survival analysis of different treatment groups. (d) aGVHD clinical scores of allogeneic recipients. (e) Appearance of mice in Group A. (f–h). Appearance of allogeneically transplanted mice (conditioned with Bu-Cy). (i-j) Appearance of allogeneically transplanted mice (conditioned with Flu-Bu). Group A: Normal BALB/c in the same condition; Group B: BALB/c injected intraperitoneally with 0.0625 mL DMSO; Group C: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg); Group D: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from BALB/c; Group E: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6; Group F: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg); Group G: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from BALB/c; Group H: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. All data are presented as mean ± s.e. for 3–7 animals in each group per time point. ∗ P < 0.05.CC BY · PubMed Central
Figure 3 Pathologic changes of aGVHD and chimerism analysis. (a) Pathologic changes of liver, spleen, intestine, and skin on day +14 after transplantation. (b) Pathologic scores for liver, spleen, intestine, and skin on day +14 after transplantation. (c) Bone marrow and spleen chimerism of allogeneic recipients on day +14 after transplantation. Group A: normal BALB/c in the same condition. Group E: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. Group H: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. All data are presented as mean ± s.e. for 3–7 animals in each group per time point. ∗ P < 0.05; NS, not significant, P > 0.05.
Figure 3 Pathologic changes of aGVHD and chimerism analysis. (a) Pathologic changes of liver, spleen, intestine, and skin on day +14 after transplantation. (b) Pathologic scores for liver, spleen, intestine, and skin on day +14 after transplantation. (c) Bone marrow and spleen chimerism of allogeneic recipients on day +14 after transplantation. Group A: normal BALB/c in the same condition. Group E: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. Group H: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. All data are presented as mean ± s.e. for 3–7 animals in each group per time point. ∗ P < 0.05; NS, not significant, P > 0.05.CC BY · PubMed Central
Figure 4 Expression of IFN-γ and IL-4 in serum on days +3, +5, +7, and +14 after allo-HSCT, proportion of CD3+ and CD19+ cells in bone marrow and CD4+ and CD8+ cells in spleen 14 days after transplantation. (a) INF-γ concentration in serum on days +3, +5, +7, and +14 after allo-HSCT. (b) IL-4 concentration in serum on days +3, +5, +7, and +14 after allo-HSCT (c) Proportion of CD4+ and CD8+ T-cells in spleen 14 days after allo-HSCT. (d, e) Proportion of CD3+ and donor-derived CD3+ T-cells and CD19+ and donor-derived CD19+ B-cells in bone marrow 14 days after allo-HSCT. Group A: normal BALB/c in the same condition. Group E: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. Group H: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. All data are presented as mean ± s.e. for 3–7 animals in each group per time point. ∗ P < 0.05; ∗∗ P < 0.01; NS, not significant, P > 0.05.
Figure 4 Expression of IFN-γ and IL-4 in serum on days +3, +5, +7, and +14 after allo-HSCT, proportion of CD3+ and CD19+ cells in bone marrow and CD4+ and CD8+ cells in spleen 14 days after transplantation. (a) INF-γ concentration in serum on days +3, +5, +7, and +14 after allo-HSCT. (b) IL-4 concentration in serum on days +3, +5, +7, and +14 after allo-HSCT (c) Proportion of CD4+ and CD8+ T-cells in spleen 14 days after allo-HSCT. (d, e) Proportion of CD3+ and donor-derived CD3+ T-cells and CD19+ and donor-derived CD19+ B-cells in bone marrow 14 days after allo-HSCT. Group A: normal BALB/c in the same condition. Group E: BALB/c conditioned with Bu (100 mg/kg)-Cy (200 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. Group H: BALB/c conditioned with Flu (500 mg/kg)-Bu (100 mg/kg) and transplanted with 2 × 107 bone marrow cells, 2 × 107 spleen cells from C57BL/6. All data are presented as mean ± s.e. for 3–7 animals in each group per time point. ∗ P < 0.05; ∗∗ P < 0.01; NS, not significant, P > 0.05.CC BY · PubMed Central

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Metadata

Authors
He, X., Ye, Y., Xu, X., Wang, J., Huang, Y., Weng, G., Zhang, M., & Guo, K.
Affiliations
  • Department of Hematology, Zhongshan Hospital of Sun Yat-Sen University and Zhongshan City People’s Hospital, Zhongshan 528403, China
  • Department of Hematology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China
ORCID
Venue
BioMed Research International
Publisher
Wiley
Published
2016
Volume
2016
Pages
1-10
ISSN
  • 2314-6133
  • 2314-6141
DOI
10.1155/2016/3071214
PMC
PMC5098055
Cited by
4
References
0
Language
en
Licence
creativecommons.org/licenses/by/4.0/ (unspecified)
Funders
  • Natural Science & Technology Major Project Foundation of Zhongshan city (20113A002, 2011106)
  • Natural Science Foundation of Guangdong province (20113A002, 2011106)

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CoChat. (2026). Citation check: Conditioning with Fludarabine-Busulfan versus Busulfan-Cyclophosphamide Is Associated with Lower aGVHD and Higher Survival but More Extensive and Long Standing Bone Marrow Damage. https://app.cochat.ai/papers/10.1155/2016/3071214
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He, X., Ye, Y., Xu, X., Wang, J., Huang, Y., Weng, G., Zhang, M., & Guo, K. (2016). Conditioning with Fludarabine-Busulfan versus Busulfan-Cyclophosphamide Is Associated with Lower aGVHD and Higher Survival but More Extensive and Long Standing Bone Marrow Damage. BioMed Research International. https://doi.org/10.1155/2016/3071214