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Immunosuppressive mechanisms of human bone marrow derived mesenchymal stromal cells in BALB/c host graft versus host disease murine models.

Robles JD, Liu YP, Cao J, Xiang Z, Cai Y, Manio M, Tang EH, Chan GC - Exp Hematol Oncol (2015)

Bottom Line: Documentation of suppression of RANTES, CCL3, CXCL9, CCR5 and CXCR3 with simultaneous decrease of donor T cell alloreactivity was demonstrated 6 days after transplantation, along with reduction of levels of inflammatory cytokines, suppression of STAT 5A/B phosphorylation, increased expression of CCR7 and increased production of nitrous oxide by hMSCs.Documentation of homing of hMSCs to lymphoid organs and target tissues was also performed.These mechanisms contribute to the current understanding of MSC mechanisms of immunosuppression and forms a comprehensive picture of how they exert immunosuppression in an in vivo model of immune dysregulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Paediatrics and Adolescent Medicine, The University of Hong Kong Li Ka Shing Faculty of Medicine, Queen Mary Hospital, 102 Pokfulam Rd., HKSAR, PRC.

ABSTRACT

Background: Mesenchymal stromal cells (MSCs) are proven to have immunosuppressive functions via various mechanisms. These mechanisms were demonstrated by administering bone marrow derived human MSCs (hMSCs) to graft versus host disease (GVHD) murine models.

Methods: BALB/c host mice were irradiated prior to receiving C57BL/6 donor T cell depleted bone marrow (TCDBM) cells (negative control) and donor CD4+ T lymphocyte with (treatment group) or without hMSCs (positive control). The presence of hMSCs in target tissues and lymphoid organs was documented by using in vivo imaging and measuring the expression of EphB2 and ephrin-B2 by RTqPCR. Survival rate and GVHD score were also monitored. Tissue sections were obtained for histopathologic analysis. Flow cytometry was used to document donor T cell alloreactivity and expression of CCR5, CXCR3 and CCR7. ELISA was utilized to determine levels of proinflammatory cytokines, RANTES (CCL5) and phosphorylated STAT 5A/B. RTqPCR was performed to quantify expression of CCL3 and CXCL9. Western blotting was performed to qualitatively measure iNOS expression.

Results: Survival rate and GVHD score improved with hMSC treatment. Pathologic changes of GVHD were abrogated. Documentation of suppression of RANTES, CCL3, CXCL9, CCR5 and CXCR3 with simultaneous decrease of donor T cell alloreactivity was demonstrated 6 days after transplantation, along with reduction of levels of inflammatory cytokines, suppression of STAT 5A/B phosphorylation, increased expression of CCR7 and increased production of nitrous oxide by hMSCs. Documentation of homing of hMSCs to lymphoid organs and target tissues was also performed.

Conclusions: These mechanisms contribute to the current understanding of MSC mechanisms of immunosuppression and forms a comprehensive picture of how they exert immunosuppression in an in vivo model of immune dysregulation.

No MeSH data available.


Related in: MedlinePlus

Detection of hMSC homing capacity. (A-B)In vivo imaging of hMSCs in GVHD host tissues. (C) mRNA levels of EphB2 and ephrin-B2 in GVHD host tissues. Lethally irradiated BALB/c host mice were given intravenous injections of 2 × 10^6. TCDBM cells and 0.25 × 10^6 CD4+ T cells from C57BL/6 donors with or without 1 × 10^6 hMSCs. hMSCs are stained with CM-Dil prior to injection to host mice. (A) Fluorescent signals from host liver (a), colon (b), lung (c)and spleen (d) 6d after transplantation without (1,2) or with (3,4) cotransplantation of hMSCs. Color bar represents signal intensity code. (B) Quantification of the signals in the liver, colon, lung and spleen. (C) Expression of EphB2 and ephrin-B2 6d after transplantation in BALB/c host spleen, colon and lung was measured by real-time RT-qPCR. Data were combined from 2 independent experiments with 2 mice in each experiment (n = 4 *p<0.05).
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Fig1: Detection of hMSC homing capacity. (A-B)In vivo imaging of hMSCs in GVHD host tissues. (C) mRNA levels of EphB2 and ephrin-B2 in GVHD host tissues. Lethally irradiated BALB/c host mice were given intravenous injections of 2 × 10^6. TCDBM cells and 0.25 × 10^6 CD4+ T cells from C57BL/6 donors with or without 1 × 10^6 hMSCs. hMSCs are stained with CM-Dil prior to injection to host mice. (A) Fluorescent signals from host liver (a), colon (b), lung (c)and spleen (d) 6d after transplantation without (1,2) or with (3,4) cotransplantation of hMSCs. Color bar represents signal intensity code. (B) Quantification of the signals in the liver, colon, lung and spleen. (C) Expression of EphB2 and ephrin-B2 6d after transplantation in BALB/c host spleen, colon and lung was measured by real-time RT-qPCR. Data were combined from 2 independent experiments with 2 mice in each experiment (n = 4 *p<0.05).

Mentions: In order to identify the earliest time we would be able to detect the presence of hMSCs in the GVHD target organs and to answer whether hMSCs home to these organs, hMSCs were stained with CM-Dil and injected into host mice at the time of transplantation. Migration patterns were observed towards the liver, colon, lungs and spleen. 6 days after transplantation, hMSCs can be localized in the liver, colon lung and spleen of GHVD host mice (Figure 1A-B).Figure 1


Immunosuppressive mechanisms of human bone marrow derived mesenchymal stromal cells in BALB/c host graft versus host disease murine models.

Robles JD, Liu YP, Cao J, Xiang Z, Cai Y, Manio M, Tang EH, Chan GC - Exp Hematol Oncol (2015)

Detection of hMSC homing capacity. (A-B)In vivo imaging of hMSCs in GVHD host tissues. (C) mRNA levels of EphB2 and ephrin-B2 in GVHD host tissues. Lethally irradiated BALB/c host mice were given intravenous injections of 2 × 10^6. TCDBM cells and 0.25 × 10^6 CD4+ T cells from C57BL/6 donors with or without 1 × 10^6 hMSCs. hMSCs are stained with CM-Dil prior to injection to host mice. (A) Fluorescent signals from host liver (a), colon (b), lung (c)and spleen (d) 6d after transplantation without (1,2) or with (3,4) cotransplantation of hMSCs. Color bar represents signal intensity code. (B) Quantification of the signals in the liver, colon, lung and spleen. (C) Expression of EphB2 and ephrin-B2 6d after transplantation in BALB/c host spleen, colon and lung was measured by real-time RT-qPCR. Data were combined from 2 independent experiments with 2 mice in each experiment (n = 4 *p<0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4440561&req=5

Fig1: Detection of hMSC homing capacity. (A-B)In vivo imaging of hMSCs in GVHD host tissues. (C) mRNA levels of EphB2 and ephrin-B2 in GVHD host tissues. Lethally irradiated BALB/c host mice were given intravenous injections of 2 × 10^6. TCDBM cells and 0.25 × 10^6 CD4+ T cells from C57BL/6 donors with or without 1 × 10^6 hMSCs. hMSCs are stained with CM-Dil prior to injection to host mice. (A) Fluorescent signals from host liver (a), colon (b), lung (c)and spleen (d) 6d after transplantation without (1,2) or with (3,4) cotransplantation of hMSCs. Color bar represents signal intensity code. (B) Quantification of the signals in the liver, colon, lung and spleen. (C) Expression of EphB2 and ephrin-B2 6d after transplantation in BALB/c host spleen, colon and lung was measured by real-time RT-qPCR. Data were combined from 2 independent experiments with 2 mice in each experiment (n = 4 *p<0.05).
Mentions: In order to identify the earliest time we would be able to detect the presence of hMSCs in the GVHD target organs and to answer whether hMSCs home to these organs, hMSCs were stained with CM-Dil and injected into host mice at the time of transplantation. Migration patterns were observed towards the liver, colon, lungs and spleen. 6 days after transplantation, hMSCs can be localized in the liver, colon lung and spleen of GHVD host mice (Figure 1A-B).Figure 1

Bottom Line: Documentation of suppression of RANTES, CCL3, CXCL9, CCR5 and CXCR3 with simultaneous decrease of donor T cell alloreactivity was demonstrated 6 days after transplantation, along with reduction of levels of inflammatory cytokines, suppression of STAT 5A/B phosphorylation, increased expression of CCR7 and increased production of nitrous oxide by hMSCs.Documentation of homing of hMSCs to lymphoid organs and target tissues was also performed.These mechanisms contribute to the current understanding of MSC mechanisms of immunosuppression and forms a comprehensive picture of how they exert immunosuppression in an in vivo model of immune dysregulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Paediatrics and Adolescent Medicine, The University of Hong Kong Li Ka Shing Faculty of Medicine, Queen Mary Hospital, 102 Pokfulam Rd., HKSAR, PRC.

ABSTRACT

Background: Mesenchymal stromal cells (MSCs) are proven to have immunosuppressive functions via various mechanisms. These mechanisms were demonstrated by administering bone marrow derived human MSCs (hMSCs) to graft versus host disease (GVHD) murine models.

Methods: BALB/c host mice were irradiated prior to receiving C57BL/6 donor T cell depleted bone marrow (TCDBM) cells (negative control) and donor CD4+ T lymphocyte with (treatment group) or without hMSCs (positive control). The presence of hMSCs in target tissues and lymphoid organs was documented by using in vivo imaging and measuring the expression of EphB2 and ephrin-B2 by RTqPCR. Survival rate and GVHD score were also monitored. Tissue sections were obtained for histopathologic analysis. Flow cytometry was used to document donor T cell alloreactivity and expression of CCR5, CXCR3 and CCR7. ELISA was utilized to determine levels of proinflammatory cytokines, RANTES (CCL5) and phosphorylated STAT 5A/B. RTqPCR was performed to quantify expression of CCL3 and CXCL9. Western blotting was performed to qualitatively measure iNOS expression.

Results: Survival rate and GVHD score improved with hMSC treatment. Pathologic changes of GVHD were abrogated. Documentation of suppression of RANTES, CCL3, CXCL9, CCR5 and CXCR3 with simultaneous decrease of donor T cell alloreactivity was demonstrated 6 days after transplantation, along with reduction of levels of inflammatory cytokines, suppression of STAT 5A/B phosphorylation, increased expression of CCR7 and increased production of nitrous oxide by hMSCs. Documentation of homing of hMSCs to lymphoid organs and target tissues was also performed.

Conclusions: These mechanisms contribute to the current understanding of MSC mechanisms of immunosuppression and forms a comprehensive picture of how they exert immunosuppression in an in vivo model of immune dysregulation.

No MeSH data available.


Related in: MedlinePlus