Publications by authors named "Songjie Cai"

25 Publications

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Overexpression of PD-1 on T cells promotes tolerance in cardiac transplantation via an ICOS-dependent mechanism.

JCI Insight 2021 Nov 9. Epub 2021 Nov 9.

Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, United States of America.

The PD-1: PD-L1 is a potent inhibitory pathway involved in immune regulation and a potential therapeutic target in transplantation. In this study, we show that overexpression of PD-1 (PD-1 Tg) on T cells promotes allograft tolerance in a fully MHC-mismatched cardiac transplant model when combined with costimulation blockade (CTLA-4-Ig). PD-1 overexpression on T cells also protected against chronic rejection in a single MHC II mismatched cardiac transplant model, while it still allowed the generation of an effective immune response against an Influenza A virus. Notably, Treg cells from PD-1 Tg mice were required for tolerance induction and presented higher ICOS expression than those from wild-type mice. Survival benefit of PD-1 Tg recipients required ICOS signaling and donor PD-L1 expression. These results indicate that modulation of PD-1 expression, in combination with a costimulation blockade, is a promising therapeutic target to promote transplant tolerance.
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http://dx.doi.org/10.1172/jci.insight.142909DOI Listing
November 2021

TAZ inhibits glucocorticoid receptor and coordinates hepatic glucose homeostasis in normal physiological states.

Elife 2021 10 8;10. Epub 2021 Oct 8.

Division of Endocrinology, Boston Children's Hospital, Harvard Medical School, Boston, United States.

The elucidation of the mechanisms whereby the liver maintains glucose homeostasis is crucial for the understanding of physiological and pathological states. Here, we show a novel role of hepatic transcriptional co-activator with PDZ-binding motif (TAZ) in the inhibition of glucocorticoid receptor (GR). TAZ is abundantly expressed in pericentral hepatocytes and its expression is markedly reduced by fasting. TAZ interacts via its WW domain with the ligand-binding domain of GR to limit the binding of GR to the GR response element in gluconeogenic gene promoters. Therefore, liver-specific TAZ knockout mice show increases in glucose production and blood glucose concentration. Conversely, the overexpression of TAZ in mouse liver reduces the binding of GR to gluconeogenic gene promoters and glucose production. Thus, our findings demonstrate that hepatic TAZ inhibits GR transactivation of gluconeogenic genes and coordinates gluconeogenesis in response to physiological fasting and feeding.
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http://dx.doi.org/10.7554/eLife.57462DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8555985PMC
October 2021

Editorial: Immune Regulation in Kidney Diseases: Importance, Mechanism and Translation.

Front Med (Lausanne) 2021 11;8:616880. Epub 2021 Feb 11.

Transplantation Research Center, Renal Division, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, United States.

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http://dx.doi.org/10.3389/fmed.2021.616880DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7928406PMC
February 2021

Follicular T cells mediate donor-specific antibody and rejection after solid organ transplantation.

Am J Transplant 2021 05 11;21(5):1893-1901. Epub 2021 Feb 11.

Transplantation Research Center, Renal Division, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.

Following solid organ transplantation, a substantial proportion of chronic allograft loss is attributed to the formation of donor-specific antibodies (DSAs) and antibody-mediated rejection (AbMR). The frequency and phenotype of T follicular helper (Tfh) and T follicular regulatory (Tfr) cells is altered in the setting of kidney transplantation, particularly in patients who develop AbMR. However, the roles of Tfh and Tfr cells in AbMR after solid organ transplantation is unclear. We developed mouse models to inducibly and potently perturb Tfh and Tfr cells to assess the roles of these cells in the development of DSA and AbMR. We found that Tfh cells are required for both de novo DSA responses as well as augmentation of DSA following presensitization. Using orthotopic allogeneic kidney transplantation models, we found that deletion of Tfh cells at the time of transplantation resulted in less severe transplant rejection. Furthermore, using inducible Tfr cell deletion strategies we found that Tfr cells inhibit de novo DSA formation but only have a minor role in controlling kidney transplant rejection. These studies demonstrate that Tfh cells promote, whereas Tfr cells inhibit, DSA to control rejection after kidney transplantation. Therefore, targeting these cells represent a new therapeutic strategy to prevent and treat AbMR.
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http://dx.doi.org/10.1111/ajt.16484DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8096660PMC
May 2021

Single-cell RNA sequencing reveals compromised immune microenvironment in precursor stages of multiple myeloma.

Nat Cancer 2020 May 27;1(5):493-506. Epub 2020 Apr 27.

Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts, 02215, USA.

Precursor states of Multiple Myeloma (MM) and its native tumor microenvironment need in-depth molecular characterization to better stratify and treat patients at risk. Using single-cell RNA sequencing of bone marrow cells from precursor stages, MGUS and smoldering myeloma (SMM), to full-blown MM alongside healthy donors, we demonstrate early immune changes during patient progression. We find NK cell abundance is frequently increased in early stages, and associated with altered chemokine receptor expression. As early as SMM, we show loss of GrK memory cytotoxic T-cells, and show their critical role in MM immunosurveillance in mouse models. Finally, we report MHC class II dysregulation in CD14 monocytes, which results in T cell suppression . These results provide a comprehensive map of immune changes at play over the evolution of pre-malignant MM, which will help develop strategies for immune-based patient stratification.
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http://dx.doi.org/10.1038/s43018-020-0053-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7785110PMC
May 2020

Regulatory T cells engineered with TCR signaling-responsive IL-2 nanogels suppress alloimmunity in sites of antigen encounter.

Sci Transl Med 2020 11;12(569)

Transplantation Research Center, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.

Adoptive cell transfer of ex vivo expanded regulatory T cells (T) has shown immense potential in animal models of auto- and alloimmunity. However, the effective translation of such T therapies to the clinic has been slow. Because T homeostasis is known to require continuous T cell receptor (TCR) ligation and exogenous interleukin-2 (IL-2), some investigators have explored the use of low-dose IL-2 injections to increase endogenous T responses. Systemic IL-2 immunotherapy, however, can also lead to the activation of cytotoxic T lymphocytes and natural killer cells, causing adverse therapeutic outcomes. Here, we describe a drug delivery platform, which can be engineered to autostimulate T with IL-2 in response to TCR-dependent activation, and thus activate these cells in sites of antigen encounter. To this end, protein nanogels (NGs) were synthesized with cleavable bis(-hydroxysuccinimide) cross-linkers and IL-2/Fc fusion (IL-2) proteins to form particles that release IL-2 under reducing conditions, as found at the surface of T cells receiving stimulation through the TCR. T surface-conjugated with IL-2 NGs were found to have preferential, allograft-protective effects relative to unmodified T or T stimulated with systemic IL-2. We demonstrate that murine and human NG-modified T carrying an IL-2 cargo perform better than conventional T in suppressing alloimmunity in murine and humanized mouse allotransplantation models. In all, the technology presented in this study has the potential to improve T transfer therapy by enabling the regulated spatiotemporal provision of IL-2 to antigen-primed T.
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http://dx.doi.org/10.1126/scitranslmed.aaw4744DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8519505PMC
November 2020

Donor myeloid derived suppressor cells (MDSCs) prolong allogeneic cardiac graft survival through programming of recipient myeloid cells in vivo.

Sci Rep 2020 08 28;10(1):14249. Epub 2020 Aug 28.

Renal Division, Transplantation Research Center, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Ave, Boston, MA, 02115, USA.

Solid organ transplantation is a lifesaving therapy for patients with end-organ disease. Current immunosuppression protocols are not designed to target antigen-specific alloimmunity and are uncapable of preventing chronic allograft injury. As myeloid-derived suppressor cells (MDSCs) are potent immunoregulatory cells, we tested whether donor-derived MDSCs can protect heart transplant allografts in an antigen-specific manner. C57BL/6 (H2K, I-A) recipients pre-treated with BALB/c MDSCs were transplanted with either donor-type (BALB/c, H2K, I-A) or third-party (C3H, H2K, I-A) cardiac grafts. Spleens and allografts from C57BL/6 recipients were harvested for immune phenotyping, transcriptomic profiling and functional assays. Single injection of donor-derived MDSCs significantly prolonged the fully MHC mismatched allogeneic cardiac graft survival in a donor-specific fashion. Transcriptomic analysis of allografts harvested from donor-derived MDSCs treated recipients showed down-regulated proinflammatory cytokines. Immune phenotyping showed that the donor MDSCs administration suppressed effector T cells in recipients. Interestingly, significant increase in recipient endogenous CD11bGr1 MDSC population was observed in the group treated with donor-derived MDSCs compared to the control groups. Depletion of this endogenous MDSCs with anti-Gr1 antibody reversed donor MDSCs-mediated allograft protection. Furthermore, we observed that the allogeneic mixed lymphocytes reaction was suppressed in the presence of CD11bGr1 MDSCs in a donor-specific manner. Donor-derived MDSCs prolong cardiac allograft survival in a donor-specific manner via induction of recipient's endogenous MDSCs.
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http://dx.doi.org/10.1038/s41598-020-71289-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7455707PMC
August 2020

High-mobility group box 1 protein antagonizes the immunosuppressive capacity and therapeutic effect of mesenchymal stem cells in acute kidney injury.

J Transl Med 2020 04 20;18(1):175. Epub 2020 Apr 20.

Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, 180 Fenglin Road, Shanghai, 200032, China.

Background: Kidney ischemia reperfusion injury (IRI) is a common cause of acute kidney injury and an unavoidable consequence of kidney transplantation and still lacks specific therapeutics. Recently, mesenchymal stem cell (MSC) has been emerging as a promising cell-based therapy for IRI in the context of transplantation. MSC negatively regulates the secretion of pro-inflammatory as well as the activation of immune cells during IRI through its unique immunosuppressive property.

Methods: We employed mice kidney IRI model and MSC cell line to monitor the IRI related checkpoints. siRNAs were utilized to knock down the potential key factors for mechanistic analysis. Statistical analysis was performed by using one-way ANOVA with Tukey's post hoc procedure by SPSS.

Results: The expression of high-mobility group box 1 protein (HMGB1) is increased in the acute phase as well as the recovery stage of IRI. Importantly, the HMGB1 upregulation is correlated with the injury severity. HMGB1 diminishes the MSC induced immunosuppressive capacity in the presence of pro-inflammatory cytokines in vitro. Toll like receptor 4 (TLR4)-mediated inducible nitric oxide synthase (iNOS) inhibition contributes to the negative effect of HMGB1 on MSCs. HMGB1-TLR4 signaling inhibition augments the therapeutic efficacy of MSCs in mice renal IRI model.

Conclusions: These findings demonstrate that HMGB1 plays a crucial role in shaping the immunoregulatory property of MSCs within the microenvironments, providing novel insights into the crosstalk between MSCs and microenvironment components, suggesting HMGB1 signals as a promising target to improve MSC-based therapy.
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http://dx.doi.org/10.1186/s12967-020-02334-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7169035PMC
April 2020

Regulatory CD8 T cells that recognize Qa-1 expressed by CD4 T-helper cells inhibit rejection of heart allografts.

Proc Natl Acad Sci U S A 2020 03 28;117(11):6042-6046. Epub 2020 Feb 28.

Division of Renal Medicine, Transplantation Research Center, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115;

Induction of longstanding immunologic tolerance is essential for survival of transplanted organs and tissues. Despite recent advances in immunosuppression protocols, allograft damage inflicted by antibody specific for donor organs continues to represent a major obstacle to graft survival. Here we report that activation of regulatory CD8 T cells (CD8 Treg) that recognize the Qa-1 class Ib major histocompatibility complex (MHC), a mouse homolog of human leukocyte antigen-E (HLA-E), inhibits antibody-mediated immune rejection of heart allografts. We analyzed this response using a mouse model that harbors a point mutation in the class Ib MHC molecule Qa-1, which disrupts Qa-1 binding to the T cell receptor (TCR)-CD8 complex and impairs the CD8 Treg response. Despite administration of cytotoxic T lymphocyte antigen 4 (CTLA-4) immunoglobulin (Ig), Qa-1 mutant mice developed robust donor-specific antibody responses and accelerated heart graft rejection. We show that these allo-antibody responses reflect diminished Qa-1-restricted CD8 Treg-mediated suppression of host follicular helper T cell-dependent antibody production. These findings underscore the critical contribution of this Qa-1/HLA-E-dependent regulatory pathway to maintenance of transplanted organs and suggest therapeutic approaches to ameliorate allograft rejection.
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http://dx.doi.org/10.1073/pnas.1918950117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7084119PMC
March 2020

Notch-1 Inhibition Promotes Immune Regulation in Transplantation Via Regulatory T Cell-Dependent Mechanisms.

Circulation 2019 09 3;140(10):846-863. Epub 2019 Jul 3.

Transplantation Research Center, Brigham & Women's Hospital, Harvard Medical School, Boston, MA (C.N.M., N.M., T.J.B., T.S., K.S., S.O., S.C., A.U., J.A., N.N., L.V.R.).

Background: Transplantation is the treatment of choice for many patients with end-stage organ disease. Despite advances in immunosuppression, long-term outcomes remain suboptimal, hampered by drug toxicity and immune-mediated injury, the leading cause of late graft loss. The development of therapies that promote regulation while suppressing effector immunity is imperative to improve graft survival and minimize conventional immunosuppression. Notch signaling is a highly conserved pathway pivotal to T-cell differentiation and function, rendering it a target of interest in efforts to manipulate T cell-mediated immunity.

Methods: We investigated the pattern of Notch-1 expression in effector and regulatory T cells (Tregs) in both murine and human recipients of a solid-organ transplant. Using a selective human anti-Notch-1 antibody (aNotch-1), we examined the effect of Notch-1 receptor inhibition in full major histocompatibility complex-mismatch murine cardiac and lung transplant models, and in a humanized skin transplant model. On the basis of our findings, we further used a genetic approach to investigate the effect of selective Notch-1 inhibition in Tregs.

Results: We observed an increased proportion of Tregs expressing surface and intracellular (activated) Notch-1 in comparison with conventional T cells, both in mice with transplants and in the peripheral blood of patients with transplants. In the murine cardiac transplant model, peritransplant administration of aNotch-1 (days 0, 2, 4, 6, 8, and 10) significantly prolonged allograft survival in comparison with immunoglobulin G-treated controls. Similarly, aNotch-1 treatment improved both histological and functional outcomes in the murine lung transplant model. The use of aNotch-1 resulted in a reduced proportion of both splenic and intragraft conventional T cells, while increasing the proportion of Tregs. Furthermore, Tregs isolated from aNotch-1-treated mice showed enhanced suppressive function on a per-cell basis, confirmed with selective Notch-1 deletion in Tregs (Foxp3Notch1). Notch-1 blockade inhibited the mammalian target of rapamycin pathway and increased the phosphorylation of STAT5 (signal transducer and activator of transcription 5) in murine Tregs. Notch-1 Tregs isolated from human peripheral blood exhibited more potent suppressive capacity than Notch-1 Tregs. Last, the combination of aNotch-1 with costimulation blockade induced long-term tolerance in a cardiac transplant model, and this tolerance was dependent on CTLA-4 (cytotoxic T-lymphocyte-associated antigen-4) signaling.

Conclusions: Our data reveal a promising, clinically relevant approach for immune modulation in transplantation by selectively targeting Notch-1.
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http://dx.doi.org/10.1161/CIRCULATIONAHA.119.040563DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722011PMC
September 2019

Cell Therapy in Solid Organ Transplantation.

Curr Gene Ther 2019 ;19(2):71-80

Transplantation Research Center, Renal Division, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Avenue, Boston, MA 02115, United States.

Transplantation is the only cure for end-stage organ failure. Current immunosuppressive drugs have two major limitations: 1) non antigen specificity, which increases the risk of cancer and infection diseases, and 2) chronic toxicity. Cell therapy appears to be an innovative and promising strategy to minimize the use of immunosuppression in transplantation and to improve long-term graft survival. Preclinical studies have shown efficacy and safety of using various suppressor cells, such as regulatory T cells, regulatory B cells and tolerogenic dendritic cells. Recent clinical trials using cellbased therapies in solid organ transplantation also hold out the promise of improving efficacy. In this review, we will briefly go over the rejection process, current immunosuppressive drugs, and the potential therapeutic use of regulatory cells in transplantation.
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http://dx.doi.org/10.2174/1566523219666190603103840DOI Listing
July 2020

Potential Roles of Siglecs in the Regulation of Allo-Immune Reaction.

Curr Protein Pept Sci 2019 ;20(8):823-828

Transplantation Research Center, Renal Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, United States.

Siglecs are mammalian sialic acid (Sia) recognizing immuno-globulin-like receptors expressed across the major leukocyte lineages, and function to recognize ubiquitous Sia epitopes on the cell surface. Many Siglecs are inhibitory receptors expressed on innate immune cells, they also have a role in maintaining B cell tolerance as well as modulating the activation of conventional and plasmocytic dendritic cells. Through these and other roles they contribute directly and indirectly to the regulation of T cell function. Siglecs have been identified to play key roles in several forms of blood cancers, autoimmune and infection deceases. So far as we know, there's no Siglecs related research works on solid organ transplantation. In this review, we describe our understanding of the potential roles of Siglecs in the regulation of immune cell function, which may be crosslinked to allo-rejection and ischemia-reperfusion injury.
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http://dx.doi.org/10.2174/1389203720666190507095759DOI Listing
September 2019

Administration of Dendritic Cells and Anti-PD-1 Antibody Converts X-ray Irradiated Tumors Into Effective In situ Vaccines.

Int J Radiat Oncol Biol Phys 2019 03 17;103(4):958-969. Epub 2018 Nov 17.

Proton Medical Research Center, University of Tsukuba, Tsukuba, Japan. Electronic address:

Purpose: Danger signals and release of tumor-specific antigens after exposure to ionizing radiation can convert an irradiated tumor into an in situ vaccine. However, radiation alone is not sufficient to induce an effective systemic immune response. In this study, we investigated whether a combination of x-ray irradiation with bone marrow-derived dendritic cells (BM-DCs) and anti-PD-1 antibody (αPD1-ab) administration can enhance both local tumor control and the systemic abscopal effect in murine subcutaneous tumor models.

Methods And Materials: B16/BL6 melanoma and Lewis lung carcinoma cells were examined for radiosensitivity and expression of H-2kd and PD-L1 before and after irradiation. The tumor cells were implanted subcutaneously in the left thigh of C57BL/6 mice as primary tumors. BM-DCs were induced from mouse bone marrow cells using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). The primary tumors were treated with 8 Gy of x-ray, followed by simultaneous intratumoral injection of BM-DCs and intraperitoneal injection of αPD1-ab. To examine the abscopal effect, the same tumor cells were also inoculated in the right thigh as metastatic tumors 4 days after the primary tumor inoculation, and only the primary tumors were treated with the same protocols. In vivo analyses of tumor growth and survival rates and in vitro analyses of splenic T-cell proliferation and interferon-γ release were performed.

Results: The triple-combination treatment of x-ray irradiation with BM-DC and αPD1-ab administration inhibited primary tumor growth and significantly extended survival time in association with significant increase of T-cell proliferation and interferon-γ release. In addition, this triple-combination treatment significantly inhibited the growth of metastatic tumors.

Conclusions: The results indicated that BM-DC and αPD1-ab administration led to the conversion of irradiated tumors into effective in situ vaccines. This combination therapy can be a promising approach to develop a novel individualized therapy for patients with solid cancers.
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http://dx.doi.org/10.1016/j.ijrobp.2018.11.019DOI Listing
March 2019

How Do Dendritic Cells Play the Role in Ischemia/Reperfusion Triggered Kidney Allograft Rejection.

Curr Gene Ther 2017 ;17(6):400-404

Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Osaka, Japan.

In deceased donors, Ischemia/Reperfusion Injury (IRI) is an important cause of allograft dysfunction. Prolonged cold and warm ischemia time leads to a high risk of early post-transplant complications, including acute and chronic rejection. Ischemia not only up-regulates inflammatory cytokines and chemokines, but also enhances the expression of MHC-class II and adhesion molecules on epithelial and dendritic cells. Moreover, the Danger Associated Molecular Patterns (DAMPs) released from stressed or dying cells, not only cause or amplify tissue inflammation and trigger tissue repair in response to IRI, but also act as adjuvants that enhance DC maturation and potentiate the adaptive immune response. In this review, we will also discuss about whether donor or recipient DCs are more important in the process of ischemia enhanced acute rejection.
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http://dx.doi.org/10.2174/1566523218666180214095956DOI Listing
April 2019

Priming of T Cells with Pulsed Dendritic Cells: Popliteal Lymph Node Assay.

Bio Protoc 2017 Sep 5;7(17):e2531. Epub 2017 Sep 5.

Division of Transplantation Immunology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo, Japan.

One-way mixed lymphocyte reaction (MLR) is a classic tool to measure how T cells react to external stimuli. However, MLR is an reaction system, which shows different response intensity compared with trails sometimes due to the lack of cytokines, tissue matrix and other immune response associated factors. The following popliteal lymph node assay (PLNA) protocol is designed to test the T cells antigen-specific reaction by using ovalbumin (OVA) specific reacted transgenic mouse OT-1 and OT-2.
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http://dx.doi.org/10.21769/BioProtoc.2531DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8413589PMC
September 2017

iPSC-Derived Regulatory Dendritic Cells Inhibit Allograft Rejection by Generating Alloantigen-Specific Regulatory T Cells.

Stem Cell Reports 2017 05 20;8(5):1174-1189. Epub 2017 Apr 20.

Division of Transplantation Immunology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan. Electronic address:

Regulatory dendritic cell (DCregs)-based immunotherapy is a potential therapeutic tool for transplant rejection. We generated DCregs from murine induced pluripotent stem cells (iPSCs), which could remain in a "stable immature stage" even under strong stimulation. Harnessing this characteristic, we hypothesized that iPS-DCregs worked as a negative vaccine to generate regulatory T cells (Tregs), and induced donor-specific allograft acceptance. We immunized naive CBA (H-2K) mice with B6 (H-2K) iPS-DCregs and found that Tregs (CD4CD25FOXP3) significantly increased in CBA splenocytes. Moreover, immunized CBA recipients permanently accepted B6 cardiac grafts in a donor-specific pattern. We demonstrated mechanistically that donor-type iPS-DCregs triggered transforming growth factor β1 secretion, under which the donor-antigen peptides directed naive CD4 T cells to differentiate into donor-specific FOXP3 Tregs instead of into effector T cells in vivo. These findings highlight the potential of iPS-DCregs as a key cell therapy resource in clinical transplantation.
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http://dx.doi.org/10.1016/j.stemcr.2017.03.020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5425686PMC
May 2017

MicroRNAs Involved in Acute Rejection and Tolerance in Murine Cardiac Allografts.

Exp Clin Transplant 2016 Aug 26;14(4):424-30. Epub 2016 May 26.

rom the Division of Transplantation Immunology National Research Institute for Child Health and Development; and the AIDS Research Center National Institute of Infectious Diseases Tokyo Japan.

Objectives: Induction of immunologic tolerance is the ultimate goal of organ transplant. To investigate the involvement of microRNA in tolerance induction after organ transplant, murine cardiac allografts were performed and the expression of microRNA in the grafts was analyzed.

Materials And Methods: Cardiac allografts were performed using C57BL/10 (H2-Kb) to CBA/N (H2-Kk) fully mismatched combination with or without eicosapentaenoic acid for tolerance induction. Ten microRNA, mir-146a, 15b, 223, 23a, 27a, 34a, 451, 101a, 101b, 148a, discovered in hepatic grafts were examined by quantitative reverse transcription polymerase chain reaction using RNA from the cardiac allografts.

Results: The administration of eicosapentaenoic acid markedly prolonged the cardiac allograft survival (median survival time > 100 days) and decreased the pathological score. Quantitative reverse transcription polymerase chain reaction revealed that mir-223 was up-regulated in accordance with pathological deterioration as compared with the expression observed in the syngeneic grafts. In contrast, the other microRNA was down-regulated. Pearson product moment correlation analysis demonstrated that the expression patterns of mir-223 and mir-146a had high or moderate positive associations between the cardiac and haptic allografts in mice.

Conclusions: The change in the microRNA expression in the allografts suggests that microRNA plays a role in the induction and/or maintenance of tolerance after allograft transplant. Our findings suggest that mir-223 may be associated with rejection while mir-146a, -15b, -23a, -27a, -34a, -451, -101a, -101b, -148a may be involved in tolerance. A superior grasp of the mechanism for rejection and tolerance observed in the murine heart allotransplant model may provide a better curative treatment strategy to mitigate allograft rejection.
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http://dx.doi.org/10.6002/ect.2015.0251DOI Listing
August 2016

Prolonged Mouse Cardiac Graft Cold Storage via Attenuating Ischemia-Reperfusion Injury Using a New Antioxidant-Based Preservation Solution.

Transplantation 2016 05;100(5):1032-40

1 Division of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan. 2 Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Osaka, Japan. 3 AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan. 4 SBI Pharmaceuticals Co., Ltd., Tokyo, Japan. 5 Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.

Background: One of the major events in ischemia-reperfusion (I/R)-induced heart injury in cardiac transplantation is the generation of reactive oxygen species. We hypothesized that a novel preservation solution called SBI-SEIIKU II (SS-II) contains 3 antioxidant reagents: L-cysteine, glycine, ascorbic acid/ascorbic acid-2-phosphate magnesium, which can block the generation of reactive oxygen species to result in a prolongation of the cold storage time via attenuating I/R injury.

Methods: C57BL/6CrSlc(B6) mice underwent syngeneic mice heterotopic heart transplantation, and the animals were derived into 3 groups: recipients with nonpreserved grafts (control group), recipients with grafts preserved in histidine-tryptophan-ketoglutarate (HTK) for 24 and 48 hours (HTK group), and recipients with grafts preserved in SS-II for 24 and 48 hours (SS-II group).

Results: After 48 hours of preservation, there were no grafts that survived in the HTK group; however, the SS-II group had a high survival rate. After 24 hours of preservation, SS-II decreased the oxidative damage, myocardial apoptosis, and the infiltration of macrophages and neutrophils in the cardiac grafts in the early phase and suppressed the development of myocardial fibrosis in long-term grafts compared with HTK.

Conclusions: The SS-II prolongs the acceptable cold storage time and protects the myocardium from I/R injury via inhibiting oxidative stress-associated damage. We believe that this novel preservation solution may be simple and safe for use in the clinical transplantation field.
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http://dx.doi.org/10.1097/TP.0000000000001079DOI Listing
May 2016

Impaired CD98 signaling protects against graft-versus-host disease by increasing regulatory T cells.

Transpl Immunol 2016 Mar 1;35:34-9. Epub 2016 Feb 1.

Division of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan. Electronic address:

Graft-versus-host disease (GvHD) is a major barrier to the broader use of allogenic hematopoietic stem cell transplantation for non-malignant clinical applications. A murine model of C57BL/6 to B6D2F1 acute GvHD was employed with T lymphocytes harboring a deletion of the CD98 heavy chain (CD98hc(-/-)) as donor cells. The CD98hc(-/-) resulted in lower responses to alloantigen stimulation in a mixed leukocyte reaction assay, and prevented the mortality associated with disease progression. The percentage of donor CD8 T lymphocytes was significantly decreased, while the percentage of Foxp3-positive regulatory T cells (Tregs) in recipients was increased by CD98hc(-/-). Decreased expression of FAS, FASL, ICOS, ICOSL, PD-1 and PD-L1 by donor CD8 T cells, and mRNA expression of cytotoxic T cell-related cytokines in the recipients were shown in those with CD98hc(-/-). Fewer infiltrated cells are found in the lungs, liver, tongue and skin of recipients with CD98hc(-/-) compared with the wild type recipients. Taken together, our data indicate that T cell-specific deletion of CD98hc can contribute to the prevention of GvHD development due to the attenuation of lymphocyte migration and by increasing the generation of Treg cells. These findings are expected to make it possible to develop novel approaches for the prevention of GvHD.
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http://dx.doi.org/10.1016/j.trim.2016.01.005DOI Listing
March 2016

Noninvasive Monitoring and Evaluation of the Renal Structure and Function in a Mouse Model of Unilateral Ureteral Occlusion Using Microcomputed Tomography.

Int Surg 2015 Jul;100(7-8):1237-43

1 Division of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan.

Mouse unilateral ureteral occlusion (UUO) is widely used as a model of renal experimental obstructive nephropathy with interstitial fibrosis. Microcomputed tomography (micro-CT) imaging has the potential to produce quantitative images. The aim of this study was to establish standard images of micro-CT for renal anatomic and functional evaluations in a mouse model of UUO. UUO was induced in adult male mice BALB/c. In total, 27 mice were used in this study. Three mice per group (a total of 6 groups) were examined with contrast-enhanced micro-CT prior to UUO (day 0) and on days 1, 3, 5, 7, 10, and 14 after UUO. In order to determine the histopathologic correlations at each point in time, contrast-enhanced micro-CT imaging was performed in the 18 remaining mice. All animals were sacrificed, and both kidneys were harvested after the final micro-CT examination. UUO resulted in hydronephrosis and changes in the renal parenchyma. The predominant alteration was substantial changes in the hemodynamics of the renal vascular system after ureteral obstruction for 24 hours or longer, which may be resulting from increased action of vasoconstrictors versus vasodilators. The renal parenchyma was significantly reduced after 1 week, and the features of the histologic changes supported the findings of the micro-CT images. In the contralateral unobstructed kidneys, the images showed a normal structure and function and the pathohistology revealed a normal histoarchitecture. Micro-CT is a useful tool for providing noninvasive monitoring and evaluating the renal structure and function.
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http://dx.doi.org/10.9738/INTSURG-D-14-00273.1DOI Listing
July 2015

Noninvasive monitoring of mouse renal allograft rejection using micro-CT.

Ann Surg Treat Res 2015 May 30;88(5):276-80. Epub 2015 Apr 30.

Division of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan.

Purpose: Acute renal graft rejection can only be definitively diagnosed by renal biopsy. However, biopsies carry a risk of renal transplant injury and loss. Micro-CT is widely used in preclinical studies of small animals. Here, we propose micro-CT could noninvasively monitor and evaluate renal location and function in a mouse kidney transplant model.

Methods: Orthotopic kidney transplantation was performed in a BALB/c -to- C57BL/6j or C57BL/6j-to- C57BL/6j mouse model. After optimizing imaging techniques, five mice were imaged with micro-CT and the findings were verified histologically.

Results: Micro-CT can monitor and evaluate renal location and function after orthotopic kidney transplantation. There were no mice deaths while renal transplants were failure.

Conclusion: We propose that graft micro-CT imaging is a new option that is noninvasive and specific, and can aid in early detection and follow-up of acute renal rejection. This method is potentially useful to improve posttransplant rejection monitoring.
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http://dx.doi.org/10.4174/astr.2015.88.5.276DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4422881PMC
May 2015

Luminal injection of hydrogen-rich solution attenuates intestinal ischemia-reperfusion injury in rats.

Transplantation 2015 Mar;99(3):500-7

1 Organ Transplant Center, National Center for Child Health and Development, Tokyo, Japan. 2 Division of Hepato-Biliary-Pancretic Surgery and Transplantation, Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. 3 Division of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan. 4 MiZ Co., Ltd., Kanagawa, Japan. 5 Department of Clinical Pathology, National Center for Child Health and Development, Tokyo, Japan.

Background: Luminal preservation of the intestine is an attractive method to locally mitigate preservation injury and ischemic-reperfusion injury in small bowel transplantation (SBT) because this method has a potential to maintain the intestinal graft integrity. Hydrogen is noted as an antioxidant material by reducing hydroxyl radicals. We hypothesized that hydrogen-containing solution can be an optimum material for luminal preservation method in SBT.

Methods: Ischemic reperfusion was induced in Lewis rats by occlusion of the supramesenteric artery and vein for 90 min. Experimental protocols were divided into four groups: sham operation group, no luminal injection (control) group, luminal injection of 5% glucose saline (GS) solution group, and luminal injection of hydrogen-rich GS (HRGS) group. Two milliliters of experimental solution was locally injected into the lumen of the intestine before declamping of vessels. Oxidative stress markers, proinflammatory cytokines, apoptosis in the crypt cells, and morphologic changes of the intestine were assessed.

Results: The production of malondialdehyde and 8-hydroxydeoxyguanosine, as oxidative stress markers, were markedly suppressed in HRGS group. The level of proinflammatory cytokines, such as inducible nitric oxide synthase and interleukin-6, was significantly inhibited in HRGS group. Crypt apoptosis was also significantly suppressed in HRGS group. Histopathologically, integrity of villus in intestine was maintained in HRGS group in comparison to the other groups.

Conclusion: Luminal injection of hydrogen-rich solution can reduce oxidative stress and consequently ameliorate ischemic-reperfusion injury. Hydrogen-containing solution can be a novel and promising luminal preservation material in SBT.
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http://dx.doi.org/10.1097/TP.0000000000000510DOI Listing
March 2015

5-Aminolevulinic acid with ferrous iron induces permanent cardiac allograft acceptance in mice via induction of regulatory cells.

J Heart Lung Transplant 2015 Feb 2;34(2):254-63. Epub 2014 Oct 2.

Division of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo. Electronic address:

Background: 5-Aminolevulinic acid (5-ALA), a precursor of heme biosynthesis, plays a fundamentally important role in aerobic energy metabolism. Heme oxygenase (HO)-1 cleaves heme to form biliverdin, carbon monoxide (CO) and iron (Fe(2+)). The anti-inflammatory properties of biliverdin and CO help to alleviate ischemia/reperfusion injury as well as acute and/or chronic allograft rejection. We investigated whether 5-ALA and Fe(2+) exerts salutary effects in the setting of organ transplantation.

Methods: An in vitro mixed-lymphocyte reaction (MLR) assay and cardiac allotransplantation model (CBA to C57BL/10) were used to evaluate the effects of 5-ALA and Fe(2+) on transplantation tolerance.

Results: Treatment with 5-ALA and sodium ferrous citrate (SFC) resulted in permanent acceptance in the murine cardiac allografts in a dose-, SFC- and HO-1-dependent manner. The number of graft-infiltrating CD8 T cells was lower and the survival response of recipient spleen T cells to donor-type alloantigens was less compared with control recipients; however, numbers of both regulatory T cells and dendritic cells were significantly increased in 5-ALA/SFC-treated recipients.

Conclusions: Our findings show that 5-ALA/SFC inhibits T-cell proliferation in response to alloantigens and an increased number of regulatory cells, resulting in permanent cardiac allograft acceptance in mice. These findings highlight the major roles of CO and/or HO-1 in inducing tolerance and suggest that 5-ALA/SFC may be a clinically effective treatment for allograft rejection.
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http://dx.doi.org/10.1016/j.healun.2014.09.037DOI Listing
February 2015

Generation and characterization of regulatory dendritic cells derived from murine induced pluripotent stem cells.

Sci Rep 2014 Feb 5;4:3979. Epub 2014 Feb 5.

1] Department of Dermatology, Huashan hospital, Fudan University, Shanghai, China [2] Division of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan.

Regulatory dendritic cells (DCregs) represent a potential therapeutic tool for assessing a variety of immune overreaction conditions; however, current approaches for generating DCregs for therapeutic purposes are limited. We attempted to generate and characterize DCregs from murine induced pluripotent stem (iPS) cells. The iPS cells co-cultured with OP9 cells displayed mesodermally differentiated flat colonies. GM-CSF drove most of the colonies exhibiting a differentiated morphology. Thereafter, cells became morphologically heterologous under the effects of TGF-β and IL-10. Most of the floating cells developed an irregular shape with areas of protrusion. The generated iPS-DCregs demonstrated high CD11b/c and low CD40, CD80, CD86 and MHC-II expressions with a high antigen uptake ability and poor T-cell stimulatory function. Importantly, iPS-DCregs showed immune responsiveness regulation effects both in vitro and in vivo and the ability to generate regulatory T-cells in vitro. Our result illustrates a feasible approach for generating functional DCregs from murine iPS cells.
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http://dx.doi.org/10.1038/srep03979DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3913921PMC
February 2014

5-Aminolevulinic acid combined with ferrous iron induces carbon monoxide generation in mouse kidneys and protects from renal ischemia-reperfusion injury.

Am J Physiol Renal Physiol 2013 Oct 31;305(8):F1149-57. Epub 2013 Jul 31.

Div. of Radiation Safety and Immune Tolerance, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan.

Renal ischemia reperfusion injury (IRI) is a major factor responsible for acute renal failure. An intermediate in heme synthesis, 5-aminolevulinic acid (5-ALA) is fundamental in aerobic energy metabolism. Heme oxygenase (HO)-1 cleaves heme to form biliverdin, carbon monoxide (CO), and iron (Fe(2+)), which is used with 5-ALA. In the present study, we investigated the role of 5-ALA in the attenuation of acute renal IRI using a mouse model. Male Balb/c mice received 30 mg/kg 5-ALA with Fe(2+) 48, 24, and 2 h before IRI and were subsequently subjected to bilateral renal pedicle occlusion for 45 min. The endogenous CO concentration of the kidneys from the mice administered 5-ALA/Fe(2+) increased significantly, and the peak concentrations of serum creatinine and blood urea nitrogen decreased. 5-ALA/Fe(2+) treatments significantly decreased the tubular damage and number of apoptotic cells. IRI-induced renal thiobarbituric acid-reactive substance levels were also significantly decreased in the 5-ALA/Fe(2+) group. Furthermore, mRNA expression of HO-1, TNF-α, and interferon-γ was significantly increased after IRI. Levels of HO-1 were increased and levels of TNF-α and interferon-γ were decreased in the 5-ALA/Fe(2+)-pretreated renal parenchyma after IRI. F4/80 staining showed reduced macrophage infiltration, and TUNEL staining revealed that there were fewer interstitial apoptotic cells. These findings suggest that 5-ALA/Fe(2+) can protect the kidneys against IRI by reducing macrophage infiltration and decreasing renal cell apoptosis via the generation of CO.
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http://dx.doi.org/10.1152/ajprenal.00275.2013DOI Listing
October 2013
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