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Cited by 10Organ and Tissue Transplantation ResearchTransplantation: Methods and OutcomesPluripotent Stem Cells Research

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Establishment of an experimental model for MHC homo-to-hetero transplantation

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Murata, Tomoki, Wada, Haruka, Otsuka, Ryo, Sasaki, Airi, Tsuji, Hyuma, Itoh, Mizuho, Eguchi, Nanami, Kawai, Tatsuo, Seino, Ken-ichiro·2020·Scientific Reports
DOI 10.1038/s41598-020-69784-4PMC PMC7421494
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Abstract

Preventing rejection is a major challenge in transplantation medicine, even when using pluripotent stem cell-derived grafts. In iPS cell (iPSC)-based transplantation, to reduce the risk of rejection, it is thought to be optimal that preparing the cells from donors whose human leukocyte antigen-haplotype are homozygous. Generally, this approach is referred to as major histocompatibility complex (MHC) homo-to-hetero transplantation, which is MHC-matched but minor antigen-mismatched. To investigate the immune response in the MHC homo-to-hetero transplantation, we established a murine experimental system in which MHC-matched but minor antigen-mismatched tissue (skin) grafts were transplanted into MHC-heterozygous recipients. Unexpectedly, only minor antigen-mismatched grafts were rejected at the same time points as rejection of fully allogeneic grafts. A vigorous anti-donor type T cell response was detected in vitro and conventional immunosuppressants targeting T cell activation had limited effects on controlling rejection. However, anti-donor antibodies were not detected only in the minor antigen-mismatched transplantation. This murine transplantation model can be used to further analyze immunological subjects for MHC homo-to-hetero iPSC-based transplantation.

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Figure 1 Combination between donor and recipient is critical for determining the extent of immune rejection in MHC-matched but minor antigen-mismatched transplantation. Auto or allogeneic skin grafts (BALB/c (MHC haplotype: d/d), C57BL/6 (b/b), CBA/N (k/k), or 129X1/Sv (b/b)) were transplanted into MHC-heterozygous recipients. The recipients were (a) C3B10F1 (k/b), (b) B6D2F1 (b/d), and (c) C3129F1 (k/b). **p < 0.01, log-rank test.
Figure 1 Combination between donor and recipient is critical for determining the extent of immune rejection in MHC-matched but minor antigen-mismatched transplantation. Auto or allogeneic skin grafts (BALB/c (MHC haplotype: d/d), C57BL/6 (b/b), CBA/N (k/k), or 129X1/Sv (b/b)) were transplanted into MHC-heterozygous recipients. The recipients were (a) C3B10F1 (k/b), (b) B6D2F1 (b/d), and (c) C3129F1 (k/b). **p < 0.01, log-rank test.CC BY · PubMed Central
Figure 2 Severe infiltration of CD3+ or Iba-1+ cells in MHC-matched but minor antigen-mismatched skin grafts. At 7 days after transplantation and until 10 days after transplantation, the skin grafts were harvested and analyzed by immunohistochemistry. This is because BALB/c grafts were too severely destroyed to be analyzed at day 10. The broken lines indicate the border of the graft tissues (G) and recipient (R). Bars: 50 µm.
Figure 2 Severe infiltration of CD3+ or Iba-1+ cells in MHC-matched but minor antigen-mismatched skin grafts. At 7 days after transplantation and until 10 days after transplantation, the skin grafts were harvested and analyzed by immunohistochemistry. This is because BALB/c grafts were too severely destroyed to be analyzed at day 10. The broken lines indicate the border of the graft tissues (G) and recipient (R). Bars: 50 µm.CC BY · PubMed Central
Figure 3 Recipient T cell response in MHC-matched but minor antigen-mismatched transplantation. (a) CFSE-labeled Thy1.2+ T cells isolated from naïve C3129F1 mice were co-cultured with bone marrow DCs derived from C3129F1, BALB/c, C57BL/6, or CBA/N mice. Four days later, the cells were stained with anti-CD4 or anti-CD8 antibodies, and then analyzed by flow cytometry. (b) To detect indirect alloantigen recognition, the CD19-negative fraction of splenocytes from C3129F1 recipient mice that had rejected skin grafts from BALB/c, C57BL/6, or CBA/N were used as responder cells. CFSE-labeled responder cells were co-cultured with 30 Gy irradiated splenocytes from C3129F1, BALB/c, C57BL/6, or CBA/N mice. Four days later, the cells were stained with anti-CD4 or anti-CD8 antibodies, and then analyzed by flow cytometry. For (a) and (b), T cell proliferation was evaluated based on CSFE reduction. Each bar shows the mean ± SD of triplicate cultures. The results are representative of three or two independent experiments. **p < 0.01, Tukey’s HSD test.
Figure 3 Recipient T cell response in MHC-matched but minor antigen-mismatched transplantation. (a) CFSE-labeled Thy1.2+ T cells isolated from naïve C3129F1 mice were co-cultured with bone marrow DCs derived from C3129F1, BALB/c, C57BL/6, or CBA/N mice. Four days later, the cells were stained with anti-CD4 or anti-CD8 antibodies, and then analyzed by flow cytometry. (b) To detect indirect alloantigen recognition, the CD19-negative fraction of splenocytes from C3129F1 recipient mice that had rejected skin grafts from BALB/c, C57BL/6, or CBA/N were used as responder cells. CFSE-labeled responder cells were co-cultured with 30 Gy irradiated splenocytes from C3129F1, BALB/c, C57BL/6, or CBA/N mice. Four days later, the cells were stained with anti-CD4 or anti-CD8 antibodies, and then analyzed by flow cytometry. For (a) and (b), T cell proliferation was evaluated based on CSFE reduction. Each bar shows the mean ± SD of triplicate cultures. The results are representative of three or two independent experiments. **p < 0.01, Tukey’s HSD test.CC BY · PubMed Central
Figure 4 Limited effect of conventional immunosuppressants to control the rejection in MHC-matched but minor antigen-mismatched transplantation. C3129F1 recipient mice were intraperitoneally administered 0.5 mg/kg/day (a) or 2.0 mg/kg/day (b) tacrolimus, or 1.0 mg/kg/day rapamycin (c). The treatment was started from the day of skin transplantation. *p < 0.05, **p < 0.01, the log-rank test.
Figure 4 Limited effect of conventional immunosuppressants to control the rejection in MHC-matched but minor antigen-mismatched transplantation. C3129F1 recipient mice were intraperitoneally administered 0.5 mg/kg/day (a) or 2.0 mg/kg/day (b) tacrolimus, or 1.0 mg/kg/day rapamycin (c). The treatment was started from the day of skin transplantation. *p < 0.05, **p < 0.01, the log-rank test.CC BY · PubMed Central
Figure 5 De novo anti-donor antibody production in MHC heterozygous recipients. (a–d) Sera were collected from recipient mice that underwent graft rejection. Thymocytes from the indicated mice were first stained with collected sera, and then further stained with PE-conjugated anti-mouse IgM antibody (a,b) or Alexa Fluor 488-conjugated anti-mouse IgG antibody (c,d). Thymocytes were analyzed by flow cytometry. The open histogram shows naïve C3129F1 mice sera staining, and filled histogram shows recipient C3129F1 sera staining. The data shown in (b,d) are representative of staining of the sera from six independent mice. Each bar shows the average mean fluorescence intensity ± SEM. **p < 0.01, Tukey’s HSD test.
Figure 5 De novo anti-donor antibody production in MHC heterozygous recipients. (a–d) Sera were collected from recipient mice that underwent graft rejection. Thymocytes from the indicated mice were first stained with collected sera, and then further stained with PE-conjugated anti-mouse IgM antibody (a,b) or Alexa Fluor 488-conjugated anti-mouse IgG antibody (c,d). Thymocytes were analyzed by flow cytometry. The open histogram shows naïve C3129F1 mice sera staining, and filled histogram shows recipient C3129F1 sera staining. The data shown in (b,d) are representative of staining of the sera from six independent mice. Each bar shows the average mean fluorescence intensity ± SEM. **p < 0.01, Tukey’s HSD test.CC BY · PubMed Central

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Research works printed but not linked: 17

  1. Ingulli, E. Mechanism of cellular rejection in transplantation. Pediatr. Nephrol.25, 61–74 (2010). · DOI · Paper page
  2. De Bueger, M. & Goulmy, E. Human minor histocompatibility antigens. Transpl. Immunol.1, 28–38 (1993). · DOI
  3. Nakatsuji, N., Nakajima, F. & Tokunaga, K. HLA-haplotype banking and iPS cells. Nat. Biotechnol.26, 739–740 (2008). · DOI
  4. Morizane, A. et al. MHC matching improves engraftment of iPSC-derived neurons in non-human primates. Nat. Commun.8, 1–12 (2017). · DOI · Paper page
  5. Kawamura, T. et al. Cardiomyocytes derived from MHC-homozygous induced pluripotent stem cells exhibit reduced allogeneic immunogenicity in MHC-matched non-human primates. Stem Cell Rep.6, 312–320 (2016). · DOI · Paper page
  6. Opelz, G. & Döhler, B. Effect of human leukocyte antigen compatibility on kidney graft survival: Comparative analysis of two decades. Transplantation84, 137–143 (2007). · DOI
  7. Jones, N. D. et al. Differential susceptibility of heart, skin, and islet allografts to T cell-mediated rejection. J. Immunol.166, 2824–2830 (2001). · DOI
  8. Neofytou, E., O’Brien, C. G., Couture, L. A. & Wu, J. C. Hurdles to clinical translation of human induced pluripotent stem cells. J. Clin. Invest.125, 2551–2557 (2015). · DOI
  9. Becker, L. E., Morath, C. & Suesal, C. Immune mechanisms of acute and chronic rejection. Clin. Biochem.49, 320–323 (2016). · DOI
  10. Vannaprasaht, S. et al. Personalized tacrolimus doses determined by CYP3A5 genotype for induction and maintenance phases of kidney transplantation. Clin. Ther.35, 1762–1769 (2013). · DOI
  11. Flechner, S. M. et al. Kidney transplantation without calcineurin inhibitor drugs: A prospective, randomized trial of sirolimus versus cyclosporine. Transplantation74, 1070–1076 (2002). · DOI
  12. Colvin, R. B. & Smith, R. N. Antibody-mediated organ-allograft rejection. Nat. Rev. Immunol.5, 807–817 (2005). · DOI
  13. Wiltshire, T. et al. Genome-wide single-nucleotide polymorphism analysis defines haplotype patterns in mouse. Proc. Natl. Acad. Sci. USA100, 3380–3385 (2003). · DOI
  14. Piazza, A. et al. Impact of donor-specific antibodies on chronic rejection occurrence and graft loss in renal transplantation: Posttransplant analysis using flow cytometric techniques1. Transplantation71, 1106–1112 (2001). · DOI
  15. Goldberg, E. H., Goble, R. & Jackman, S. H. Monoclonal antibodies defining the skin-selective alloantigens, Skn. Immunogenetics31, 393–395 (1990). · DOI
  16. Zhang, Q. & Reed, E. F. Non-MHC antigenic targets of the humoral immune response in transplantation. Curr. Opin. Immunol.22, 682–688 (2010). · DOI
  17. Sasaki, H. et al. New immunosuppressive cell therapy to prolong survival of induced pluripotent stem cell-derived allografts. Transplantation99, 2301–2310 (2015). · DOI

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Metadata

Authors
Murata, T., Wada, H., Otsuka, R., Sasaki, A., Tsuji, H., Itoh, M., Eguchi, N., Kawai, T., & Seino, K.-I.
Affiliations
  • Hokkaido University (JP)
  • Massachusetts General Hospital (US)
Corresponding author
Seino, Ken-ichiro
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2020-08-11
Volume
10
Issue
1
Pages
13560-13560
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-020-69784-4
PMC
PMC7421494
OpenAlex
W3048669947
Cited by
10
References
1
Citations by year
2025: 3 · 2024: 1 · 2023: 4 · 2022: 2
FWCI
1.27
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Hokkaido University Collection of Scholarly and Academic Papers (Hokkaido University) (submittedVersion, cc-by)
  • Institutional Repositories DataBase (IRDB) (submittedVersion, cc-by)
Funders
  • Ministry of Education, Culture, Sports, Science and Technology
  • Japan Agency for Medical Research and Development
  • TERUMO LIFE SCIENCE FOUNDATION
Topics
  • Health Sciences › Medicine › Transplantation › Organ and Tissue Transplantation Research
  • Health Sciences › Medicine › Surgery › Transplantation: Methods and Outcomes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
graft rejection, skin grafts, immunosuppressants, anti-donor antibodies, murine transplantation model, HLA haplotype matching
MeSH terms
Animals, Female, Graft Rejection, Graft Survival, Histocompatibility Antigens, Isoantibodies, Male, Mice, Inbred BALB C, Mice, Inbred CBA, Mice, Inbred C57BL, Models, Theoretical, Tissue Donors, Transplantation, Heterologous, Transplantation, Homologous, Skin Transplantation, Mice

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