Curing autoimmune diabetes in mice with islet and hematopoietic cell transplantation after CD117 antibody-based conditioning
Preksha Bhagchandani, Stephan A. Ramos, Bianca Rodriguez, Xueying Gu, Shiva Pathak, Yuqi Zhou, Yujin Moon, Nadia Nourin, Charles A. Chang, Jessica Poyser, Brenda J. Velasco, Weichen Zhao, Hye-Sook Kwon, Richard Rodriguez, Diego M. Burgos, Mario A. Miranda, Everett Meyer, Judith A. Shizuru, Seung K. Kim
Preksha Bhagchandani, Stephan A. Ramos, Bianca Rodriguez, Xueying Gu, Shiva Pathak, Yuqi Zhou, Yujin Moon, Nadia Nourin, Charles A. Chang, Jessica Poyser, Brenda J. Velasco, Weichen Zhao, Hye-Sook Kwon, Richard Rodriguez, Diego M. Burgos, Mario A. Miranda, Everett Meyer, Judith A. Shizuru, Seung K. Kim
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Research Article Autoimmunity Endocrinology

Curing autoimmune diabetes in mice with islet and hematopoietic cell transplantation after CD117 antibody-based conditioning

Abstract

Mixed hematopoietic chimerism after allogeneic hematopoietic cell transplantation (HCT) promotes tolerance of transplanted donor-matched solid organs, corrects autoimmunity, and could transform therapeutic strategies for autoimmune type 1 diabetes (T1D). However, development of nontoxic bone marrow conditioning protocols is needed to expand clinical use. We developed a chemotherapy-free, nonmyeloablative (NMA) conditioning regimen that achieves mixed chimerism and allograft tolerance across MHC barriers in NOD mice. We obtained durable mixed hematopoietic chimerism in prediabetic NOD mice using anti–CD117 monoclonal antibody, T cell depleting antibodies, JAK1/2 inhibition, and low-dose total body irradiation prior to transplantation of MHC-mismatched B6 hematopoietic cells, preventing diabetes in 100% of chimeric NOD:B6 mice. In overtly diabetic NOD mice, NMA conditioning followed by combined B6 HCT and islet transplantation durably corrected diabetes in 100% of chimeric mice without chronic immunosuppression or graft-versus-host disease (GVHD). Chimeric mice remained immunocompetent, as assessed by blood count recovery and rejection of third-party allogeneic islets. Adoptive transfer studies and analysis of autoreactive T cells confirmed correction of autoimmunity. Analysis of chimeric NOD mice revealed central thymic deletion and peripheral tolerance mechanisms. Thus, with NMA conditioning and cell transplantation, we achieved durable hematopoietic chimerism without GVHD, promoted islet allograft tolerance, and reversed established T1D.

Authors

Preksha Bhagchandani, Stephan A. Ramos, Bianca Rodriguez, Xueying Gu, Shiva Pathak, Yuqi Zhou, Yujin Moon, Nadia Nourin, Charles A. Chang, Jessica Poyser, Brenda J. Velasco, Weichen Zhao, Hye-Sook Kwon, Richard Rodriguez, Diego M. Burgos, Mario A. Miranda, Everett Meyer, Judith A. Shizuru, Seung K. Kim

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Figure 2

Mixed chimerism prevents autoimmune diabetes and establishes donor-specific islet tolerance.

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Mixed chimerism prevents autoimmune diabetes and establishes donor-speci...
(A) Autoimmune diabetes development curves of prediabetic NOD:B6 mixed chimeric mice (n = 19 from 4 independent experiments), conditioned controls (n = 10), and naive NOD mice (n = 15). P < 0.0001 for chimeras vs. naive NOD mice, P < 0.01 for chimeras vs. conditioned controls using log-rank (Mantel-Cox) test. P < 0.0001 for chimeras vs. naive NOD, P < 0.05 for chimeras vs. conditioned controls using Fisher’s exact test. P < 0.05 and no significance for conditioned controls vs. naive NOD mice using Mantel-Cox and Fisher’s exact test, respectively. (B) Nonfasting blood glucose of NOD:B6 chimeras (red) compared with naive NOD mice (blue) and conditioned controls (green). Dotted line indicates normoglycemic threshold (200 mg/dL). (C) Percentage of different stages of insulitis in individual islets and representative histology of pancreas from NOD:B6 mice 20 weeks after HCT (n = 14) compared with naive NOD mice (n = 5) and conditioned controls (n = 10). ****P < 0.0001. Statistical analyses done using χ2 test. (D) Representative pancreatic histology of NOD:B6 chimeras at 20 weeks after HCT stained for insulin, CD45.1 (host), and CD45.2 (donor) or B220 and CD3 or CD49b, CD11b, and CD3 (n = 3-6). Pink arrowheads indicate CD45.1+ immune cells. (E) Experimental transplantation schematic. Prediabetic NOD:B6 mixed chimeras received B6 islets in the left kidney and FVB islets in the opposite kidney at 14 weeks after HCT. (F) FVB and B6 islet grafts 2 weeks after islet transplantation in mixed chimeras stained for insulin and CD3 or CD45 (n = 3). (G) B6 islet grafts 14 weeks after islet transplantation in mixed chimeras stained for insulin and CD3 or CD45 (n = 4). (C–G) Scale bars = 200 μm (C, D, F, and G); 20 μm (D, insets). HCT, hematopoietic cell transplantation.