Modeling immune responses to autologous and allogeneic human stem cell–derived islet grafts in vivo
Camillo Bechi Genzano, Giorgia Zanetti, Qian Du, Daniel Traum, Deeksha Lahori, Grant M. Downes, Sakshi A. Bhatele, Xiaolan Ding, Kyle D. Apley, Rebuma Firdessa Fite, Matthew Ishahak, Enrique Eduardo Sanchez-Castro, Jeffrey R. Millman, Yiming Luo, Klaus H. Kaestner, Cory Berkland, Dieter Egli, Megan Sykes, Remi J. Creusot
Camillo Bechi Genzano, Giorgia Zanetti, Qian Du, Daniel Traum, Deeksha Lahori, Grant M. Downes, Sakshi A. Bhatele, Xiaolan Ding, Kyle D. Apley, Rebuma Firdessa Fite, Matthew Ishahak, Enrique Eduardo Sanchez-Castro, Jeffrey R. Millman, Yiming Luo, Klaus H. Kaestner, Cory Berkland, Dieter Egli, Megan Sykes, Remi J. Creusot
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Research Article Endocrinology Immunology

Modeling immune responses to autologous and allogeneic human stem cell–derived islet grafts in vivo

Abstract

Stem cell–derived β cells offer a promising approach for type 1 diabetes (T1D) treatment. However, the processes of graft infiltration and rejection by immune cells remain poorly understood in humans. In this study, autologous or allogeneic stem cell–derived islets (SC-islets) were transplanted in human immune system mice and analyzed 14 to 18 weeks later. Imaging mass cytometry revealed unique characteristics of SC-islet grafts, including a high percentage of glucagon+ cells and the presence of cysts and CD57+ enterochromaffin cells, features not typically observed in endogenous or transplanted allogeneic primary pancreatic islets. Allogeneic SC-islet grafts exhibited heavy immune infiltration, cell proliferation, and pro-fibrotic processes, whereas autologous grafts showed minimal infiltration and little fibrosis. In some mice, autologous T cells expressing islet antigen-reactive (IAR) T cell receptors (TCRs) were adoptively transferred. Three weeks after transfer, autologous grafts injected with IAR-TCR+ T cells showed negligible immune infiltration, even though IAR-TCR+ T cells were detected in the spleen. Under the conditions tested, human SC-islet grafts were not rejected by an autologous immune system, even in the presence of autoreactive T cells, pointing to several limitations that remain to be addressed for a model of spontaneous autologous SC-islet infiltration and destruction.

Authors

Camillo Bechi Genzano, Giorgia Zanetti, Qian Du, Daniel Traum, Deeksha Lahori, Grant M. Downes, Sakshi A. Bhatele, Xiaolan Ding, Kyle D. Apley, Rebuma Firdessa Fite, Matthew Ishahak, Enrique Eduardo Sanchez-Castro, Jeffrey R. Millman, Yiming Luo, Klaus H. Kaestner, Cory Berkland, Dieter Egli, Megan Sykes, Remi J. Creusot

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Figure 2

Human immune system and stem cell–derived islet reconstitution.

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Human immune system and stem cell–derived islet reconstitution. (A and B...
(A and B) Human immune system reconstitution in cohort 1 (A) and 2 (B); human chimerism is measured as the percentage of human CD45+ cells among total CD45+ cells. Each symbol represents an individual animal. In A, a hollow triangle represents an allogeneic mouse not injected with polyclonal T cells. (C) CD34– nonadherent fetal liver cells reprogramming into iPSCs using Klf4, Oct3/4, Sox2, and c-Myc transcription factors delivered via the nonintegrating Sendai virus, with examples of iPSC colonies. Scale bar: 200 µm. (D) iPSC-to-β cell differentiations performed in the study. (E) Circulating C-peptide levels in autologous (cohort 1 and 2) and allogeneic grafts. In autologous cohort 1 and 2, n = 2/3 and n = 3/12 mice were injected with IAR-TCR+ T cells that did not infiltrate the grafts, respectively, and n = 5/6 allogeneic mice were injected with additional in vitro activated polyclonal T cells from a mouse with the same HIS. In E, the asterisk indicates the level of significance (*P < 0.05) when comparing the percentage of circulating C-peptide in allogeneic and autologous cohort 2 at weeks 12 and 15 after β cell transplantation. (F) Correlation between percentage of C-peptide+ cells in islet before transplant and circulating human C-peptide levels 12 and 15 weeks later. In A, B, and E, data show the mean ± SEM. In F, different groups are represented with the same symbols as E, and each symbol represents the mean of the group. Statistical analysis was performed using a mixed-effects model with Geisser-Greenhouse correction and Tukey’s post hoc test for E and a simple linear regression for F. C-1, cohort 1; C-2, cohort 2.