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Clinical Trial
. 2017 Nov;17(11):2945-2954.
doi: 10.1111/ajt.14415. Epub 2017 Aug 14.

Polyclonal Regulatory T Cell Therapy for Control of Inflammation in Kidney Transplants

S Chandran  1 , Q Tang  2   3 , M Sarwal  2 , Z G Laszik  4 , A L Putnam  3 , K Lee  2 , J Leung  2 , V Nguyen  2 , T Sigdel  2 , E C Tavares  2 , J Y C Yang  2 , M Hellerstein  5 , M Fitch  5 , J A Bluestone  3 , F Vincenti  1   2
Affiliations
Clinical Trial

Polyclonal Regulatory T Cell Therapy for Control of Inflammation in Kidney Transplants

S Chandran et al. Am J Transplant. 2017 Nov.

Abstract

Early subclinical inflammation in kidney transplants is associated with later graft fibrosis and dysfunction. Regulatory T cells (Tregs) can reverse established inflammation in animal models. We conducted a pilot safety and feasibility trial of autologous Treg cell therapy in three kidney transplant recipients with subclinical inflammation noted on 6-month surveillance biopsies. Tregs were purified from peripheral blood and polyclonally expanded ex vivo using medium containing deuterated glucose to label the cells. All patients received a single infusion of ~320 × 106 (319, 321, and 363.8 × 106 ) expanded Tregs. Persistence of the infused Tregs was tracked. Graft inflammation was monitored with follow-up biopsies and urinary biomarkers. Nearly 1 × 109 (0.932, 0.956, 1.565 × 109 ) Tregs were successfully manufactured for each patient. There were no infusion reactions or serious therapy-related adverse events. The infused cells demonstrated patterns of persistence and stability similar to those observed in non-immunosuppressed subjects receiving the same dose of Tregs. Isolation and expansion of Tregs is feasible in kidney transplant patients on immunosuppression. Infusion of these cells was safe and well tolerated. Future trials will test the efficacy of polyclonal and donor alloantigen-reactive Tregs for the treatment of inflammation in kidney transplants.

Keywords: cellular biology; clinical trial; immune regulation; immunobiology; immunosuppressant - polyclonal preparations; kidney transplantation/nephrology; protocol biopsy; rejection: subclinical; translational research/science.

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Conflict of interest statement

DISCLOSURE

The authors of this manuscript have conflicts of interest to disclose as described by the American Journal of Transplantation. F.V. has received research grants from Novartis, Immucor and Genentech. J.A. B. is an unpaid adviser in a phase 2 trial of Treg therapy for type 1 diabetes sponsored by Caladrius Biosciences. J.A. B and Q.T. have a patent pending on polyTregs. The other authors have no conflicts of interest to disclose.

Figures

Figure 1
Figure 1. Study design and subject schedule of events
Blood for Treg manufacturing was drawn at day −14, and Treg infusion was given on day 0. Subjects were seen for follow-up assessments on day 4, 7, 14 and 28, and at months 3, 6 and 12. Laboratory assessments were done weekly after the first week, and monthly after the first month.
Figure 2
Figure 2. Treg manufacturing
Parameters of Treg manufacturing for the three kidney transplant patients in this study - TASKp (pilot Treg adoptive therapy for subclinical kidney transplant inflammation, n=3) (solid symbols) are compared with those obtained from type 1 diabetes patients (T1D) enrolled in a separate trial (n=14) (12)(open symbols). Identical standard operating procedures were followed in the two trials. Mann Whitney test was used to calculate p values and determine the statistical significance between the two groups.
Figure 3
Figure 3. Graft function before and after Treg infusion
The arrow indicates the day of infusion (day 0) 3A. Estimated GFR by MDRD equation for all subjects 3B. Urinary protein excretion for all subjects.
Figure 4
Figure 4. Treg tracking in vivo
4A. Persistence of infused Tregs. Tregs were expanded ex vivo in the presence of deuterated glucose, which achieves ~60% enrichment of deuterium in the DNA of expanded Tregs and allows tracking of the infused Tregs after infusion. On the indicated days after Treg infusion, Tregs were isolated using FACS on indicated days and deuterium enrichment in the genomic DNA of purified Tregs were determined using gas chromatography and mass spectrometry. The enrichment values from the 3 kidney patients in this study (solid lines) are compared with those obtained from type 1 diabetes patients (dotted lines) who received similar dose of Tregs in a separate trial. 4B. Stability of infused Tregs. Peripheral blood CD4 cells outside The CD25+CD127lo/− Treg gate (non-Tregs) are sorted into three fractions based CD45RO and CD62L expression. Deuterium levels in these cell subsets were monitored for potential destabilization of Tregs into non-Tregs.
Figure 5
Figure 5. Graft inflammation on kidney biopsies at baseline and post-Treg infusion
CD45 (leukocyte common antigen) immunohistochemical staining was done on kidney biopsy sections at baseline and at 2 weeks and 6 months post-Treg infusion. Representative slides from each subject are shown. The positive cells stain brown.
Figure 6
Figure 6. Donor specific antibody (DSA) levels in subject 3 pre- and post-Treg infusion
Day 0 refers to the day of Treg infusion. Antibodies were already present prior to the infusion of Tregs and continued rising post-infusion. Biopsies were done on day 14, day 103 and day 180 post-infusion. MFI=mean fluorescent intensity
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References

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