A robust, good manufacturing practice-compliant, clinical-scale procedure to generate regulatory T cells from patients with amyotrophic lateral sclerosis for adoptive cell therapy

Cytotherapy. 2016 Oct;18(10):1312-24. doi: 10.1016/j.jcyt.2016.06.012. Epub 2016 Aug 3.

Abstract

Regulatory T cells (Tregs) play a fundamental role in the maintenance of self-tolerance and immune homeostasis. Defects in Treg function and/or frequencies have been reported in multiple disease models. Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder affecting upper and lower motor neurons. Compelling evidence supports a neuroprotective role for Tregs in this disease. Indeed, rapid progression in ALS patients is associated with decreased FoxP3 expression and Treg frequencies. Thus, we propose that strategies to restore Treg number and function may slow disease progression in ALS. In this study, we developed a robust, Good Manufacturing Practice (GMP)-compliant procedure to enrich and expand Tregs from ALS patients. Tregs isolated from these patients were phenotypically similar to those from healthy individuals but were impaired in their ability to suppress T-cell effector function. In vitro expansion of Tregs for 4 weeks in the presence of GMP-grade anti-CD3/CD28 beads, interleukin (IL)-2 and rapamcyin resulted in a 25- to 200-fold increase in their number and restored their immunoregulatory activity. Collectively, our data facilitate and support the implementation of clinical trials of adoptive therapy with ex vivo expanded and highly suppressive Tregs in patients with ALS.

Keywords: amyotrophic lateral sclerosis; good manufacturing practice; rapamycin; regulatory T cells.

MeSH terms

  • Adoptive Transfer / methods
  • Adoptive Transfer / standards*
  • Amyotrophic Lateral Sclerosis / immunology
  • Amyotrophic Lateral Sclerosis / pathology*
  • Case-Control Studies
  • Cell Separation* / methods
  • Cell Separation* / standards
  • Cell- and Tissue-Based Therapy / methods
  • Cell- and Tissue-Based Therapy / standards*
  • Cells, Cultured
  • Guideline Adherence / standards
  • Humans
  • Immune Tolerance
  • Interleukin-2 / metabolism
  • Primary Cell Culture* / methods
  • Primary Cell Culture* / standards
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / pathology*

Substances

  • Interleukin-2