Home › Microbiology › Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant
Microbiology JoVE (Open Access) Citable · DOI

Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant

DOI: 10.3791/2736-v
What you'll learn
  • ✓ Nucleofect dendritic cells with multivirus peptide-encoding mRNA
  • ✓ Stimulate donor T cells against virus-pulsed antigen-presenting cells
  • ✓ Expand multivirus-specific CTLs using G-Rex bioreactor technology
  • ✓ Manufacture GMP-compliant donor-derived T cells for HSCT recipients
Protocol

Biopharma Insights A rapid, simple and cost-effective protocol for the generation of donor-derived multivirus-specific CTLs (rCTL) for infusion to allogeneic hematopoietic stem cell transplant (HSCT) recipients at risk of developing CMV, Adv or EBV infections. This manufacturing process is GMP-compliant and should ensure the broader implementation of T-cell immunotherapy beyond specialized centers.

Difficulty
advanced
Total time
~10–14 days (including expansion phase)
Biosafety
BSL-2

Steps

1
Nucleofect dendritic cells with viral peptide mRNA

Transfect donor-derived dendritic cells with mRNA encoding CMV, Adenovirus, and EBV epitopes using nucleofection technology. This generates antigen-presenting cells that display multivirus peptides on MHC molecules.

▶ 01:49
2
Co-culture T cells with nucleofected dendritic cells

Incubate donor-derived T cells with peptide-pulsed dendritic cells to stimulate antigen-specific T cell activation and proliferation against CMV, Adv, and EBV epitopes.

▶ 04:01
3
Expand multivirus-specific T cells in G-Rex bioreactor

Culture activated multivirus-specific CTLs in G-Rex expansion vessels to generate large numbers of polyfunctional, antigen-specific effector T cells suitable for clinical infusion.

▶ 06:18
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