Manufacture of GMP-compliant functional adenovirus-specific T-cell therapy for treatment of post-transplant infectious complications

Horlock, Claire; Skulte, Amanda; Mitra, Arindam; Stansfield, Alka; Bhandari, Shristi; Ip, Winnie; Qasim, Waseem; Lowdell, Mark W.; Patel, Shreenal; Friedetzky, Anke; Purbhoo, Marco A.; Newton, Katy

doi:10.1016/j.jcyt.2016.06.009

Cited by 13 publications

(7 citation statements)

References 24 publications

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“…ADV-specific T cells were generated as recently described [18]. Briefly, peripheral blood mononuclear cells (…”

Section: Manufacture Of Adv-specific T Cellsmentioning

confidence: 99%

“…The product was administered as a single intravenous infusion following allogeneic HSCT D+28 or later on PCR evidence of ADV viraemia (two consecutive ADV level >1000 copies/mL). Previous data from Cytovir ® ADV manufactured from healthy adult donors demonstrated low, but detectable, levels of alloreactivity in the T-cell product [18]. In order to minimise risk of GvHD, an initial cell dose of 1x10 4 CD3+/kg was chosen, with 1x10 5 CD3+/kg as subsequent dose if required.…”

Section: Adv-specific T Cell Infusionmentioning

confidence: 99%

“…An initial cell dose of 1x 10 4 CD3+/kg was chosen as a safe starting dose with provision for 10-fold escalation if required. Characterisation of manufactured products along with ADV specificity has been recently published [18].…”

Section: Adv-specific T Cell Infusionmentioning

confidence: 99%

“…Immune reconstitution was traced by lymphocyte subsets (CD3 + CD4 + , CD3 + CD8 + and CD3 -/CD16+CD56 + ) and chimerism studies performed by short tandem repeat variability as described previously [17]. IFNγ enzyme-linked immunospot assay (ELISpot) was performed at 30, 90 and 180 days post infusion when counts were sufficient [18].…”

Section: Patient Monitoringmentioning

confidence: 99%

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Multicenter phase 1/2 application of adenovirus-specific T cells in high-risk pediatric patients after allogeneic stem cell transplantation

Silva

Gaspar

et al. 2018

Cytotherapy

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“…ADV-specific T cells were generated as recently described [18]. Briefly, peripheral blood mononuclear cells (…”

Section: Manufacture Of Adv-specific T Cellsmentioning

confidence: 99%

Section: Adv-specific T Cell Infusionmentioning

confidence: 99%

Section: Adv-specific T Cell Infusionmentioning

confidence: 99%

Section: Patient Monitoringmentioning

confidence: 99%

See 2 more Smart Citations

Multicenter phase 1/2 application of adenovirus-specific T cells in high-risk pediatric patients after allogeneic stem cell transplantation

Silva

Gaspar

et al. 2018

Cytotherapy

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show abstract

“…The data showing the transition of the relevant small T cell component in the starting mixed leucocyte material to its expansion to the final product gives a possible multi‐parameter analysis for quality control as part of a batch manufacturing record. This cytometric phenotype and analysis approach is sufficiently adaptable and inclusive that it would be suitable for phenotypical and functional assay of other cellular therapies, including virus‐specific and genetically modified T cell products [ 24 , 34 , 35 , 36 , 37 ].…”

Section: Discussionmentioning

confidence: 99%

Cytometric analysis of T cell phenotype using cytokine profiling for improved manufacturing of an EBV-specific T cell therapy

Cooper

Kowalczuk

Wilkie

et al. 2021

Clinical and Experimental Immunology

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Adoptive immunotherapy using Epstein-Barr Virus (EBV)-specific T cells is a potentially curative treatment for patients with EBV-related malignancies where other clinical options have proved ineffective. We describe improved good manufacturing practice (GMP)compliant culture and analysis processes for conventional lymphoblastoid cell line (LCL)-driven EBV-specific T cell manufacture, and describe an improved phenotyping approach for analysing T cell products. We optimized the current LCL-mediated clinical manufacture of EBV-specific T cells to establish an improved process using xenoproteinfree GMP-compliant reagents throughout, and compared resulting products with our previous banked T cell clinical therapy. We assessed effects of changes to LCL:T cell ratio in T cell expansion, and developed a robust flow cytometric marker panel covering T cell memory, activation, differentiation and intracellular cytokine release to characterize T cells more effectively. These data were analysed using a t-stochastic neighbour embedding (t-SNE) algorithm. The optimized GMP-compliant process resulted in reduced cell processing time and improved retention and expansion of central memory T cells. Multiparameter flow cytometry determined the optimal protocol for LCL stimulation and expansion of T cells and demonstrated that cytokine profiling using interleukin (IL)-2, tumour necrosis factor (TNF)-α and interferon (IFN)-γ was able to determine the differentiation status of T cells throughout culture and in the final product. We show that fully GMP-compliant closed-process culture of LCL-mediated EBV-specific T cells is feasible, and profiling of T cells through cytokine expression gives improved characterization of start material, in-process culture conditions and final product. Visualization of the complex multi-parameter flow cytometric data can be simplified using t-SNE analysis.

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Prevention and Treatment of Infectious Complications in Haploidentical SCT

Hassler

Lehrnbecher

Bader

et al. 2017

Stem Cell Biology and Regenerative Medicine

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Manufacture of GMP-compliant functional adenovirus-specific T-cell therapy for treatment of post-transplant infectious complications

Cited by 13 publications

References 24 publications

Multicenter phase 1/2 application of adenovirus-specific T cells in high-risk pediatric patients after allogeneic stem cell transplantation

Multicenter phase 1/2 application of adenovirus-specific T cells in high-risk pediatric patients after allogeneic stem cell transplantation

Cytometric analysis of T cell phenotype using cytokine profiling for improved manufacturing of an EBV-specific T cell therapy

Prevention and Treatment of Infectious Complications in Haploidentical SCT

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