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Gianpietro Dotti G Center for Cell and Gene Therapy Baylor College of Medicine, Houston TX Clinical Experience at Baylor College of Medicine with T Lymphocytes.

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Presentation on theme: "Gianpietro Dotti G Center for Cell and Gene Therapy Baylor College of Medicine, Houston TX Clinical Experience at Baylor College of Medicine with T Lymphocytes."— Presentation transcript:

1 Gianpietro Dotti G Center for Cell and Gene Therapy Baylor College of Medicine, Houston TX Clinical Experience at Baylor College of Medicine with T Lymphocytes Genetically Modified Using Retroviral Vectors

2 - Moloney-derived retroviral vector (SFG from Mulligan R.) - Packaging cell line Phoenix-Eco original cells from ATCC generation of a master cell bank under GMP conditions - Packaging cell line PG13 original cells from ATCC generation of a master cell bank under GMP conditions Type of vector and packaging cell lines

3 - Multiple infection of PG13 cells using Eco-pseudotyped retroviral particles (transient supernatant) - Generation of single cell clone from infected PG13 cells - Expansion of the single cell clone to make a master cell bank (MCB) - Production and collection of supernatant from the MCB Generation of stable packaging cell line

4 - Manufacturing of the single cell clone under GLP conditions - Expansion of the master cell bank (MCB) and production of the retroviral supernatant under GMP conditions Manufacturing

5 - 9 Packaging cell lines generated and tested - RCR detection negative in all cases for both Ecotropic and Galv envelopes (co-culture assay) - 18 lots of supernatant produced and tested - RCR detection negative in all cases for GalV envelope (co-culture assay) Packaging cell lines

6 IND Transgene Cell product Disease # CAR.GD2 Activated T cells vs Neuroblastoma EBV-CTLs 19 patients treated. 38 T-cell/CTL products infused all negative by co-culture assay. Clinical protocols

7 IND Transgene Cell product Disease #13296 CAR.CD19.CD28 vs Activated T cells CD19+ Lymphoma CAR.CD19 #13902 CAR.CD19.CD28 vs Activated T cells vs CD19+ Lymphoma CAR.CD19 EBV-CTLs #13958 CAR.CD19.28 Virus-specific CTLs CD19+ Lymphoma 8 patients treated (6 in # and 2 in #). 16 T-cell/CTL products infused. 6 negative by co-culture assay. 8 pending and 2 not needed (frozen within 4 days of culture). Clinical protocols

8 IND TransgeneCell product Disease #14050 CAR.HER2.CD28 Activated T cells Osteosarcoma #14036 CAR.HER2.CD28 and EBV-CTLs Lung cancer TGFbRIIdcyt (DNRII) #14050 CAR.HER2.CD28. CMV-specific CTLs Glioblastoma 4 patients treated (2 in #14050 and 2 in #14036). 4 T-cell/CTL products infused. 1 negative by co-culture assay. 3 pending. Clinical protocols

9 IND Transgene Cell product Disease #12830 TGF RIIdcyt (DNRII) LMP-2a-CTLs EBV+ Lymphoma 4 patients treated. 4 CTL products infused. All negative by co-culture assay. Clinical protocols

10 IND Transgene Cell product Disease # icaapase9.2A. CD19 allodepleted T cells hematological malignancies/haplo HSC transplant 4 patients treated. 4 T-cell products infused. All negative by co-culture assay. Clinical protocols

11 92 lines infused to 73 patients 2 did not require assay 8 results pending 82 lines negative RCR co-culture assay for GalV envelope T/CTL line infused summary

12 - 73 patients infused (follow up from 1 month to 15 years) samples analyzed - All RCR negative by Q-PCR (GalV envelope) Follow up patient samples

13 - 34 patients infused with gene marked EBV-specific CTLs - All 34 CTL lines negative by co-culture assay follow up samples all negative by PCR assay (samples continue to be archived from 2002 but no longer tested) Previous experience at St. Jude Childrens Research Hospital (Gene marking) (Gene marking)

14 - T cell/CTLs modified with retroviral vectors are consistently RCR negative - RCR assay based on co-culture is time consuming and extremely expensive limiting the possibility to extend the use of promising immunotherapy approaches - Based on the current experience samples can be stored and analyzed for RCR contamination in case of severe adverse events Conclusions and Suggestions


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