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Safety and feasibility of virus-specific T cells derived from umbilical cord blood in cord blood transplant recipients

Allistair A. Abraham, Tami D. John, Michael D. Keller, C. Russell Y. Cruz, Baheyeldin Salem, Lauren Roesch, Hao Liu, Fahmida Hoq, Bambi J. Grilley, Adrian P. Gee, Hema Dave, David A. Jacobsohn, Robert A. Krance, Elizabeth. J. Shpall, Caridad A. Martinez, Patrick J. Hanley and Catherine M. Bollard
This article has an Erratum 3(16):2453

Data supplements

Article Figures & Data

Figures

  • Figure 1.

    Schematic of CB-VST manufacture in ACT-CAT and ACTCAT2 clinical trials. Eighty percent of the CB unit was used as the CB transplant graft, and 20% was used to expand the VST. From the 20% fraction, 5 to 10 million peripheral blood mononuclear cells (PBMCs) were used for EBV lymphoblastoid cell line (LCL) generation; the remaining cells were used to generate dendritic cells (DCs) or used as the source of T cells. DCs were matured and transduced with the Ad5f35-pp65 vector (ACT-CAT) on day 5 and/or cocultured with overlapping viral peptides (PepMix) (ACT-CAT2), then cocultured with the nonadherent fraction (containing T cells) on day 7 in the presence of IL-7, IL-12, and IL-15. At the second and third stimulation, EBV-transformed B cells were transduced with the same Ad5f35-pp65 vector 2 days before the T-cell stimulation or pulsed with PepMix the day of stimulation, which was performed in the presence of IL-15 (second stimulation) and IL-2 (third stimulation).

  • Figure 2.

    VST product characteristics. (A) Cell yields after manufacture using APCs either transduced with the Ad5f35-pp65 vector (ACT-CAT) or cocultured with overlapping viral peptides (ACT-CAT2). (B) VST products demonstrated multivirus specificity as measured by IFN-γ ELISPOT assay in response to CMV, EBV, and adenovirus antigen. Data for 19 VST products is represented as the number of SFCs per 100 000 after subtracting the values of the actin negative control. (C) A CD4+ T-cell predominant phenotype was observed with similar effector memory (CD45RA/CD62L) and central memory (CD45RA/CD62L+) percentages in the VST products. Memory phenotype data were not available for products P0020, P0035, P0039, P005, and P0067 because of limited product availability for analysis after infusion.

  • Figure 3.

    T-cell persistence and viral load in patients treated with CB-VSTs. (A) Detection of CMV-specific T cells in the peripheral blood of patient P0039 (red bars) and the CMV viral load (green line). Detection of CMV-, EBV-, and adenovirus-specific T cells at baseline compared with 3 months after VST infusion in the viral treatment group (B) and the prophylaxis group (C).

  • Figure 4.

    Detection and persistence of unique TCRs over time using TCR sequencing. (A-C) Shown are unique clonotypes that were present in the VST cell product but not present before VST cell infusion. (D) TCR-Vβ diversity over time in patient P2891 in CD45RA+ (naïve) and CD45RA (memory) T-cell populations. (E) Persistence of unique T-cell clones over time in all patients with available data. (F) Unique T-cell clone contribution at 1 month after VST infusion separated by whether patients had detectable virus within the first month of infusion (reactivation) or not (no reactivation).

Tables

  • Table 1.

    Demographics of patients receiving CB-derived VST infusions

    Patient IDDiagnosisAge, ySexWeight, kgConditioning regimenHLA match (of 6)Immunosuppression before VSTRecipient CMV serostatusTNCs from UCB, × 107TNCs/kg of UCB fraction used for primary transplant
    ACT-CAT
     P2891Fanconi anemia5M13.0Flu/Cy/TBI5CSA,MMF, steroidsPositive1619.9
     P3010ALL2M11.0Flu/Cy/TBI5CSA, MMFPositive19113.9
     P3275SCID1M5.2Bu/Cy/Flu6CSA, MMF, steroidsPositive12118.8
     P3317Myelofibrosis2M9.7Bu/Cy/Flu5CSA, MMF, steroidsPositive22518.6
     P3457JMML2M12.3Bu/Cy/Flu5CSA, MMFPositive18512.0
     P3531SCID1M6.4Bu/Cy/Flu5CSA, MMF, steroidsPositive12916.0
     P3555SCID1M8.0Bu/Cy/Flu6CSA, MMF, steroidsPositive11011.0
     P3923AML1F7.0Bu/Cy/Flu5CSA, MMF, steroidsPositive10211.7
     P3626JMML1M9.0Bu/Cy/Flu5CSA, MMFPositive20918.6
    ACT-CAT2
     P0020AML3F16.7Bu/Cy5CSA, MMF, steroids, ECPPositive1034.9
     P0035Hurler’s syndrome1.1F8.8Bu/Flu/ATG/ rituximab4CSA, steroidsPositive25222.9
     P0039Infant ALL1.9M12.2Flu/Cy/TBI6CSA, MMF, steroidsPositive674.4
     P0067Sickle cell disease8F19.8Flu/Mel/Thio/alemtuzumab/HU6CSA, MMF, infliximab, steroidsPositive1094.4
     P0123Sickle cell disease4M16.6Flu/Mel/Thio/alemtuzumab/HU5CSA, tacro, MMF, steroidsNegative1979.5
    • ALL, acute lymphocytic leukemia; AML, acute myeloid leukemia; ATG, anti-thymocyte globulin; Bu, busulfan; CSA, cyclosporin A; Cy, cyclophosphamide; ECP, extracorporeal photopheresis; F, female; Flu, fludarabine; HLA, human leukocyte antigen; HU, hydroxyurea; JMML, juvenile myelomonocytic leukemia; M, male; Mel, melphalan; MMF, mycophenolate mofetil; SCID, severe combined immunodeficiency; tacro, tacrolimus; TBI, total body irradiation; Thio, thiotepa; TNC, total nucleated cells; UCB, umbilical cord blood.

  • Table 2.

    Patient infection/reactivation status and outcome

    PatientIndicationPrior therapyViral course after VSTStatus within first year
    P0123Adenovirus low-level viremia (<200 copies per mL) and positive upper respiratory tractCidofovirResolution (with brincidofovir)Alive, GVHD resolved at 1 y
    P0067CMV 2239 cells per mLFoscarnet/cidofovir/ganciclovirCMV viremia resolved by 6 mo but developed CMV retinitis 6 mo after infusionAlive, CMV retinitis at 6 mo treated with third-party T cells and intravitreal ganciclovir with complete resolution, GVHD resolved at 1 y
    P0039CMV low-level viremia (<1000 copies per mL)Foscarnet/cidofovir/ganciclovirResolutionLeukemia relapse after 90 d, in remission after CAR T-cell therapy
    P0035CMV low-level viremia (<1000 copies/mL) and positive stoolFoscarnetResolution (with valganciclovir)Alive, quiescent GVHD on cyclosporine at 1 y
    P2891CMV viremia (100 copies/mL)Foscarnet, CMV immunoglobulinResolution (with ganciclovir + 2 additional CB-VST infusions)Alive, GVHD resolved at 1 y
    P3626EBV in bone marrow (3500 copies/mL)Cidofovir for previous adenoviremiaFivefold decline in bone marrow EBV at 5 mo, negative in blood at 1 y, no PTLDAlive, MDS + monosomy 7 at <1 y resolved at 2 y. No GVHD, off immunosuppression at 1 y
    P3457EBV viremia (109 copies/mL)NonePersistent viremia but no PTLDDied after second transplant for relapsed leukemia
    P3010ProphylaxisGanciclovir prophylaxisNo reactivation requiring therapy, no diseaseAlive, no GVHD, off immunosuppression at 1 y
    P3275ProphylaxisNoneNo reactivation requiring therapy, no diseaseAlive, no GVHD, off immunosuppression at 1 y
    P3317ProphylaxisGanciclovir prophylaxisNo reactivation requiring therapy, no diseaseRelapse/AML progression <1 y after HCT, deceased day +284.
    P3531ProphylaxisValganciclovir prophylaxisNo reactivation requiring therapy, no diseaseAlive, no GVHD, off immunosuppression at 1 y
    P3555ProphylaxisNoneNo reactivation requiring therapy, no diseaseAlive, resolved GVHD at 1 y
    P3923ProphylaxisNoneNo reactivation requiring therapy, no diseaseAlive, no GVHD, off immunosuppression at 1 y
    P0020Prophylaxis/past history of adenovirus in stoolCidofovirAdenovirus reactivation in stool at 3 wk and then blood 3 mo after VSTs; given second CB-VST infusion and continued with cidofovirAlive, quiescent GVHD on cyclosporine at 1 y
    • CAR, chimeric antigen receptor; HCT, hematopoietic stem cell transplantation; MDS, myelodysplastic syndrome; PTLD, posttransplantation lymphoproliferative disorder.