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“Mini” bank of only 8 donors supplies CMV-directed T cells to diverse recipients

Ifigeneia Tzannou, Ayumi Watanabe, Swati Naik, Rachel Daum, Manik Kuvalekar, Kathryn S. Leung, Caridad Martinez, Ghadir Sasa, Mengfen Wu, Adrian P. Gee, Robert A. Krance, Stephen Gottschalk, Helen E. Heslop and Bilal Omer

Data supplements

Article Figures & Data

Figures

  • Figure 1.

    Characteristics of generated CMVST lines and degree of matching with screened subjects. (A) T-cell expansion of CMVSTs achieved over a 20-day period based on cell counting using trypan blue exclusion (n = 8). Phenotype of the expanded CMVST lines on the day of cryopreservation (mean ± standard error of the mean [SEM], n = 8) (B) and frequency of antigen-specific T cells as determined by IFN-γ ELISPOT assay (C) after overnight stimulation of CMVSTs with IE1 and pp65 antigen-spanning PepMixes. Results are reported as SFCs per 2 × 105 VSTs plated. CMVST lines with a total of ≥30 SFCS per 2 × 105 were considered to be positive (n = 8). (D) Number of matching HLA antigens (of 8 total) of CMVST lines identified for clinical use with recipient HLA of screened patients (n = 29).

  • Figure 2.

    Treatment outcomes in individual patients infected with CMV. Depiction of plasma CMV viral loads in patients 2 weeks before (viral load level closest to week −2), immediately before (pre), and after (post) infusion (weeks 2, 4, and 6) of CMVSTs. Arrows indicate infusion time points.

  • Figure 3.

    Frequency of CMV-specific T cells in vivo. (A) Frequency of CMVSTs in the peripheral blood before (pre) infusion and after (post) infusion, as measured by using the IFN-γ ELISPOT assay after overnight stimulation with IE1 and pp65 viral PepMixes. Results are expressed as SFCs per 5 × 105 input cells (mean ± SEM, n = 10). (B) Persistence of infused CMVSTs in individual patients. Frequency of T cells in peripheral blood as measured by using the IFN-γ ELISPOT assay after stimulation with epitope-specific CMV peptides with restriction to HLA antigens exclusive to the CMVST line or shared between the recipient and the CMVST line.

Tables

  • Table 1.

    Characteristics of generated VST lines

    VST line compound no. CMV specificity, SFC/1 × 105 IE1CMV specificity, SFC/1 × 105 pp65CD3, %CD4, %CD8, %CD56, %CD45RO+/CD62L+, %CD45RO+/CD62L, %HLA-AHLA-BHLA-DRHLA-DQNo. of patients screened*No. of patients treated
    6790127118697.8174.2319.483.8875.4516.3302,3315,4407,1302,0643
    679861280598.7917.7575.734.0540.344.8302,0240,5204,0803,0364
    6802113135499.665.2092.821.6969.7527.5111,2335,5701,0703,0510
    680882798699.7712.5983.183.1074.0920.1302,2440,5204,1303,0641
    6814639257399.6828.2569.850.9941.5655.7802, 2408,1401, 0302,0511
    682370071799.3910.9986.491.5147.5948.5911,6807,3503,0702,0231
    683412872599.7715.4082.902.2764.6432.7202,2415,3504,0903,0361
    6838205.521199.755.5787.468.7654.5036.4202,3013,3507,0802,0610
    • * Indicates how frequently the VST lines were determined to be the most suitable line for a screened patient.

  • Table 2.

    Patient demographic characteristics

    Patient ID no.SexAge, yEthnicityRaceDiagnosisType of transplantR/D CMV serostatusNo. of infusionsDays posttransplant
    3910Male12Non-HispanicAfrican AmericanSickle cell anemiaMRDNegative/positive161
    3944Female45HispanicWhiteAMLUCBPositive/negative1197
    3976Male13HispanicWhiteALLMUDPositive/positive246
    3762Female10HispanicWhiteHLHMMUDPositive/negative1161
    3967Male51Non-HispanicWhiteAMLUCBPositive/negative1365
    4091Male70Non-HispanicWhiteCTCLHaploPositive/positive1215
    4115Female3Non-HispanicWhiteFanconi anemiaMUDPositive/positive1105
    4170Male3Non-HispanicAfrican AmericanSickle cell anemiaMRDNegative/positive176
    4134Female16Non-HispanicWhiteSCIDMUDPositive/positive1218
    4201Male11Non-HispanicWhiteAnaplastic large-cell lymphomaMUDPositive/negative370
    • ALL, acute lymphoblastic leukemia; AML, acute myeloid leukemia; CTCL, cutaneous T-cell lymphoma; Haplo, haploidentical; HLH, hemophagocytic lymphohistiocytosis; MMUD, mismatched unrelated donor; MUD, matched unrelated donor; MRD, matched related donor; R/D, recipient/donor; SCID, severe combined immunodeficiency; UCB, umbilical cord blood.

  • Table 3.

    Patient clinical characteristics

    Patient ID no.Infused VSTsHLA matchingViral diseaseReduction in immuno-suppressionPrior antiviral(s)Duration prior antiviral therapyAntiviral drug intoleranceCMV resistanceChange in antiviral post-VSTResponse by 6 wkOverall outcome
    3910C67902/8NoNoGanciclovir17NoNoNoCRSustained CR
    3944C67984/8NoNoLeflunomide94AKI, cytopeniaYesNoCRSustained CR
    3976C67986/8NoYes, reduction in tacrolimus between wk 4 and 6Foscarnet, ganciclovir14Renal tubulopathyYes (U97 mutation)NoCRSustained CR after second infusion
    3762C68085/8CMV retinitisNoNoneUnable to toleratePoor graft functionNot doneYes, ganciclovir administered for 1 wkCRSustained CR
    3967C68232/8CMV colitis (diarrhea)NoFoscarnet211AKIYes (U97 mutation)NoPRDied of renal failure and AdV infection
    4091C67905/8NoNoFoscarnet, cidofovir, leflunomide29AKINot doneNoCRDied of renal failure
    4115C68345/8NoNoFoscarnet24AKINot doneNoCRSustained CR
    4170C67903/8NoNoGanciclovir, foscarnet42PancreatitisNoNoCRSustained CR
    4134C67985/8CMV colitis (diarrhea)NoFoscarnet, ganciclovir17NoNot doneNoPRSustained CR
    4201C68143/8NoYes, tacrolimus taper initiated before VST infusionFoscarnet, ganciclovir41NoNoNoPRSustained CR after third infusion
    • AdV, adenovirus; AKI, acute kidney injury; PR, partial response.

  • Table 4.

    GVHD preinfusion and postinfusion

    Patient ID no.Prior GVHDBaselineGVHD Rx/PPx at infusionaGVHDcGVHD
    3910NoneNoneCyclosporineNoneNone
    3944NoneNoneTacrolimusNoneNone
    3976NoneNoneTacrolimusNoneNone
    3762NoneNoneNoneNoneNone
    3967GI grade IINoneSirolimusNoneNone
    4091GI, skin grade IINoneTacrolimusNoneNone
    4115NoneNoneNoneNoneNone
    4170NoneNoneTacrolimusNoneNone
    4134GI grade IIINoneNoneNoneNone
    4201NoneNoneTacrolimusNoneNone
    • aGVHD, acute GVHD; cGVHD, chronic GVHD; GI, gastrointestinal; PPx, prophylaxis; Rx, treatment.