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Idiotype-pulsed dendritic cell vaccination for B-cell lymphoma: clinical and immune responses in 35 patients

John M. Timmerman, Debra K. Czerwinski, Thomas A. Davis, Frank J. Hsu, Claudia Benike, Zheng Mei Hao, Behnaz Taidi, Ranjani Rajapaksa, Clemens B. Caspar, Craig Y. Okada, Adrienne van Beckhoven, Tina Marie Liles, Edgar G. Engleman and Ronald Levy

Article Figures & Data

Figures

  • Fig. 1.

    Complete regression of lymphoma after idiotype-pulsed dendritic cell vaccination.

    CT images of the chest of patient D9 show multiple enlarged left axillary lymph nodes that normalized 11 months after vaccination. Arrows indicate sites of disease before vaccination. Similarly sized lymph nodes in the periaortic and iliac regions also completely regressed.

  • Fig. 2.

    Idiotype specificity of the humoral anti-Id response after vaccination with Id-KLH–pulsed dendritic cells.

    Prevaccine or postvaccine sera from 2 patients (D31 in panel A, D29 in panel B) were serially diluted and incubated on plates coated with the patients' autologous tumor Id (relevant Id) or with Id proteins from 3 other patient's tumors (irrelevant Id). Binding of IgG anti-Id antibodies was detected by anti–human IgG antibodies conjugated to HRP, and expressed as optical density (O.D.). Panel A: ● indicates prevaccine sera on relevant Id (D31); ▪, postvaccine sera on relevant Id (D31); ▵, postvaccine sera on irrelevant Id no. 1; ×, postvaccine sera on irrelevant Id no. 2; Embedded Image , postvaccine sera on irrelevant Id no. 3. Panel B: ● indicates prevaccine sera on relevant Id (D29); ▪, postvaccine sera on relevant Id (D29); ▵, postvaccine sera on irrelevant Id no. 1; ×, postvaccine sera on irrelevant Id no. 2; and Embedded Image , postvaccine sera on irrelevant Id no. 3.

  • Fig. 3.

    Flow cytometric analysis of tumor-specific cell staining by sera from patients vaccinated with Id-KLH–pulsed dendritic cells.

    Tumor cells from patients D31 and D29 (upper and lower sets of panels, respectively) were incubated with autologous postvaccine serum or that of the other patient as a control. Bound anti-Id IgG antibodies were detected by anti-IgG-PE (y-axis), and tumor B cells (IgM+) were counterstained by anti-IgM–FITC (x-axis). Each patient's serum specifically recognizes only the autologous tumor cells but not the control, irrelevant tumor cells.

  • Fig. 4.

    Idiotype-immune serum induces tyrosine phosphorylation specifically in tumor B cells.

    (A) Detection of tyrosine phosphorylation in tumor cells by Western blotting. Tumor cells from patient D31 were incubated with the indicated sera and then lysed. Solubilized proteins were separated by SDS-PAGE and transferred to nitrocellulose. The blot was stained with antiphosphotyrosine antibody 4G10. (B) Detection of tyrosine phosphorylation in tumor cells by flow cytometry. Tumor cells from patient D31 were treated with the indicated sera, permeabilized, and stained with antibody 4G10 coupled to FITC. Phosphotyrosine content was then quantitated in aliquots of cells counterstained with either phycoerythrin-labeled anti-CD20 or anti-CD3.

  • Fig. 5.

    Regression of lymphomatous pleural effusion after vaccination with Id-KLH–pulsed dendritic cells.

    Despite chemotherapy, a left-sided pleural effusion persisted in patient D29 and remained cytologically positive before vaccination. Near-complete resolution is noted 9 months after completing vaccination with Id-KLH–pulsed dendritic cells.

  • Fig. 6.

    Regression of tumors after booster vaccination with Id-KLH + chemical adjuvant.

    Patient D12 was given Id-pulsed DC vaccination during first remission after chemotherapy but relapsed 15 months later. Subsequent to the development of widespread disease 18 months later, she was given injections of Id-KLH protein plus a chemical adjuvant. (A) CT images of the pelvis show that bulky left-sided pelvic lymph nodes have completely regressed 4 months after booster vaccinations. (B) Regression of axillary (top panels) and inguinal (bottom panels) lymph nodes after booster vaccinations. Arrows indicate sites of disease before vaccination.

  • Fig. 7.

    Tumor-specific cytolysis by peripheral blood lymphocytes derived from patient D12 before and after a repeat series of Id-KLH vaccinations (third course of vaccinations).

    Prevaccine and postvaccine PBMC effectors were restimulated using 2 weekly cycles of CD40L-activated autologous tumor cells plus interleukin-2. After 14 days, the ability of effectors to lyse unmodified, cryopreserved tumor cells was assessed in a 4-hour51Cr release assay. Purified autologous normal peripheral blood B cells serve as negative control targets. The percentage specific lysis is expressed as the mean of triplicate values. Results are representative of 3 independent experiments. ▴ indicates postvaccine course no. 3 versus tumor targets; ▵ indicates prevaccine course no. 3 versus tumor targets; ● indicates postvaccine course no. 3 versus normal B cells; and ○ indicates prevaccine course no. 3 versus normal B cells.

Tables

  • Table 1.

    Characteristics and responses of initial patients with measurable disease (n = 10)

    PatientAge/
    sex
    HistologyPrior therapiesSites of diseaseTotal DCs given*Anti-Id responsesClinical response, time to progressionCurrent status, subsequent therapies, months since DC vaccine
    T cellAb
    D1 59/FFMChl, ChlVBParacardiac, mediastinal69 × 106 +CR, 44 monthsRD after CHOP, rituximab, XRT, fludarabine, repeat DC vaccine, 94 mos
    D2 44/FFMChl, CVPPeri-aortic, para-iliac22 × 106 +PR, 12 monthsNED after Id-KLH/SAF,1-153 fludarabine, 83 mos
    D3 34/FF/DSCCVP, fludarabineBM14 × 106 + Cleared BM PCR signalNED 79 mos after DC vaccine without subsequent therapies
    D4 45/MFSCCVP, fludarabineCervical, axillary, peri-aortic, iliac, inguinal16 × 106 +PD at 10 mosNED after MINE, rituximab, 69 mos
    D550/FFMCVP, fludarabinePeri-aortic, BM23 × 106 +PD at 7 mosNED after Id-KLH/SAF,1-153 rituximab, 65 mos
    D647/MFSCChl, CHOPPeri-aortic, spleen, mesenteric22 × 106 +PD at 12 mosRD after Id-KLH/SAF,1-153 rituximab, XRT, 63 mos
    D743/FFSCCHOPAxillary, mesenteric27 × 106 +PD at 9 mosMRD after rituximab, 58 mos
    D840/MFSCCVP, XRT, CEP, fludarabineAxillary, peri-aortic10 × 106 + PD at 5 mosNED after Id-KLH/SAF,1-153 57 mos
    D943/FFSCChl, Id-KLH/SAFAxillary, iliac, peri-aortic13 × 106 CR, 57 mosPD at single site 57 mos after DC vaccine
    D1051/FFMCVP, fludarabine, CHOPAxillary, spleen12 × 106 PD at 7 mosNED after Id-KLH/SAF,1-153 tositumomab, 47 mos
    • FSC indicates follicular small cleaved cell; FM, follicular mixed small cleaved and large cell; F/DSC, follicular and diffuse small cleaved cell; Chl, chlorambucil; ChlVB, chlorambucil, vincristine, and bleomycin; CVP, cyclophosphamide, vincristine, and prednisone; CHOP, cyclophosphamide, adriamycin, vincristine, and prednisone; CEP, cyclophosphamide, etoposide, and prednisone; Id-KLH/SAF, Id protein coupled to KLH plus Syntex adjuvant formulation; XRT, radiation therapy; MINE, mesna, ifosfamide, mitoxantrone, etoposide; PD, progressive disease; NED, no clinical evidence of disease; MRD, minimum residual disease; RD, residual disease; Ab, antibody; BM, bone marrow.

    • * Total number of DCs infused over the course of 4 vaccinations.

    • Previously reported in original pilot study.22

    • IgM, but not IgG anti-Id antibodies detectable in serum.

    • F1-153 Revaccinated with Id-KLH plus SAF adjuvant. See Table 3.

  • Table 2.

    Characteristics and immune responses of patients vaccinated in first remission after chemotherapy (n = 25)

    PatientAge/sexHistology, stageChemotherapy, no. cyclesPrevaccine tumor statusDCs pulsed withNo. DCs*Anti-Id responseFollow-up after chemotherapy
    T cellAb
    D1165/MFM, IVACHOP × 4RDId33 × 106 SD, 51 mos
    CVP/F × 4
    D1244/FFSC, IVACVP × 8MRDId9 × 106 +PD at 15 mos, NED at 62 mos
    D1353/FFSC, IIIACVP × 8RDId19 × 106 NED, 52 mos
    D1460/MFSC, IIIACVP × 6NEDId9 × 106 NED, 53 mos
    D1530/MFSC, IVBCVP × 8MRDId19 × 106 +SD, 47 mos
    D1634/MFSC, IIIACVP × 6MRDId11 × 106 +NED, 52 mos
    D1744/FFSC, IVACVP/F × 8MRDId17 × 106 NED, 56 mos
    D1852/FFSC, IVAChl × 6MRDId17 × 106 +NED, 43 mos
    D1960/FFSC, IVACVP × 8MRDId14 × 106 SD, 46 mos
    D2048/MFSC, IVACVP × 6PDId18 × 106 PD at 9 mos, alive at 50 mos
    D2138/MFM, IVACVP × 6MRDId8 × 106 +PD at 25 mos, dead at 32 mos2-153
    D2240/MFSC, IVACVP × 8MRDId19 × 106 PD at 35 mos, alive at 44 mos
    D2355/FFSC, IVACVP × 8RDId-KLH15 × 106 SD, 39 mos
    D2447/FFM, IVACVP × 6NEDId-KLH16 × 106 +NED, 38 mos
    D2530/MFSC, IVACVP × 6RDId-KLH10 × 106 PD at 18 mos, alive at 37 mos
    D2656/MFM, IVACVP × 7NEDId-KLH16 × 106 ++NED, 45 mos
    D2750/FFM, IVACVP × 6PDId-KLH21 × 106 PD at 9 mos,2-153 alive at 35 mos
    D2861/MFM, IVACVP × 6NEDId-KLH10 × 106 +PD at 21 mos, alive at 36 mos
    D2950/MFSC, IVBCVP × 8RDId-KLH12 × 106 +MRD, 28 mos
    D3050/FFM, IVACVP × 6MRDId-KLH20 × 106 SD, 24 mos
    CHOP × 2
    D3154/FFSC, IVACVP × 8NEDId-KLH10 × 106 +NED, 24 mos
    D3243/FFSC, IVACVP × 8MRDId-KLH15 × 106 PD at 16 mos, alive at 25 mos
    D3355/MFM, IIIACVP × 7RDId-KLH21 × 106 SD, 23 mos
    CHOP × 4
    D3453/MFSC, IVACVP × 6RDId-KLH35 × 106 +SD, 25 mos
    CHOP × 2
    D3547/MFSC, IVACVP × 8MRDId-KLH28 × 106 +PD at 28 mos, alive at 33 mos
    • FSC indicates follicular small cleaved cell; FM, follicular mixed small cleaved and large cell; F/DSC, follicular and diffuse small cleaved cell; CHOP, cyclophosphamide, adriamycin, vincristine, and prednisone; CVP, cyclophosphamide, vincristine, and prednisone; CVP/F, CVP alternating with fludarabine; Chl, chlorambucil; RD, residual disease; MRD, minimum residual disease; SD, stable disease; PD, progressive disease; NED, no clinical evidence of disease; Ab, antibody.

    • * Total number of DCs infused over the course of 4 vaccinations.

    • IgM, but not IgG anti-Id antibodies detectable in serum.

    • Revaccinated with Id-KLH plus SAF adjuvant. See Table 3.

    • F2-153 Histologic transformation to diffuse large cell lymphoma.

  • Table 3.

    Characteristics and responses of patients revaccinated with Id-KLH protein + SAF1

    PatientInterval from DC to Id-KLH vaccinationSites of diseaseAnti-Id responseClinical response, time to progression
    T cellAb
    D218 mosCervical, iliac, peri-aortic+PR, 14 mos
    D516 mosSkin, cervical, axillary, inguinalPD
    D616 mosParotid, axillary, mesenteric, peri-aorticPD
    D8 8 mosAxillary, peri-aortic3-150 CR, 48+ mos
    D1022 mosSkin, peri-aortic, axillary, inguinalPD
    D123-151 33 mosCervical, axillary, iliac, inguinal, BM+CRu, 16 mos
    49 mos3-152 Cervical, BM+CR, 1+ mos
    • CR indicates complete response; CRu, complete response unconfirmed (normalization of radiographs, equivocal findings in bone marrow); PR, partial response; PD, progressive disease; BM, bone marrow.

    • F3-150 IgM, but not IgG anti-Id antibodies detectable in serum.

    • F3-151 Adjuvant = ISAF (“incomplete” SAF1, without threonyl MDP component).

    • F3-152 Repeat course of Id-KLH protein + ISAF vaccinations (vaccine course 3).