Supplementary MaterialsSupp_data. median follow up of 17.4 months, 9 patients reached
June 13, 2019
Supplementary MaterialsSupp_data. median follow up of 17.4 months, 9 patients reached PFS12. In these patients (responders, 37.5%), DC vaccination induced a significant, persistent activation of NK cells, whose increased response was significantly associated with prolonged survival. CD8+ T cells underwent Dabrafenib reversible enzyme inhibition rapid expansion and priming but, after the first administration of adjuvant TMZ, failed to generate a memory status. Resistance to TMZ was associated with robust expression of the multidrug resistance protein ABCC3 in NK but not CD8+ T cells. The negative effect of TMZ on the formation of T cell-associated antitumor memory deserves consideration Dabrafenib reversible enzyme inhibition in future clinical trials including immunotherapy. = 0.02 and = 0.03 respectively). The median PFS was 17.2 months (CI 95% 7.9C28.3) in the presence of MGMT methylation and 10.2 months in the other cases (CI 95% 7.9C14.7) (Fig.?1D). The median OS was 32.8 months (95%CI 20.2C33.9) in patients with MGMT methylation and 17.8 months in the others (CI 95% 14.4C22.6) (Fig.?1E). Exemplificative MRI of one responder and one non-responder (patients number 12 Rabbit polyclonal to ADORA3 and 17, respectively) are shown in Fig.?2. Open in a separate window Figure 2. Exemplificative MRI. Patient 17. Top, T2 weighted images (w.i.); bottom, contrast enhanced T1 w.i. (small box, pre-contrast T1 w.i.). (A) Pre-surgery, Dec 5, 2012 (necrotic lesion, GBM). (B). Post-surgery: Dec 6, 2012 (blood presence). (C-E) Immunotherapy: Jan, Mar and May 2013 (progressively enhancing lesion). Patient 12. Top, T2 w.i., bottom, contrast-enhanced T1 w.i. (F) Pre-surgery May 14, 2012 (necrotic lesion, GBM). (G) Post-surgery May 18, 2012 (scarce blood). (H-O) Immunotherapy: H-L (Aug 2012, Nov 2012, Jan 2013) show enhancing lesion. (M) subsequently reduced (M, Mar 2013) suggesting pseudoprogression. (N) subsequent stabilization (May 2014). (O) and disease progression (July 2014). Vaccine safety and adverse events. Three intradermal injections of mature DC loaded with autologous whole tumor lysate were administered before adjuvant TMZ, 4 further injections were performed during adjuvant TMZ (Fig.?1A). The treatment was well tolerated. One patient stopped treatment before disease progression due to pulmonary embolism. One patient died before progression because of deep venous thrombosis and pulmonary embolism. One case of grade 5 disseminated intravascular coagulation (DIC) was reported. Five cases of partial seizures, 8 convulsions and 1 myositis were also recorded. nonserious skin reactions included itching, erythema, urticaria and temporary inflammation at the injection site. A list of adverse events occurred during immunotherapy with relative grades is provided in Table?S1. Radio-chemotherapy and adjuvant chemotherapy affect CD8+ T, CD4+ T and NK cell counts. TMZ-induced lymphodepletion has been associated with expansion of a specific anti-tumor immune response.13 We measured the absolute lymphocyte counts (ALCs) at leukapheresis (the basal time point), during and at the end of the treatment. Basal ALCs were 1,000 cells per L peripheral blood in 22/24 patients at leukapheresis, and dropped significantly after RT-TMZ (from 1710.9 753.9 to 726.0 276.3, 0.0001) (Fig.?3A). RT-TMZ induced significant lymphopenia ( Dabrafenib reversible enzyme inhibition 1000 lymphocytes/microl) in 19/24 patients (79%). In 6/19 patients, the ALCs were 500 after RT-TMZ. Open in a separate window Figure 3. Radiotherapy and chemotherapy impact on CD8+, CD4+ T and NK cell counts. (A) Absolute lymphocyte counts in the peripheral blood of patients before (leukapheresis = leuka) and after (first vaccination = I vacc) RT-TMZ. (B-D) CD8+, CD4+ T and NK cell absolute counts before and after RT-TMZ. (E-G) Time course of CD8+ T cell count, CD4+ T cell count and NK cell count measured by flow cytometry Dabrafenib reversible enzyme inhibition of responder (left, n = 9) and non-responder (right, n = 15) patients over the treatment (* 0.01, ** 0.005, *** 0.0005 vs. 1st vaccine; underlined asterisk * vs. leukapheresis). Data are presented as mean SEM. RT-TMZ decreased CD8+ T cell (499.8 318.9 before RT-TMZ to 279.9 165.4, = 0.004), CD4+ T cell (from 708.0 371.8 to 312.6 107.7, 0.0005) and NK cell counts (from 88.0 72.9 to 45.7 37.9, = 0.02) (Fig.?3B-D). We also investigated the absolute count of CD8+ T, CD4+ T and NK cell subsets in the peripheral blood of all patients before, during and after immunotherapy..