vaccination of melanoma patients with dendritic cells loaded with allogeneic apoptotic-necrotic melanoma cells

Conditions: Melanoma

Summary: background: sixteen melanoma patients (1 stage iic, 8 stage iii, and 7 stage iv) were treated in a phase i study to evaluate safety and immune responses, with a vaccine (dc/apo-nec) composed of autologous dendritic cells (dcs) loaded with a mixture of apoptotic/necrotic melanoma cell lines (apo-nec). methods: pbmc were obtained from leukapheresis and dcs were generated from monocytes cultured in the presence of gm-csf and il-4 in serum-free medium. immature dcs (idcs) were loaded with gamma-irradiated apo-nec cells and injected id without adjuvant. cohorts of four patients were given four vaccines with 5, 10, 15, or 20 x106 dc/apo-nec per vaccine, two weeks apart. results: the vaccine was well tolerated in all patients. toxicity to vaccine was mild, and the toxicity-limiting dose has not been reached. we found that 42.3 ±13.7 % melanoma patients´ idcs were able to phagocyte apo-nec cells wich induced dcs maturation, as evidenced by increased expression of cd83, cd80, cd86, hla class i and ii compared to idcs. also, after phagocytosis, a 75.2 ±16 % reduction in dextran-fitc endocytosis was observed compared to idcs. ccr7 was upregulated upon apo-nec phagocytosis in dcs from all patients and accordingly in vitro dc/apo-nec cells were able to migrate towards mip-3 beta. the dth score increased significatively in the patients after the first vaccination and slightly decreased by the fourth vaccine (mann-whitney test, p\<0.05). for patient #1 a positive dth reaction was detected to her own tumor irradiated cells. the presence of cd8+ t lymphocytes specific to gp100 and melan a/mart-1 ags were studied by tetramers binding in hla-a\*0201 patients (7 /15 patients) before and after vaccination. two patients who remain ned increased ags their specific t lymphocytes after vaccination. no humoral responses to apo-nec cells were detected. with a mean follow-up of 44.5 months post-surgery, the stage iic pt is ned, 7/8 stage iii pts are ned and 7/7 stage iv patients have progressed. conclussions: we conclude that dc/apo-nec vaccine is well tolerated, it induces specific immunity against melanoma ags and in stage iii patients it may prolong disease-free survival, affording protection from relapse in an adjuvant setting.


Eligibility criteria:
inclusion criteria: * histologically confirmed cutaneous melanoma stages iib, iic, iii or iv (ajcc) * pts with minimal or non-detectable disease (ned) after surgery as asserted by cat scans. melanoma pts with unknown primary tumor site could be included in the study * life expectancy \> 6 months * ages:from 15 to 60 years * performance status (ecog) 0 or 1 * pts with stage iii disease had to be previously treated with ifn-alpha, and either finished the treatment or suspended it due to disease progression, toxicity or other clinical reasons. alternatively, those pts who had not started ifn-alpha within six months after surgery could be included in this study * a suitable venous access for the leukapheresis procedure * laboratory eligibility criteria included: hemoglobin \> 10 gr %; wbc count \> 4800/mm3, platelets \> 150.000/mm3, total and direct bilirubin, serum oxalacetic transaminase and glutamic pyruvic transaminase \< 1.5 fold the upper normal value; ldh \< 450 mu/ml * absence of pregnancy, with serum βhcg determined one week before vaccination in premenopausal women * serum creatinine \< 1.4 mg % * no chemotherapy, radiotherapy or any biological treatments during the previous month * no concurrent medication with corticosteroids or nsaids * l no active brain metastases m- normal ecg * all pts gave written informed consent before inclusion in the study. exclusion criteria: * ocular melanoma or melanoma of mucosa * active brain metastases * other previous carcinoma (with the exeption of cervical or in situ basal cells carcinoma adequately treated) * pregnant or breast-feeding women * cardiac arythmia, severe heart disease. * bacterial, mycotic or viral serious infections ( \> grade 2 according to nci common toxicity criteria) * hiv, b or c hepatitis previous infection * primary or secondary immunodeficiencies * other diseases that require treatment with regular corticoids or non steroids anti-inflammatory drugs or cox-2 inhibitors

Keywords: melanoma dendritic cells apoptotic/necrotic cells therapeutic vaccine