RESUMO
Effective eradication of established tumor and generation of a lasting systemic immune response are the goals of cancer immunotherapy. The objective of this phase IB study was to assess the safety and toxicity of treatment to metastatic tumor underlying the skin with the DNA encoding interleukin-12 (IL-12). This treatment strategy allowed the patient's own tumor to serve as a source of autologous antigen in the tumor microenvironment. We proposed that IL-12 protein produced by the transfected cells would result in the generation of both a local and systemic antitumor response. The tumor was treated with either three or six intratumoral injections of plasmid containing IL-12 DNA. This treatment strategy resulted in no significant local or systemic toxicity. The treatment did not result in an increase in serum IL-12 protein. The size of the treated lesion decreased significantly (greater than 30%) in five of the 12 patients. However, nontreated subcutaneous lesions or other disease did not decrease in size.
Assuntos
DNA de Neoplasias/administração & dosagem , Vetores Genéticos/administração & dosagem , Interleucina-12/administração & dosagem , Interleucina-12/genética , Melanoma/terapia , Plasmídeos/genética , Adulto , Idoso , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/terapia , DNA de Neoplasias/efeitos adversos , Feminino , Vetores Genéticos/efeitos adversos , Humanos , Injeções Intralesionais , Interleucina-12/efeitos adversos , Neoplasias Renais/genética , Neoplasias Renais/terapia , Masculino , Melanoma/genética , Melanoma/secundário , Pessoa de Meia-IdadeRESUMO
The primary objective of this phase I study was to determine the safety of an autologous tumor vaccine given by intradermal injection of lethally irradiated granulocyte-macrophage colony-stimulating factor (GM-CSF) gene-transfected autologous melanoma and sarcoma cells. Secondary objectives included validation of the gene delivery technology (particle-mediated gene transfer), determining the host immune response to the tumor after vaccination, and monitoring patients for evidence of antitumor response. Sixteen patients were treated with either of two different doses of GM-CSF-treated tumor cells. One patient received treatment with both doses of tumor cells. No treatment-related local or systemic toxicity was noted in any patient. Patients administered 100% treated cells (i.e., with a preparation of tumor cells that had all been exposed to GM-CSF DNA transfection) had a more extensive lymphocytic infiltrate at the vaccine site than did patients given 10% treated cells (a preparation of tumor cells in which 10% had been exposed to GM-CSF transfection) or nontreated tumor. The generation of a systemic immune response to autologous tumor by a delayed-type hypersensitivity response to the intradermal placement of nontransfected tumor cells was noted in one patient. One patient had a transient partial response of metastatic tumor sites. The entire procedure, from tumor removal to vaccine placement, was accomplished in less than 6 hr in all patients. Four of 17 patient tumor preparations produced greater than 3.0 ng of GM-CSF per 10(6) cells per 24 hr in vitro. The one patient with greater than 30 ng of GM-CSF per 10(6) cells per 24 hr in vitro had positive DTH, a significant histologic inflammatory response, and clinically stable disease. This technique of gene transfer was safe and feasible, but resulted in clinically relevant levels of gene expression in only a minority of patients.
Assuntos
Biolística , Vacinas Anticâncer/uso terapêutico , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fatores Imunológicos/genética , Melanoma/terapia , Sarcoma/terapia , Neoplasias de Tecidos Moles/terapia , Vacinas de DNA/uso terapêutico , Vacinas Anticâncer/administração & dosagem , DNA Complementar/genética , Relação Dose-Resposta Imunológica , Estudos de Viabilidade , Ouro , Humanos , Hipersensibilidade Tardia/imunologia , Injeções Intradérmicas , Leiomiossarcoma/patologia , Leiomiossarcoma/terapia , Lipossarcoma/patologia , Lipossarcoma/terapia , Melanoma/patologia , Melanoma/secundário , Segurança , Sarcoma/patologia , Neoplasias de Tecidos Moles/patologia , Resultado do Tratamento , Células Tumorais Cultivadas/transplante , Vacinas de DNA/administração & dosagemRESUMO
The genomic clones encoding lignin peroxidase isozyme H8 and two closely related genes were isolated from Phanerochaete chrysosporium BKM-1767, and their nucleotide sequences were determined. The positions and approximate lengths of introns were found to be highly conserved in all three clones. Analysis of homokaryotic derivatives indicated that the three clones are not alleles of the same gene(s).
Assuntos
Basidiomycota/genética , Clonagem Molecular , Genes Fúngicos , Isoenzimas/genética , Peroxidases/genética , Sequência de Aminoácidos , Sequência de Bases , Basidiomycota/enzimologia , Southern Blotting , Códon , Dados de Sequência Molecular , Homologia de Sequência do Ácido NucleicoRESUMO
Sequence analysis of Chinese hamster V79 lung fibroblast cDNA clones, which code for UV radiation-inducible transcripts, revealed that many of the clones corresponded to metallothioneins (MTs) I and II. A third cDNA clone, DDIU4, was found also to code for a similar-size UV-inducible transcript which was unrelated to MT by both sequence analysis and kinetics of induction. MTI and MTII RNAs rapidly increased in V79 cells within 1 h after UV irradiation, and maximum induction was seen by 4 h. This rapid induction of MT RNA by UV irradiation was not observed in human fibroblasts. MTI and MTII were coordinately induced in both time course and dose-response experiments, although the induction of MTII, up to 30-fold, was three to four times greater than that of MTI. The induction of MT did not appear to be a general stress response, since no increase occurred after exposure to X rays or H2O2.
