dinitrobenzenes and Neoplasms

We have devised a novel approach to active immunotherapy based on modification of autologous cancer cells with the hapten, dinitrophenyl (DNP). The treatment program consists of multiple intradermal injections of DNP-modified autologous tumor cells mixed with BCG. Administration of DNP-vaccine to patients with metastatic melanoma induces a unique reaction - the development of inflammation in metastatic masses. Histologically, this consists of infiltration of T lymphocytes, most of which are CD8+. These T cells usually produce gamma interferon in situ. Moreover, they represent expansion of T cell clones with novel T cell receptor structures. Occasionally, administration of DNP-vaccine results in partial or complete regression of measurable metastases. The most common site of regression has been small lung metastases. Administration of DNP-vaccine to patients in the post-surgical adjuvant setting produces a more striking clinical effect. We have treated 214 patients with clinically evident stage III melanoma who had undergone lymphadenectomy. With a median follow-up time of 4.4 years (1.8-10.4 years) the 5-year overall survival (OS) rate is 47% (one nodal site = 51%, two nodal sites = 33%). These results appear to be comparable to those obtained with high dose interferon. More recent studies suggest that this therapeutic approach is also applicable to ovarian cancer. There appear to be no insurmountable impediments to applying this approach to much larger numbers of patients or to developing it as a standard cancer treatment.

Topics: Antigens, Neoplasm; Autoantigens; Cancer Vaccines; Combined Modality Therapy; Dinitrobenzenes; Female; Haptens; Humans; Inflammation; Lymphocytes, Tumor-Infiltrating; Neoplasm Metastasis; Neoplasms; Ovarian Neoplasms

We have previously developed a universal chimeric antigen receptor (CAR), which recognizes dinitrophenyl (DNP) and can redirect T and NK cells to target cancer and HIV antigens using DNP-conjugated antibodies as adaptor molecules. However, the DNP-antibody conjugates are generated by random modification, which may not be optimal for this modular system. Here, we report the development of enhanced adaptor molecules by site-specific DNP modification. We use the genetic code expansion technology to generate single-chain fragment variable (scFv) antibodies with site-specific DNP. We compare four anti-CD19 scFv mutants and find that the one with DNP at the flexible peptide linker between V

Topics: Antigens, CD19; Antigens, Neoplasm; Cell Line, Tumor; Dinitrobenzenes; Humans; Immunotherapy, Adoptive; Neoplasms; Receptors, Chimeric Antigen; Single-Chain Antibodies; T-Lymphocytes

The development of more efficient photosensitizers with minimal damage to surrounding normal tissues has been a valuable and challenging subject during photodynamic therapy (PDT). Herein, a stimuli-activated porphyrinic photosensitizer (PEG-TPP-DNB; PEG = poly(ethylene glycol); TPP = 5,10,15,20-tetraphenylporphyrin; DNB = 2,4-dinitrobenzene) with capabilities of fluorescence and, remarkably, singlet oxygen quenching was prepared successfully for photodynamic therapy with high efficiency and biosecurity. The amphiphilic PEG-TPP-DNB could be self-assembled into nanomicelles in aqueous media and dissociated in response to reductive thiol such as glutathione. Meanwhile, the fluorescence and singlet oxygen generation of porphyrinic photosensitizer would be activated to regenerate. Moreover, the intracellular uptake and localization effectively confirmed the redox-responsive and activated behavior of PEG-TPP-DNB micelles. The cytotoxicity in vitro revealed that the micelles had low dark toxicity and great phototoxicity, and in vivo bioimaging and antitumor evaluation further indicated that the micelles possessed selective tumor imaging and targeted PDT antitumor effect as well as low systemic toxicity. Overall, this tumor microenvironment-activated photosensitizer system may provide a useful strategy for precise photodynamic therapy.

Topics: Cell Proliferation; Dinitrobenzenes; Humans; Micelles; Neoplasms; Oxidation-Reduction; Oxygen; Photochemotherapy; Photosensitizing Agents; Polyethylene Glycols; Porphyrins; Surface-Active Agents

Cellular imaging technologies employing metallic surface-enhanced Raman scattering (SERS) tags have gained much interest toward clinical diagnostics, but they are still suffering from poor controlled distribution of hot spots and reproducibility of SERS signals. Here, we report the fabrication and characterization of high narrow nanogap-containing Au@Au core-shell SERS nanoparticles (GCNPs) for the identification and imaging of proteins overexpressed on the surface of cancer cells. First, plasmonic nanostructures are made of gold nanoparticles (∼15 nm) coated with gold shells, between which a highly narrow and uniform nanogap (∼1.1 nm) is formed owing to polyA anchored on the Au cores. The well controlled distribution of Raman reporter molecules, such as 4,4'-dipyridyl (44DP) and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), are readily encoded in the nanogap and can generate strong, reproducible SERS signals. In addition, we have investigated the size-dependent SERS activity of GCNPs and found that with the same laser wavelength, the Raman enhancement discriminated between particle sizes. The maximum Raman enhancement was achieved at a certain threshold of particle size (∼76 nm). High narrow nanogap-containing Au@Au core-shell SERS tags (GCTs) were prepared via the functionalization of hyaluronic acid (HA) on GCNPs, which recognized the CD44 receptor, a tumor-associated surface biomarker. And it was shown that GCTs have a good targeting ability to tumour cells and promising prospects for multiplex biomarker detection.

Topics: Cell Line, Tumor; Dinitrobenzenes; Gold; Humans; Hyaluronan Receptors; Metal Nanoparticles; Neoplasm Proteins; Neoplasms; Pyridines; Spectrum Analysis, Raman

Epidemiological and toxicological studies point to a potential carcinogenic effect of dinitrotoluene (DNT), particularly with respect to renal and urothelial cancer.. The cohort comprised all men born between 1920 and 1974 (n = 16,441) who were gainfully employed between 1953 and 1990 in one of two underground copper mines in Mansfeld, Saxony-Anhalt, former German Democratic Republic, and who were followed up for cancer incidence, 1961-2005. Incident cancer cases were identified by record linkage with the Common Cancer Registry of the New Laender. Standardized incidence ratios (SIR) were calculated with the general population of Saxony-Anhalt as the reference.. Standardized incidence ratios for all cancers were not significantly elevated in the cohort (SIR = 1.04; 95 % confidence intervals (CI) 0.96-1.14). We found an increase in lung cancer (SIR = 1.29; 1.13-1.46), but not in kidney cancer (SIR = 1.01; 95 % CI 0.79-1.27) or bladder cancer (SIR = 1.04; 95 % CI 0.82-1.30). Standardized incidence ratios stratified by duration of employment with DNT exposure indicated moderately increased risks for kidney and bladder cancer in cohort members with longer exposure.. The SIR analysis of workers in the copper mining industry in comparison with the general population of Saxony-Anhalt overall did not indicate increased risks for renal or bladder cancer. However, results by years of exposure to DNT suggested weakly increased risks for outcomes of a priori interest, bladder and kidney cancer. A subsequent case-cohort analysis including expert assessment of DNT exposure and identification of additional cancer cases from a network of pathology institutes will provide further insight into a potential etiologic role of DNT in renal and urothelial cancer.

Topics: Adult; Aged; Aged, 80 and over; Air Pollutants, Occupational; Carcinogens; Cohort Studies; Copper; Dinitrobenzenes; Germany; Humans; Incidence; Kidney Neoplasms; Male; Middle Aged; Mining; Neoplasms; Occupational Diseases; Occupational Exposure; Urinary Bladder Neoplasms

We have designed and evaluated UTX-12 as a novel fluorescent pH probe for tumor hypoxia imaging. UTX-12 consists of a p-nitro benzyl moiety, which is a latent hypoxia-selective leaving group activated by nitro reduction, directly linked to SNARF. Although UTX-12 itself is colorless and non-fluorescent in aqueous solution, nitro reduction triggers the release of SNARF which has well-characterized long wavelength absorption and fluorescence that is sensitive to pH. The resultant SNARF, released intracellularly by enzymatic reduction of UTX-12, allows measurement of pH by pH-dependent dual emission shifts. UTX-12 showed clear differences in fluorescence behavior between hypoxic and aerobic conditions in liver microsomes and inside V79 cells. These data are confirmation that UTX-12 is biologically reduced inside tumor cells and the released SNARF should monitor intracellular pH of tumor cells selectively with reduced background signal.

Topics: Animals; Benzopyrans; Cell Hypoxia; Cell Line; Diagnostic Imaging; Dinitrobenzenes; Drug Design; Fluorescent Dyes; Humans; Hydrogen-Ion Concentration; Microsomes, Liver; Neoplasms; Oxidation-Reduction

Here we described novel interactions of the mammalian selenoprotein thioredoxin reductase (TrxR) with nitroaromatic environmental pollutants and drugs. We found that TrxR could catalyze nitroreductase reactions with either one- or two-electron reduction, using its selenocysteine-containing active site and another redox active center, presumably the FAD. Tetryl and p-dinitrobenzene were the most efficient nitroaromatic substrates with a k(cat) of 1.8 and 2.8 s(-1), respectively, at pH 7.0 and 25 degrees C using 50 muM NADPH. As a nitroreductase, TrxR cycled between four- and two-electron-reduced states. The one-electron reactions led to superoxide formation as detected by cytochrome c reduction and, interestingly, reductive N-denitration of tetryl or 2,4-dinitrophenyl-N-methylnitramine, resulting in the release of nitrite. Most nitroaromatics were uncompetitive and noncompetitive inhibitors with regard to NADPH and the disulfide substrate 5,5'-dithiobis(2-nitrobenzoic acid), respectively. Tetryl and 4,6-dinitrobenzofuroxan were, however, competitive inhibitors with respect to 5,5'-dithiobis(2-nitrobenzoic acid) and were clearly substrates for the selenolthiol motif of the enzyme. Furthermore, tetryl and 4,6-dinitrobenzofuroxan efficiently inactivated TrxR, likely by alkylation of the selenolthiol motif as in the inhibition of TrxR by 1-chloro-2,4-dinitrobenzene/dinitrochlorobenzene (DNCB) or juglone. The latter compounds were the most efficient inhibitors of TrxR activity in a cellular context. DNCB, juglone, and tetryl were highly cytotoxic and induced caspase-3/7 activation in HeLa cells. Furthermore, DNCB and juglone were potent inducers of apoptosis also in Bcl2 overexpressing HeLa cells or in A549 cells. Based on these findings, we suggested that targeting of intracellular TrxR by alkylating nitroaromatic or quinone compounds may contribute to the induction of apoptosis in exposed human cancer cells.

Topics: Animals; Apoptosis; Binding Sites; Caspase 3; Caspase 7; Caspases; Cell Line, Tumor; Dinitrobenzenes; Dithionitrobenzoic Acid; Enzyme Activation; Enzyme Inhibitors; Humans; Molecular Structure; Naphthoquinones; Neoplasms; Nitrogen Compounds; Oxidation-Reduction; Proto-Oncogene Proteins c-bcl-2; Sulfhydryl Reagents; Superoxides; Thioredoxin-Disulfide Reductase

It is often believed that small interfering RNA (siRNA) is at least 10-fold more effective than the single-stranded antisense oligonucleotide for silencing the same target gene in the same cells. In view of the recent discovery that the RNA-induced silencing complex (RISC) contains only a single-stranded RNA (ssRNA) molecule and can be reconstituted using single-stranded antisense RNA, such a large difference in efficacy seems puzzling. One possible reason is that hybridization protects siRNA from hydrolysis by endogenous RNase activity until it is incorporated in the RISC, whereas ssRNA is rapidly hydrolyzed. Because the single-stranded poly-2'-O-(2,4-dinitrophenyl)-RNA (DNP-ssRNA) is both RNase resistant and membrane permeable, we synthesized homologous native siRNAs, DNP-siRNAs, native ssRNAs, and DNP-ssRNAs and made a comparative study of their efficacies for inhibiting the growth of two cancer cell lines with different overexpressed target genes under equivalent experimental conditions. It was found that the efficacy of antisense DNP-ssRNA is higher than that of the corresponding siRNA and that the efficacy of native siRNA for inhibiting cell growth can also be enhanced from 2-fold to 6-fold by replacing the native strands of RNA in siRNA with homologous DNP-RNA. Thermal denaturation data show that the hybridization affinity of the DNP-RNA/RNA duplex is higher than that of the native RNA/RNA duplex. Western blotting analysis of A549 cells treated with antisense DNP-ssRNAs containing single mismatching bases shows that the gene silencing by antisense DNP-ssRNA is as sequence specific as that by siRNA. The observed large enhancement of inhibition efficacy of native RNAs by DNP derivatization should be advantageous for both gene silencing studies and therapeutic applications.

Topics: Blotting, Western; Cell Line, Tumor; Cell Proliferation; Dinitrobenzenes; Growth Inhibitors; Humans; Inhibitory Concentration 50; Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA; Sensitivity and Specificity; Substrate Specificity

A novel approach to active immunotherapy has been devised based on modification of autologous cancer cells with the hapten, dinitrophenyl (DNP). This technology is being developed by AVAX Technologies as a treatment for melanoma under the brand name, M-Vax(TM). The treatment program consists of multiple intradermal injections of DNP-modified autologous tumour cells mixed with Bacille Calmette-Guérin (BCG). DNP-vaccine administration to patients with metastatic melanoma induces a unique reaction - the development of inflammation in metastatic masses. The inflammation is mediated by IFN-gamma-producing T-lymphocytes, some of which represent expansion of novel clones. Following DNP-vaccine treatment, almost all patients develop delayed-type hypersensitivity (DTH) to autologous, DNP-modified melanoma cells; approximately half also exhibit DTH to autologous, unmodified tumour cells. The toxicity of the vaccine is mild, consisting mainly of papules or pustules at the injection sites. Clinical trials have been conducted in two populations of melanoma patients: stage IV with measurable metastases and clinical stage III patients, rendered tumour-free by lymphadenectomy. In 83 patients with measurable metastases, there were 11 antitumour responses: two complete responses (CRs), four partial responses (PRs) and five mixed. Both CRs and two of four PRs occurred in patients with lung metastases. In 214 stage III patients the 5-year overall survival rate was 46% (one nodal site = 48%, in-transit metastases = 50%, two nodal sites = 36%). In both populations, the induction of DTH to unmodified autologous tumour cells was associated with significantly longer survival. This technology is applicable to other human cancers and clinical trials have been initiated with ovarian adenocarcinoma. There appear to be no insurmountable impediments to applying this approach to much larger numbers of patients or to developing it as a standard cancer treatment.

Topics: Animals; BCG Vaccine; Cancer Vaccines; Combined Modality Therapy; Dinitrobenzenes; Haptens; Humans; Immunotherapy; Melanoma; Neoplasms

An analysis of the mortality experience of workers exposed to dinitrotoluene (DNT) was conducted to test the hypothesis that DNT exposure is associated with an increased risk of cancers of the liver and biliary tract. A total of 4,989 workers exposed to DNT and 7,436 unexposed workers who had worked for at least 5 months at the study facility between January 1, 1949 and January 21, 1980, were included in this investigation. Workers were considered exposed if they had worked at least 1 day on a job with probable exposure to DNT. The vital status as of December 31, 1982, was successfully ascertained for approximately 97% of these workers. Standardized mortality ratios (SMRs) were estimated based upon comparisons with the US population using a modified life-table program. In addition, standardized rate ratios (SRRs) were computed based upon direct comparisons between the DNT and the internal unexposed cohort. An excess of hepatobiliary cancer was observed among workers exposed to DNT in this study. The rate ratio for hepatobiliary cancer was 2.67 (six cases observed) based upon comparison with the US population (SMR = 2.67, 95% CI = 0.98, 5.83), and 3.88 based upon comparison using the internal unexposed referent group (SRR = 3.88, 95% CI = 1.04, 14.41). This study failed to demonstrate an exposure-response relationship between duration of DNT exposure and hepatobiliary cancer mortality. Our study was limited by the small number of workers with long duration of exposure to DNT, and by the lack of quantitative information on exposure to DNT and other chemicals.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Air Pollutants, Occupational; Bile Duct Neoplasms; Carcinogens; Carcinoma, Hepatocellular; Cause of Death; Cohort Studies; Dinitrobenzenes; Humans; Life Tables; Liver Neoplasms; Male; Middle Aged; Neoplasms; Occupational Diseases; Occupational Exposure; Retrospective Studies; Survival Analysis

Immune memory to DNP-Ficoll, a nonmitogenic, thymic-independent (TI-2) antigen, was demonstrated in several inbred strains of mice. Direct and indirect splenic plaque-forming cell responses were measured in mice given a secondary challenge with DNP-Ficoll and in appropriate control mice. Strong IgM memory, but no IgG memory, was observed in SJL/J, AKR/J, and C58/J mice. C57L/J mice gave both a strong IgM memory response and a relatively strong IgG memory response to DNP-Ficoll. C57BL/6N, C57BR/cdJ, and A/HeJ mice were unable to mount significant IgM or IgG memory responses to this antigen under an identical immunization schedule. These results are indicative of marked genetic variation in mice in the capacity for B memory cell expression. They also identify a provocative but unexplained association between positive IgM memory responses to DNP-Ficoll in SJL/J, AKR/J, and C58/J mice and a propensity to develop lymphoreticular neoplasia. This association is observed when there is no clear evidence for IgG memory.

Topics: Animals; Antigens; B-Lymphocytes; Dinitrobenzenes; Female; Ficoll; Guinea Pigs; Hemolytic Plaque Technique; Immunologic Memory; Mice; Mice, Inbred A; Mice, Inbred AKR; Mice, Inbred C57BL; Mitogens; Neoplasms; Rabbits

Topics: Adult; Aged; Colitis, Ulcerative; Dinitrobenzenes; Female; Hepatitis, Alcoholic; Humans; Immunity, Cellular; Male; Methods; Middle Aged; Neoplasms; Streptokinase; Tuberculin Test

Topics: Animals; Antigens, Neoplasm; Autoantibodies; BCG Vaccine; Cell Membrane; Dinitrobenzenes; Dog Diseases; Dogs; Immunotherapy; Male; Mammary Glands, Animal; Neoplasms; Nitrobenzenes

Topics: BCG Vaccine; Dinitrobenzenes; Humans; Immunity, Cellular; Immunotherapy; Neoplasms