Alkyl dimethyl benzyl ammonium chloride (ADBAC) and didecyl dimethyl ammonium chloride (DDAC) are common quaternary ammonium compounds used as disinfectants in households, medical, and restaurant settings. They cause occupational skin and respiratory hazards in humans, and developmental and reproductive toxicity in mice. They also cause increased secretions of proinflammatory cytokines in cell lines and vaginal inflammation in porcine models; but have not been evaluated for developmental immunotoxicity. We assessed immunotoxicity in-vitro with J774A.1 murine macrophage cell line by analyzing cytokine production and phagocytosis; and evaluated developmental immunotoxicity in CD-1 mice by analyzing antibody production. Additionally, because of the associations between gut microbiome dysbiosis and immune disease, we monitored changes in the microbiome as a result of ADBAC+DDAC exposure. Production of cytokines TNF-alpha and IL-6 increased at low ADBAC+DDAC concentrations, and IL-10 decreased in the murine macrophages with ADBAC+DDAC exposure. The phagocytic function of macrophages was also severely decreased. ADBAC+DDAC altered the mouse microbiome by decreasing the relative abundance of Bacteroides and increases in Clostridia in F0 and F1 generations. IgG primary and secondary responses were altered in F1 male mice; and IgA and IgM production were decreased in secondary response in F2 male mice. Since ADBAC+DDAC show signs of immunotoxicity in mice, further studies are needed to reassess risk for human exposure as ADBAC+DDAC may be contributing to immune disease. / Master of Science

Principal objectives of my research were to: (1) report for the first time that coelomocytes are able to reduce NBT dye and confirm the presence of lysozyme-like activity in earthworm; (2) develop a standard methodology for determination of NBT reduction and lysozyme-like activity in earthworms; (3) compare NBT reduction and lysozyme-like activity in earthworms with those of murine and human cells and fluids; and (4) demonstrate the sensitivity of earthworm NBT reduction and lysozyme-like activity as the assays using matrics in refuse-derived fuel fly ash (RDFF) and CuSO4.

The Murray-Darling basin is the largest river system in Australia with significant economic, social, recreational and cultural value. It supplies water for drinking and agriculture to a large inland area of the eastern and southern states of Australia. It is also the ultimate sink for many environmental contaminants that result from human activities within the catchment. Aquatic organisms live intimately with their environment and may be continuously exposed to these contaminants through the water column or the food chain. Some chemicals are bioaccumulated and biomagnified in tissue to reach high body burdens. Populations of native fish species within the Murray-Darling basin have been in decline since human settlement, yet little is known about the lethal and sublethal effects of environmental pollutants on native freshwater fish and many of the Australian water quality guidelines are based on data from exotic fish species. Researchers have correlated levels of pollution with immune dysfunction and an increased incidence of disease amongst wildlife populations. Many of the pollutants of the Murray-Darling basin have known immunotoxicity in both mammals and exotic fish species. The immune system is a sensitive target organ because, in order to maintain integrity, it requires constant renewal through the rapid proliferation and differentiation of cells. Efforts to increase numbers of native fish in the wild have led to an aquaculture industry that produces fingerlings for the restocking of waterways. In more recent years, this industry has matured and now produces table-size native freshwater fish for local and international markets. Although the industry has researched areas of reproduction, nutrition and stocking, there is little understanding of the immunology or immunotoxicology of Australian freshwater fish. This research project investigated the immunology of three large native fish species (i.e. 2 Murray cod, golden perch and silver perch), which are the basis of the native freshwater aquaculture industry. Additionally, a small fish species native to the basin (i.e. crimsonspotted rainbowfish) was studied as an alternative to the use of large fish. Of the four species, Murray cod possessed characteristics that made it an excellent candidate for ecoimmunotoxicity testing.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a highly toxic environmental pollutant and is well known for its immunotoxic effects, particularly on the thymus. The exact mechanism by which TCDD induces thymic atrophy is not known. In the current study, we investigated whether TCDD triggers apoptosis in the thymocytes and whether Fas and Fas ligand play a role in TCDD-mediated immunotoxicity. Administration of a single dose of TCDD at 0.1, 1, 5 or 50 mg/kg body weight intraperitoneally into C57BL/6 +/+ mice caused a significant dose-dependent decrease in the thymic cellularity; whereas, in the C57BL/6 lpr/lpr (lpr) (Fas-deficient) and C57BL/6 gld/gld (gld) (Fas ligand-defective) mice, TCDD failed to induce a decrease in thymic cellularity at doses of 0.1-5 mg/kg body weight. In the lpr and gld mice, thymic atrophy was seen only at 50 mg/kg body weight of TCDD. Significant apoptosis was detected within 8-12 hours after injection in the wild type mice, whereas, in the lpr and gld mice apoptosis could not be detected. Upon culturing the thymocytes from TCDD-treated mice for 24 hours in vitro, the wild-type cells showed increased apoptosis when compared to the control; whereas, similar cells from lpr and gld mice did not show apoptosis. Furthermore, TCDD-treatment caused significant alterations in the expression of surface molecules on the thymocytes in the wild-type mice and minimal changes in the lpr or gld mice. Sera from TCDD-treated wild-type mice also exhibited increased levels of soluble Fas ligand. Also, TCDD-induced apoptosis was inhibited both in vitro and in vivo by caspase inhibitors and other inhibitors of apoptosis. Together, the current study demonstrates that TCDD-induced apoptosis plays an important role in thymic atrophy caused by TCDD in vivo. Furthermore, phenotypic changes in the density of thymocyte surface molecules may serve as a useful biomarker for chemical toxicity involving apoptosis. The current study also demonstrates that Fas-Fas ligand interactions play an important role in the induction of apoptosis and immunotoxicity by TCDD. / Ph. D.

Inhibited immune responses have been observed following occupational, inadvertent, or therapeutic exposure to chemically diverse xenobiotics. In the present studies, preliminary data were generated showing limited but significant systemic immunotoxicity following low-level topical exposure to the pyrethroid insecticide, permethrin (formerly not considered an immunotoxicant). Permethrin was applied to the shaved dorsal interscapular region of C57Bl/6N mice at doses of 0.5, 1.5, or 5.0 μl/day. The highest of these doses was approximately equal to 215 μg/kg/day, which is about seven times the estimated daily human exposure in individuals wearing permethrin treated clothing for insect protection. Mice were thus exposed to permethrin daily for 10 or 30 consecutive days, or every other day for 7 or 14 exposures. Body weight was not affected by the treatment. However thymic weight was decreased and splenic weight increased 2 days after termination of the topical exposure. Histopathology of immune organs showed no significant changes. Splenic macrophages showed significantly depressed chemiluminescent responses up to 10 days following termination of exposure, but macrophage phagocytic activity was not affected. Cell surface markers of thymocytes, splenocytes and bone marrow cells were not affected. Antibody production as shown by plaque forming cell (PFC) assay decreased significantly at 10 days after dosing termination. Taken together, these data indicate that low-level topical permethrin exposure may produce systemic immunotoxicity. / Master of Science

The immune system is a target of arsenic toxicity. Epidemiological data have shown that arsenic exposure is associated with characteristics of immunosuppression. Human B-lymphoblastoid cell lines (LCL) were used as an in vitro model of immune cell targeting by arsenic to investigate the mechanism of arsenic-induced cytotoxicity, which could provided insight into the mechanism underlying arsenic-induced immunotoxicity leading to the immunosuppression observed in humans. In LCL arsenite-induced cytotoxicity was not associated with apoptosis, but associated with hallmarks of autophagy, a cell stress-responsive process that facilitates the removal of cellular components through lysosome-mediated degradation. At environmentally relevant concentrations, arsenite-induced toxicity resulted in a decrease in cell proliferation that was correlated with hallmarks of autophagy including expansion of acidic vesicles, global induction of lysosomal gene expression, increased flux of the autophagosome marker LC3-II, and increased enzymatic activity of the lysosomal hydrolase cathepsin D. Investigation of the upstream cellular damage leading to the induction of autophagy revealed that arsenite induces proteotoxic damage leading to an accumulation of protein aggregates that may be targeted to the lysosome for degradation. In addition, global gene expression data showed an enrichment of ER stress responsive genes after arsenite exposure. Further evaluation of global gene expression data indicated that the global induction of lysosomal genes occurs before the activation of ER stress genes, suggesting that the induction of autophagy may occur before the generation of ER stress. To investigate the effect of arsenite-induced proteotoxicity and autophagy on normal immune function, the ability of LCL to process and present exogenous antigens onto MHC class II molecules was evaluated. Arsenite decreased antigen presentation of the exogenous antigen HSA. This decrease was associated with decreased lysosomal degradation of the model substrate DQ-Ova, suggesting that arsenite is disrupting lysosome-mediated degradation. In addition, arsenite exposure was associated with an increase in MHC class II protein aggregates, which could render them unavailable to bind peptide fragments. Through the identification that arsenite induces proteotoxicity and autophagy in LCL, it provides novel insight into the mechanisms of arsenic-induced immunotoxicity that could lead to a better understanding of the mechanisms underlying arsenic-induced immunosuppression observed in humans.

Several biomarkers (cell viability and phagocytosis) based on earthworm (Lumbricus terrestris) immune cells (coelomocytes), together with whole-worm mortality (LC/LD50's), were used to assess a bioremediation attempt to reduce pentachlorophenol (PCP) toxicity in a former wood processing hazardous waste site (HWS).

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Graphene oxide (GO) is a single carbon layer, oxygen bearing graphene derivative, containing hydroxyl and carboxyl groups. Graphene oxide nanoparticles (GONPs) are promising nanomaterials for a variety of applications such as electrochemical analysis, adsorption of biomolecules, biosensors and drug and vaccine delivery systems. While these newly engineered nanoparticles hold great potential for developments in industry and medicine, the widespread use of these material will inevitably result in GO residues in the environment where they could possibly pose a risk to human and wildlife health. Interaction of the nanoparticles and biota can affect numerous biological processes. In humans they can affect any of the physiological systems such as the immune, endocrine, reproductive and cardiovascular systems. Although studies have indicated that GO exposure cause increased reactive oxygen species in cells, they mechanisms whereby GO act on the cell are still poorly understood. A few studies have investigated the effects of GONP and other graphene nanoparticle derivatives on the immune system. The aim of this study was to investigate the in vitro effects of GONPs on the immune system by the exposure of the murine macrophage cell line, RAW 264.7, to different concentrations of GONPs.

Resumo: Escassas informações são encontradas na literatura a respeito do potencial imunotóxico do Diuron, um herbicida derivado da uréia, utilizado no Brasil e no mundo nas lavouras de café, cana-de-açúcar, algodão, abacaxi, citros, alfafa e trigo. Desta forma, o presente estudo teve por objetivo investigar a toxicidade do Diuron sobre o sistema imunológico. Para isso, foram avaliados parâmetros gerais de toxicidade, bioquímicos, hematológicos, histopatologia de órgãos linfohematopoiéticos e fenotipagem de linfócitos T CD4, T CD8 e B de ratos Wistar machos. Foram utilizados dois protocolos experimentais: I - toxicidade aguda (28 dias) e II - toxicidade subcrônica (90 dias) sendo os animais expostos ao Diuron por via oral (adicionado à ração), nas concentrações de 125, 1250 e 2500 ppm. A exposição ao Diuron resultou em diminuição do ganho de peso corpóreo médio nos grupos 1250 e 2500 ppm aos 28 e 90 dias acompanhada pela redução do consumo de ração. O peso relativo do baço e dos rins foi maior nas três concentrações de Diuron aos 28 dias, e apenas em 2500 ppm, aos 90 dias. O peso relativo do fígado foi maior em 1250 e 2500 ppm aos 28 dias e apenas em 2500 ppm aos 90 dias. Aos 28 dias, os níveis de albumina e proteína total foram maiores nos três grupos expostos e os níveis de creatinina e uréia aumentaram apenas no grupo 2500 ppm. A análise histológica revelou que o Diuron, especialmente na maior concentração levou à depleção de polpa branca no baço, associada a redução do número de linfócitos T CD4+ e aumento de hematopoiese extramedular, deposição de hemosiderina e hiperplasia eritróide na polpa vermelha. Não foram observadas alterações hematológicas importantes. Com relação à análise fenotípica das subpopulações de linfócitos, observamos uma redução de células CD4+... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Few information is available regarding immunotoxic potential of Diuron, a phenylurea herbicide widely used in Brazil and around the world on sugar cane, cotton, coffee, pineapple, citros, alfafa and wheat crops.The objective of the present study was to evaluate the toxicity of Diuron in the immune system. General parameters of toxicity were evaluated and biochemistry, hematology, histologic aspects of lymphohematopoietic organs and CD4+, CD8+ and CD45RA+ splenic lymphocytes subpopulations of male Wistar rats. Two experimental protocols were used: I Acute toxicity (28 days) and II Sub-Chronic toxicity (90 days) and the animals were treated with 125 ppm, 1250 ppm and 2500 ppm of Diuron by feeding. The experimental groups treated with Diuron at 1250 ppm and 250o ppm, showed decreased body weight gain after 28 and 90 days of treatment, corresponding with the reduced food intake. The relative weight of the spleen and kidneys were higher at the three concentrations in the end of 28 days, and only at 2500 ppm after 90 days. The relative weight of the liver was higher at 1250 ppm and 2500 ppm to the 28 days, and only at 2500 ppm to the 90 days of treatment. After 28 days, the albumin and total protein serum levels were higher at the three treated groups and the creatine and urea higher only at 2500 ppm of Diuron. The treatment with Diuron at higher doses causes the depletion of the lymphoid white pulp and increases the hemosiderin deposition in the red pulp. We didn't observed significant hematological alterations between the treated and control groups. Regarding fenotipic analysis of splenic lymphocytes, it was observed a decrease in CD4+ subpopulations at 1250 ppm and 2500 ppm in the end of 28 days, and only at 2500 ppm after 90 days of treatment compared with the control groups values. Concluding, Diuron had a systemic toxic effect and also on lymphohematopoietic organs. / Orientador: Ana Lúcia Tozzi Spinardi-Barbisan / Coorientador: Luís Fernando Barbisan / Banca: Carla Adriene da Silva Franchi / Banca: Isis Machado Hueza / Mestre

Made available in DSpace on 2014-06-11T19:27:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-26Bitstream added on 2014-06-13T20:17:33Z : No. of bitstreams: 1 domingues_a_me_botfm.pdf: 594143 bytes, checksum: 583e125c7a9e0c3c3a4f6516dabc6d61 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Escassas informações são encontradas na literatura a respeito do potencial imunotóxico do Diuron, um herbicida derivado da uréia, utilizado no Brasil e no mundo nas lavouras de café, cana-de-açúcar, algodão, abacaxi, citros, alfafa e trigo. Desta forma, o presente estudo teve por objetivo investigar a toxicidade do Diuron sobre o sistema imunológico. Para isso, foram avaliados parâmetros gerais de toxicidade, bioquímicos, hematológicos, histopatologia de órgãos linfohematopoiéticos e fenotipagem de linfócitos T CD4, T CD8 e B de ratos Wistar machos. Foram utilizados dois protocolos experimentais: I - toxicidade aguda (28 dias) e II - toxicidade subcrônica (90 dias) sendo os animais expostos ao Diuron por via oral (adicionado à ração), nas concentrações de 125, 1250 e 2500 ppm. A exposição ao Diuron resultou em diminuição do ganho de peso corpóreo médio nos grupos 1250 e 2500 ppm aos 28 e 90 dias acompanhada pela redução do consumo de ração. O peso relativo do baço e dos rins foi maior nas três concentrações de Diuron aos 28 dias, e apenas em 2500 ppm, aos 90 dias. O peso relativo do fígado foi maior em 1250 e 2500 ppm aos 28 dias e apenas em 2500 ppm aos 90 dias. Aos 28 dias, os níveis de albumina e proteína total foram maiores nos três grupos expostos e os níveis de creatinina e uréia aumentaram apenas no grupo 2500 ppm. A análise histológica revelou que o Diuron, especialmente na maior concentração levou à depleção de polpa branca no baço, associada a redução do número de linfócitos T CD4+ e aumento de hematopoiese extramedular, deposição de hemosiderina e hiperplasia eritróide na polpa vermelha. Não foram observadas alterações hematológicas importantes. Com relação à análise fenotípica das subpopulações de linfócitos, observamos uma redução de células CD4+... / Few information is available regarding immunotoxic potential of Diuron, a phenylurea herbicide widely used in Brazil and around the world on sugar cane, cotton, coffee, pineapple, citros, alfafa and wheat crops.The objective of the present study was to evaluate the toxicity of Diuron in the immune system. General parameters of toxicity were evaluated and biochemistry, hematology, histologic aspects of lymphohematopoietic organs and CD4+, CD8+ and CD45RA+ splenic lymphocytes subpopulations of male Wistar rats. Two experimental protocols were used: I Acute toxicity (28 days) and II Sub-Chronic toxicity (90 days) and the animals were treated with 125 ppm, 1250 ppm and 2500 ppm of Diuron by feeding. The experimental groups treated with Diuron at 1250 ppm and 250o ppm, showed decreased body weight gain after 28 and 90 days of treatment, corresponding with the reduced food intake. The relative weight of the spleen and kidneys were higher at the three concentrations in the end of 28 days, and only at 2500 ppm after 90 days. The relative weight of the liver was higher at 1250 ppm and 2500 ppm to the 28 days, and only at 2500 ppm to the 90 days of treatment. After 28 days, the albumin and total protein serum levels were higher at the three treated groups and the creatine and urea higher only at 2500 ppm of Diuron. The treatment with Diuron at higher doses causes the depletion of the lymphoid white pulp and increases the hemosiderin deposition in the red pulp. We didn't observed significant hematological alterations between the treated and control groups. Regarding fenotipic analysis of splenic lymphocytes, it was observed a decrease in CD4+ subpopulations at 1250 ppm and 2500 ppm in the end of 28 days, and only at 2500 ppm after 90 days of treatment compared with the control groups values. Concluding, Diuron had a systemic toxic effect and also on lymphohematopoietic organs.