Phenotypical and Functional Analysis of Intraepithelial Lymphocytes from Small Intestine of Mice in Oral Tolerance

In this work, we evaluated the effects of administration of OVA on phenotype and function of intraepithelial lymphocytes (IELs) from small intestine of transgenic (TGN) DO11.10 and wild-type BALB/c mice. While the small intestines from BALB/c presented a well preserved structure, those from TGN showed an inflamed aspect. The ingestion of OVA induced a reduction in the number of IELs in small intestines of TGN, but it did not change the frequencies of CD8+ and CD4+ T-cell subsets. Administration of OVA via oral + ip increased the frequency of CD103+ cells in CD4+ T-cell subset in IELs of both BALB/c and TGN mice and elevated its expression in CD8β+ T-cell subset in IELs of TGN. The frequency of Foxp3+ cells increased in all subsets in IELs of BALB/c treated with OVA; in IELs of TGN, it increased only in CD25+ subset. IELs from BALB/c tolerant mice had lower expression of all cytokines studied, whereas those from TGN showed high expression of inflammatory cytokines, especially of IFN-γ, TGF-β, and TNF-α. Overall, our results suggest that the inability of TGN to become tolerant may be related to disorganization and altered proportions of inflammatory/regulatory T cells in its intestinal mucosa

Erratum to “Phenotypical and Functional Analysis of Intraepithelial Lymphocytes from Small Intestine of Mice in Oral Tolerance”

[No abstract available]201

Evaluation Of In Vitro Anti-inflammatory Effects Of Crude Ginger And Rosemary Extracts Obtained Through Supercritical Co2 Extraction On Macrophage And Tumor Cell Line: The Influence Of Vehicle Type.

Numerous plants from have been investigated due to their anti-inflammatory activity and, among then, extracts or components of ginger (Zingiber officinale Roscoe) and rosemary (Rosmarinus officinalis L.), sources of polyphenolic compounds. 6-gingerol from ginger rhizome and carnosic acid and carnosol from rosemary leaves present anti-tumor, anti-inflammatory and antioxidant activities. However, the evaluation of the mechanisms of action of these and other plant extracts is limited due to their high hydrophobicity. Dimethylsulfoxide (DMSO) is commonly used as a vehicle of liposoluble materials to mammalian cells in vitro, presenting enhanced cell penetration. Liposomes are also able to efficiently deliver agents to mammalian cells, being capable to incorporate in their structure not only hydrophobic molecules, but also hydrophilic and amphiphilic compounds. Another strategy is based on the use of Pluronic F-68, a biocompatible low-foaming, non-ionic surfactant, to disperse hydrophobic components. Here, these three delivery approaches were compared to analyze their influence on the in vitro anti-inflammatory effects of ginger and rosemary extracts, at different concentrations, on primary mammalian cells and on a tumor cell line. Ginger and rosemary extracts free of organic solvents were obtained by supercritical fluid extraction and dispersed in DMSO, Pluronic F-68 or liposomes, in variable concentrations. Cell viability, production of inflammatory mediators and nitric oxide (NO) release were measured in vitro on J774 cell line and murine macrophages primary culture stimulated with bacterial lipopolysaccharide and interferon-γ after being exposed or not to these extracts. Ginger and rosemary extracts obtained by supercritical CO2 extraction inhibited the production of pro-inflammatory cytokines and the release of NO by peritoneal macrophages and J774 cells. The delivery vehicles influenced the anti-inflammatory effects. Comparatively, the ginger extract showed the highest anti-inflammatory activity on the tumor cell line. Controversially, rosemary extract dispersed on DMSO induced a more significant IL-1 and TNF-α reduction than ginger extract in primary macrophages. Amongst the tested delivery vehicles, DMSO was the most suitable, presenting reduced cytotoxicity, followed by Pluronic F-68 and liposomes, provably due to differences in their form of absorption, distribution and cellular metabolism. Co-administration of liposomes and plant extracts may cause death of macrophages cells and induction of NO production. It can be concluded that some of the beneficial effects attributed to extracts of ginger and rosemary may be associated with the inhibition of inflammatory mediators due to their high antioxidant activity. However, these effects were influenced by the type of delivery vehicle.1539

Oral tolerance and OVA-induced tolerogenic dendritic cells reduce the severity of collagen/ovalbumin-induced arthritis in mice

AbstractDietary proteins play an important role in the regulation of systemic immune response, in a phenomenon known as oral tolerance (OT). To evaluate the effects of OT on a murine model of type II collagen (CII) plus ovalbumin (OVA)-induced arthritis (CIA), mice were fed with OVA either before or after CIA induction. OT significantly reduced the paw edema and synovial inflammation, as well as serum levels of anti-CII, the ex vivo proliferation and inflammatory cytokine production by spleen cells from CIA mice. The frequencies of Foxp3+ and IL-10+ cells were higher, whereas IFNγ+ cells and IL-17+ cells were lower, among gated CD4+ spleen T cells from tolerized CIA mice than in those from non-tolerized CIA mice. Adoptive transfer of tolerogenic dendritic cells (DCs) before CIA induction mimics the effects observed in the OT. We demonstrate here that bystander suppression induced by OT can modify the course of CIA and tolerogenic DCs play a role this phenomenon

Adoptive transfer of dendritic cells expressing CD11c reduces the immunological response associated with experimental colitis in BALB/c mice.

Introduction In addition to conventional therapies, several new strategies have been proposed for modulating autoimmune diseases, including the adoptive transfer of immunological cells. In this context, dendritic cells (DCs) appear to be one of the most promising treatments for autoimmune disorders. The present study aimed to evaluate the effects of adoptive transfer of DCs obtained from both na?ve and ovalbumin (OVA)-tolerant mice on the severity of TNBS induced colitis and analyze the eventual protective mechanisms. Methods and results To induce oral tolerance, BALB/c mice were fed 4mg/mL OVA solution for seven consecutive days. Spleen DCs were isolated from tolerant (tDC) and na?ve (nDC) mice, and then adoptively transferred to syngeneic mice. Three days later, colitis was induced in DC treated mice by intrarectal instillation of 100?g2,4,6-trinitrobenzenesulfonic acid (TNBS) dissolved in 50% ethanol. Control subjects received only intrarectal instillation of either TNBS solution or a vehicle. Five days later, mice from all groups were euthanized and examined for physiological and immunological parameters. Regarding the phenotype, we observed that the frequencies of CD11+ MHC II+ and CD11+ MHCII+ CD86+ cells were significantly lower in DCs isolated from tolerant mice than in those from naive mice. However, pretreatment with both types of DCs was able to significantly reduce clinical signs of colitis such as diarrhea, rectal prolapse, bleeding, and cachexia, although only treatment with tDCs was able to prevent weight loss from instillation of TNBS. In vitro proliferation of spleen cells from mice treated with either type of DCs was significantly lower than that observed in splenic cell cultures of na?ve mice. Although no significant difference was observed in the frequencies of Treg cells in the experimental groups, the frequency of Th17+CD4+cellsand the secretion of IL-17 were more reduced in the cultures of spleen cells from mice treated with either type of DCs. The levels of IL-9 and IFN-? were lower in supernatants of cells from mice treated with nDCs. Conclusion The results allow us to conclude that the adoptive transfer of cells expressing CD11c is able to reduce the clinical and immunological signs of drug-induced colitis. Adoptive transfer of CD11c+DC isolated from both naive and tolerant mice altered the proliferative and T cell responses. To the best of our knowledge, there is no previously published data showing the protective effects of DCs from na?ve or tolerant mice in the treatment of colitis

Isolamento e caracterização parcial de uma fração antigenica da superficie do trypanosoma cruzi (Chagas, 1909)

Orientador : H. A. RangelDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: Foram realizadas experiências com o sentido de verificar a possibilidade de se extrair determinantes antigênicos da superfície do Trypanosoma cruzi, através da ação da proteinase existente no sedimento dos lisados das formas epimastigotas do parasito. Os resultados mostraram que pode ser extraída uma fração antigênica (FAD), quando esses sedimentos são incubados em condições propícias à ação da proteinase (PBS, pH 7.2; 37ºCº por 1 hora). O teor de proteína e de polissacáride da fração antigênica era aumentado quando se utilizava ativadores da ação enzimática (37ºC; 2-Me) e se encontrava diminuído quando se utilizava inibidores dessa ação (0ºC; IAc), indicando que a enzima facilita a obtenção dos constituintes da superfície. Através de provas de imunoeletroforese cruzada, foi demonstrado que a enzima presente nesses sedimentos é antigenicamente idêntica à anteriormente isolada por Araújo (1979). A análise em gel de poliacrilamida, mostrou que as frações obtidas em condições que favoreciam a ação da proteinase, eram constituídas por três componentes de baixo PM (Rm : 0.5, 0.6, e 0.9, respectivamente), ao passo que as frações obtidas na presença do inibidor ou a 0ºC eram constituídas principalmente por componentes de alto PM. Em ambos os casos, as frações obtidas inibiam as reações de aglarinação dos soros anti-epimastigotas (a-EL), indicando que as diferentes molecular, de alto e de baixo PM, encaminham os mesmos determinantes antigênicos... Observação: O resumo, na íntegra, poderá ser visualizado no texto completo da tese digitalAbstract: Not informed.MestradoImunologiaMestre em Ciências Biológica

Obtenção e estudo de anticorpos monoclonais anti Trypanosoma Cruzi Chagas, 1909

Orientador : Humberto de Araujo RangelTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: O presente trabalho teve três objetivos principais: (i) estudar a resposta imune humoral específica de camundongos F1 (CBA x C57 B1/10) infectados com a cepa Y do T.cruzl, (ii) produzir anticorpos monoclonais anti-T.cruzl, através da tecnologia de hibridomas, utilizando-se para as fusões células de mieloma SP2/0 e células esplênicas de camundongos F1 cronicamente infectados com o parasita, (iii) caracterizar os anticorpos monoclonais obtidos quanto a sua reatividade para os antígenos expressos nos diferentes estágios do T.cruzi ... O resumo, na íntegra, poderá ser visualizado no texto completo da tese digitalAbstract: Not informed.DoutoradoImunologiaDoutor em Ciências Biológica

NK1.1+ cells and T-cell activation in euthymic and thymectomized C57BL/6 mice during acute Trypanosoma cruzi infection

Cardillo, Fabíola; Mengel Junior, Jose Orivaldo “Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta à informação no documento”.Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2017-07-05T14:17:49Z No. of bitstreams: 1 Cardillo F NK1.1+ cells and T-cell....pdf: 194793 bytes, checksum: 33cb19d87590fc6f0383da2c0cc9b020 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2017-07-06T12:30:15Z (GMT) No. of bitstreams: 1 Cardillo F NK1.1+ cells and T-cell....pdf: 194793 bytes, checksum: 33cb19d87590fc6f0383da2c0cc9b020 (MD5)Made available in DSpace on 2017-07-06T12:30:15Z (GMT). No. of bitstreams: 1 Cardillo F NK1.1+ cells and T-cell....pdf: 194793 bytes, checksum: 33cb19d87590fc6f0383da2c0cc9b020 (MD5) Previous issue date: 2002FAPESP, CNPqUniversity of São Paulo. Institute of Biomedical Sciences-IV. Department of Immunology. São Paulo, SP, BrazilUniversity of Sao Paulo. Faculty of Ribeirão Preto. Department of Pharmacology. Ribeirão Preto, SP, BrazilUniversity of Campinas. Institute for Biology. Department of Microbiology and Immunology. Campinas, SP, BrazilUniversity of São Paulo. Institute of Biomedical Sciences-IV. Department of Immunology. São Paulo, SP, BrazilUniversity of São Paulo. Faculty of Medicine of Ribeirão Preto. Department of Morphology. Ribeirão Preto, SP, SP, BrasiUniversity of São Paulo. Institute of Biomedical Sciences-IV. Department of Immunology. São Paulo, SP, BrazilNatural killer (NK) cells may provide the basis for resistance to Trypanosoma cruzi infection, because the depletion of NK1.1 cessla causes high levels of parasitemia in Young C57B1/6 mice infected with T. cruzi. Indeed, NK1.1 cells have been implicated in the early production of large amounts of interferon (IFN)-y, na importante cytokine in host resistance. The NK.1 marker is also expressed on special subpopulations of T cells. Most NK1.1+ T cells are of thymic origin, and their constant generation may be prevented by thymectomy. This procedure, by itself, decreased parasitemia and increased resistance in young mice. However, the depletion of NK1.1+ cells by the chronic administration of a monoclonal antibody (MoAb) (PK-136) did not increase the parasitemia or mortality in thymectomized C57B1/6 mice infected with T. cruzi (Tulahuen strain). To study the cross-talk between NK1.1+ cells and conventional T cells in this model, we examined the expression. of activation/memory markers (CD45RB) on splenic CD4+ and CD8+ T cells from Young euthymic or thymectomized mice with or without depletion of NK1.1+ cells and also in aged mice during acute infection. Resistance to infection correlated with the amount of CD4+ T cells that are already acitivated at the moment of infection, as judged by the number of splenic CD4+ T cells expressing CD45RB-. In addition, the specific antibody response to T cruzi antigens was precocious and na accumulation of immunoglobulin (Ig)M with little isotype switch occurred in euthymic mice depleted of NK1.1+ cells. The data presented here suggest that NK1.1+ cells have importante regulatory functions in euthymic, but not in thymectomized mice infected with T. cruzi. These regulatory functions include a helper activity in the generation of effector or activated/mewmory T cells