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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Activation antigens in the proliferation and differentiation of normal and malignant human leucocytes

Barnett, David January 1991 (has links)
No description available.
2

Investigating Antigen Presentation by Inactivated Lymphocytic Choriomeningitis Virus and by Baculovirus Encoding the LCMV-Nucleoprotein

Spence, Tara 03 September 2009 (has links)
Professional antigen presenting cells (pAPCs) process and present antigens on their cell surface in association with MHC class I molecules through two general pathways: direct or cross-presentation. The process of antigen presentation by pAPCs to naïve T cells resulting in their proliferation and differentiation into activated cytotoxic lymphocytes (CTLs) is called T cell priming. In these studies, we examine the cross-presentation of antigens from two non-replicating viruses: inactivated Lymphocytic Choriomeningitis virus (LCMV), and recombinant baculovirus encoding the LCMV nucleoprotein (NP). Since effective activation of pAPCs is essential for efficient priming of CD8+ T cells and CTL activation, and because infection with inactivated viruses generally induces an extremely poor level of CTL activation, we examined the activation state of pAPCs by measuring their cytokine profiles following infection to help further delineate their involvement in the CTL response to inactivated viruses. Our results indicate a pro-inflammatory cytokine mRNA upregulation in pAPCs in response to the inactivated virus, similar to the cytokine profiles subsequent to live LCMV infection, but to a lesser extent. In these studies, we also examined CTL activation following infection with inactivated LCMV and bAc-NP. We have demonstrated that the presentation of antigens from inactivated LCMV and bAC-NP results in a low level of CTL activation in vivo, though there is an undetectable level of CTL activation in vitro, in comparison to activation following infection with the live virus. Ultimately, the characterization of the cytokine profiles of pAPCs and the CD8+ T cell profiles induced in response to inactivated LCMV or the baculovirus derived NP may lead to a better understanding of how cross-presentation of these viral antigens may occur. This information may be applied to enhance the process of pAPC activation and T cell priming, for the induction of more effective cellular immune responses and the generation of stronger protective immunity. / Thesis (Master, Microbiology & Immunology) -- Queen's University, 2009-09-02 15:30:13.883
3

Assessment of humoral and cellular immune responses of the RTS,S/AS02D malaria vaccine candidate administered to infants living in a malaria endemic area in Mozambique

Aide, Pedro Carlos Paulino 12 April 2010 (has links)
MSc (Med), Faculty of Health Sciences, University of the Witwatersrand, 2009 / Background: RTS,S candidate malaria vaccine has been shown to be highly immunogenic in children and infants, but the protective immune mechanisms still remain to be clearly elucidated. It is believed that RTS,S elicits a strong neutralizing humoral immune response directed against surface-exposed sporozoite proteins and cell mediated immune (CMI) responses characterized by predominantly CD4+ Th1 cells. The objective of this study was to investigate humoral and cell-mediated immune responses to the RTS,S/AS02D malaria vaccine and its association with protection against infection and disease by P. falciparum. Methodology and Principal Findings: This secondary data analysis from data of a phase I/IIb randomized, double-blind, controlled trial, included 154 healthy infants living in rural Mozambique, previously immunized with RTS,S/AS02D candidate malaria vaccine or the control Engerix-B™ vaccine. Antibodies against circumsporozoite protein (CSP) and hepatitis-B surface antigen (HBsAg) were measured with a standard ELISA. Fresh blood intracellular staining assay was performed to evaluate the expression of IL-2 and IFN-γ by CD4+ and CD8+ cells in response to in vitro stimulation of specific peptides. Data was evaluated for association with the risk of malaria detected by both active and passive case detection of infection over a period of 6 months post dose 3. Anti-HBs antibody geometric mean titers declined from 10,082 mIU/mL one month post Dose 3 to 2,751 mIU/mL at 12 months post Dose 3 in the RTS,S/AS02D group; anti-HBs v geometric mean titers were 392.4 mIU/mL and 263.9 mIU/mL, respectively in the Engerix- BTM group. Anti-CSP antibody geometric mean titers declined from 199.9 EU/mL one month post Dose 3 to 7.3 EU/mL at 12 months post Dose 3 in the RTS,S/AS02D group. Median stimulation indices of HBs-specific IL-2 and IFN-γ producing CD8+ T cells was higher in the RTS,S/AS02D group than in control group (Wilcoxon rank sum p-values for IFN-γ = 0.015, for IL-2 = 0.030) at 10.5 weeks post immunization. Median stimulation indices of anti-CSP specific IFN-γ producing CD8+ T cells at the same time point was 1.13 (IQR: 0.79 - 1.67; p=0.029). For specific IL-2-producing CD4+ T cells, the median SI was 1.14 (IQR: 0.74 – 1.60, p=0.043) at 10.5 weeks post dose three. The reduction in hazards of malaria infection were 18.3 % (95% CI: -267.9 – 81.8, p=0.793) and -12.0 % (95% CI: -295 – 68.2, p=0.86) for specific IL-2 CD4+ stimulation indices; For specific CD8+ IFN-γ stimulation indices the hazards were -103.6% (95% CI: -690.9 – 47.6; p=0.305) and 48.8% (95% CI: -97.0 – 86.7; p=0.33) at four and 10.5 weeks post immunization respectively. Conclusion: The RTS,S/AS02D vaccine was immunogenic and has elicited detectable levels of CSP specific cell mediated responses. No evidence of association was found between the antibodies anti-CSP and specific cell mediated responses and the risk of malaria.
4

Caracterização da resposta imune celular induzida pela imunização experimental com antígenos recombinantes de  Plasmodium vivax / Characterization of the cellular immune response induced by experimental immunization with recombinant antigens of Plasmodium vivax.

Casale, Patricia Ostermayer Athayde 22 November 2013 (has links)
A malária continua sendo um dos maiores problemas de saúde pública no mundo e apesar dos muitos esforços no combate a doença, as estratégias disponíveis hoje ainda não foram capazes de controlar com eficiência sua disseminação global. Por isso o desenvolvimento de uma vacina é de extrema importância. Devido a complexidade do ciclo do parasita, consideramos que uma formulação vacinal eficaz para a indução de resposta imune protetora contra o P. vivax deverá conter regiões de antígenos de vários estágios do parasita. Dentre os vários antígenos da fase sanguínea do plasmódio identificados, o Antígeno 1 de Membrana Apical (AMA1), a porção C-terminal da Proteína 1 de Superfície do Merozoíto (MSP119) e as proteínas da família MSP3 têm se mostrado promissores. Portanto, o objetivo deste estudo foi caracterizar a resposta imune celular induzida em camundongos pela imunização experimental com antígenos de P. vivax candidatos a vacina, tendo como foco principal a combinação das proteínas AMA1, MSP119 e MSP3β. Inicialmente, as imunizações experimentais com a combinação dos 3 antígenos foram realizadas na presença de Adjuvante Incompleto de Freund (AIF) em camundongos C57BL/6. Os camundongos imunizados não apresentaram respostas significativas com proliferação e secreção de IFN-γ por linfócitos T CD4+ e CD8+ após estimulação in vitro com as proteínas recombinantes ou peptídeos sintéticos baseados na proteína AMA1. Por outro lado, o adjuvante Poly (I:C) mostrou-se mais eficiente na indução de resposta proliferativa por linfócitos T CD4+, quando testado com a proteína MSP119-PADRE. Assim, selecionamos este adjuvante para dar continuidade às imunizações experimentais com a combinação de antígenos. Por ELISPOT, foi possível demonstrar que a co-administração dos 3 antígenos foi capaz de estimular células produtoras de IFN-γ em resposta à todos os antígenos testados. Também foi nosso objetivo avaliar a produção de IFN-γ após imunização de linhagens distintas de camundongos com uma proteína quimérica recentemente produzida em nosso laboratório (AMA1-MSP119). Em C57BL/6, a produção de IFN-γ foi significativamente maior no grupo imunizado com a proteína de fusão, quando comparado com o grupo que recebeu a co-adminstração dos dois antígenos, mostrando que a utilização de proteínas fusionadas pode solucionar possíveis problemas de competição antigênica. Em conjunto, nossos dados demonstram que a utilização de múltiplos antígenos fusionados, ou co-administrados, pode ser uma estratégia promissora de vacinação contra a malária. / Malaria remains as a major public health problem worldwide. In spite of the efforts for prevention, the strategies available today have not succeeded to control its global distribution. Therefore, the development of a vaccine is extremely important. Due to the complexity of the parasite cycle, we consider that an effective vaccine formulation to induce protective immune response against P. vivax should contain regions of antigens from several stages of the parasite. Among the different blood stage antigens of Plasmodium identified, the Apical Membrane Antigen 1 (AMA1), the C-terminal portion of the Merozoite Surface Protein 1 (MSP119) and MSP3 family proteins have shown promising results. Therefore, the aim of this study was to characterize the cellular immune response induced in mice by experimental immunization with antigens from P. vivax vaccine candidates, focusing mainly on the combination of proteins AMA1, MSP119 and MSP3β. Initially, the experimental immunizations with the combination of the three antigens were performed in the presence of Incomplete Freund\'s Adjuvant (IFA) in C57BL/6 mice. Immunized mice displayed no significant proliferative responses and secretion of IFN-γ by CD4+ and CD8+ T lymphocytes after in vitro stimulation with recombinant proteins or synthetic peptides based on the AMA1 protein. In contrast, Poly (I:C) adjuvant was more efficient inducing CD4+ T lymphocytes proliferative response to the MSP119-PADRE protein. Thus, we selected this adjuvant to continue the experimental immunizations with the different antigen combinations. The ELISPOT results demonstrated that co-administration of the 3 antigens stimulated IFN-γ producing cells in response to all antigens tested. We also assessed the production of IFN-γ after immunization of distinct mouse strains with a chimeric protein produced recently in our laboratory (AMA1-MSP119). In C57BL/6, IFN-γ production was significantly higher when we used spleen cells from mice immunized with the fusion protein, when compared with cells from mice that received the co-administration of two antigens, showing that the use of fused proteins can solve possible antigenic competition problems. Together, our data demonstrate that the use of multiple fused or co-administered antigens may be a promising strategy for malaria vaccination.
5

Perfil da resposta imune celular em pacientes infectados pelo HIV com leishmaniose ou tuberculose

Gois, Luana Leandro January 2015 (has links)
Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2016-02-04T16:10:12Z No. of bitstreams: 1 Luana Leandro Gois Perfil... 2015.pdf: 11133579 bytes, checksum: 84c1592269b916a72d1e76ba863403c7 (MD5) / Approved for entry into archive by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2016-02-04T16:10:24Z (GMT) No. of bitstreams: 1 Luana Leandro Gois Perfil... 2015.pdf: 11133579 bytes, checksum: 84c1592269b916a72d1e76ba863403c7 (MD5) / Made available in DSpace on 2016-02-04T16:10:24Z (GMT). No. of bitstreams: 1 Luana Leandro Gois Perfil... 2015.pdf: 11133579 bytes, checksum: 84c1592269b916a72d1e76ba863403c7 (MD5) Previous issue date: 2015 / Fundação Oswaldo Cruz, Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / A infecção pelo HIV promove a redução do número de linfócitos T CD4+ e, consequentemente, o surgimento de doenças oportunistas. A leishmaniose visceral e a tuberculose são comumente reconhecidas como doenças oportunistas importantes e associadas ao óbito de indivíduos infectados por HIV. Ambos os patógenos, Leishmania e Mycobacterium tuberculosis (Mtb) infectam cronicamente macrófagos. A imunidade protetora associada a estas infecções envolve linfócitos Th1 produtores de IFN-g. O prejuízo na resposta imune celular causado pelo HIV perturba a resposta imune contra estes patógenos. Não são bem determinadas quais alterações imunológicas causadas pelo HIV promovem o prejuízo na resposta imune específica contra a Leishmania spp. e Mtb, induzindo o desenvolvimento de formas atípicas e graves destas infecções. Deste modo, esta tese teve como objetivo descrever o perfil da resposta imune celular aos antígenos de Leishmania spp. ou Mtb em pacientes infectados com HIV. Para tal., foram recrutados pacientes infectados por HIV e com diagnóstico de leishmaniose (HIV/LV) e tuberculose (HIV/TB). Indivíduos não infectados por HIV e diagnóstico de leishmaniose (LV) ou tuberculose (TB) forma incluídos como controles. Foram avaliadas a linfoproliferação e a frequência das subpopulações de memória dos linfócitos T CD4+ em resposta aos antígenos solúveis de Leishmania spp. (SLA). Igualmente, foram avaliadas a linfoproliferação, a frequência das subpopulações de memória dos linfócitos T CD4+ e CD8+, o perfil de funcional de linfócitos T CD4+ e CD8+ produtores de citocinas e a atividade citotóxica de linfócitos T CD8+ e células NK em resposta ao purificado protéico derivado (PPD) do M. bovis. Duas revisões sistemáticas da literatura que abordam a associação entre estas infecções e a síndrome inflamatória de reconstituição imune em indivíduos infectados por HIV após terapia antiretroviral foram realizadas. Os linfócitos T CD4+ e CD8+ dos pacientes HIV-LV não apresentavam resposta proliferativa ao SLA e houve redução na frequência de subpopulações de linfócitos T CD4+, a qual foi restaurada após o tratamento para leishmaniose visceral. Nos pacientes HIV-TB foi igualmente observada ausência de resposta proliferativa de linfócitos T CD4+ e CD8+ e de células produtoras de IFN-g em resposta ao PPD. Além disso, o HIV promoveu a redução da atividade degranulativa de células NK, o que contribui para o descontrole da infecção e desenvolvimento de TB ativa. Nos pacientes HIV-TB, HAART foi capaz de induzir uma recuperação parcial de células específicas produtoras de IFN-g, bem como da proliferação em resposta ao PPD. Em conjunto, os resultados desta tese sugerem que a infecção pelo HIV induz alterações na resposta celular de memória central e efetora contra patógenos intracelulares oportunistas. Essas alterações são parcialmente restauradas no curso da HAART. / The HIV-infection promotes reduced number of CD4+ T-lymphocytes and manifestation of opportunistic diseases. Visceral leishmaniasis and tuberculosis are commonly known as main opportunistic infections and are associated with mortality in HIV-infected individuals. Both pathogens, Leishmania and Mycobacterium tuberculosis (Mtb), infect macrophages. The protect immune response involve T-lymphocytes help 1 (Th1) and producing of IFN-g. The impairment of cellular immune response caused by HIV disrupts the immune response against these pathogens. It is unclear which immunological alterations caused by HIV infection promote the damage in specific cellular immune response against Leishmania and Mtb and induces the development of atypical and severe forms. Thus, this thesis aimed to describe the profile of the cellular immune response to Leishmania antigens or Mtb in HIV infected patients. To this end, were recruited HIV infected patients with visceral leishmaniasis (HIV/VL) and HIV infected patients with active tuberculosis (HIV/TB). Moreover, HIV uninfected individuals with VL or TB were also included as controls. Lymphoproliferation and frequency of memory CD4+ T-lymphocyte subsets in response to soluble Leishmania antigen (SLA) were evaluated. Also were evaluated lymphoproliferation, frequency of memory CD4+ and CD8+ T-lymphocyte subsets, functional profile of cytokines producing CD4+ and CD8+ T-lymphocytes and cytotoxic activity of CD8+ Tlymphocytes and NK cells in response to purified protein derivative (PPD) of M. bovis. Two systematic reviews of the literature concerning the association between leishmaniasis and tuberculosis with the inflammatory immune reconstitution syndrome (IRIS) in HIV-infected individuals after antiretroviral therapy were done. The absence of proliferative response to SLA of CD4+ and CD8+ T-lymphocytes and the reduced frequency of memory CD4+ T-lymphocyte subsets were observed in HIV/VL patients. The frequency of memory CD4+ T-lymphocyte subset was restored after treatment for visceral leishmaniasis. In HIV/TB patients was also observed absence of proliferative response to PPD of CD4+ and CD8+ T-lymphocytes and of PPD-specific IFN-g producing cells. In addition, HIV infection promotes the reduction in degranulative activity of NK cells which contributes to the survival of Mtb into macrophages and development of active TB. In HIV/TB patients, HAART was able to induce a partial recovery of PPD-specific IFN-g producing cells and of lymphoproliferation in response to PPD. Taken together, the results suggest HIV infection induces changes in cellular immune response of central and effector memory against opportunistic intracellular pathogens. These alterations are partially restored in the after HAART. Keywords: HIV,
6

Avaliação imunoistoquímica da musculatura estriada esquelética em cães com leishmaniose visceral /

Gomes, Ana Amélia Domingues. January 2009 (has links)
Orientadora: Mary Marcondes / Banca: Raimundo Souza Lopes / Banca: Vera Lúcia Fonseca de Camargo Neves / Resumo: A leishmaniose visceral pode ser incluída como uma das causas de miopatia inflamatória em cães, entretanto, pouco se sabe sobre a patogênese da doença no sistema muscular, sendo incriminada muitas vezes apenas à natureza catabólica da enfermidade. O objetivo deste estudo foi avaliar, por meio de imunoistoquímica, a presença de formas amastigotas de Leishmania sp, linfócitos T (CD3+), macrófagos e IgG nos músculos tríceps braquial, extensor carpo radial, bíceps femoral e gastrocnêmio de 23 cães naturalmente acometidos por leishmaniose visceral. Dentre os 92 músculos avaliados,11 (12%) apresentaram marcação antigênica para formas amastigotas de Leishmania sp, 35 (38,1%) para linfócitos T (CD3+), 29 (31,5%) para macrófagos e 14 (12%) para IgG. Os resultados obtidos permitiram concluir que em cães com leishmaniose visceral apresentam imunomarcação para formas amastigotas de Leishmania sp., linfócitos T CD3+, macrófagos e IgG, sugerindo a participação direta do parasito e de uma resposta imune celular e humoral na fisiopatogenia da lesão muscular. / Abstract: Visceral leishmaniasis may be included as a cause of inflammatory myophathy in dogs, however, little is known about the pathogenesis of the disease in the muscular system, which is frequently associated with the catabolic nature of the illness. The purpose of this study was investigate, through immunohistochemistry, the presence of amastigote forms of Leishmania sp, T lymphocytes (CD3+), macrophages and IgG in the muscle triceps brachial, extensor carpi radialis, biceps femoris and gastrocnemius of 23 dogs with visceral leishmaniasis. Among 92 evaluated muscles, 11 (12%) presented antigenic marking for amastigote forms of Leishmania sp., 35 (38,1%) for T lymphocites (CD3+), 29 (31,5%) for macrophages and 14 (12%) for IgG. The results of the present experiment led to the conclusion that in dogs with visceral leishmaniasis there may be a straight participation of the parasite and of cellular and humoral immune response in the ethiopatogeny of the muscular injury. / Mestre
7

Caracterização da resposta imune celular induzida pela imunização experimental com antígenos recombinantes de  Plasmodium vivax / Characterization of the cellular immune response induced by experimental immunization with recombinant antigens of Plasmodium vivax.

Patricia Ostermayer Athayde Casale 22 November 2013 (has links)
A malária continua sendo um dos maiores problemas de saúde pública no mundo e apesar dos muitos esforços no combate a doença, as estratégias disponíveis hoje ainda não foram capazes de controlar com eficiência sua disseminação global. Por isso o desenvolvimento de uma vacina é de extrema importância. Devido a complexidade do ciclo do parasita, consideramos que uma formulação vacinal eficaz para a indução de resposta imune protetora contra o P. vivax deverá conter regiões de antígenos de vários estágios do parasita. Dentre os vários antígenos da fase sanguínea do plasmódio identificados, o Antígeno 1 de Membrana Apical (AMA1), a porção C-terminal da Proteína 1 de Superfície do Merozoíto (MSP119) e as proteínas da família MSP3 têm se mostrado promissores. Portanto, o objetivo deste estudo foi caracterizar a resposta imune celular induzida em camundongos pela imunização experimental com antígenos de P. vivax candidatos a vacina, tendo como foco principal a combinação das proteínas AMA1, MSP119 e MSP3β. Inicialmente, as imunizações experimentais com a combinação dos 3 antígenos foram realizadas na presença de Adjuvante Incompleto de Freund (AIF) em camundongos C57BL/6. Os camundongos imunizados não apresentaram respostas significativas com proliferação e secreção de IFN-γ por linfócitos T CD4+ e CD8+ após estimulação in vitro com as proteínas recombinantes ou peptídeos sintéticos baseados na proteína AMA1. Por outro lado, o adjuvante Poly (I:C) mostrou-se mais eficiente na indução de resposta proliferativa por linfócitos T CD4+, quando testado com a proteína MSP119-PADRE. Assim, selecionamos este adjuvante para dar continuidade às imunizações experimentais com a combinação de antígenos. Por ELISPOT, foi possível demonstrar que a co-administração dos 3 antígenos foi capaz de estimular células produtoras de IFN-γ em resposta à todos os antígenos testados. Também foi nosso objetivo avaliar a produção de IFN-γ após imunização de linhagens distintas de camundongos com uma proteína quimérica recentemente produzida em nosso laboratório (AMA1-MSP119). Em C57BL/6, a produção de IFN-γ foi significativamente maior no grupo imunizado com a proteína de fusão, quando comparado com o grupo que recebeu a co-adminstração dos dois antígenos, mostrando que a utilização de proteínas fusionadas pode solucionar possíveis problemas de competição antigênica. Em conjunto, nossos dados demonstram que a utilização de múltiplos antígenos fusionados, ou co-administrados, pode ser uma estratégia promissora de vacinação contra a malária. / Malaria remains as a major public health problem worldwide. In spite of the efforts for prevention, the strategies available today have not succeeded to control its global distribution. Therefore, the development of a vaccine is extremely important. Due to the complexity of the parasite cycle, we consider that an effective vaccine formulation to induce protective immune response against P. vivax should contain regions of antigens from several stages of the parasite. Among the different blood stage antigens of Plasmodium identified, the Apical Membrane Antigen 1 (AMA1), the C-terminal portion of the Merozoite Surface Protein 1 (MSP119) and MSP3 family proteins have shown promising results. Therefore, the aim of this study was to characterize the cellular immune response induced in mice by experimental immunization with antigens from P. vivax vaccine candidates, focusing mainly on the combination of proteins AMA1, MSP119 and MSP3β. Initially, the experimental immunizations with the combination of the three antigens were performed in the presence of Incomplete Freund\'s Adjuvant (IFA) in C57BL/6 mice. Immunized mice displayed no significant proliferative responses and secretion of IFN-γ by CD4+ and CD8+ T lymphocytes after in vitro stimulation with recombinant proteins or synthetic peptides based on the AMA1 protein. In contrast, Poly (I:C) adjuvant was more efficient inducing CD4+ T lymphocytes proliferative response to the MSP119-PADRE protein. Thus, we selected this adjuvant to continue the experimental immunizations with the different antigen combinations. The ELISPOT results demonstrated that co-administration of the 3 antigens stimulated IFN-γ producing cells in response to all antigens tested. We also assessed the production of IFN-γ after immunization of distinct mouse strains with a chimeric protein produced recently in our laboratory (AMA1-MSP119). In C57BL/6, IFN-γ production was significantly higher when we used spleen cells from mice immunized with the fusion protein, when compared with cells from mice that received the co-administration of two antigens, showing that the use of fused proteins can solve possible antigenic competition problems. Together, our data demonstrate that the use of multiple fused or co-administered antigens may be a promising strategy for malaria vaccination.
8

Assessment Of Immune Protective Capacity Of The Recombinant Iron-superoxide Dismutase (fesod) From Bordetella Pertussis

Apak, Aycan 01 December 2011 (has links) (PDF)
Whooping cough (pertussis) is a highly contagious acute respiratory disease caused by the strict human pathogen Bordetella pertussis, a gram-negative coccobacillus. The worldwide mass-vaccination was started in 1940s and to date, a number of whole-cell (Pw) and acellular pertussis vaccine (Pa) formulations were developed. Yet the current vaccines are incapable of providing sustained, lifelong immunity and eliminating subclinical infections, which pose a threat especially for unimmunized infants as well as adolescents and adults. Thus, finding new protein candidates with high immune protective capacities is necessary to enhance the clinical efficacy of current acellular pertussis (Pa) vaccines. In this study, iron-superoxide dismutase (FeSOD) protein was investigated for its capacity of conferring protectivity as well as stimulating humoral and cellular responses against B. pertussis infection in a mouse model. For this purpose, sodB gene, which encodes iron-superoxide dismutase FeSOD protein, was amplified from the genomic DNA of the universal B. pertussis strain &lsquo / Tohama I&rsquo / and sequentially cloned to pGEM&reg / -T subcloning and pET-28a(+) expression vectors. Afterwards sodb/pET28a(+) construct was introduced to E. coli BL21(DE3) cells and the gene was overexpressed therein via IPTG induction. The expressed FeSOD protein was then purified by affinity chromatography and its previously reported immunogenicity was confirmed by Western blot. After filter-sterilization, the protein was adsorbed to alum [Al(OH)3] adjuvant and introduced to BALB/c twice at three weeks intervals intraperitoneally at a concentration of 20 &mu / g purified FeSOD protein/mouse. Another group of mice were immunized in tandem with heat-inactivated whole-cell suspension of B. pertussis. Ten days after the second immunization, mice were intranasally challenged with the local &lsquo / Saadet&rsquo / strain of B. pertussis. Next the lungs of groups of mice were excised, homogenized and plated as serial dilutions on days 5, 8 and 14 post-challenge, and viable lung CFU counts were carried out. Whole cell immunization conferred complete bacterial clearance following B. pertussis intranasal infection while FeSOD immunization failed to attain such protection. In addition to the protectivity assay, ELISA was performed to assess the humoral (i.e. IgG) immune response triggered upon FeSOD- and whole-cell immunizations and a statistically significant increase in anti-FeSOD IgG production was observed in FeSOD-immunized group. Finally, cellular immune response was tested via cytokine (IFN-&gamma / ) assay, in which spleens of mice were excised, splenocytes were cultured and the level of IFN-&gamma / production upon FeSOD addition to the cultures was measured via ELISA. This test showed that whole-cell immunization triggered IFN-&gamma / production at significant levels while FeSOD-immunization did not / indicating the failure of alum-adsorbed FeSOD immunization in inducing cell-mediated immune response.
9

Avaliação da resposta imune celular na coinfecção por HIV e Leishmania

Gois, Luana Leandro January 2011 (has links)
Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2012-07-23T21:28:41Z No. of bitstreams: 1 Luana Leandro Gois Avaliação da resposta imune celular....pdf: 1029778 bytes, checksum: 6ffe55d1ff3bccef874a467c3f7cec99 (MD5) / Made available in DSpace on 2012-07-23T21:28:41Z (GMT). No. of bitstreams: 1 Luana Leandro Gois Avaliação da resposta imune celular....pdf: 1029778 bytes, checksum: 6ffe55d1ff3bccef874a467c3f7cec99 (MD5) Previous issue date: 2011 / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, Brasil / A Leishmania é considerada um patógeno oportunista em indivíduos infectados pelo HIV. Os mecanismos imunopatogênicos pelos quais o HIV e a Leishmania interagem não estão bem esclarecidos. Assim como, não está definido de que forma a infecção pelo HIV provoca alterações na resposta imune celular específica à Leishmania, o que conduz às apresentações atípicas e disseminadas da leishmaniose descritas nos pacientes coinfectados. O objetivo desta dissertação foi avaliar a resposta imune celular dos pacientes coinfectados por HIV e Leishmania, mais especificamente avaliar a resposta Th1 e perfil das subpopulações de linfócitos T de memória. Para tal, foi avaliada a resposta linfoproliferativa ao antígeno solúvel de Leishmania (SLA) e a proporção das subpopulações de memória central e efetora dos linfócitos T CD4+ específicos para Leishmania por citometria de fluxo. O índice de divisão celular dos linfócitos T CD4+ e CD8+ após o estímulo com SLA dos pacientes coinfectados foi estatisticamente menor em comparação com os pacientes infectados apenas por Leishmania. A resposta proliferativa dos linfócitos T CD4+ ao SLA foi observada em 25 % dos pacientes coinfectados, enquanto que em 12 % dos pacientes coinfectados foi observada proliferação dos linfócitos T CD8+ em resposta ao SLA. Entretanto, em todos os pacientes infectados apenas por Leishmania foi notada a linfoproliferação em resposta ao SLA. As proporções de linfócitos T CD4+ de memória central e efetora específicos para Leishmania foram similares entre os pacientes coinfectados e os pacientes infectados apenas por Leishmania. Entretanto, o número absoluto dos linfócitos T CD4+ de memória central e efetora foi significantemente menor nos pacientes coinfectados em comparação aos pacientes infectados apenas por Leishmania. Os resultados demonstram um prejuízo funcional na imunidade celular específica dos pacientes coinfectados. / The Leishmania is opportunistic pathogens in HIV-1-infected individuals. The immunopathogenic mechanisms by which HIV and Leishmania adversely affect each other are unclear. Furthermore, the impact of HIV-1 on Leishmania-specific cellular immune response leads to atypical and disseminated lesions as described in co-infected patients. The aim of this dissertation is to evaluate the cellular immune response of HIV and Leishmania co-infected patients, specifically to evaluate the profile Th1 and the memory CD4+ T-cell subset. The proliferative response of CD4+ and CD8+ T-cells to soluble Leishmania antigens (SLA) and the frequency memory Leishmania-specific CD4+ T-cell were performed flow cytometry. The median of cell division index of CD4+ and CD8+ T-cells after stimulation with SLA from co-infected patients was significantly lower than in patients infected Leishmania solely. A proliferative response of CD4+ T-cells after stimulation with SLA was observed in 25 % of co-infected patients, while in 12 % of co-infected patients had a proliferative response in the CD8+ T-cells to SLA. In contrast, both CD4+ and CD8+ T-cells subset from all patients infected with Leishmania solely proliferated in response to SLA. The proportion of Leishmania-specific central and effector memory CD4+ T-cells were similar between the coinfected patients and patients infected Leishmania solely. However, the median of absolute number of Leishmania-specific central and effector memory CD4+ T-cells from co-infected patients were significantly lower compared to patients infected Leishmania solely. These results demonstrate the impairment in cellular immune response.
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Avaliação imunoistoquímica da musculatura estriada esquelética em cães com leishmaniose visceral

Gomes, Ana Amélia Domingues [UNESP] 18 February 2009 (has links) (PDF)
Made available in DSpace on 2014-06-11T19:23:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-18Bitstream added on 2014-06-13T19:30:23Z : No. of bitstreams: 1 gomes_aad_me_jabo.pdf: 566624 bytes, checksum: 460e92b269ccf96c5bd806ac86bfd00f (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A leishmaniose visceral pode ser incluída como uma das causas de miopatia inflamatória em cães, entretanto, pouco se sabe sobre a patogênese da doença no sistema muscular, sendo incriminada muitas vezes apenas à natureza catabólica da enfermidade. O objetivo deste estudo foi avaliar, por meio de imunoistoquímica, a presença de formas amastigotas de Leishmania sp, linfócitos T (CD3+), macrófagos e IgG nos músculos tríceps braquial, extensor carpo radial, bíceps femoral e gastrocnêmio de 23 cães naturalmente acometidos por leishmaniose visceral. Dentre os 92 músculos avaliados,11 (12%) apresentaram marcação antigênica para formas amastigotas de Leishmania sp, 35 (38,1%) para linfócitos T (CD3+), 29 (31,5%) para macrófagos e 14 (12%) para IgG. Os resultados obtidos permitiram concluir que em cães com leishmaniose visceral apresentam imunomarcação para formas amastigotas de Leishmania sp., linfócitos T CD3+, macrófagos e IgG, sugerindo a participação direta do parasito e de uma resposta imune celular e humoral na fisiopatogenia da lesão muscular. / Visceral leishmaniasis may be included as a cause of inflammatory myophathy in dogs, however, little is known about the pathogenesis of the disease in the muscular system, which is frequently associated with the catabolic nature of the illness. The purpose of this study was investigate, through immunohistochemistry, the presence of amastigote forms of Leishmania sp, T lymphocytes (CD3+), macrophages and IgG in the muscle triceps brachial, extensor carpi radialis, biceps femoris and gastrocnemius of 23 dogs with visceral leishmaniasis. Among 92 evaluated muscles, 11 (12%) presented antigenic marking for amastigote forms of Leishmania sp., 35 (38,1%) for T lymphocites (CD3+), 29 (31,5%) for macrophages and 14 (12%) for IgG. The results of the present experiment led to the conclusion that in dogs with visceral leishmaniasis there may be a straight participation of the parasite and of cellular and humoral immune response in the ethiopatogeny of the muscular injury.

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