Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 7 uppsatser.

Xiao Wei. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 95-109). / Abstracts in English and Chinese. / ABSTRACT --- p.i / ABSTRACT (CHINESE VERSION) --- p.iii / ACKNOWLEDGEMENTS --- p.v / TABLE OF CONTENTS --- p.vii / LIST OF TABLES --- p.ix / LIST OF FIGURES --- p.x / LIST OF ABBREVIATIONS --- p.xii / Chapter Chapter One --- Introduction --- p.1 / Chapter 1.1 --- Discovery of HHV8 --- p.2 / Chapter 1.2 --- Biology of HHV8 --- p.3 / Chapter 1.2.1 --- Morphology --- p.3 / Chapter 1.2.2 --- Classification --- p.3 / Chapter 1.2.3 --- Cell culture --- p.4 / Chapter 1.2.4 --- Latent and lytic life cycle --- p.4 / Chapter 1.2.5 --- Genome organisation --- p.5 / Chapter 1.2.6 --- Strain variability --- p.6 / Chapter 1.3 --- Disease association of HHV8 --- p.8 / Chapter 1.3.1 --- Kaposi's sarcoma --- p.8 / Chapter 1.3.2 --- Other associated diseases --- p.10 / Chapter 1.4 --- Tropism of HHV8 --- p.11 / Chapter 1.5 --- Epidemiology of HHV8 --- p.13 / Chapter 1.5.1 --- Geographic distribution --- p.13 / Chapter 1.5.2 --- Age distribution --- p.15 / Chapter 1.5.3 --- HIV-infected individuals without Kaposi's sarcoma --- p.16 / Chapter 1.5.4 --- Bone marrow transplant and blood transfusion recipients --- p.19 / Chapter 1.5.5 --- Sexually transmitted disease clinic attendees --- p.18 / Chapter 1.5.6 --- HIV-negative intravenous drug users --- p.19 / Chapter 1.6 --- Transmission of HHV8 --- p.20 / Chapter 1.7 --- Methods for HHV8 antibody detection --- p.23 / Chapter 1.7.1 --- Antibodies against latent antigens --- p.23 / Chapter 1.7.2 --- Antibodies against lytic antigens --- p.24 / Chapter 1.7.3 --- Comparison between assays --- p.25 / Chapter 1.8 --- Project design --- p.26 / Chapter Chapter Two --- Methods and Materials --- p.29 / Chapter 2.1 --- Study I. Seroprevalence of HHV8 in Hong Kong --- p.30 / Chapter 2.1.1 --- Study population --- p.30 / Chapter 2.1.2 --- Source of HHV8 antigens --- p.32 / Chapter 2.1.3 --- Positive and negative controls --- p.34 / Chapter 2.1.4 --- Procedures for immunofluorescence assay --- p.34 / Chapter 2.1.5 --- Statistical methods --- p.36 / Chapter 2.2 --- Study II. Prevalence of HHV8 DNA in cervical scrapes --- p.36 / Chapter 2.2.1 --- Study population --- p.36 / Chapter 2.2.2 --- Preparation of cell lysate --- p.38 / Chapter 2.2.3 --- PCR amplification for human beta-actin gene --- p.38 / Chapter 2.2.4 --- PCR for HHV8 DNA --- p.41 / Chapter 2.2.5 --- Statistical methods --- p.45 / Chapter 2.3 --- Study III. HHV8 ORF 26 nucleotide sequence determination --- p.45 / Chapter 2.3.1 --- Study samples --- p.45 / Chapter 2.3.2 --- Sequencing method --- p.46 / Chapter 2.3.3 --- Nucleotide sequence data analysis --- p.49 / Chapter Chapter Three --- Result --- p.51 / Chapter 3.1 --- Study I. Seroprevalence of HHV8 in Hong Kong populations --- p.52 / Chapter 3.1.1 --- Seroprevalence of HHV8 in reference group --- p.52 / Chapter 3.1.2 --- HIV-negative intravenous drug users --- p.55 / Chapter 3.1.3 --- HIV-positive individuals without Kaposi's sarcoma --- p.57 / Chapter 3.1.4 --- Transfusion-dependent patients --- p.59 / Chapter 3.2 --- Study II. Prevalence of HHV8 DNA in cervical samples --- p.61 / Chapter 3.2.1 --- Optimisation of beta-actin PCR --- p.61 / Chapter 3.2.2 --- Optimisation of HHV8 ORF 26 PCR --- p.61 / Chapter 3.2.3 --- Analytical sensitivity of HHV8 nested PCR --- p.67 / Chapter 3.2.4 --- General female population --- p.67 / Chapter 3.2.5 --- Sexually transmitted disease clinic attendees --- p.73 / Chapter 3.3 --- Study III. Sequence variation of HHV8 ORF 26 fragment --- p.76 / Chapter Chapter Four --- Discussion --- p.84 / Chapter 4.1 --- Study I. Seroprevalence of HHV8 in Hong Kong populations --- p.85 / Chapter 4.1.1 --- Distribution of HHV8 in reference group --- p.86 / Chapter 4.1.2 --- HIV-positive individuals without Kaposi's sarcoma --- p.87 / Chapter 4.1.3 --- HIV-negative intravenous drug users --- p.88 / Chapter 4.1.4 --- Transfusion dependent patients --- p.88 / Chapter 4.2 --- Study II. Prevalence of HHV8 DNA in cervical samples --- p.89 / Chapter 4.2.1 --- General female population --- p.90 / Chapter 4.2.2 --- Sexual transmitted disease clinic attendees --- p.90 / Chapter 4.3 --- Study III. HHV8 ORF 26 nucleotide sequence variation --- p.91 / Chapter 4.3.1 --- Sequence variability --- p.91 / Chapter 4.3.2 --- HHV8 ORF26 genotyping --- p.91 / Chapter 4.3.3 --- Comparison with HHV8 isolates from other populations --- p.92 / Chapter 4.4 --- Conclusions --- p.93 / REFERENCES --- p.95

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2002. / Härtill 6 uppsatser.

Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2008. / Härtill 4 uppsatser.

Thesis (Ph.D. in Microbiology & Immunology) -- University of Colorado Denver, 2007. / Typescript. Includes bibliographical references (leaves 200-212).

Le premier but de mon projet de doctorat a ete la caracterisation moleculaire de l'interface entre les recepteurs activateurs des cellules Natural Killer (NK) et de leurs ligands exprimes dans les cellules infectees ainsi que l'implication de cette interaction sur la reponse a l'infection par le cytomegalovirus (MCMV). / J'ai tire un avantage de la variation naturelle au sein des membres lies aux recepteurs Ly49C ainsi que de la disponibilite des structures cristallines des Ly49 afin de comprendre les determinants moleculaires des interactions Ly49H-m157 et egalement identifier les residus des acides amines qui permettent de discriminer entre les recepteurs qui se lient et ceux qui ne se lient pas a m157. Mes decouvertes suggerent que le "site 2" du contact entre le CMH de classe I et Ly49 n'est visiblement pas implique dans la liaison avec m157. Au contraire, les residus localises au niveau de l'interface homodimere-recepteur seraient probablement critiques a la reconnaissance fonctionnelle de la glycoproteine m157. Notre approche fonctionnelle et de modelisation tridimensionnelle suggerent que l'architecture du dimere Ly49H est cruciale pour l'accessibilite de m157 mais non pour les molecules de CMH de classe I et relient la variabilite dans l'homodimerisation des Ly49 a la reconnaissance directe des produits pathogeniques. / Un autre mecanisme de la reponse de l'hote contre MCMV provient de l'etude de la souche de souris MA/My, laquelle, malgre l'absence du gene Ly49h ainsi que la proteine pour laquelle il code, y est hautement resistant. Des etudes anterieures ont demontre qu'une interaction epistatique entre un gene issu du groupe des genes Ly49 sur le chromosome 6 et le CMH (H2) sur le chromosome 17 est associee avec la resistance au virus. Utilisant une methode de co-culture de cellules reportrices NFAT-GFP exprimant les recepteurs activateurs Ly49 et de fibroblastes primaires infectes, j'ai montre que le recepteur activateur Ly49P des cellules NK reconnait specifiquement les cellules infectees par MCMV et que cette reconnaissance depend de la presence de l'haplotype H2k. Ce signal etait bloque par l'utilisation des anticorps anti-H2-D k mais non par anti-H2-Kk. Ces resultats indiquent l'existence d'un nouveau mecanisme des cellules NK implique dans la resistance au MCMV, lequel depend de l'interaction fonctionnelle entre le recepteur Ly49P et la molecule du MHC de classe I, H2-Dk, dans les cellules infectees par MCMV. Comme contribution directe de ce travail, nous avons demontre que la resistance chez MA/My est au moins partiellement dependante des interactions entre le recepteur Ly49P et la molecule H2-Dk modifiee par le virus dans les cellules infectees. Comme MCMV regule negativement l'expression des molecules du CMH de classe I, j'ai confirme la presence de H2-Dk dans les cellules infectees par l'utilisation d'un virus MCMV recombinant portant un gene rapporteur GFP. En permutant la plateforme peptidique de liaison, les domaines transmembranaires et intracellulaires entre les molecules H2-Db et H2-D k, j'ai demontre que la plateforme peptidique de liaison est critique pour la reconnaissance des cellules infectees. Par le criblage d'un panel de mutants MCMV portant des genes impliques dans l'evasion immune, j'ai demontre que l'infection de fibroblastes par le MCMV depourvu du gene m04 (Deltam04) abolit totalement l'activation de Ly49P. (Abstract shortened by UMI.)

Orientador: Sandra Helena Alves Bonon / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T11:41:57Z (GMT). No. of bitstreams: 1 Catalan_DanielThome_M.pdf: 2829035 bytes, checksum: 10145f8b164a7e70fc037a905f6026f6 (MD5) Previous issue date: 2012 / Resumo: Pacientes com doenças oncológicas possuem grandes períodos de neutropenia grave e são susceptíveis a complicações infecciosas. O tratamento com drogas quimioterápicas, radioterapia e em alguns casos cirurgia promovem depleção do sistema imunológico e exposição a microrganismos durante o processo cirúrgico. Durante o período de tratamento e em média até um ano após o término de terapia os episódios de febre são freqüentes e na maioria dos casos o agente etiológico não é conhecido. O tratamento faz-se então com administração de antibióticos de grande espectro, com a finalidade de realizar o tratamento empírico para vários tipos de microrganismos e a febre é chamada como febre de origem desconhecida. Através dos meios convencionais de diagnóstico uma pequena porcentagem de agentes etiológicos é identificada, e em muitos estudos não ultrapassa 25% dos episódios de febre. As hemoculturas normalmente são negativas e o tempo do diagnostico é prolongado. Dada a urgência nos cuidados com os pacientes oncológicos pediátricos a busca por um diagnóstico rápido com alta sensibilidade e especificidade se faz necessário. O objetivo deste estudo foi identificar a infecção ativa causada pelos herpesvírus (EBV, CMVH, HHV-6 e HHV-7) em pacientes oncológicos pediátricos com episódios de febre através das técnicas de Antigenemia para o CMV e N-PCR para o EBV, CMVH, HHV-6 e HHV- 7. Foram coletadas 169 amostras de soro para realização da N-PCR e 100 amostras de sangue total para realização do teste de antigenemia provenientes de 62 pacientes. Infecção ativa pelo Epstein-Barr vírus detectada pela N-PCR no soro ocorreu em 3/169 amostras (1,8%) em 3/62 pacientes diferentes (4,8%). Infecção ativa pelo CMVH foi detectada pela N-PCR em 16/169 amostras (9,5%) e em 12/62 (19,4%) pacientes; N-PCR no soro para detectar infecção ativa pelo HHV-6 ocorreu em 27/169 amostras (16,0%) e destas, 23/62 pacientes (37,1 %) foram acometidos. Infecção ativa causada pelo HHV-7 detectada pela N-PCR no soro ocorreu em 24/169 (14,8%) das amostras coletadas e em 18/62 (29,0%) pacientes. Para o teste de antigenemia para CMVH foram encontrados 5/100 (5,0%) de positividade nas amostras testadas e em 5/62 (8,1%) dos pacientes. Esses resultados mostram que a infecção ativa causada por pelo menos um dos herpesvírus EBV, CMV, HHV-6 e HHV-7 é frequente nessa população representando em nosso estudo 71/169 (42,0%) dos casos de febre e ocorreu em 62 pacientes portadores de neoplasia. Estudos futuros serão necessários com a inclusão de outros microrganismos para avaliar a etiologia da febre nos pacientes oncológicos pediátricos, assim como a introdução de novas técnicas de biologia molecular como a PCR quantitativa em tempo real / Abstract: Patients with infiltration cancer cells in bone marrow, solids tumors and treatment with chemotherapy promote high depletion of immune system activity. Most episodes with febrile neutropenia are treated with empiric broad-spectrum antibacterial therapy without identifying the site of infection or agent, such as fever unknown origin (FUO). Through the conventional diagnostic methods a small number of etiologic agents are identified, blood cultures and urine culture are usually negative and time-consuming to identify the source of infection for the clinic usually is difficult, nevertheless only identify bacteria and fungi. They are identified and confirmed by culture in only 25-30% of the cases, in other 15-25% of patients with fever bacterial or fungi infections are suspected on clinical findings and approximately 50% of cases are classified and treated as FUO. Given the urgency of care to cancer patients and fever need rapid diagnosis of infections in these patients, the search for new laboratory tests in order to promote high accuracy in identification of pathogens in these patients is important. The aim of this study is identify active infections caused by herpesvirus EBV, CMV, HHV-6 and HHV-7 in episodes of fever of pediatric oncology patients. 169 samples of whole blood and serum collected from patients with episodes of fever were analyzed using N-PCR methods and cytomegalovirus was analyzed by antigenemia and N-PCR. Active infection caused by Epstein-Barr virus occurred in 3/169 (1,8%)of samples and 3/62 (4,8%) of the patient; cytomegalovirus 16/169 (9,5%) of samples and 12/62 (19,4%) of the patient; HHV-6 27/169 (16,0%) of templates and 23/62 (37,1%) of the patient; HHV-7 24/169 (14,8%) of sample and 18/62 (29,1%) of the patient. This results shows that active infections caused by EBV, CMV, HHV-6 and HHV-7 are frequent in children receiving cancer treatment, representing high prevalence this cases of fever, with or without neutropenia. New assay are need with real time PCR for to better the diagnostic of the fever episodes / Mestrado / Clinica Medica / Mestre em Clinica Medica

No description available.

Thesis (Ph.D. in Microbiology) -- University of Colorado Denver, 2007. / Typescript. Abstract available online via ProQuest Digital Dissertations. Includes bibliographical references (leaves 207-223).