No description available.

Les protéines solubles sont généralement faiblement immunogènes, ce qui constitue unelimite pour le développement de vaccins sous unitaires à base de protéines. Mes travaux de thèseont eu pour objectif de décrypter certains mécanismes moléculaires et cellulaires qui contribuent àl’immunogénicité et d’en tirer partie pour développer des approches originales permettantd’améliorer la capacité des protéines à déclencher la réponse immunitaire. Pour cela, j’aiprincipalement utilisé le transactivateur transcriptionnel (Tat) du VIH-1. J’ai montré quel’oligomérisation de Tat permet à un mécanisme de collaboration B-TH-2 d’induire la réponseimmunitaire en absence d’adjuvant. J’ai identifié le déterminant minimal responsable de l’effet etmontré qu’il confère la propriété adjuvante à d’autres antigènes. J’ai ensuite montré que laprésentation aux cellules T restreinte aux CMH I et CMH II est accrue lorsque les protéines sontdotées de la capacité à lier des sucres sulfatés d’expression ubiquitaire: les héparanes sulfate. Cestravaux ont permis de définir de nouvelles approches pour améliorer l’immunogénicité de protéinessusceptibles d’être intégrées dans des préparations vaccinales. / Soluble proteins are usually poorly immunogenic, which is a limit to the development ofsubunit vaccines based on proteins. My thesis work aimed to decipher some molecular and cellularmechanisms that contribute to the immunogenicity and to exploit them to develop innovativeapproaches to improve the ability of proteins to trigger the immune response. For this purpose, Imainly used the transcriptional transactivator (Tat) of HIV-1. I showed that the oligomerization of Tatenables a B-TH-2 collaborative mechanism to induce the immune response in the absence ofadjuvant. I identified the minimum region determining the effect and showed that it confers the selfadjuvantingproperty to other antigens. In the second part of my work, I showed that the MHC I andMHC II restricted presentation to T cells is increased when the proteins are endowed with the abilityto bind ubiquitous sulfated polysaccharides: heparan sulfates. This work helped to define newapproaches to improve the immunogenicity of proteins that are likely to be included in vaccinepreparations.

Les cellules Natural Killer (NK) sont des effecteurs cytotoxiques impliqués dans la réponse immune contre les infections et les tumeurs. Pendant ma thèse j’ai étudié la fonctionnalité des cellules NK humaines en réponse à des lignées cellulaires de carcinome rénal à cellules claires (RCC) et de mélanome métastatique, deux tumeurs immunogènes. Nos résultats montrent que certaines mutations de VHL augmentent la susceptibilité des lignées RCC à la lyse NK. La perte de fonction de VHL corrèle avec une expression membranaire diminuée des molécules HLA-I par les lignées RCC mutées pour VHL. Chez les patients atteints de mélanome métastatique de stade IV, nous avons décrit un phénotype particulier des NK sanguines (NKp46dim/NKG2Adim) qui leur confère une forte activité antitumorale. Après traitement des patients par chimiothérapie, la fonctionnalité NK était réduite et le phénotype modifié. Pour étudier les cellules NK infiltrant les mélanomes, nous avons mis au point des conditions expérimentales pour caractériser les cellules NK de ganglions métastatiques de patients de stade III. Nos résultats préliminaires montrent que, par rapport aux ganglions sains, les NK des ganglions métastatiques présentent un phénotype altéré et un potentiel fonctionnel diminué. Nos résultats suggèrent que d’une part l’immunogénicité dépendante des oncogènes et d’autre part les altérations NK induites par la tumeur et/ou par la chimiothérapie sont des facteurs importants à considérer dans le choix des protocoles d’immunothérapie basés sur les cellules NK. / Natural Killer cells are cytotoxic lymphocytes involved in the immune response against tumours and infections. We investigated the NK-mediated functions in response to clear-cell renal cell carcinoma (RCC) and metastatic melanoma, two human immunogenic tumours. We showed that certain VHL mutations increased RCC cell susceptibility to NK lysis. VHL loss of function correlated with lower expression levels of membrane HLA-I molecules on VHL-mutated RCC and a decreased triggering of inhibitory NK receptors compared to RCC with a functional VHL. In stage IV melanoma patients, we showed that blood NK cells displayed a unique NKp46dim/NKG2Adim phenotype and high lytic potential towards melanoma cells. Following chemotherapy, NK cell function was reduced and the phenotype modulated. To study melanoma-infiltrating NK cells, we have set up experimental conditions to characterise NK cells in metastatic LNs from stage III melanoma patients. Our preliminary data show that, compared to normal LNs, NK cells from metastatic LNs are altered. Our findings suggest that oncogenic-dependent immunogenicity, tumour-associated NK alterations and chemotherapy are important factors that must be taken into account in the choice of immunotherapeutic protocols based on NK cells.

abstract: Biomarkers find a wide variety of applications in oncology from risk assessment to diagnosis and predicting and monitoring recurrence and response to therapy. Developing clinically useful biomarkers for cancer is faced with several challenges, including cancer heterogeneity and factors related to assay development and biomarker performance. Circulating biomarkers offer a rapid, cost-effective, and minimally-invasive window to disease and are ideal for population-based screening. Circulating immune biomarkers are stable, measurable, and can betray the underlying antigen when present below detection levels or even no longer present. This dissertation aims to investigate potential circulating immune biomarkers with applications in cancer detection and novel therapies. Over 600,000 cancers each year are attributed to the human papillomavirus (HPV), including cervical, anogenital and oropharyngeal cancers. A key challenge in understanding HPV immunobiology and developing immune biomarkers is the diversity of HPV types and the need for multiplexed display of HPV antigens. In Project 1, nucleic acid programmable protein arrays displaying the proteomes of 12 HPV types were developed and used for serum immunoprofiling of women with cervical lesions or invasive cervical cancer. These arrays provide a valuable high-throughput tool for measuring the breadth, specificity, heterogeneity, and cross-reactivity of the serologic response to HPV. Project 2 investigates potential biomarkers of immunity to the bacterial CRISPR/Cas9 system that is currently in clinical trials for cancer. Pre-existing B cell and T cell immune responses to Cas9 were detected in humans and Cas9 was modified to eliminate immunodominant epitopes while preserving its function and specificity. This dissertation broadens our understanding of the immunobiology of cervical cancer and provides insights into the immune profiles that could serve as biomarkers of various applications in cancer. / Dissertation/Thesis / Doctoral Dissertation Molecular and Cellular Biology 2018

Les protéines lipidées purifiées à partir d’extraits de pathogènes ou produites de façon recombinante ont démontré leur pouvoir immunogène. Malheureusement, ce type de protéines est généralement difficile à purifier et/ou à produire. Afin de résoudre ces difficultés, l’objectif de cette thèse a été de mettre au point un procédé de lipidation chimique d’antigènes protéiques d’intérêt vaccinal, présents naturellement sous forme lipidée mais produits de façon recombinante sous forme non lipidée. La lipidation chimique a ainsi été réalisée sur 2 protéines, à savoir rTbpB-dl provenant de N. meningitidis et rNWMN_A issue de S. aureus, par des lipopeptides synthétiques de structure minimale, Pam2CAG et Pam3CAG, ciblant respectivement les récepteurs TLR2/6 et TLR2/1. Ces lipopeptides ont été préalablement fonctionnalisés par des groupes réactifs aux fonctions thiols (maléimide ou bromoacétyle) afin de réaliser le couplage chimique. Des analyses physico-chimiques et biochimiques ont démontré que la modification des protéines antigéniques a bien été réalisée. La lipidation de rTbpB-dl et rNWMN_A par les lipopeptides a induit in vitro l’activation des cellules immunitaires murines et humaines via TLR2 et a également augmenté in vivo l’immunogénicité de ces protéines recombinantes, en présence ou non d’un adjuvant. De plus, ces protéines lipidées ont joué in vivo le rôle d’adjuvant en augmentant l’immunogénicité d'une protéine antigénique co-administrée. Notre procédé de lipidation chimique a été simple, rapide, de faible coût et répétable. Au final, ce procédé pourrait s’appliquer sur des antigènes protéiques d’intérêt vaccinal et de faible immunogénicité provenant de différents pathogènes et/ou sur des antigènes lipoprotéiques rencontrant des problèmes de production/purification. Il pourraitreprésenter un choix pertinent dans la mise au point et dans le développement de candidatsvaccins, en présence ou non d’adjuvant(s) et/ou d’autre(s) antigène(s) d’intérêt. / Lipidated proteins, purified from pathogens extracts or produced recombinantly, have demonstrated their immunogenic power. Unfortunately, this type of proteins is generally difficult to purify and/or to produce. To solve these difficulties, the objective of this thesis was to develop a process of chemical lipidation of protein antigens of vaccine interest, which are present naturally inlipidated form but produced recombinantly in non-lipidated form. The chemical lipidation was carried out on two proteins, namely rTbpB-dl from N. meningitidis and rNWMN_A from S. aureus, by minimum structure synthetic lipopeptides, Pam2CAG and Pam3CAG targeting receptors TLR2/6 and TLR2/1 respectively. These lipopeptides have been functionalized beforehand with thiolreactive groups (maleimide or bromoacetyl) for performing chemical coupling. Physicochemical and biochemical analysis have shown that the modification of the antigenic proteins has been achieved. The lipidation of rTbpB-dl and rNWMN_A by these lipopeptides induced in vitro activation of murine and human immune cells through TLR2. This lipidation also increased in vivo immunogenicity (mainly humoral) of the recombinant protein, in the presence or absence of an adjuvant. Furthermore, these lipidated proteins acted in vivo as an adjuvant by increasing immunogenicity of a co-administered antigen protein. Our process of chemical lipidation was simple, rapid, low-cost and repeatable. Taken together, this process could apply to antigens of protein nature, of vaccine interest and poorly immunogenic from different pathogens and/or to lipoprotein antigens encountering production/purification problems. It could be a relevant choice for the development of candidate vaccines, in the presence or absence of adjuvant(s) and/or other antigen(s) of interest.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-20T12:49:28Z No. of bitstreams: 2 Dissertação - Alex Jesus de Carvalho - 2014.pdf: 1685689 bytes, checksum: 5b4f38809e4a3fe6252bba20b25d949b (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-20T12:58:32Z (GMT) No. of bitstreams: 2 Dissertação - Alex Jesus de Carvalho - 2014.pdf: 1685689 bytes, checksum: 5b4f38809e4a3fe6252bba20b25d949b (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-04-20T12:58:33Z (GMT). No. of bitstreams: 2 Dissertação - Alex Jesus de Carvalho - 2014.pdf: 1685689 bytes, checksum: 5b4f38809e4a3fe6252bba20b25d949b (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-06-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The Gram-positive bacterium Staphylococcus saprophyticus, one of the coagulasenegative staphylococci, is the second most common causative agent of urinary tract infection, affecting mainly sexually active women. Staphylococcus saprophyticus can cause acute diseases as pyelonephritis, sepsis, nephrolithiasis, endocarditis, urethritis, epididymitis and prostatitis. This work aims to identify Staphylococcus saprophyticus surface proteins by using a proteolytic shaving approach, a methodology that was established to identify surface-exposed protein domains by tripsinization of intact cells. The peptides obtained were treated by trypsin, reduced, alkylated and identified by nano-chromatography using a nanoACQUITY UPLCTM system (Waters) coupled to a SYNAPT Q-TOF mass spectrometer (Waters). The homology analysis was performed using the software ProteinLynx 2.3 (Waters). Through the shaving, it was possible to identify 219 proteins, many of them, described as virulence factors. Of total, 01 is cell wall protein, 09 are extracelular proteins, 19 are membrane proteins and 190 are citoplasmatic proteins. Besides of the lysis process, the presence of cytoplasmic proteins on cell surface can be due to the activity of export pathways not yet identified and many of these proteins can be proteins with moonlighting function, in other words, proteins that plays more of one function, it can, in this case, plays functions on S. saprophyticus cell surface related to bacterial virulence. The main proteins with moonlighting function include metabolic enzymes of the glycolytic pathway; enzymes of other metabolic pathways, such as, glyoxalate cycle; chaperones and proteins related with the proteic folding. The prediction of cellular localization was performed through LocateP database. The results of this research help to elucidate the strategies and machineries used by proteins during the adhesion, infection and proliferation, leading us to understand the interaction between the pathogenic bacteria S. saprophyticus and the human host. The knowledge about the proteins present on the cell surface is of extreme importance, because many of these proteins represent targets to new drugs, therapeutic antibodies or vaccines, since the pathogen cell surface is the first to contact with the host cells during the infection process. / A bactéria Gram-positiva Staphylococcus saprophyticus, uma das bactérias estafilococos coagulase negativa, é o segundo agente mais comum causador de infecções do trato urinário, afetando principalmente mulheres sexualmente ativas. S. saprophyticus pode causar doenças agudas como pielonefrite, sepse, nefrolitíase, endocardite, uretrite, epididimite e prostatite. Este trabalho teve como objetivo identificar proteínas de superfície de S. saprophyticus pela abordagem de shaving proteolítico, uma metodologia que foi estabelecida para identificar proteínas que possuem domínios proteicos na superfície celular utilizando a tripsinização de células intactas. Posteriormente, os peptídeos obtidos foram tripsinizados, reduzidos, alquilados e identificados através de nano-cromatografia utilizando um sistema nanoACQUITY UPLCTM (Waters) acoplado a um espectrômetro de massas SYNAPT Q-TOF (Waters). Com isso foi possível identificar 219 proteínas, muitas delas descritas como fatores de virulência. Do total, 01 proteína é de parede celular, 09 extracelulares, 19 de membrana e 190 citoplasmáticas. Além do processo de lise, a presença de proteínas citoplasmáticas na superfície celular pode ser devida à atividade de vias de exportação ainda não identificadas e muitas dessas proteínas podem ser proteínas com função moonlighting, ou seja, proteínas que desempenham mais de uma função, podendo, neste caso, desempenhar funções na superfície de S. saprophyticus relacionadas à virulência bacteriana. As principais proteínas com função moonlighting incluem enzimas metabólicas da via glicolítica; enzimas de outras vias metabólicas, tais como, ciclo do glioxalato; chaperonas e proteínas relacionadas com o dobramento proteico. A predição de localização celular foi realizada com o banco de dados LocateP. Os resultados desta pesquisa contribuíram na elucidação das estratégias e maquinarias utilizadas pelas proteínas durante a adesão, infecção e proliferação, levando-nos a compreender a interação entre a bactéria patogênica S. saprophyticus e o hospedeiro humano. O conhecimento acerca das proteínas presentes na superfície celular é de extrema importância, visto que muitas dessas proteínas representam alvos para novas drogas, anticorpos terapêuticos ou vacinas, uma vez que a superfície celular do patógeno é a primeira a entrar em contato com as células do hospedeiro durante o processo de infecção.

Introdução: As infecções pelo papiloma vírus (HPV), geralmente são assintomáticas e transitórias, porém, em indivíduos com doenças autoimune, onde há um comprometimento da imunidade causado por mecanismo inerentes a própria doença e pelo tratamento com drogas imunossupressoras, o risco destas infecções evoluírem para lesões metaplásicas e neoplásicas é alto. A forma mais eficiente de prevenção deste tipo de desfecho é a vacinação contra HPV. Assim, a indicação desta vacina seria fundamental para a proteção destes pacientes, porém, devido aos fatores citados, pressupõem-se que a resposta poderia ser menos eficiente. Objetivo: Avaliar a imunogenicidade da vacina contra HPV quadrivalente em pacientes com Lúpus Eritematoso Sistêmico Juvenil (LESJ) e Dermatomiosite juvenil (DMJ). Métodos: Estudo prospectivo de intervenção multicêntrico, onde foram incluídas 328 meninas e adolescentes (9-20 anos) em 13 centros oriundos de diferentes regiões do Brasil. Porém apenas 246 indivíduos preencheram os critérios de inclusão. O protocolo do estudo previa a aplicação de 3 doses da vacina (0,2 e 6 meses). Os grupos foram divididos em: Grupo A (paciente 206 portadoras de LESJ); Grupo B (pacientes 40 portadoras de DMJ); Grupo C (controle 41 indivíduos saudáveis). A avaliação sorológica foi realizada nos momentos 0, 3, 7 e 12 meses do início do estudo e para mensurar os níveis de anticorpos produzidos para subtipos HPV 16 e HPV18 foi utilizado o imunoensaio Luminex. Foram considerados respondedoras as pacientes ou controles que apresentassem anti-HPV 16 acima do cutt-off= 9 mMU/ml e para antiHPV 18 acima de 13 mMU/ml. Resultados: 246 meninas e adolescentes completaram o protocolo do estudo, sendo 206 pacientes do grupo A, 40 pacientes do grupo B e 41 controles grupo C. Ao todo tivemos 39 indivíduos que não responderam a vacina, sendo 23 do grupo A,8 grupo B e 8 grupo C. E 82 descontinuidades do estudo por motivos diversos como: receio a reações a vacina, mudança de cidade, gravidez e óbito. Na análises comparativas feitas entre os grupos. Não foi observado nenhum fator de risco que influenciou na não resposta a vacina dentre as variáveis consideradas: faixa etária (9-13, 14- 20 anos), número de doses (2 ou 3), tipo e atividade da doença de base, além das várias drogas utilizadas no tratamento, analisadas individual e em associações. Conclusão: A vacinação contra HPV parece ser imunogênica para o grupo de pacientes com LESJ ou DMJ, porém parte delas apresentam uma resposta parcial à vacina, com resposta efetiva a apenas um dos sorotipos avaliados (16 ou 18). Nenhum fator de risco foi encontrado que possa ter influenciado na não resposta a vacina. / Introduction: Papillomavirus (HPV) infections are generally asymptomatic and transient, but in individuals with autoimmune diseases whom used to have a compromise of the immunity caused by mechanisms inherent to the disease itself in addition to the treatment with immunosuppressive drugs. Thus, the risk of these infections progress for metaplastic and neoplastic lesions is high in these individuals. Vaccination is essential for the protection of these patients, however because the characteristics of their immune system may be less effective. The most efficient way to prevent this type of outcome is vaccination against HPV. Thus, the indication of this vaccine would be fundamental for the protection of these patients, however due to the compromise in the immune system, it is possible to assumed that the response could be less efficient. Objective: To evaluate the immunogenicity of the quadrivalent HPV vaccine in patients with Juvenile Systemic Lupus Erythematosus (JSLE) and Juvenile Dermatomyositis (JDM). Methods: A multicenter prospective study, including 328 girls and adolescents (9-20 years) in 13 centers from different regions of Brazil. The study protocol provided for the application of 3 doses of the vaccine (0.2 and 6 months). During the study blood collections and serological evaluation was performed at time points 0, 3, 7 and 12 months after the start of the study in 206 SLEJ patients (group A), 40 JDM patients (group B) and 41 healthy controls (group C). To evaluate the levels of antibodies produced for HPV 16 and HPV18 subtypes the Luminex immunoassay was used. Patients or controls with anti-HPV 16 cutt-off = 9 anti-HPV 18 = 13 were considered as responders. RESULTS: Of the 328 girls and adolescents included, 246 girls and adolescents completed the study protocol, being 206 patients in group A, 40 patients in group B and 41 controls in group C. In all we had 39 individuals who did not respond to the vaccine, being 23 of group A, 8 group B and 8 group C. E 82 study discontinuities for various reasons such as: fear of reactions to the vaccine, city change, pregnancy and death. In the comparative analyzes made between the groups. No risk factor was observed that influenced the non-response to the vaccine among the variables considered: age range (9-13, 14-20 years), number of doses (2 or 3), type and activity of the underlying disease, Of the various drugs used in the treatment, analyzed individually and in associations. 287 completed the study. Conclusion: the vaccine anti-HPV seems to be immunogenic in JLES and JDM patient, however a percentage of patients have showed a partial response to the vaccine, just a one of the serotypes 16 or 18. No risk factor was found that May have influenced the nonresponse to the vaccine.

No description available.

Introdução: As infecções pelo papiloma vírus (HPV), geralmente são assintomáticas e transitórias, porém, em indivíduos com doenças autoimune, onde há um comprometimento da imunidade causado por mecanismo inerentes a própria doença e pelo tratamento com drogas imunossupressoras, o risco destas infecções evoluírem para lesões metaplásicas e neoplásicas é alto. A forma mais eficiente de prevenção deste tipo de desfecho é a vacinação contra HPV. Assim, a indicação desta vacina seria fundamental para a proteção destes pacientes, porém, devido aos fatores citados, pressupõem-se que a resposta poderia ser menos eficiente. Objetivo: Avaliar a imunogenicidade da vacina contra HPV quadrivalente em pacientes com Lúpus Eritematoso Sistêmico Juvenil (LESJ) e Dermatomiosite juvenil (DMJ). Métodos: Estudo prospectivo de intervenção multicêntrico, onde foram incluídas 328 meninas e adolescentes (9-20 anos) em 13 centros oriundos de diferentes regiões do Brasil. Porém apenas 246 indivíduos preencheram os critérios de inclusão. O protocolo do estudo previa a aplicação de 3 doses da vacina (0,2 e 6 meses). Os grupos foram divididos em: Grupo A (paciente 206 portadoras de LESJ); Grupo B (pacientes 40 portadoras de DMJ); Grupo C (controle 41 indivíduos saudáveis). A avaliação sorológica foi realizada nos momentos 0, 3, 7 e 12 meses do início do estudo e para mensurar os níveis de anticorpos produzidos para subtipos HPV 16 e HPV18 foi utilizado o imunoensaio Luminex. Foram considerados respondedoras as pacientes ou controles que apresentassem anti-HPV 16 acima do cutt-off= 9 mMU/ml e para antiHPV 18 acima de 13 mMU/ml. Resultados: 246 meninas e adolescentes completaram o protocolo do estudo, sendo 206 pacientes do grupo A, 40 pacientes do grupo B e 41 controles grupo C. Ao todo tivemos 39 indivíduos que não responderam a vacina, sendo 23 do grupo A,8 grupo B e 8 grupo C. E 82 descontinuidades do estudo por motivos diversos como: receio a reações a vacina, mudança de cidade, gravidez e óbito. Na análises comparativas feitas entre os grupos. Não foi observado nenhum fator de risco que influenciou na não resposta a vacina dentre as variáveis consideradas: faixa etária (9-13, 14- 20 anos), número de doses (2 ou 3), tipo e atividade da doença de base, além das várias drogas utilizadas no tratamento, analisadas individual e em associações. Conclusão: A vacinação contra HPV parece ser imunogênica para o grupo de pacientes com LESJ ou DMJ, porém parte delas apresentam uma resposta parcial à vacina, com resposta efetiva a apenas um dos sorotipos avaliados (16 ou 18). Nenhum fator de risco foi encontrado que possa ter influenciado na não resposta a vacina. / Introduction: Papillomavirus (HPV) infections are generally asymptomatic and transient, but in individuals with autoimmune diseases whom used to have a compromise of the immunity caused by mechanisms inherent to the disease itself in addition to the treatment with immunosuppressive drugs. Thus, the risk of these infections progress for metaplastic and neoplastic lesions is high in these individuals. Vaccination is essential for the protection of these patients, however because the characteristics of their immune system may be less effective. The most efficient way to prevent this type of outcome is vaccination against HPV. Thus, the indication of this vaccine would be fundamental for the protection of these patients, however due to the compromise in the immune system, it is possible to assumed that the response could be less efficient. Objective: To evaluate the immunogenicity of the quadrivalent HPV vaccine in patients with Juvenile Systemic Lupus Erythematosus (JSLE) and Juvenile Dermatomyositis (JDM). Methods: A multicenter prospective study, including 328 girls and adolescents (9-20 years) in 13 centers from different regions of Brazil. The study protocol provided for the application of 3 doses of the vaccine (0.2 and 6 months). During the study blood collections and serological evaluation was performed at time points 0, 3, 7 and 12 months after the start of the study in 206 SLEJ patients (group A), 40 JDM patients (group B) and 41 healthy controls (group C). To evaluate the levels of antibodies produced for HPV 16 and HPV18 subtypes the Luminex immunoassay was used. Patients or controls with anti-HPV 16 cutt-off = 9 anti-HPV 18 = 13 were considered as responders. RESULTS: Of the 328 girls and adolescents included, 246 girls and adolescents completed the study protocol, being 206 patients in group A, 40 patients in group B and 41 controls in group C. In all we had 39 individuals who did not respond to the vaccine, being 23 of group A, 8 group B and 8 group C. E 82 study discontinuities for various reasons such as: fear of reactions to the vaccine, city change, pregnancy and death. In the comparative analyzes made between the groups. No risk factor was observed that influenced the non-response to the vaccine among the variables considered: age range (9-13, 14-20 years), number of doses (2 or 3), type and activity of the underlying disease, Of the various drugs used in the treatment, analyzed individually and in associations. 287 completed the study. Conclusion: the vaccine anti-HPV seems to be immunogenic in JLES and JDM patient, however a percentage of patients have showed a partial response to the vaccine, just a one of the serotypes 16 or 18. No risk factor was found that May have influenced the nonresponse to the vaccine.

L’infection au papillomavirus humain (HPV) est connue pour être le principal facteur causal d’une série de maladies aussi bien bénignes (condylomatose ano-génitale, papillomatose lyringée, et autres) que malignes (cancer du col de l’utérus, certains cancers ORL, et autres). Deux vaccins prophylactiques (Gardasil® et Cervarix®) sont sur la marché depuis à peu près quatre ans pour prévenir cette infection. Le présent travail de thèse comportait trois objectifs principaux : i) étudier in-silico l’immunogénicité du vaccin Gardasil® ; ii) étudier in-silico l’histoire naturelle d’une infection à HPV et iii) évaluer in-silico le potentiel de l’hypothèse thérapeutique suivante : l’administration intramusculaire du vaccin Gardasil® chez des patients atteints d’une papillomatose laryngée induirait un effet bénéfique car l’arrivée des immunoglobulines au tissu affecté empêcherait l’HPV de compléter son cycle de vie et, par conséquent, la maladie de se propager. Les principales conclusions sont : i) pour qu’une papillomatose laryngée ne s’étende pas il faudrait, d’après nos simulations, que le taux d’IgGs sériques soit maintenu au-dessus de 200 mMU/mL ; ii) pour rester, sur une période de 10 ans, le plus longtemps possible au-dessus de ce seuil (d´effet thérapeutique), en administrant la quantité minimale de vaccin, il faudrait, d’après nos simulations, suivre le protocole suivant : l’immunisation de base (à 0, 2 et 6 mois), suivie de trois rappels successifs tous les six mois jusqu’au 24ème mois, suivis d’un rappel 18 mois plus tard ; iii) par ailleurs, il semble inutile (voire contreproductif), d’après nos simulations, de modifier le schéma traditionnel de base (0-2-6 mois) / Two prophylactic vaccines have demonstrated to prevent infections with the human papillomavirus (HPV). Thus, they have been in the market for the last four years, or so. The three main objectives of the present project were: i) to study in-silico the immunogenicity of one of these vaccines (Gardasil®); ii) to study in-silico the natural history of an HPV infection, and iii) to assess in-silico the potential of the following therapeutic hypothesis : the intramuscular administration of Gardasil® to patients already suffering from a recurrent respiratory papillomatosis would result in a better prognosis thanks to the fact that the HPV-specific immunoglobulins that would bathe the affected tissue would impede the virus to complete its life cycle and, therefore, the disease to progress. The main conclusions are: i) according to our simulations, the minimum serum IgG titer required for hampering the progression of a recurrent respiratory papillomatosis would be 200 mMU/mL ; ii) in order to keep, within a time window of ten years, the anti-HPV IgG titer over the just-mentioned therapeutic-effect threshold, the biggest possible fraction of time and through the administration of the smallest possible number of booster doses, it would be necessary, according to our simulations, to adopt the following vaccination schedule: the basic three doses (at months 0, 2 and 6), followed by three successive booster doses, every six months, until reaching the 24th month, followed by a late final booster dose, 18 months later. iii) incidentally, it would seem to be inappropriate, according to our simulations, to modify the original initial vaccination schedule (at months 0, 2 and 6)