Ο καρκίνος του τραχήλου της μήτρας (ΚΤΜ), αποτελεί παγκοσμίως έναν από τους συχνότερα εμφανιζόμενους καρκίνους στο γυναικείο πληθυσμό. Ο ΚΤΜ επιφέρει σημαντική ψυχολογική, κοινωνική και οικονομική επιβάρυνση στους ασθενείς και στα συστήματα υγείας. Η κυριότερη αιτία εμφάνισης της νόσου συσχετίζεται με τη μόλυνση από τον ιό των ανθρώπινων θηλωμάτων – HPV (Human papilloma virus). O HPV συγκαταλέγεται στους πιο συχνά σεξουαλικ ώςμεταδιδόμενους ιούς. Ένα από τα μεγαλυτερα βήματα της δημόσιας υγείας για την προφύλαξη του πληθυσμού έναντι του HPV και κατ’ επέκταση του καρκίνου του τραχήλου της μήτρας, είναι η εισαγωγή του εμβολίου στο Εθνικό Πρόγραμμα Εμβολιασμού. Στα παρακάτω κεφάλαια (θεωρητικό μέρος) γίνεται εκτενής περιγραφή της ιστοπαθολογίας του ΚΤΜ και της βασικής βιολογίας του HPV, καθώς επίσης και του μηχανισμού δράσης του εμβολίου έναντι του ιού. Εν συνεχεία, στο ειδικό μέρος της παρούσας διπλωματικής εργασίας παρατίθενται στοιχεία που αφορούν την ευαισθητοποίηση και την αποδοσχή του πληθυσμού για τον προληπτικό εμβολιασμό έναντι του ιού HPV. / -

published_or_final_version / Paediatrics and Adolescent Medicine / Master / Master of Philosophy

DNA vaccines in larger animals, such as horses, are generally less effective and elicit significantly weaker immune responses, than in small animal model systems. To provide optimal protection against pathogenic microorganisms, the induction of both humoral and cellular immune responses from DNA vaccination may be necessary. One limitation to DNA immunization in the horse is the difficulty in generating high levels of antigen-specific antibody and CTL responses. Previous work in the laboratory has demonstrated that expression constructs containing native sequences encoding the surface unit (SU) envelope glycoprotein (pCiSU) of the Equine Infectious Anemia Virus (EIAV) are ineffective at stimulating immune responses in the horse. This was attributed to an unusual codon-usage bias of the EIAV genome that significantly limits the expression of SU sequences. Optimizing the codon usage of pCiSU (pSYNSU) in DNA vaccines stimulated low-titer immune responses in inoculated ponies. Another plausible explanation for the reduced effectiveness of these DNA vaccines may be transfection deficiency and low level expression elicited by plasmid vectors in the horse. These studies investigated if the addition of a cationic polymer, deacylated polyethyleneimine (PEI), and/or codon optimized molecular immune-stimulatory cytokines could augment the relatively weak immunogenicity of pSYNSU in DNA vaccination of horses/ponies. Administration of DNA in formulation with PEI resulted in the robust production of very long-lived humoral (15 months after vaccination) responses and induced cell-mediated IFN-y responses five days after immunization.. Additionally, the co-expression of a family of IL-15 cytokines expanded the repertoire of T cell recognition to SU-specific peptides, in terms of lymphoproliferation. DNA vaccination incorporating one IL-15 family member, IL-15 (SSLSS) significantly enhanced serum antibody levels of IgGA and IFN-y mRNA expression levels. These responses were distinctly different from results seen with vaccinates that received „naked‟ pSYNSU DNA vaccines. It is evident from these vaccine studies that PEI can enhance DNA vaccine-elicited antibody and CTL-associated responses in the horse and IL-15 (SSLSS) can dramatically augment these responses. These results demonstrate an important role for PEI in promoting the longevity of immune responses to genetic immunization, which has not been reported previously in any large animal model.

The HPV vaccine represents an important step in the primary prevention of cervical cancer, yet uptake rates for the vaccine remain below what is needed to establish "herd immunity" from the virus. While many studies have examined both psychosocial and communication factors affecting HPV vaccination decisions, this study adopts a unique approach to understand the communication environment within which this health decision happens, such as the many and sometimes conflicting messages about vaccine efficacy and safety guiding young women's decisions. Using the message convergence framework, this project identifies how further study of converging and diverging messages in the communication environment in which young women make their vaccination decision can extend research in considering optimal communication strategies to enhance demand for HPV vaccination. In Study 1, 39 unvaccinated women participated in qualitative interviews and were asked questions in order to understand the important elements of the HPV vaccination communication environment that affected their decision (i.e., common sources and content of messages, how they discussed these messages "interacting" and influencing their decision). Study 2 builds on the findings of Study 1 by employing an experimental design to test different message convergence conditions on women's intent to vaccinate (e.g., what happens when a doctor and a family member give conflicting information and recommendations about HPV vaccination?). Three hundred and nine unvaccinated women were randomly assigned to one of nine experimental message conditions and then assessed on behavioral intentions. Support was found for the message convergence framework. This project represents the first formal testing of the message convergence framework and the first time it has been used in the health context. The findings from these studies are discussed in terms of the implications for future cervical cancer research and prevention campaigns, as well as the utility of the message convergence framework for other health communication research topics in which researchers are seeking to better understand and consider the communication environment when designing health behavior interventions.

Les tiques sont des arthropodes ectoparasites obligatoires qui se nourrissent sur une grande variété de vertébrés sur une large partie du globe. Au cours de leur repas, les tiques sécrètent dans leur salive de nombreux facteurs leur permettant de contourner bon nombre des défenses de l’hôte. Bien que la littérature rapporte beaucoup d’informations au sujet des effets du repas de la tique sur l’hôte, la nature des facteurs actifs exprimés par les glandes salivaires de la tique est peu connue. Au cours d’anciens travaux au sein du laboratoire, le crible de deux banques d’ADN complémentaires - issues de la rétro-transcription des ARN messagers synthétisés par les glandes salivaires de la tique Ixodes ricinus – a permis l’identification de 27 protéines dont l'expression est spécifiquement induite ou régulée positivement pendant le repas sanguin de la tique I. ricinus. Parmi ces protéines, la protéine Seq24, induite au cours du repas sanguin, présente la capacité de moduler les immunités innée et acquise de l’hôte. En conséquence, la protéine Seq24 a été nommée Iris pour « Ixodes ricinus Immunosuppressor ». Au cours de la présente étude, notre but fût de caractériser le rôle d’Iris et de déterminer son importance dans le repas sanguin de la tique I. ricinus. La protéine Iris appartient à la famille des inhibiteurs de sérine protéases et présente une homologie significative avec l’inhibiteur d’élastase de leucocytes. Une analyse in silico a confirmé qu’Iris présentait la structure des serpines, et notamment le RCL (Reactive Center Loop), boucle responsable de l’activité anti-protéasique. Comme attendu (sur base de l’analyse in silico), Iris inhibe de manière spécifique l’activité de plusieurs sérine protéases, et en particulier l’élastase de leucocyte. Ces tests effectués, nous avons essayé de comprendre quel(s) pouvai(en)t être le(s) rôle(s) d’Iris dans l’accomplissement du repas sanguin de la tique, c’est à dire dans la lutte contre les différents systèmes de défenses de l’hôte. Tout d’abord, des tests ont démontré la capacité d’Iris à inhiber les mécanismes de l’hémostase. Des tests sur du plasma et du sang complet ont montré qu’Iris allonge le temps de fibrinolyse, la voie intrinsèque de la coagulation et l’adhésion plaquettaire. L’utilisation de mutants a également démontré que si les deux premières activités sont dépendantes du RCL, et donc d’un mode de fonctionnement anti-protéolytique, l’adhésion plaquettaire est indépendante de ce système. Ce résultat met en évidence l’existence d’autres sites actifs, isolés par analyse in silico, nommés Receptor Binding Domain (RBD). Un travail antérieur du laboratoire avait permis d’indiquer la capacité de la protéine recombinante Iris semi-purifiée à inhiber la production de TNF-a, d’IL-6, et d’IL-8 (cytokines pro-inflammatoires) ainsi que l’IFN-g par des PBMCs (Peripherical Blood Mononuclear Cells) humaines. Ces résultats ont été confirmés avec de la protéine purifiée. Des analyses complémentaires ont démontré qu’un mutant d’Iris - dépourvu d’activité anti-protéasique - conserve l’activité pro-inflammatoire. Là encore, ce mécanisme semble impliquer un ou plusieurs RBD. L’utilisation d’anticorps dirigés contre ces zones a permis de déterminer le domaine d’interaction (aa : 105-120) impliqué dans cette fonction. D’autre part, une analyse par FACS a permis de démontrer qu’Iris interagit uniquement avec les cellules d’origine monocytaire. Enfin, nous avons également analysé l’importance d’Iris au cours du repas sanguin de la tique par une approche vaccinale. Les résultats observés indiquent que 30 % des tiques nourries sur des lapins immunisés par la protéine rIris ne survivent pas au repas.

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.

A quantitative competitive binding "triple sandwich" enzyme immunoassay was used to evaluate pathogen/class-specific antibody responses in Holstein-Friesian calves vaccinated against Clostridium perfringens B-toxin at various ages postpartum. Vaccination of dams at six weeks and again at two weeks prepartum increased pathogen-specific antibody levels in their colostrum and respective calf's serum. Calves initially vaccinated at three days produced both a primary and secondary pathogen-specific antibody response, whereas calves initially vaccinated at 12 and 21 days produced only secondary responses. Maternally-derived antibodies were found to suppress neonatal antibody production following primary immunization. They were also found to influence secondary humoral immune responses, although in a diminished capacity. Pathogen-specific IgG and IgM concentrations in dams' sera and colostra were found related to subsequent pathogen-specific IgG and IgM neonatal serum concentrations. Only pathogen-specific IgA in dams' colostra was correlated to neonatal levels, possibly owing to a different origin and role of this immunoglobulin class. All class-specific colostral immunoglobulin levels were related to subsequent neonatal concentrations. Based on results from this experiment, it is recommended that calves be vaccinated at three days postpartum with a booster administered at 63 days.

Objective: To look at the Red Cross and the Red Crescent societies integrated campaigns between 2002 and 2006 with free distribution of insecticide treated nets (ITN)that have taken place and its contribution to the Millennium Development Goals(MDG) and the Abuja target.  Method: Review of surveys, evaluations and reports from the International Federation of Red Cross and Red Crescent integrated campaigns. Published articles up to 2007 have been accessed from electronic databases Medline, PubMed, the Cochrane Library and website`s from WHO, UNICEF, GFATM , and related articles available from international organisations web sites in addition to informal discussions and meetings with key stakeholders. Results: The integrated vaccination and free distribution of long lasting insecticidal nets (LLINs) achieved a rapid, high and equal LLIN coverage among all wealth quintiles. The MDG and Abuja target for ITN coverage at household level were reached within a week giving a unique opportunity for a significant reduction in malaria incidences, morbidity and mortality. The ITN possession remained higher than utilisation, but utilisation increased if a follow up visit, ensuring nets being hung and properly used, had taken place at household level post campaign. Conclusion: Large scale free distribution of LLINs  bridge the equity gap between poor and rich and increased the use rate among children under five and pregnant women. The low utilisation versus possession remains a challenge and thus a “minimum standard” of a two phased strategy is recommend to reach maximum impact and the MDG; Phase one preparing for pre campaign data, logistical planning and distribution while phase two should focus on a post campaign Keep Up program providing health education at household level to ensure proper net hanging and use. / <p>ISBN 978-91-85721-42-9</p>

This thesis explores the events and controversies surrounding the smallpox epidemic that hit Muncie, Indiana, in the summer and fall of 1893. The disease struck 150 individuals and left 22 dead, but it also raised broad questions regarding the authority of local and state public health officials to force vaccination upon citizens. Following recent historiographical trends that interpret anti-vaccinationist sentiment in Progressive-Era America as an important part of the political dialog, it argues that anti-vaccinationists in connection with the Muncie epidemic were not simply anti-modem, but had reasonable concerns as to the safety of smallpox vaccination and the government's authority to enforce it. / Department of History

The unsafe use of needles propagates cross infections with bloodborne pathogens and reduces the positive impact of vaccinations on global health. While a plethora of needle-free injection devices exist, the reformulation of protein-based vaccines is largely empirical and costly, which presents a barrier to their widespread clinical application. This thesis contributes to the identification of approaches that facilitate rapid vaccine reformulation and enhance the immunogenicity of needle-free dry-powder vaccines with the help of novel antigen delivery platforms. We hypothesised that the thermodynamic stabilisation of diphtheria toxin mutant 197 (CRM197), a glycoconjugate vaccine carrier protein, may enhance its structural preservation during spray-freeze-drying (SFD), and that its formulation in either soluble, surface-adsorbed, or nanoparticle form impacts the elicited immune response. Differential scanning fluorimetry was used to study the effect of excipients on the thermal stability of CRM197. Dry-powder formulation of CRM197 used i) encapsulation into a thermodynamically stabilising excipient matrix by SFD, ii) surface-immobilisation via physisorption onto a novel potassium-doped hydroxyapatite (kHA) carrier microparticle formed by molten salt synthesis, and iii) chemical conjugation and surface presentation on amphiphilic block copolymer nanoparticles that were incorporated into SFD-powders (SFD-NP). The structural integrity of CRM197 was assessed by size separation in addition to various spectral and thermal analysis methods. The immunogenicity of dry-powder CRM197 formulations was subsequently tested in vivo. The results suggest that the thermodynamic stability of CRM197 in solution does not ensure its structural stability during SFD. While needle-free dermal vaccination with kHA-adsorbed CRM197 induced comparable antibody titres to conventional IM injection of alum-adjuvanted CRM197, needle-free SFD and SFD-NP powders were less immunogenic. The highest mean IgG titre and most balanced Th1/Th2 response was achieved with nanoparticle-conjugated CRM197 by IM, which outperformed the current clinical standard. Therefore, future vaccine design should combine thermodynamic and kinetic stability screening, and place special emphasis on the delivery and structural presentation of the antigen to the immune system.