La protection contre certains agents infectieux ainsi que le traitement de cancers nécessite l'induction d'une réponse cellulaire. Le développement de vaccins induisant une réponse T CD8 efficace est donc essentiel. La présentation croisée de l'antigène est importante pour l'activation de lymphocytes T CD8 spécifiques et de nombreux facteurs participent au développement d'une réponse lymphocytaire T efficace. Nous nous sommes intéressés à deux d'entre eux : la voie d'immunisation et la séquence d'aministration de l'antigène et d'un adjuvant. Nous avons développé une technique d'enrichissement des lymphocytes T CD8 spécifiques d'un antigène, ce qui a permis une étude précise de la réponse T CD8 endogène. Nous avons ensuite étudié l'influence de la voie d'immunisation sur l'efficacité de la réponse T CD8. Nous avons observé que l'injection intradermique d'un antigène cellulaire induit une réponse T CD8 plus tardive, comparée à une administration par voie systémique. Cependant, la réponse T CD8 induite par une injection locale de l'antigène est plus efficace. Puis, nous avons évalué l'influence de l'administration d'un adjuvant - le poly I:C connu pour induire la production d'interférons (IFN) de type I - en parallèle de celle de l'antigène. Nous avons montré que le moment optimal d'administration de l'adjuvant dépend de la voie d'immunisation. De plus, il existe une durée limitée durant laquelle l'adjuvant induit des effets positifs sur l'activation des lymphocytes T CD8. Nous avons identifié plusieurs effets du poly I:C et des IFN de type I sur les cellules du système immunitaire

Les vecteurs nanoparticulaires comme systèmes de relargage contrôlé pour des applications vaccinales font l'objet d'intenses recherches, notamment dans le domaine du VIH1. Une approche novatrice consiste à co-administrer des molécules immuno-stimulatrices avec les antigènes d'intérêt, afin d'amplifier le recrutement et l'activation des cellules dendritiques (DCs). Un tel vecteur vaccinal stimulerait l'intensité de la réponse immunitaire et une immunité au niveau des muqueuses vaginales et anales pourrait être obtenue après vaccination. Des nanoparticules de poly(acide lactique) (NP-PLA) ou de chitosane/sulfate de dextrane (NP-CSD) ont été utilisées comme véhicules et adjuvants de protéines du VIH1, gp140 et p24. Le poly(I:C), ligand de TLR3 est la molécule immuno-stimulatrice retenue pour ses propriétés adjuvantes. Les NP-PLA et NP-CSD présentent un potentiel équivalent pour l'adsorption de protéines. Par contre, si les NP-CSD permettent l'adsorption du poly(I:C) (95%), elle est moins reproductible sur les NP-PLA. Pour chaque formulation, la capacité à induire in vitro la maturation des DCs a été évaluée en suivant les marqueurs CD25, CD80, CD83, par cytométrie en flux. L'adsorption de poly(I:C) sur les NP-PLA ou les NP-CSD amplifie les capacités de maturation de ces nanoparticules, un effet synergique étant observé avec les NP-CSD. Nos travaux montrent que la co-adsorption d'un TLR ligand, avec des antigènes protéiques du VIH sur des nanoparticules biodégradables, est possible et confère à la formulation vaccinale un effet immuno-stimulant in vitro. In vivo, les formulations vaccinales contenant du poly(I:C) induisent de très forts taux d'anticorps sériques chez la souris.

The ground work for this research thesis was completed seven years ago when the Dow Pharmaceutical Company selected Muncie, Indiana to test their new rubella vaccine. In the Fall of 1970, 311 seronegative elementary school children were innoculated with HPV-77, DK12 rubella vaccine. Of the 302 children who seroconverted, 98 were available for follow-up six years post innoculation. The participant's sera drawn six years previously was recovered in order to determine a six week and six year post innoculation titer on paired specimens. All 98 children, when retested, has measurable antibody. The geometric mean titer six years post innoculation was 55; a 14% decrease from the geometric titer of 64 six weeks after innoculation. At the six year point in time, there was at least a 9.2% incidence of subclinical reinfection, detected by a fourfold rise in the HI-titer, within the study group.Ball State UniversityMuncie, IN 47306

La infecció pel virus de la immunodeficiència humana (VIH) constitueix una de les epidèmies més importants actualment. El VIH es caracteritza per la seva elevada variabilitat genètica al llarg de la seva història natural. Les múltiples variants existents dins del mateix hoste, el que es coneix com quasiespècies, constitueixen una font d'adaptabilitat del virus, ja que permeten la disponibilitat d'una variant d'escapament davant una possible pressió selectiva en l'ambient (resposta immune, fàrmacs antiretrovirals, etc) . Malgrat els grans avenços en el desenvolupament de tractaments eficaços, actualment no ha estat possible l'eradicació de la infecció. El tractament antiretroviral de gran activitat (TARGA) ha disminuït significativament la morbi-mortalitat associada a la infecció pel VIH, però, no aconsegueix l'eliminació del virus. Conseqüència directa d'això és que, en suspendre'l, el virus reapareix en sang perifèrica, i caldrà mantenir el tractament de per vida, amb els seus costos tant a nivell d'efectes secundaris com econòmics. Davant aquesta situació, ha estat necessari el disseny de teràpies alternatives com les teràpies immunomediades (interrupcions estructurades del tractament o vacunes terapèutiques), enfocades a estimular el sistema immunològic del pacient per poder suspendre el TARGA de forma definitiva o, almenys, durant períodes variables de temps. Avui en dia no es coneixen amb exactitud els efectes que pugui tenir la variabilitat del VIH en les teràpies immunomediades, i l'interès del nostre estudi parteix d'aquesta premissa. Per això, l'objectiu d'aquesta tesi és analitzar, per primera vegada, l'evolució del virus en pacients sotmesos a vacunació terapèutica (en el nostre cas, una vacuna terapèutica de cèl•lules dendrítiques polsades amb el virus autòleg inactivat per calor). / La infección por el virus de la inmunodeficiencia humana (VIH) constituye una de las epidemias más importantes actualmente. El VIH se caracteriza por su elevada variabilidad genética a lo largo de su historia natural. Las múltiples variantes existentes dentro del mismo huésped, lo que se conoce como cuasiespecie, constituyen una fuente de adaptabilidad del virus, pues permiten la disponibilidad de una variante de escape frente una posible presión selectiva en el ambiente (respuesta inmune, fármacos antiretrovirales, etc). A pesar de los grandes avances en el desarrollo de tratamientos eficaces, actualmente no ha sido posible la erradicación de la infección. El tratamiento antirretroviral de gran actividad (TARGA) ha disminuido significativamente la morbi-mortalidad asociada a la infección por el VIH, no obstante, no consigue la eliminación del virus. Consecuencia directa de ello es que, al suspenderlo, el virus reaparece en sangre periférica, siendo necesario mantener el tratamiento de por vida, con sus costes tanto a nivel de efectos secundarios como económicos. Frente esta situación, ha sido necesario el diseño de terapias alternativas como las terapias inmunomediadas (interrupciones estructuradas del tratamiento o vacunas terapéuticas), enfocadas a estimular el sistema inmunológico del paciente para poder suspender el TARGA de forma definitiva o, al menos, durante períodos variables de tiempo. Hoy en día no se conocen con exactitud los efectos que pueda tener la variabilidad del VIH en las terapias inmunomediadas, y el interés de nuestro estudio parte de esta premisa. Por ello, el objetivo de esta tesis es analizar, por primera vez, la evolución del virus en pacientes sometidos a vacunación terapéutica (en nuestro caso, una vacuna terapéutica de células dendríticas pulsadas con el virus autólogo inactivado por calor). / Human Immunodeficiency Virus (HIV) infection is one of the largest epidemics today. HIV is characterized by high genetic variability throughout its natural history. Multiple variants exist within the same host, which is known as quasispecies. These quasispecies are a source of adaptability of the virus, and allow the availability of alternative escape variants against a possible selective pressure in the environment (immune response, antiretroviral drugs, etc). Despite the great advances in the development of effective treatments, now has not been possible to eradicate the infection. Antiretroviral therapy (HAART) has significantly decreased morbidity and mortality associated with HIV infection, however, it fails to eliminate the virus. Direct consequence is that, when HAART is stopped, virus rebound in peripheral blood, being necessary to maintain treatment for life, with its costs both in terms of side effects and economic burden. Facing this situation, it has been necessary to design alternative therapies such as immune-mediated therapies (structured treatment interruptions or therapeutic vaccines), aimed to stimulate the patient's immune system to permanently remove HAART or, at least, for varying periods of time. Today, effects of the variability of HIV in immune-mediated therapies are not exactly known , and the interest of our study is based on this premise. Therefore, the objective of this thesis is to analyze, for the first time, the evolution of HIV in patients undergoing therapeutic vaccination (in our case, a therapeutic vaccine based on dendritic cells pulsed with autologous heat-inactivated virus).

Background: Following the publication of Andrew Wakefield’s article claiming a link between Autism and the MMR vaccine in 1998, the U.K. and U.S. experienced a decline in vaccination rates. Combating the anti-vaccine messages highlighted by the media are the medical providers, who are consistently reported as an influential source of information for parental vaccine decision making. Despite efforts of the medical and public health community, some developed countries have seen a resurgence of vaccine preventable diseases. Purpose: This study seeks to examine parental vaccination concern in a private pediatric practice in metropolitan Atlanta. Methods: A questionnaire was created by the PI to assess parental vaccination concerns, including items to assess parental feelings toward the providers and nurses regarding preventative care. Data was analyzed in SPSS version 19.0. The study was approved by the IRB at Georgia State University. Results: A total of 283 participant responses were included in the sample. Overall vaccine adherence was 96.1% (272). However, a large minority of participants who were considered to have vaccine concerns were identified: 40.3% (114) of participants responded yes to at least one vaccine hesitation item. Conclusion: Vaccine adherence in a private pediatric practice remains high. However some parents continue to have vaccination concerns and may be at risk for deviating from the vaccine schedule. Using qualitative methods to obtain parental beliefs may provide a deeper understanding of parental decisions to aid in the development of public health education programs. The feasibility of collecting data at a private pediatric practice is discussed.

Syfte: Syftet med denna studie var att jämföra BVC-sjuksköterskors och föräldrars upplevelse av vaccinationstillfället samt skattning av barns smärta i form av skriktid vid två typer av vaccinationstekniker, dubbelvaccinering respektive en injektion i taget, när barnet ska ha två sprutor vid samma vaccinationstillfälle. Metod: Studien är en jämförande deskriptiv studie med kvantitativ ansats. Från både Stockholm och Uppsala valdes 50 barn ut konsekutivt varav 25 barn från Stockholms län samt 25 barn från Uppsala län. Datainsamling skedde med hjälp av enkäter samt mätning av skriktid. Dataanalysen genomfördes med hjälp av chitvåtest samt t-test. Resultat: Det fanns signifikanta skillnader vad gäller både BVC-sjuksköterskors och föräldrars upplevelser av vaccinationsproceduren. Skillnader kunde även ses såväl hos både föräldrar (p=0,020) och BVC-sjuksköterskor (p=0,032) när de fick skatta sin upplevelse av barnets smärta i samband med vaccinationen. Skriktiden visade inte någon signifikant skillnad (p=0,051) mellan de olika vaccinationsteknikerna. Slutsats: Resultatet av denna studie visade att barn som fick dubbelvaccinering upplevdes ha mindre ont när deras smärta skattades av både föräldrar och BVC-sjuksköterskor. Barn som fick sprutorna samtidigt skrek/grät generellt kortare tid än de barn som fick en spruta i taget, skillnaden visade dock ingen signifikans. Flertalat föräldrar i denna studie föredrog dubbelvaccinering. / Aim: The aim of this study was to compare child health center nurses' and parents' experience of the immunization and the child's estimated pain in terms of cry duration when two types of vaccination techniques were used, simultaneous versus sequential immunization injections, when the child are given two shots at the same occasion. Methods: The study is a comparative descriptive study with quantitative approach. From both Stockholm and Uppsala 50 children were selected consecutively with 25 children from the Stockholm and 25 children from the Uppsala. Data collection was done by means of questionnaires and measurement of cry duration. Data analysis was performed using the chi-square test and t-test. Results: There were significant differences from both nurses' and parents' perceptions of the vaccination procedure. Differences could also be seen in terms of the child's pain when both parents' (p=0,020) and nurses' (p=0,032) rated the child's pain associated with the immunization. Cry duration did not show a significant difference (p=0,051) between the two vaccination techniques. Conclusions: The results of this study showed that the children who received two vaccinations simultaneously were experienced by both parents’ and nurses’ to have less pain. Children that received the two injections at the same time cried less in general then the children that received the two injections one by one, the difference was not however significant. Most of the parents in this study preferred that the injections should be given at the same time.

Mitt syfte med denna uppsats är att undersöka varför och på vilket sätt individer söker sig till antroposofi. Jag vill främst göra detta genom att undersöka och beskriva attityder hos individer som har valt att gå till en antroposofisk mödra- och barnavårdcentral. Individer som inte på ett enkelt sätt definierar sig som antroposofer, eller ens som associerade med den organisationen, men som trots allt ändå väljer antroposofisk mödravård. Syftet är att lyfta fram individens val, tankar och förhållningsätt till antroposofin och till mörda- och barnavårdcentralen.

Hintergrund: Patienten mit chronischen entzündlichen Erkrankungen sind aufgrund ihrer Grundkrankheit, aber auch durch die häufig notwendige immunsuppressive Therapie gefährdet, an einer impfpräventablen Infektionskrankheit zu erkranken. In einer Stichprobe sollte der Impfstand bei Patienten mit chronisch-entzündlichen Darmerkrankungen (CED) erhoben werden. Besondere Beachtung galt Vorbehalten der Patienten gegen die empfohlenen Schutzimpfungen. Methoden: Wir baten 203 Patienten mit CED (davon 57% Mb. Crohn, 63% weiblich; medianes Alter 36 Jahre), die im letzten Jahr keine Impfberatung erhalten hatten, einen Fragebogen mit 38 Fragen zu beantworten. Zudem wurden alle Impfnachweise erfasst und mit den aktuellen Empfehlungen der Ständigen Impfkommission abgeglichen. Die Befragung erfolgte vom 1.4. bis 30.9.2009. Ergebnisse: Nur 83% der Patienten hatten einen Impfausweis. Es fanden sich erhebliche Impfdefizite; so wurden in den letzten zehn Jahren nur 67% der Patienten gegen Tetanus und 21% gegen Pertussis geimpft, 28% nahmen die Impfung gegen die saisonale Grippe 2008 wahr und nur 9% wurden jemals gegen Pneumokokken geimpft. Im Subgruppenvergleich von Patienten mit TNF-Blockern (n=39) mit denjenigen Patienten, die noch nie eine immunsuppressive Dauertherapie erhielten (n=67), zeigten sich keine Unterschiede. 80% der Patienten wären bereit, alle offiziell empfohlenen Schutzimpfungen durchführen zu lassen. 22% aller Patienten gaben an, Schutzimpfungen zu vermeiden, weil sie Nebenwirkungen befürchteten, 15% weil das Immunsystem „nicht intakt“ ist und 9% befürchten eine Verschlimmerung der CED durch eine Impfung. Schlussfolgerungen: Der Impfstand in der untersuchten Stichprobe war unzureichend. Es fand sich insbesondere eine deutliche Diskrepanz zwischen der hohen Bereitschaft der Patienten, Schutzimpfungen durchführen zu lassen, und dem tatsächlichen Impfstand. Unsere Daten legen die Notwendigkeit einer erhöhten ärztlicher Wachsamkeit für Impflücken bei immunsuppressiv behandelten Patienten nahe. / Background: Patients with chronic inflammatory diseases are at increased risk for vaccine preventable infectious diseases. This is caused by the inflammatory state itself as well as often necessary immunosuppressive therapy. In a random sample, we investigated whether patients with inflammatory bowel disease (IBD) are sufficiently vaccinated. Special attention was spent to arguments for vaccine refusal. Methods: Between 1.4.2009 and 30.9.2009, we asked 203 consecutive IBD patients (thereof 57% Crohn’s disease, 63% female; median age 36 years), who got no vaccination advise within the last year to answer a questionnaire with 38 questions. As well, the vaccination cards were adjusted with the official recommendations. Results: Only 83% of patients had a vaccination card. We recognized substantial vaccination deficiencies. Within the past 10 years, only 67% of patients had tetanus and 21% had pertussis vaccination. Only 28% had an influenza vaccination in 2008 and only 9% were ever immunized against pneumococcus. A subgroup analysis of patients with TNF-blockers (n=39) with patients, who never had immunosuppressive therapy (n=67) revealed no difference. 80% of all patients are willing to adhere to all officially recommended vaccinations. 22% and 15% of patients stated that they avoid vaccinations as they afraid side effects or as they assess their immune system as not intact. Nine per cent feared a worsening of IBD after vaccination. Conclusions: In this random sample, the adherence to vaccination recommendations was low. We observed a marked difference between the willingness of IBD patients for immunizations and the realized vaccinations. Our data suggest that an increased medical awareness for vaccination deficiencies in immunosuppressed patients is mandatory.

<p><p>The receptor-recognized form of &alpha;₂-macroglobulin (&alpha;₂M*) targets antigens (Ag) to professional Ag-presenting cells (APCs) for rapid internalization, processing, and presentation. When employed as an Ag delivery vehicle, &alpha;₂M* amplifies major histocompatibility complex (MHC) class II presentation as demonstrated by increased antibody (Ab) titers. Recent evidence, however, suggests that &alpha;₂M*-encapsulation may also enhance Ag-specific cytotoxic T lymphocyte (CTL) immunity. In these studies, we demonstrate that &alpha;₂M*-delivered Ag (ovalbumin, OVA) enhances the production of specific <italic>in vitro</italic> and <italic>in vivo</italic> CTL responses. <br><p>Murine splenocytes expressing a transgenic T cell receptor (TCR) specific for CTL peptide OVA_257-264 (SIINFEKL) demonstrated up to 25-fold greater IFN-&gamma; and IL-2 secretion when treated <italic>in vitro</italic> with &alpha;₂M*-OVA compared to soluble OVA. The frequency of IFN-&gamma; -producing cells was increased ~15-fold as measured by ELISPOT. Expansion of the OVA-specific CD8<super>+</super> T cells, as assayed by tetramer binding and [<super>3</super>H]thymidine incorporation, and cell-mediated cytotoxicity, as determined by a flow cytometric assay, were also significantly enhanced by &alpha;₂M*-OVA. Furthermore, CTL responses were observed at Ag doses tenfold lower than those required with OVA alone. <br><p>We also observed enhanced humoral and CTL responses by naïve mice following intradermal immunization with &alpha;₂M*-OVA. These &alpha;₂M*-OVA-immunized mice displayed increased protection against a subcutaneously implanted OVA-expressing tumor, as demonstrated by delayed tumor growth and prolonged animal survival. The anti-tumor response observed with &alpha;₂M*-mediated Ag delivery was comparable to that of an accepted vaccine adjuvant (CpG 1826) and appeared superior to a cell-based vaccine technique. <br><p>To further understand the mechanism underlying this enhanced CTL immunity, the subsets of professional APCs capable of cross-presenting &alpha;₂M*-encapsulated Ag were investigated. Although both dendritic cells (DCs) and macrophages appear to stimulate some degree of cross-priming in response to &alpha;₂M*-encapsulated Ag, CD8<super>+</super>CD4<super>-</super> and CD8<super>-</super>CD4<super>+</super> DCs appear to do so with the greatest efficiency. The implications of this finding to the ongoing debate regarding the relative contributions of APC subsets to Ag cross-presentation and the determinants of which cells cross-present with high efficiency are discussed. <br><p>These observations demonstrate that &alpha;₂M*-mediated Ag delivery promotes cross-presentation resulting in enhanced Ag-specific CTL immunity. Considered in the context of previous work, these results support &alpha;₂M* as an effective Ag delivery system that may be particularly useful for vaccines based on weakly immunogenic subunits or requiring dose sparing.</p> / Dissertation

No description available.