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La candidose oropharyngée (COP) constitue l’infection fongique opportuniste la plus fréquente chez les patients infectés au VIH-1. Malgré la profonde immunosuppression causée par le VIH-1, l’infection à Candida albicans demeure confinée au niveau de la muqueuse buccale sans dissémination aux organes profonds. La souris transgénique (Tg) CD4C/HIVMut exprimant le génome tronqué du VIH-1 présente, suite à l’inoculation orale de C. albicans, une COP chronique reproduisant fidèlement l’infection chez les patients séropositifs. Cette souris Tg a donc été utilisée afin de déterminer si les macrophages contribuent au confinement de C. albicans à la muqueuse buccale. Cette étude a permis de démontrer que i) les macrophages sont recrutés aux muqueuses buccale et gastrique en réponse au champignon malgré l’expression du transgène, ii) les macrophages de ces souris Tg présentent une polarisation vers un phénotype d’activation alternative et iii) la production de monoxyde d’azote par les macrophages des souris Tg n’est pas requise pour limiter la prolifération de Candida à la muqueuse buccale et pour restreindre sa dissémination aux organes profonds. Les macrophages ne semblent donc pas directement responsables de l’établissement de l’infection chronique à Candida chez la souris Tg CD4C/HIVMut. / Oropharyngeal candidiasis (OPC) is the most frequent opportunistic fungal infection among HIV-infected patients. Despite the profound immunosuppression caused by HIV-1, Candida albicans infection is limited to the oral epithelium and rarely disseminates to deep organs. The CD4C/HIVMut transgenic (Tg) mice, which expresses the truncated HIV-1 genome, developed a chronic OPC after oral inoculation with C. albicans that closely reproduces infection in seropositive patients. Here, we used this Tg mouse to investigate the contribution of macrophages in limiting candidiasis to the oral mucosa. This study shows that i) macrophages are recruited to the oral and gastric mucosa in response to C. albicans despite transgene expression, ii) the macrophages of this Tg mouse exhibited a polarization toward an alternatively activated phenotype and iii) nitric oxide production by these macrophages is dispensable for limiting chronic oral carriage and for preventing systemic dissemination of the fungi in these Tg mice. Overall, these result indicate that macrophage do not directly determine the susceptibility to chronic carriage of Candida in these CD4C/HIVMut Tg mice.

Introduction: Oropharyngeal tumours belong to the most common malignancies situated in the head and neck area. In the past, their incidence has been associated with classical risk factors (smoking and alcohol use). In the recent years, the increase of oropharyngeal carcinomas resulting from HPV (human papilloma virus) infection incidence has been observed. The change of their aetiology also implies a number of clinical differences, the most important being a significantly better survival prognosis in patients with HPV+ carcinomas. Aims: The main goal of this thesis was to characterize correctly patients with oropharyngeal carcinomas and to identify different specifics of nursing intensive care for these patients. Methods: The quantitative prospective study involved 30 patients following their surgical treatment of oropharyngeal carcinoma, hospitalised in an intensive care unit. The patients were divided into 3 groups: HPV+ non-smokers, HPV+ smokers, HPV- smokers. Results: The study confirmed the demographic differences between individual patient groups. The HPV+ non-smokers group is characterized by a lower age in average, which is significantly reflected in a reduced duration of hospital stay. In addition, opioid administration rate, infection rates and postoperative complications incidences are...

La candidose oropharyngée (COP) constitue l’infection fongique opportuniste la plus fréquente chez les patients infectés au VIH-1. Malgré la profonde immunosuppression causée par le VIH-1, l’infection à Candida albicans demeure confinée au niveau de la muqueuse buccale sans dissémination aux organes profonds. La souris transgénique (Tg) CD4C/HIVMut exprimant le génome tronqué du VIH-1 présente, suite à l’inoculation orale de C. albicans, une COP chronique reproduisant fidèlement l’infection chez les patients séropositifs. Cette souris Tg a donc été utilisée afin de déterminer si les macrophages contribuent au confinement de C. albicans à la muqueuse buccale. Cette étude a permis de démontrer que i) les macrophages sont recrutés aux muqueuses buccale et gastrique en réponse au champignon malgré l’expression du transgène, ii) les macrophages de ces souris Tg présentent une polarisation vers un phénotype d’activation alternative et iii) la production de monoxyde d’azote par les macrophages des souris Tg n’est pas requise pour limiter la prolifération de Candida à la muqueuse buccale et pour restreindre sa dissémination aux organes profonds. Les macrophages ne semblent donc pas directement responsables de l’établissement de l’infection chronique à Candida chez la souris Tg CD4C/HIVMut. / Oropharyngeal candidiasis (OPC) is the most frequent opportunistic fungal infection among HIV-infected patients. Despite the profound immunosuppression caused by HIV-1, Candida albicans infection is limited to the oral epithelium and rarely disseminates to deep organs. The CD4C/HIVMut transgenic (Tg) mice, which expresses the truncated HIV-1 genome, developed a chronic OPC after oral inoculation with C. albicans that closely reproduces infection in seropositive patients. Here, we used this Tg mouse to investigate the contribution of macrophages in limiting candidiasis to the oral mucosa. This study shows that i) macrophages are recruited to the oral and gastric mucosa in response to C. albicans despite transgene expression, ii) the macrophages of this Tg mouse exhibited a polarization toward an alternatively activated phenotype and iii) nitric oxide production by these macrophages is dispensable for limiting chronic oral carriage and for preventing systemic dissemination of the fungi in these Tg mice. Overall, these result indicate that macrophage do not directly determine the susceptibility to chronic carriage of Candida in these CD4C/HIVMut Tg mice.

The Gastroesophageal Reflux Disease (GERD) and the symptoms of the Pharyngeal Bolus can make the subject show complaints of impairments of swallowing. The goal of this study was to analyze the dynamics of swallowing in subjects that showed GERD and/or Pharyngeal Bolus through videofluoroscopy in young adults and old people. The reports of 34 subjects between 18 and 85 years old from both genders from the Radiology and Image Service of the Gastroenterological Institute of São Paulo were analyzed and categorized in groups: G1 - with GERD; G2 - with Pharyngeal Bolus; G3 - with Pharyngeal Bolus and GERD; and in sub-groups GA - elderly and GB - adults, with an average age of 74.9 and 43.8 years respectively, in order to study if the effects of ageing would represent an impact on the swallowing physiology. The analyses of the characteristics of the oral an pharyngeal phases of swallowing were carried out in percentages through the spreadsheets and based on the Ott et al. scales, (1996) to the oropharyngeal swallowing dysphagia, in slight, moderate and severe dysphagia -, and based on the Martin-Harris et al. scale, (2007) to the pharyngeal swallowing. The analysis of the pharyngeal phase was realized on the percentages of the most evident characteristics and also with the examination of the statistics of dependant variables. The chi-square test was used, with the level of significance p < 0,05 among the groups, so as to analyze a possible association among the variables. The results showed delay in the control and transportation of the food bolus and intraoral stasis with higher percentage on G3 and in tongue base on G2. There was a predominance of early pharyngeal swallowing in vallecula and piriform recesses in all groups. The laryngeal penetration occurred in all groups, but the incidence was higher in G1 and 1 subject breathed. The cleaning maneuvers were performed in all of them, with the most evident DS in higher levels to the pudding consistency. The harshness degree ranged from normal swallowing to slight dysphagia for all. The study of the relations among the characteristics of the pharyngeal phase presented significant results (p < 0,05) to the relation of the variables beginning of pharyngeal swallowing with laryngeal penetration and the beginning place of the pharyngeal swallowing with the harshness degree of the oropharyngeal dysphagia in young adults. / A Doença do Refluxo Gastroesofageano (DRGE) e o sintoma do Glóbus Faríngeo podem levar o sujeito a apresentar queixas de distúrbios da deglutição. O objetivo do estudo foi analisar a dinâmica da deglutição em sujeitos que apresentaram DRGE e / ou Glóbus Faríngeo por meio da videofluoroscopia em adultos jovens e idosos. Foram interpretados laudos de 34 sujeitos com idade entre 18 e 85 anos, de ambos os sexos, do Serviço de Radiologia e Imagem do Instituto Gastroenterológico de São Paulo, caracterizados como: G1 - Grupo com DRGE; G2 - Grupo com Glóbus Faríngeo; G3 - Grupo com Glóbus Faríngeo e DRGE; e criados os subgrupos GA adultos idosos e GB - adultos jovens com média de idade de 74,9 e 43,8 anos respectivamente, para estudar se os efeitos da idade apresentariam impacto na fisiologia da deglutição. As análises das características das fases oral e faríngea da deglutição foram realizadas em percentuais através de planilhas e baseadas nas escalas de Ott et al. (1996) para disfagias orofaríngeas em disfagia leve, moderada e grave e na escala de Martin-Harris et al. (2007) para a deglutição faríngea. A análise da fase faríngea foi realizada diante dos percentuais das características de maior evidência e analisada a estatística de variáveis dependentes. Foi utilizado o teste Qui - quadrado, com nível de significância p < 0,05 nos grupos, para avaliar possível associação entre as variáveis. Os resultados mostraram atraso no controle e transporte do bolo alimentar e estase intraoral com maior percentual no G3 e em base de língua para o G2. Houve predomínio de início de deglutição faríngea em valécula e recessos piriformes para todos os grupos. A penetração laríngea ocorreu em todos grupos, mas a maior incidência foi em G1. Nas manobras de limpeza a deglutição seca apresentou maior evidência em níveis mais elevados para a consistência pudim. O grau de severidade variou entre deglutição normal e disfagia leve para todos os grupos estudados. O estudo da relação entre as características da fase faríngea apresentou resultados significativos (p < 0,05) para a relação das variáveis início de deglutição faríngea com penetração laríngea e com grau de severidade das disfagias orofaríngeas para todos os grupos.

Introduction: Human papillomavirus (HPV) is changing the prevalence, survival and treatment paradigms in oropharyngeal squamous cell carcinoma (OPSCC). Improved survival of patients with HPV positive compared to HPV negative OPSCC has led to trials of treatment de-escalation. Current HPV detection methods are imprecise, therefore standardised assessment of transcriptionally active HPV in OPSCC is required. Furthermore, the differences in immune characteristics and/or the hypoxia response/effects could explain observed differences in prognosis between HPV positive and negative OPSCC. Rigorous HPV detection and subsequent biomarker evaluation should provide additional information required before introduction of treatment de-escalation in broad patient groupings. Methods: The study cohort was 218 patients with OPSCC who received radiotherapy with curative intent. HPV status was determined on pre-treatment, formalin-fixed paraffin-embedded blocks using: 1) polymerase chain reaction (PCR); 2) in-situ hybridisation (ISH) and 3) immuno-histochemistry (IHC). QuantiGene multiplex assay was designed to detect mRNA of reference sequences of the common high-risk HPV types (16, 18, 33, 35, 45, 52 and 58). HPV detection methods were compared with mRNA quantification. Multimarker IHC of immune cell markers using chromogenic and fluorescent staining was performed, analysed and compared with single marker IHC using automated multispectral image analysis. A validated multiplex IHC method was used for a) chromogenic (CD3, CD4, CD8, and FoxP3) and b) fluorescent (CD8, CD68 and PD1/PD-L1) evaluation in tumour and stroma compartments. Single marker IHC was used to investigate tumour hypoxia markers (HIF-1α and CA-IX) in HPV positive and negative OPSCC. Results: p16 IHC and ISH were the most sensitive and specific, respectively, for classifying HPV status. The combination of the three tests had the highest positive/negative predictive values compared with QuantiGene mRNA detection. Multiplex validation showed that, for serial sections up to 6 μm apart, there were highly significant correlations (P<0.0001) between single and multiplex counts for both chromogenic and fluorescent IHC. Overall there was less variation in cell counts with fluorescent staining when compared to chromogenic staining. Multiplex IHC of TILs in HPV positive and negative OPSCC showed higher infiltration in both tumour and stromal areas of CD3+CD4+ and CD3+CD8+ T cells but not CD4+FoxP3 Tregs in HPV positive compared with HPV negative OPSCC. Only CD3+CD8+ stromal and not tumour area infiltration was associated with increased survival (P=0.02). PD-L1 expression was higher in HPV negative OPSCC and this was related to macrophage (CD68) expression of PD-L1. In HPV negative tumours infiltration with CD68+PD-L1 was associated with a good prognosis. HPV negative patients had higher expression of HIF-1α but not CA-IX. High expression of both markers was associated with a poor prognosis irrespective of HPV status. Conclusions: There are other prognostic factors operating in the larger subdivision of HPV positive and negative OPSCC. Precise HPV detection and inclusion of other prognostic factors is required before treatment de-escalation is used. Expression of immune inhibitory factors (PD1/PD-L1) alone without contextualisation with immune cell density is insufficient for patient prognostication and potential selection for therapy using immune checkpoint inhibitors. Hypoxia modification of radiotherapy should be explored in both HPV positive and negative OPSCC.

Only a few individual cells within less than 5% of all primary tumors form the cell lines commonly used in cancer research. These growth bottlenecks result in cell lines that are often poor models of primary tumors. Co-culture of primary tumor-derived cells with an irradiated mouse fibroblast feeder layer and ROCK inhibitor, known as the Georgetown Method, offers a way to culture over 80% of tumor-derived cells in vitro to create more representative tumor cell models. In our studies, we optimized the Georgetown Method to culture head and neck cancer cells, including oropharyngeal squamous cell carcinoma, and investigated its mechanism of conditionally immortalizing cells in culture. Differential trypsinization and regular feeder layer replacement were found to significantly improve the efficacy of immortalizing co-cultured cells at both atmospheric and physiological oxygen levels. Medium conditioned by irradiated fibroblasts can also substitute for direct co-culture with a feeder layer. The Georgetown Method was found to maintain low levels of p16 in co-cultured cells, suggesting a potential mechanism by which the Georgetown Method prevents differentiation and senescence. Our ability to culture over 80% of primary tumor-derived cells allows us to test the translational value of tumor-derived cell cultures and xenografts using BH3 profiling. Conditioned medium simplifies maintenance of cell cultures and will also allow us to perform high-throughput screens without the need to separate tumor-derived cells from the fibroblast feeder layer. The Georgetown Method provides opportunities to expand small tissue specimens for future diagnostics, therapeutics, and biobanking.

Bakgrund. Vid tumörresektion i de orofaryngeala vävnaderna, utförs mandibulotomi för att skapa åtkomst och möjliggöra kirurgiskt avlägsnande. Allvarliga komplikationer har rapporterats i litteraturen och bidragande orsaker är den kirurgiska tekniken, strålningsterapin samt en inskränkt näringstillförsel till mandibelkroppen.Syfte. Syftet med denna studie var att undersöka om bevarande av periost vid utförande av mandibulotomi, leder till färre patienter med komplikationer relaterade till mandibulotomin..Metod. Patienter som genomgick mandibulotomi mellan 2003-2012 vid Skånes Universitetssjukhus granskades retrospektivt. Trettiosex patienter inkluderades fördelade på 18 i en supraperiostal- (locking plates) och 18 i en subperiostal grupp (non-locking plates). Den kliniska uppföljningen var 12 månader och den röntgenologiska varierade mellan 10 till 17 månader efter mandibulotomin. Komplikationerna delades in i mindre allvarliga; abscess, fistel och gingival nekros samt mer allvarliga; osteoradionekros, ben- och plattblotta, non-union och mikrovaskulärlambåinfektion.Resultat. Antalet patienter med komplikationer under 12 månaders uppföljning var 14 (38,9 %). Subperiostala gruppen hade en incidens på 9 fall medan den supraperiostala hade en incidens på 5. Antalet patienter som uppvisade komplikationer vid 12 månaders uppföljning var 9 (25 %), med endast en patient representerad i den supraperiostala gruppen.Slutsats. Enligt denna retrospektiva studie har patienter, som genomgått mandibulotomi med supraperiostal placering av osteosyntesplattorna, färre persisterande komplikationer av allvarlig grad efter ett år, jämfört med en subperiostal placering. Det förefaller därför fördelaktigt att använda supraperiostal placeringsteknik, när man applicerar osteosyntesplattor efter mandibulotomi på strålade patienter. / Background. Mandibulotomy is frequently used to gain access at resection of tumours in the oropharyngeal space. Severe complications have been reported in the literature related to the surgical technique, radiation and nutritional deficiency of the mandibular body.Objective. The purpose of this study is to investigate if preserving the periosteum, while performing a mandibulotomy, leads to fewer patients with complications related to the mandibulotomy.Method. Patients undergoing mandibulotomy between 2003 and 2012 at Skåne University Hospital were reviewed retrospectively. Thirty six patients were included, 18 in the supraperiosteal group (locking plates) and 18 patients in the subperiosteal group (non-locking plates). Clinical follow up was 12 months and radiographic follow up was 10 to 17 months post-surgery. Complications were divided into minor; abscess, fistula, gingival necrosis and major; osteoradionecrosis, bone and plate exposure, non-union and micro vascular flap infection.Results. The summation of patients with complications during the 12 months follow up was 14 (38 %). The subperiosteal group was represented in 9 cases and the supraperiosteal group in 5. The total number of patients with complications persisting at 12 months after the operation were 9 (25 %), with only one patient represented in the supraperiosteal group.Conclusion. Fewer patients, undergoing mandibulotomy with the supra- compared to the subperiosteal placement of the osteosynthetic plates, showed persisting complications at one year postoperatively (p <0.05). It seems to be preferable to use a supraperiosteal fixation method when placing the osteosynthetic plates after a mandibulotomy in irradiated patients.

La candidose oro-pharyngée (COP) est l’infection opportuniste la plus répandue chez les patients infectés au VIH-1. Un modèle de COP chez la souris transgénique (Tg) exprimant une partie du génome du VIH-1 (CD4C/HIVMutA) est maintenant disponible. Grâce à ce modèle, il est possible d’étudier les perturbations quantitatives et fonctionnelles des macrophages exprimant les gènes nef, rev et env du VIH-1 dans le contexte d’une COP. Cette étude démontre que la présence du transgène n’influence pas le pourcentage des macrophages dans la muqueuse buccale et le petit intestin, malgré le fait que la charge buccale de C. albicans soit significativement plus élevée chez les souris Tg. Cependant, l’expression du transgène cause une diminution de la production de H2O2 par les macrophages, ainsi que l’augmentation de la production de la cytokine proinflammatoire IL-6 et de la chimiokine MCP-1. / Oro-pharyngeal candidiasis (OPC) is the most common opportunistic infection in HIV-1 infected patients. An OPC model using transgenic mice (CD4C/HIVMutA) expressing selected genes of the HIV-1 genome is now available. Using this model, it is now possible to study potential quantitative and functional disturbances in macrophages expressing the nef, rev and env genes of HIV-1 in the context of OPC. This study shows that transgene expression does not affect quantitative percentage values of macrophages in the oral mucosa and the small intestine, although burdens of C. albicans loads are increased in Tg mice. Transgene expression does induce diminished H2O2 production in macrophages, while increasing production of the proinflammatory cytokine IL-6 and the chemokine MCP-1.