The eosinophil cationic protein (ECP) is usually associated with the eosinophil granulocyte. In this thesis the presence and production of this protein has been studied in two other cells. The circulating monocyte was found to contain ECP mRNA and small amounts of ECP, one thousand times less than that found in the eosinophil. The production decreased by differentiation of the myelomonoblastic cell line U937 into a macrophage phenotype. Submucosal lung macrophages did not stain for ECP and alveolar macrophages did not contain ECP mRNA. The circulating neutrophil contains ECP at a level hundred fold less than the eosinophil. We found that the protein is located to the primary granules of the neutrophil but could detect no ECP mRNA in the cell. It was shown in vitro that the protein was taken up by the cell and partly transported to the primary granules. The uptake did not seem to be receptor mediated. Upon stimulation of the neutrophils, ECP previously taken up, was re-secreted. The ECP protein is heterogeneous both to molecular characteristics and to function. To evaluate if a genetic component is involved, the ECP gene was analysed in 70 individuals. Three single nucleotide polymorphisms (SNP´s) were found, denoted 277(C>T), 434(G>C) and 562(G>C). The two first were located to the mature peptide-coding region and would change the amino acids, arg45cys and arg97thr. The prevalence of the most common SNP, 434, was evaluated in two eosinophil-related diseases, allergy/asthma and Hodgkin Lymphoma (HL). Forty-three HL patients were evaluated and it was found that the 434GG was significantly more prevalent in patients having nodular sclerosis (NS) as compared to other histologies (p=0.03). Erythrocyte sedimentation rate was also related to the 434GG genotype (p=0.009). In 209 medical students 434GG was more common (p=0.002) in those who indicated allergy. The genotype was unrelated to the production of IgE antibodies to allergens. In analysis of 76 subjects with asthma it was found that the 434GG genotype was significantly more common among allergic asthmatics (p=0.04). Asthma and HL-NS are characterized by fibrosis and eosinophils and ECP has been suggested in fibrosis development.

Lymphomas are a heterogeneous group of neoplasias originating from B- or T-lymphocytes. In this thesis, we determined the genetic and immunophenotypic characterization of DLBCL and their prognostic impact. Moreover, genomic alterations associated with the transformation to DLBCL from Hodgkin lymphoma (HL) and follicular lymphoma (FL) were elucidated. In order to outline the impact of cytogenetic as well as immunophenotypic prognostic markers in DLBCL, we firstly studied a series of 54 DLBCL tumors using comparative genomic hybridization (CGH) and we identified several frequently occurring chromosomal imbalances. Loss of 22q was more often found in the diagnostic tumors with a more advanced clinical stage, while gain of 18q21 was more commonly identified in relapses. Secondly, we correlated the expression patterns of CD10, bcl-6, IRF-4 and bcl-2 with clinical parameters in a series of 173 de novo DLBCL patients. Patients with a germinal center (GC) phenotype displayed a better survival than the non-GC group. Expression of bcl-6 and CD10 was correlated with a better survival while bcl-2 expression was associated with a poor prognosis. In approaching the HL transformation, two novel B-cell lines (U-2932 and U-2940), derived from patients with DLBCL following HL, were characterized. Interestingly, a translocation with materials from 2q and 7q as well as loss of material on 6q was found in both cell lines. For FL transformation, we assessed chromosomal alterations in a panel of 28 DLBCL patients with a previous history of FL. The DLBCL tumors displayed more chromosomal imbalances compared to FL tumors. Loss of 6q16-21 and gain of 7pter-q22 were more commonly found in the DLBCL counterparts, suggesting the chromosomal location of putative genes that may be involved in the transformation process.

Mast cell (MC) accumulation around tumours is an old observation gaining new relevance due to the multifaceted nature of MCs and their many roles in immunity, beyond allergy. Knowledge about tumour specific recruitment of, and interactions with, MCs is needed to unravel the function of their presence. This study investigates the participation of mast cells in the tumourigenesis of Hodgkin lymphoma (HL), a tumour with many inflammatory features. We report that MC recruitment into HL lymphomatous tissue is possibly due to the production of CCL5/RANTES by malignant Hodgkin and Reed-Sternberg (HRS) cells. In addition, increased levels of IL-9, a cytokine implicated in mast cell heterogeneity and as an autocrine growth factor for HRS cells, were found in HL patient sera and correlate with negative prognostic factors. The ubiquitous expression of CD30 by HRS cells has been implicated in HL tumour development. In HL tissue MCs were found to be the predominant CD30 ligand (CD30L) expressing cells, and through CD30L/CD30 engagement they induced a proliferative response in HRS cells. This interaction proved to be bi-directional as it induced a degranulation-independent de novo synthesis of a specific set of chemokines in MCs, including IL-8. This novel trigger of MC activation is suggested to be of importance also in atopic dermatitis (AD) and psoriasis since increased numbers of CD30L and IL-8 positive MCs were detected along with increased expression of CD30. Data presented in this study supports a specific recruitment of MCs into HL tumours and co-operative interactions between HRS cells and MCs. Our identification of reversed signalling via CD30L as a novel MC trigger provides a mechanism behind leukocyte infiltration and chronic development in diseases associated with CD30 and MCs, such as HL, AD and psoriasis.

INTRODUCCIÓ: Les neoplàsies limfoides agrupen un conjunt de malalties que, tot i compartir certes característiques comunes, presenten una gran heterogeneïtat en la seva biologia i manifestacions clíniques. En la present tesi s´han estudiat els mecanismes moleculars implicats en el desenvolupament i progressió de diferents entitats que s´inclouen dins del grup dels Limfomes No-Hodgkin de cèl·lula B (NHL, de l´anglès Non-Hodgkin Lymphoma), especialment el limfoma de cèl·lules del mantell (MCL, de l´anglès Mantle Cell Lymphoma) i la leucèmia limfàtica crònica (CLL, de l´anglès Chronic Lymphocytic Leukemia), els quals són limfomes de cèl·lula B petita. MATERIAL I MÈTODES: En aquest treball s´han emprat una gran diversitat de tècniques experimentals, de les que destaquen la hibridació genòmica comparada (CGH, de l´anglès Comparative Genomic Hybridization) per estudiar les alteracions cromosòmiques dels tumors, la reacció en cadena de la polimerasa (PCR, de l´anglès Polymerase Chain Reaction) i la cromatografia líquida desnaturalitzant (DHPLC, de l´anglès Denaturing High Perfomance Liquid Chromatography) per determinar l´adquisició d´alteracions en el material genètic a nivell de DNA; així com la PCR quantitativa a temps real (qPCR) i la realització de microarrays d´oligonucleòtids d´alta densitat per analitzar l´expressió gènica a nivell de mRNA.RESULTATS I CONCLUSIONS: L´estudi de gens implicats en la proliferació de MCL ha permès la construcció d´un model predictor de la supervivència dels pacients basat en l´expressió dels cinc gens RAN, MYC, TNFRSF10B, POLE2 i SLC29A2. Aquest model es pot aplicar tant en material congelat com en teixits biològics fixats amb formol i inclosos en parafina (FFPE, de l´anglès Formalin-Fixed and Paraffin-Embedded tissue), podent ésser útil en la presa de decisions terapèutiques1. També s´ha estudiat el gen MDM2, implicat en les vies de resposta al dany del DNA. S´ha observat que, a part de les alteracions del gen p53, l´augment de l´expressió gènica de MDM2 es correlaciona directament amb una disminució de la supervivència en MCL. En alguns casos aquest augment de l´expressió pot donar-se per guany del locus genòmic del gen, mentre que la presència del SNP309 al seu promotor no es relaciona amb els canvis d´expressió (2).La desregulació dels mecanismes implicats en la mort cel·lular programada o apoptosi també juga un paper clau en els processos de tumorogènesi. S´ha observat que els receptors de mort TNFRSF10A i TNFRSF10B es troben poc mutats en neoplàsies limfoides, indicant que la seva alteració no és rellevant per explicar la resistència a l´apoptosi observada en aquests tipus de limfomes. Tot i això, la presència del polimorfisme A1322G al domini de mort de TNFRSF10A s´associa amb un augment de risc a patir MCL i CLL; mentre que la presència del polimorfisme C626G al domini d´unió a lligand de TNFRSF10A sembla jugar un paper protector en MCL; suggerint un possible paper de dits polimorfismes en la resistència a la mort mediada per TRAIL (3).Finalment, l´estudi global de les alteracions que participen en els fenòmens de progressió clínica primerenca de la CLL ha permès determinar la modulació de l´expressió d´un petit grup de gens (58) que participen en diferents vies cel·lulars. Destaca un subconjunt de gens que actuen en la inhibició de l´adhesió i motilitats cel·lulars, els quals presenten una disminució de la seva expressió. També s´ha observat que la progressió clínica de les CLL s´associa, en alguns casos, amb la inactivació de certs gens supressors de tumors. Per altra banda, l´evolució primerenca d´aquest tips de leucèmia sembla implicar poca inestabilitat cromosòmica (4).ARTICLES GENERATS PER LA PRESENT TESI:(1). Hartmann E, Fernandez V, Moreno V, Valls J, Hernandez L, Bosch F, Müller-Hermelink HK, Ott G, Rosenwald A, Campo E. A five-gene model to predict survival in mantle cell lymphoma and its application to formalin-fixed paraffin-embedded tissue. J Clin Oncol, under revision (Journal impact factor 2006: 13.598).(2). Hartmann E, Fernandez V, Stoecklein H, Hernández L, Campo E, Rosenwald A. Increased MDM2 expression is associated with inferior survival in mantle cell lymphoma, but not related to the MDM2 SNP309, Haematologica, 92:574-575 (Journal impact factor 2006: 5.032).(3). Fernandez V, Jares P, Bea S, Salaverria I, Guino E, de Sanjose S, Colomer D, Ott G, Montserrat E, Campo E. Frequent polymorphic changes but not mutations of TRAIL receptors DR4 and DR5 in mantle cell lymphoma and other B-cell lymphoid neoplasms. Haematologica. 2004 Nov;89(11):1322-31 (Journal impact factor 2006: 5.032).(4). Fernandez V, Jares P, Salaverria I, Giné E, Beà S, Aymerich M, Colomer D, Villamor N, Bosch F, Montserrat E, Campo E. Gene expression profile and genomic changes in disease progression of early-stage chronic lymphocytic leukemia. Haematologica, accepted (Journal impact factor 2006: 5.032). / "ANALYSIS OF THE MOLECULAR MECHANISMS IMPLIED IN THE DEVELOPMENT AND PROGRESSION OF SMALL B CELL LYMPHOMAS" INTRODUCTION:Different molecular mechanisms implied in the development and progression of distinct subtypes of small B cell lymphomas that belong to Non-Hodgkin´s Lymphomas (NHL) have been studied in the present work, specially focusing on Mantle Cell Lymphoma (MCL) and Chronic Lymphocytic Leukemia (CLL).MATERIALS AND METHODS:Comparative genomic hybridization (CGH) has been aplied to study the chromosomal alterations of the tumors. Polymerase chain reaction (PCR) and denaturing high performance liquid chromatography (DHPLC) techniques have been performed to detect nucleotide alterations in certain genes; whereas real time quantitative PCR (qPCR) and high density oligonucleotide microarrays have been used to analyse gene expression.RESULTS AND DISCUSSION:The gene expression model composed of the five genes RAN, MYC, TNFRSF10B, POLE2 and SLC29A2 allows the survival prediction of MCL patients with widely disparate clinical outcome and is superior to the immunohistochemical marker Ki-67. The predictor is applicable to fresh frozen and formalin-fixed paraffin-embedded (FFPE) tumor samples. On the other side, the study of the MDM2 gene has shown that increased gene expression correlates with inferior survival in MCL. MDM2 overexpression is associated with copy number gains of the MDM2 locus in single tumors, but not with the recently reported MDM2 promoter SNP309.The study of the death receptors TNFRSF10A and TNFRSF10B has shown that mutations in these genes are uncommon in lymphoid neoplasms, but the presence of certain TNFRSF10A polymorphisms (A1322G in the death domain and C626G in the extracellular binding domain) can contribute to the pathogenesis of these malignancies. The study of early clinical progression in CLL has allowed the identification of a significant modulation of the gene expression of 58 genes, with a particular downregulation of genes that are inhibitors of cell adhesion and motility. On the other side, our results indicate that clinical progression of early stage CLL is associated with karyotype evolution and inactivation of certain tumor suppressor genes.

Linfomas são cânceres que se iniciam a partir da transformação maligna de um linfócito no sistema linfático. Os linfomas são divididos em duas categorias principais: os linfomas de Hodgkin e todos os outros linfomas, denominados linfomas não-Hodgkin (LNH). Os pacientes com LNH são comumente tratados com radioterapia apenas ou combinada com quimioterapia utilizando-se de anticorpo monoclonal anti-CD20, principalmente o rituximab (MabThera&reg). O uso de anticorpos monoclonais (Acm) conjugados à quelantes bifuncionais radiomarcados com radionuclídeos metálicos ou lantanídeos é uma realidade de tratamento para portadores de LNH pelo princípio de radioimunoterapia (RIT). Este estudo concentrou-se nas condições de conjugação do anticorpo monoclonal rituximab (MabThera&reg) com grupamentos quelantes bifuncionais DOTA e DTPA. Na marcação dos Acm conjugados com lutécio-177, foram estudadas as condições de pré-purificação do Acm, condições de conjugação, determinação de número de quelantes acoplados à molécula do anticorpo, purificação do anticorpo conjugado, radiomarcação do anticorpo conjugado, com lutécio-177, purificação do anticorpo marcado, a ligação específica in vitro dos compostos marcados às células Raji, e distribuição biológica em camundongos BALB/c sadios. As três metodologias empregadas na pré-purificação do anticorpo (diálise, cromatografia de exclusão molecular com coluna Sephadex G-50 e ultrafiltração) demonstram-se eficientes e proporcionaram recuperação da amostra superior a 90%. A metodologia de ultrafiltração foi considerada a mais simples e prática, podendo ser aplicada a procedimentos rotineiros de produção de radiofármacos. Além disso, proporcionou a recuperação final de amostra de 97% em microlitros. Nas conjugações do anticorpo com os quelantes DOTA e DTPA em razões molares diferentes do Acm:quelante, observou-se número de grupamentos quelantes acoplados à molécula do Acm proporcional à razão molar estudada. Quando foi avaliada a influência de condições diferentes de conjugação no número de quelantes acoplados à molécula do Acm, não foram observadas diferenças significativas, com resultados de pureza radioquímica (PR) inferior a 80% em todas as condições estudadas. Na comparação de métodos de purificação do Acm conjugado, a abordagem inédita apresentada neste estudo, na qual a cromatografia de exclusão molecular foi combinada com a ultrafiltração resultou em maior eficiência na purificação e preservação da estrutura do anticorpo. Nos estudos de radiomarcação do anticorpo conjugado com DOTA e DTPA, os imunoconjugados de DTPA apresentaram, de forma geral, maior eficiência de marcação com resultados reprodutíveis quando comparados com os imunoconjugados de DOTA, considerando-se as diferentes razões molares utilizadas. As metodologias cromatográficas empregadas no controle de pureza radioquímica do composto radiomarcado proporcionaram a discriminação das diferentes espécies radioquímicas no meio de marcação. A metodologia de purificação do composto conjugado e radiomarcado utilizada proporcionou a obtenção de compostos com alta pureza radioquímica, 97,4&plusmn;1,3% (DOTA 1:50) e 98,7&plusmn;0,2% (DTPA 1:50). Nos estudos de ligação específica às células tumorais Raji, o anticorpo conjugado com quelante DTPA nas razões molares de 1:50 e 1:20 apresentaram perfil semelhante de ligação, com aumento da porcentagem de ligação específica proporcional à concentração celular, enquanto que o imunoconjugado na razão molar de 1:10 apresentou alta porcentagem de ligação não específica. Os resultados obtidos nos estudos de biodistribuição in vivo do anticorpo conjugado e radiomarcado nem sempre se mostraram compatíveis com a biodistribuição de anticorpos radiomarcados íntegros. No caso do quelante DOTA, o imunoconjugado obtido a partir da razão molar 1:20, apresentou melhores características de biodistribuição. No caso do quelante DTPA, a razão molar utilizada pareceu refletir diretamente no clareamento sanguíneo do anticorpo e todas as razões molares utilizadas apresentaram instabilidade in vivo. / Lymphomas are malignancies or cancers that start from the malign transformation of a lymphocyte in the lymphatic system. Lymphomas are divided in two major categories: Hodgkin lymphoma and non-Hodgkin lymphoma (NHL). Patient with NHL are generally treated with radiotherapy alone or combined with immunotherapy using monoclonal antibody rituximab (MabThera&reg). Currently, monoclonal antibodies (Mab) conjugated with bifunctional chelate agents and radiolabeled with metallic or lanthanides radionuclides are a treatment reality for patients with NHL by the principle of radioimmunotherapy (RIT). This study focused on the conditions of conjugation of Acm rituximab (MabThera&reg) with bifunctional chelating agents DOTA and DTPA and labeling with 177-luthetium. Various parameters were studied: method of Acm purification, conditions of Acm conjugation and the determination of the number of chelate coupled to the Acm, the purification of the conjugated Mab, labeling conditions with lutetium-177, purification of the radiolabeled immunoconjugate, radiochemical purity (RP), in vitro specific binding determination to Raji cells (Human Burkitt) and biological distribution performed in normal BALB/c mouse. The three methodologies employed in pre-purification of Acm (dialysis, size exclusion chromatograph and ultrafiltration) demonstrated to be efficient; they provided sample recovery exceeding 90%. However, the methodology of ultrafiltration resulted in greater sample recovery and in microliters. The number of chelate attached to the Mab molecule was proportional to the molar ratio studied. When the influence of different conditions of conjugation in the number of chelate bounded to the Mab was studied, no notable differences were observed. The RP < 80% was observed in all the methods applied. Purification of the conjugated antibody by different methods showed that the innovative combination of Sephadex and ultrafiltration methods resulted in higher efficiency of purification. The optimized conditions for purification of the conjugated antibody preserved the protein integrity. Radiolabelling studies of DOTA and DTPA immunoconjugated showed that DTPA derivatives presented, in general, radiochemical yield superior than DOTA conjugated Mab, considering the different molar ratios studied. The chromatographic methods employed in the RP determination were efficient to separate the different radiochemical species presented in the reaction medium. The methodology used in the purification of the labeled Mab resulted in labeled compounds with high radiochemical purity, 97.4&plusmn;1.3% (DOTA 1:50) and 98.7&plusmn;0.2% (DTPA 1:50). Considering specific cell binding assays (Raji cells), the Mab conjugated to DTPA at 1:50 and 1:20 molar ratios presented similar results, and the percent of cell binding were proportional to the cell concentration, whereas the cell binding for 1:10 molar ratio showed high percent of nonspecific cell binding. The results of in vivo biodistribution studies of labeled Mab not always were compatible with the biodistribution of intact radiolabelled antibody. The DOTA immunoconjugated produced at 1:20 molar ratio, showed better performance in biodistribution studies. In the case of DTPA immunoconjugated, the blood clearance seems to be influenced by the molar ratio applied and the immunoconjugated produced with DTPA chelate at different molar ratio resulted in high in vivo instability compounds.

Abstract Hodgkin lymphoma (HL) is among a heterogeneous group of lymphomas. Over 80% of all patients can be cured with chemo- and radiotherapy. HL has become a model to study long-term effects of radio- and chemotherapy, because of the excellent prognosis. There are a significant number of patients who suffer or die because of the treatment-related long-term toxicity. The aim of this work was to discover new possible biological factors to predict poor prognosis and offer new aspects to individualize patient treatment in a convenient manner in HL. The retrospective study involved HL patients uniformly treated in 1997–2015. Immunohistochemistry was used to determine the expression of various biological markers, including oxidative stress markers 8-hydroxydeoxyguanosine (8-OHdG) and nitrotyrosine and the antioxidant enzymes manganese superoxide dismutase (MnSOD) as well as peroxiredoxins (Prx II, Prx III, Prx V, Prx VI) in HL patient samples. Using immunohistochemistry, we also evaluated expression of hypoxia-inducible factors (HIF-1&#945;, HIF-2&#945;), prolyl hydroxylase domain enzymes (PHD1, PHD2, PHD3), the epigenetic regulator lysine (K)-specific demethylase 4 (KDM4A, KDM4B, KDM4D) as well as sirtuins (SIRT1, SIRT4, SIRT6), the DNA-repair proteins Human Rap1 interacting factor 1 (Rif1) and O6-alkylguanine DNA alkyltransferase (MGMT) from representative classical Hodgkin lymphoma (cHL) patient samples. Low-level expression of 8-OHdG was associated with poorer relapse-free survival (RFS) in advanced-stage HL and a high extent of MnSOD predicted early relapse in the whole HL cohort. Strong expression of PHD1, KDM4B and KDM4D predicted dismal RFS in radiotherapy-treated cHL patients. The results also showed that strong expression of HIF-1&#945;, SIRT6 and Rif1, and SIRT6 together with Rif1, were associated with prolonged RFS, especially in advanced-stage radiotherapy-treated cHL patients. In multivariate analysis, PHD1, MnSOD, 8-OHdG and Rif1 separately and together with SIRT6 were statistically significant predictors of RFS. The results reflect the significance of the studied biomarkers in HL, especially in radiotherapy-treated patients. This might be beneficial when individualizing treatment strategies, avoiding overtreatment and controlling long-term treatment-related toxicity. Further research, however, is needed to confirm these preliminary findings. / Tiivistelmä Hodgkinin lymfooma (engl. HL) kuuluu heterogeeniseen imukudossyöpien eli lymfoomen ryhmään. Yli 80 % lymfoomapotilaista voidaan parantaa solunsalpaaja- ja sädehoidon avulla. Hyvän ennusteen takia HL- tutkimuksen tärkeä painopiste on säde- ja solunsalpaajahoidon pitkän ajan haittavaikutukset. Huomattava määrä potilaista kärsii tai jopa kuolee hoitoon liittyvistä pitkäaikaishaitoista johtuen. Tämän tutkimuksen tarkoituksena oli löytää uusia mahdollisia biologisia tekijöitä, jotka ennakoisivat taudin huonoa ennustetta ja samalla antaa uusia näkökulmia HL potilaiden hoidon yksilöllistämiseen. Tämä retrospektiivinen tutkimus käsitti vuosina 1997-2015 samanlaisesti hoidettuja Hodgkinin lymfooma -potilaita. Immunohistokemiallisilla värjäyksillä määritettiin biologisten merkkiaineiden, mukaan lukien oksidatiivisen stressin markkereiden 8- hydroksideoksiguanosiinin (8-OHdG) ja nitrotyrosiinin, sekä antioksidanttientsyymien mangaanisuperoksidi-dismutaasin (MnSOD) sekä peroksiredoksiinien (Prx II, Prx III, Prx V, Prx VI) ilmentymistä HL -potilasnäytteissä. Määrittelimme myös immunohistokemiallisilla värjäyksillä epigeneettisten säätelijöiden lysiinin spesifisen demetylaasientsyymin 4 (KDM4A, KDM4B, KDM4D) sekä sirtuiinien (SIRT1, SIRT4, SIRT6), hypoksiaa indusoivien tekijöiden (HIF-1&#945;, HIF-2&#945;), prolyylihydroksylaasientsyymien (PHD1, PHD2, PHD3) ja DNA:ta korjaavien proteiinien Rap1 vaikuttuvan tekijä 1 (Rif1) ja O6-metyyliguaniini-DNA metyylitransferaasin (MGMT) ilmentymistä edustavissa klassista Hodgkinin lymfoomaa sairastavien potilaiden (engl. cHL) näytteissä. Heikko 8-OHdG värjäytyminen ennusti ennenaikaista taudin uusiutumaa levinneessä HL:ssa ja korkea MnSOD ilmaantuvuus ennusti ennenaikaista taudin uusiutumaa koko HL -ryhmässä. Sädehoidetuilla cHL potilailla voimakas PHD1, KDM4B ja KDM4D värjäytyminen ennusti ennenaikaista taudin uusiutumaa. Tulokset osoittivat myös, että erityisesti sädehoidetuilla levinneen taudin cHL potilailla voimakas HIF-1&#945;, SIRT6, Rif1 ja SIRT6 yhdessä Rif1:n kanssa oli yhteydessä pidentyneeseen uusiutumavapaaseen aikaan. Monimuuttuja-analyysissä PHD1, MnSOD, 8-OHdG ja Rif1 itsenäisenä ja yhdessä SIRT6 kanssa ennustivat tilastollisesti merkitsevästi taudin ennenaikaista uusiutumaa. Tulokset osoittavat näiden eri biomarkkereiden merkittävyyden HL:ssä, erityisesti sädehoitoa saaneilla potilailla. Tuloksista voi olla hyötyä, kun hoitokäytäntöjä yksilöidään, mikä voisi helpottaa välttämään liiallista hoitoa ja hallitsemaan pitkäaikaisiin hoitoihin liittyviä haittoja. Näiden alustavien havaintojen vahvistamiseksi tarvitaan kuitenkin lisätutkimuksia.

La survie des lymphomes non hodgkiniens (LNH) en population générale s'est améliorée durant la dernière décennie. Pourtant, des disparités persistent, suggérant le rôle de certains facteurs comme les facteurs socio-économiques et des inégalités dans l’accès ou la qualité des soins.Entre 2002 et 2008, 1798 LNH diffus à grandes cellules B et folliculaire ont été diagnostiqués dans 3 registres spécialisés en hématologie (Basse Normandie, Côte d’Or et Gironde). De fortes inégalités territoriales entre les 3 zones registres sont observées concernant le lieu de leur prise en charge, les délais d'initiation au traitement ou la survie, mais aucune association entre le score de défavorisation (EDI) et ces trois critères n'a été mise en évidence.La survie s'est améliorée durant la période d'étude. Cette tendance positive peut être expliquée par l’usage de l'immunothérapie en 1ère ligne. La zone géographique de diagnostic,la spécialité médicale (onco-hématologie vs autres) sont indépendamment associées à une meilleure survie à 5 ans quel que soit l’âge. Enfin, l'amélioration de la survie chez les patients âgés (75-84 ans en particulier) peut s'expliquer par un bilan initial plus complet (TEP scan entrainant un « upstaging ») suivi de traitements plus agressifs.Le temps de déplacement était associé au lieu de prise en charge et à la survie, avec un pronostic défavorable des patients résidant à plus de 15 minutes du centre de référence le plus proche.Malgré les avancées thérapeutiques, de nombreux facteurs non biologiques peuvent affecter le pronostic des patients atteints de LNH. L'expertise des équipes prenant en charge ces maladies semble primordiale pour obtenir une prise en charge optimale. / Due to the addition of innovative treatment, survival of non-Hodgkin lymphoma (NHL) increased during the last decade. Nevertheless, disparities persist, suggesting the role of certain factors as socio-economic factors and disparities in the access or the quality of healthcare.Between 2002 and 2008, 1798 Diffuse Large B-cell (DLBCL) and follicular lymphomas werediagnosed in 3 hematological malignancies specialized registries (Basse-Normandie, Côted'Or and Gironde). Important territorial disparities between the 3 registries areas were observed regarding the place of care, the delay of treatment initiation or the survival whatever the age while there was no association with the deprivation score (EDI).The survival improved during the study period. This positive trend could be explained by the use of immunotherapy as 1st line therapy. The geographical area where the patient was diagnosed, the medical specialization (onco-hematology vs others departments) are independently associated with a better 5-years survival whatever the age.Finally, the improvement of the survival in elderly (especially 75-84 years) could be explained by better work up (higher TEP scan use leading to "upstaging") and thus to use more aggressive therapies. Also, the travel time was associated with the place of care and the survival, with a poorer prognosis for patients living more than 15 min from the closest reference center. Despite therapeutic advances, various non biological factors can affect the prognosis ofpatients with lymphomas. The notion of lymphoma-specific expertise seems to be essential to achieve optimal DLBCL care management and reopen the debate of centralization of NHLpatients care in hematology/oncology departments.

Introduction:Acute Lymphoid Leukemia (ALL) is the most frequent cancer in young people and, if analyzed together with Non-Hodgkin Lymphoma (NHL), we find that they are responsible for at least one third of all cases of childhood cancer. Present-day therapeutic protocols include high doses of glucocorticoids (GC), drugs associate with high potential for elevating intraocular pressure (IOP) and, consequently provoking damage to the fibers of the optic nerve fibers, a pathology classified as cortisone glaucoma. In genetically susceptible patients, ocular hypertension normally occurs some weeks into the use of a steroid but is generally reversible with the suspension of its use. However, depending on the levels of ocular pressure and the duration of ocular hypertension, it can result in optic neuropathology and, in extreme cases, blindness. Since ALL and NHL are oncological disorders with elevated potential for cure of young people with have high life expectancy, the identification of eventual long-term treatment complications could give support to a delineation still lacking in scientific literature, that is, an ophthalmological protocol for these cases. Objective: The aim of this study was to evaluate the behavior of intraocular pressure in pediatric patients treated with GC for the acute lymphoproliferative neoplasias that are most common during childhood and adolescence.Methods: A systematic review of the theme was carried out, followed by a descriptive, prospective study of children and adolescents of both sexes who were diagnosed with ALL and NHL, and who were registered for beginning chemotherapeutic treatment at the Dr. Oswaldo Leite Oncology Center of Sergipe. The inclusion criteria were: diagnosis of ALL or NHL-T confirmed by immunophenotyping of bone marrow or peripheral blood samples (ALL), or immunohistochemistry of material obtained by open biopsy (NHL); age less than 19; no previous chemotherapy; absence of previous diagnosis compatible with glaucoma or any other disorder envolving change in intraocular pressure; no systemic use of GC in the six months preceding diagnosis of ALL or NHL. Patients whose evaluation of IOP might not have been technically adequate, as well as those who expired during the follow-up period, were excluded. Intraocular pressure was measured before treatment (D0), on the eighth (D8), the fourteenth (D14) and twentieth (D28) treatment day. The IOP results above 21 mm Hg were considered to be ocular hypertension. Results: Results of the systematic review indicate the need for new studies, for the review found a total of only three published articles whose results varied between total control of ocular pressure and visual function, to irreversible blindness. The results of our field research involved 15 patients, two of them with ocular hypertension, and with a statistically significant difference of measurements of IOP between D0 vs D8 and D0 vs D14 (p=0.013). Conclusion: The possibility of silent ocular hypertension, with the consequent risk of irreversible blindness, indicates the need to assess the introduction of a protocol for verification of IOP in patients recently diagnosed with ALL and NHL, including weekly exams, at least until the complete cessation of GC use. / Introdução: Leucemia Linfóide Aguda (ALL) é o câncer mais comumente encontrado entre os jovens e, se analisada em conjunto com o Linfoma não-Hodgkin (NHL), são responsáveis por pelo menos um terço dos casos de câncer infantil. Protocolos terapêuticos atuais incluem altas doses de glicocorticóides (GC), droga associada com alto potencial para elevar a pressão intraocular (IOP) e, consequentemente, provocar danos às fibras do nervo óptico, patologia classificada como glaucoma cortisônico. A hipertensão ocular geralmente ocorre com algumas semanas de uso de GC em pacientes geneticamente susceptíveis, mas é geralmente reversível com a descontinuação do tratamento. Entretanto, dependendo dos níveis pressóricos oculares e do tempo de elevação, pode resultar em neuropatia óptica e, em situações extremas, em cegueira. Por serem a ALL e o NHL doenças oncológicas com potencial elevado de cura, em indivíduos jovens com elevada expectativa de vida, a identificação de eventuais complicações de longo prazo decorrentes do tratamento poderá subsidiar o delineamento de um protocolo oftalmológico para esses casos, ainda inexistente na literatura científica. Objetivo: O objetivo deste estudo foi avaliar o comportamento da pressão intraocular em pacientes pediátricos portadores das mais frequentes neoplasias linfoproliferativas agudas da infância e adolescência, e que são tratados com GC. Métodos: Foi feita uma revisão sistemática sobre o tema estudado, seguida por um estudo descritivo, prospectivo, em crianças e adolescentes de ambos os sexos, com diagnóstico de ALL e NHL, matriculados para início de tratamento quimioterápico no Centro de Oncologia de Sergipe Dr. Oswaldo Leite. Os critérios de inclusão foram: diagnóstico de ALL ou NHL-T, confirmada por imunofenotipagem de amostra de medula óssea ou sangue periférico (ALL) ou imuno-histoquímica de material obtido por biópsia aberta (NHL); idade menor de 19 anos; sem quimioterapia anterior; ausência de diagnóstico prévio compatível com glaucoma ou doença anterior relacionada a qualquer mudança na pressão intra-ocular; não uso sistêmico de GC nos seis meses anteriores ao diagnóstico da ALL ou NHL. Pacientes cuja avaliação da pressão intraocular (PIO) pode não ter sido tecnicamente adequada e os que faleceram durante o período de seguimento foram excluídos. Realizaram-se medidas de pressão intraocular antes do tratamento (D0), no oitavo (D8), décimo quarto (D14) e vigésimo (D28) dias de tratamento. Os resultados da PIO acima de 21 mm de Hg foram considerados como hipertensão ocular Resultados: Os resultados da revisão sistemática apontaram para necessidade de novos estudos, limitando-se a um total de três publicações de relatos de casos envolvendo sete pacientes, com resultados variando de total controle da pressão ocular e conservação da função visual, até cegueira irreversível. Os resultados da pesquisa de campo envolveram 15 pacientes, com dois casos de hipertensão ocular e com diferença estatisticamente significativa entre as médias de PIO entre D0 vs D8 e D0 vs D14 (p = 0,013). Conclusão: A possibilidade de hipertensão ocular silenciosa, com o consequente risco de cegueira irreversível, indica a necessidade de se avaliar a introdução de um protocolo para verificação da IOP em pacientes jovens recentemente diagnosticados com ALL e NHL, incluindo exames semanais, pelo menos até a retirada completa do GC.

No description available.

Le Lymphome à Cellules du Manteau (LCM) est une entité récemment identifiée qui se caractérise par la translocation génétique t(11 ;14)(q13 ;q32) et compte pour 2 à 10 % de tous les Lymphomes non-Hodgkiniens. Avec une survie médiane entre 3 et 5 ans après le diagnostic, le LCM est une pathologie agressive et malgré les récentes avancées thérapeutiques, peu d’informations sont disponibles concernant ses facteurs pronostiques. Certaines études ont analysé le rôle des caractéristiques clinicopathologiques et des nouvelles stratégies thérapeutiques, mais on connait peut le rôle des facteurs environnementaux et du mode de vie sur le pronostic des patients atteints de LCM. Entre 2008 et 2012, le groupe LYSA a mené en France deux essais cliniques prospectifs multicentriques : LM manteau 2010 SA "RiBVD" (NCI01457144) et Manteau 2007 SJ "LyMa" (NCT00921414). Après une comparaison de ces patients avec les patients de population générale, l’effet de facteurs socioéconomiques et des habitudes de vie sur la survie des patients a été étudié à l’aide d’un questionnaire qualitatif administré à tous les volontaires après le diagnostic. Nos résultats suggèrent qu’un faible niveau d’éducation, un indice de masse corporelle élevé et de la consommation d’alcool sont associés à un risque de décès plus élevé chez les patients atteints de LCM. Toutefois, l’étude de tels facteurs et de leur influence sur un sous-type de LNH aussi rare requiert des échantillons d’étude de taille plus importante. L’élargissement des critères d’inclusion des patients dans les essais cliniques permettrait de sélectionner davantage de patients mais aussi des patients mieux représentatifs de la population générale. Enfin, l’intégration systématique de ce type de questionnaire dans les protocoles d’essais cliniques serait aussi un atout majeur. / Mantle Cell Lymphoma (MCL) is a recently defined entity, typically characterised by the genetic translocation t(11 ;14)(q13 ;q32) and counting for 2 - 10% of all non-Hodgkin Lymphomas. With a median survival between 3 and 5 years after diagnosis, MCL is an agressive disease and despite the recent therapeutic advances little in know about its prognostic factors. Some studies had investigated clinicopathological features and new treatment strategies, but there is a lack of knowledge regarding the impact of lifestyle and environnemental factors on outcome of MCL patients. From 2008 to 2012, the LYSA Group conducted in France two prospective multi center clinical trials on MCL : LM manteau 2010 SA "RiBVD" (NCI01457144) and Manteau 2007 SJ "LyMa" (NCT00921414). After a comparison of these patients with population-based data, socioeconomic factors, lifestyle factors and their influence on survival had been investigated through a qualitative survey administrated to each volunteer after diagnosis. Our findings suggest that low educational attainment, low body body mass index and alcohol consumption are associated with a higher risk of death in MCL. However, to investigate lifestyle factors in this rare NHL subtype, larger studies should be carried out. Clinical trial inclusion criteria must be widen to select more patients and patients more representative of general population. Implementation of these epidemiological studies in clinical practice should be considered.