Thesis (Ph. D.)--University of St Andrews, February 2009. / Restricted until 23rd February 2012.

Orientador: Manoel Barros Bértolo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-16T11:36:12Z (GMT). No. of bitstreams: 1 Bonfiglioli_Rubens_D.pdf: 4535259 bytes, checksum: c162b3d3293837dabf58f540050f6b46 (MD5) Previous issue date: 2009 / Resumo: Este estudo prospectivo analisou a epidemiologia, clínica e perfil genético de 102 pacientes brasileiros com Artrite Psoriásica. A associação do complexo maior de histocompatibilidade (MHC) de classe I, e os alelos do HLA-B27 com aquelas variáveis foram avaliados e comparados com sadios controles, HLA-B27 positivos, compondo um grupo de 111 indivíduos. A predominância foi do sexo masculino (59,8%), raça caucasóide (89,2%) e HLA-B27 negativos (79,4%). Oligoartrite assimétrica (62,7%) foi o subgrupo de Artrite Psoriásica mais observado, seguido pela forma espondilítica (16,7%) e poliarticular (15,7%). O sexo masculino e o subgrupo dos espondilíticos foram estatisticamente mais associados ao HLA-B27, e o subgrupo oligoarticular ao HLA-B27 negativo. Entre os 21 pacientes com Artrite Psoriásica e HLA-B27 positivos existiu uma significante prevalência do HLA-B*2705 (90,5%), similar ao observado no grupo controle (80,2%); HLA-B*2703 e HLA-B*2707 foram estatisticamente associados ao grupo controle / Abstract: This prospective study analyzed the epidemiologic, clinical and genetic profile of 102 Brazilian patients with psoriatic arthritis (PsA). The association of the major histocompatibility complex (MHC) class I and the HLA-B27 alleles with these variants was outlined, and compared to a control healthy HLA-B27 positive group of 111 individuals. There was a predominance of male gender (59.8%), Caucasian race (89.2%) and negative HLA-B27 (79.4%) patients. Asymmetric oligoarthritis (62.7%) was the most frequently observed clinical PsA subgroup, followed by spondylitis (16.7%) and polyarthritis (15.7%). Male gender and the spondylitis subgroup were statistically associated to the positive HLAB27 and the oligoarthritis subgroup was associated to the negative HLA-B27. Among the 21 HLA-B27 positive PsA patients, there was a significant prevalence of the HLA-B*2705 allele (90.5 %), similar to that observed in the control group (80.2%); HLA-B*2703 and HLA-B*2707 were statistically associated to the control group. Other antigens such as HLA- B07 (14 patients), HLA-B08 (14 patients) and HLA-B44 (13 patients) among others, were found in HLA-B27-negative patients / Doutorado / Clinica Medica / Doutor em Clínica Médica

The MHC class I allele, HLA-B27, is strongly associated with a group of inflammatory arthritic conditions collectively known as spondyloarthropathies (SpA). Ankylosing spondylitis (AS) shows the strongest association with 90-95 % of patients being HLA-B27 positive. The relationship between HLA-B27 and SpA has been known for over 30 years, however despite ongoing research, the reason for this association has not yet been elucidated. In more recent years, research has focused on intrinsic properties of the HLA-B27 allele, in particular its propensity to misfold, forming homodimers. It has been proposed that these homodimers could be associated with the disease process through the activation of an ER stress response known as the unfolded protein response (UPR), or through aberrant recognition at the cell surface. We have investigated whether the expression of HLA-B27 is associated with the activation of the UPR. We have studied the expression of BiP, and the cleavage of XBP1 and ATF6 using stable and transiently expressing cell lines. We have also investigated the formation of non-B27 homodimers using a human cell line stably expressing HLA-B8, and finally we have studied the expression of homodimers in exosomes, small immunomodulatory vesicles released from numerous cell types. The results presented here lead us to conclude that in vitro studies of the UPR are complicated, prone to a number of technical issues, and may therefore not be appropriate for gaining information that would be of significant use when comparing to the real disease scenario. Our data suggest that non-B27 dimers may be strongly influenced by both the overexpression of MHC class I heavy chains and also the redox environment within the cell. We have isolated a novel fully folded, beta-2m-associated, MHC class I homodimer in exosomes and have detected a novel HLA-A and HLA-B mixed heavy chain dimer. Our results suggest that these dimers form through interactions between the cysteine residues in the cytoplasmic tail and that these dimers form in exosomes because they contain lower levels of the important antioxidant glutathione when compared to whole cells. Together, these results define a new MHC class I structure present on exosomes at significant levels, which could potentially influence immune recognition by both antigen-specific T cell receptors and NK family receptors. The data also poses questions about whether these novel structures, when they involve HLA-B27, could influence the pathogenesis of spondyloarthropathies.

<p>Uvod: Pojam &quot;sekretor&rdquo; ili &bdquo;nesekretor&quot; se odnosi na sposobnost pojedinca da luči antigene krvnih grupa ABO sistema u telesnim tečnostima. Određivanje ABO krvne grupe i sekretornog statusa, testom inhibicije hemaglutinacije pljuvačke i Lewis fenotipa na eritrocitima su važni u kliničkoj i forenzičkoj medicini, u odnosu na etiopatogenezu mnogih bolesti. Nesekretorstvo ABO krvnogrupne supstance je udruženo sa če&scaron;ćom pojavom autoimunih inflamatornih oboljenja među kojima su i seronegativne spondiloartropatije. Veća učestalost seronegativnih spondiloartropatija među osobama koje imaju HLA-B27 antigen predstavlja polaznu osnovu stanovi&scaron;ta da genetski faktori u kombinaciji sa faktorima sredine utiču na pojavu seronegativnih spondiloartropatija u genetski predisponiranih individua. Teza istražuje značaj testa inhibicije hemaglutinacije plijuvačke i određivanja ABO i Lewis fenotipa na eritrocitim u dijagnostici seronegativnih spondiloartropatija. Ciljevi i hipoteze: Cilj istraživanja je bio da se utvrdi učestalost nesekretora i sekretora ABO krvnogrupne supstance i Lewis fenotipa u grupi obolelih od seronegativnih spondiloartropatija i izvr&scaron;iti poređenje rezultata u odnosu na grupu zdravih ispitanka. Pretpostavljeno je da postoji značajno veći broj nesekretora ABO krvnogrupne supstance u obolelih od seronegativnih spondiloartropatija u odnosu na zdrave ispitanike. Pretpostavljeno je i da postoji značajno veća učestalost nesekretora ABO krvnogrupne supstance u obolelih od seronegativnih spondiloartropatija sa negativnim HLA-B27 antigenom. Pretpostavljeno je da kod osoba obolelih od seronegativnih spondiloartropatija dolazi do promene Lewis fenotipa na eritrocitima u odnosu na sekretorni status u pljuvačci. Metode: Sprovedena je longitudinalna prospektibvna studija. Ispitanici stariji od &scaron;est godina oba pola podeljeni su u dve randomizovane grupe. Eksperimentalnu grupu sačinjavalo je 110 ispitanika sa dijagnozom oboljenja iz grupe seronegativnih spondiloartropatija. Kontrolnu grupu sačinjavalo je 103 dobrovoljna davaoca krvi, bez dijaghnoze oboljenja iz grupe seronegativnih spondiloartropatija. Ispitanicima kontrolne i eksperimentalne grupe određena je pripadnosti ABO krvnogrupnom sistemu, sekretorni status i Lewis fenotip, dok je osobama eksperimentalne grupe određen i fenotip HLA-B27. Uključujući kriterijumi osim navedenog bili su da ispitivane osobe ženskog pola nisu trudinice i da ispitanici obe grupe nisu primali transfuziju krvi tri meseca pre uključivanja u istraživanje. Rezultati: Rezultati istraživanja ukazuju da nesekretori ABO krvnogrupne supstance imaju 1,63 puta veći rizik (veću učestalost), oboljevanja od seronegativnih spondiloartropatija u odnosu na zdravu populaciju ispitanika, kao i da smanjena ekspresija Lewis (b) antigena predstavlja doprinoseći faktor razvoja seronegativnih spondiloartropatija. Ustanovljeno je da pod uticajem seronegativnih spondiloartropatija dolazi do izmene Lewis fenotipa na eritrocitima obolelih. Verovatnoća dokazivanja oboljenja iz grupe seronegativnih spondilartropatija među obolelima veća je za 11% kod nesekretora fenotipa HLA-B27- u odnosu na obolele nesekretore fenotipa HLA-B27+. Zaključak: Sekretorni status i Lewis fenotip predstavljaju zasebne dijagnostičke biohemijske markere nezavisne od HLA-B27 antigena koji doprinose ranijem otkrivanju osoba koje imaju predispoziciju razvoja oboljenja iz grupe seronegativnih spondiloartropatija.</p> / <p>Introduction:The term secretory state referes to ability of individual to secrete ABO blood group antigens in body fluids. Determination of the ABO blood group antigens and secretory status by hemagglutionation inhibition test using saliva as well as Lewis phenotype on erythrocytes are important in clinical and forensic medicine, in relation to the etiopathogenesis of many diseases. Non secretory status of ABO blood groupantegens is related with higher incidence of autoimmune inflammatory disease which include seronegative spondyloarthropathyes. Increased frequency of seronegative spondyloarthropathies among people who have the HLA-B27 antigen is starting point of the view that genetic factors in combination with environmental factors influence the occurence of seronegative spondyloarthropathies in genetically predisposed individuals. The tesis explores the significance of the hemagglutionation inhibition test of saliva and determination of ABO and Lewis antigens in diagnostics of seronegativespondyloarthropathyes. Goals and hypothesis: The aim of this study was to determine the frequency of non secretors and secretors of ABO blood group antigens and Lewis phenotype in a group of patient with seronegative spondyloarthropathies and make comparsion to the healthy examined group. It was assumed that there is a significantly higher number of non secretors of ABO blood group antigens among the patients with seronegative spondyloarthropathies compared to healthy examined persons. It was assumed that there is a significantly higher number of non secretors of ABO blood group antigens among the patients with seronegative spondyloarthropathies who do not have HLA-B27 antigen. It was assumed that in patients with seronegative spondyloarthropathies Lewis phenotype on erythrocits can be changed in relation to the secretory status in the saliva. Methods: We performed a prospective longitudinal study. Respondants older than six years of both gender were divided into two randomized groups. Experimental group is consisted of 110 patients diagnosed with seronegative spondyloarthropathy. The control group consisted of 103 blood donors who did not have diagnosed disease from the group of seronegative spondyloarthropathies. To the subjects of the control and experimental groups was determined ABO blood group antigens, secretory status and Lewis phenotype, while to the subjects of experimental group also was designated HLA-B27 phenotype. Including criteria other than the above were that among female respondants were not pregnant, and that both groups of respondants did not receive a blood transfusion three months before joining the study. Results: The resuts of the study showed that non secretors of ABO blood group antigens have a 1,63 times higher rise (higher incidence) of developing disease from the group of seronegative spondyloarthropathies compared to a healthy population of subjects, as well as that decreased expression of Lewis antigens (b) represents a contributing factor for development of seronegativespondyloarthropathies. It was found that under the influence of seronegative spondyloarthropathies there are changes in Lewis phenotype on erythrocytes of patients. It was found that under the influence of seronegative spondyloarthropathies there are changes in Lewis phenotype on erythrocytes of patients. Probability for confirmation seronegative spondyloarthropathies is higher for 11% among non secretors who have HLA-B27 negative phenotype in comparison to non secretors who have HLA-B27 positive phenotype. Conclusion: Secretory status and Lewis phenotype are separate diagnostic biochemical markers independent of HLA-B27 antigen that contribute to the early detection of people who have a predisposition of the disease from the group of seronegative spondyloarthropathies.</p>