Orientador: Claudio Lucio Rossi / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-10T14:37:04Z (GMT). No. of bitstreams: 1 Nascimento_FernandaSantos_M.pdf: 1195114 bytes, checksum: 8f7292945661258264c33482af428d55 (MD5) Previous issue date: 2008 / Resumo: A toxoplasmose, uma zoonose com ampla distribuição mundial, causada pelo parasita intracelular obrigatório Toxoplasma gondii, é geralmente adquirida por meio da ingestão de cistos ou oocistos viáveis do parasita, presentes, respectivamente, em carne crua ou mal cozida e no solo, alimento ou água contaminados com fezes de gatos infectados. A toxoplasmose pode ser altamente debilitante, e ocasionalmente fatal, em crianças infectadas no útero e em receptores de transplante. O diagnóstico de infecção aguda primária em mulheres grávidas é geralmente baseado em testes sorológicos, visto que, na grande maioria dos casos, a toxoplasmose não é reconhecida clinicamente. A longa persistência dos anticorpos IgM em algumas pessoas e a dificuldade para demonstrar soroconversão ou aumento significativo da concentração de anticorpos específicos, têm complicado a interpretação dos testes sorológicos, quando se suspeita de infecção aguda. Com relação à infecção toxoplámica em pacientes transplantados, em muitos casos o status sorológico do doador não é conhecido e a pesquisa periódica de anticorpos anti-T. gondii no receptor raramente é realizada. O objetivo do primeiro estudo foi determinar o valor da demonstração dos anticorpos IgA anti-T.gondii para o diagnóstico da fase aguda da infecção toxoplásmica. Nossos resultados mostraram que os anticorpos IgA são detectados com alta freqüência em amostras de soros obtidas de mulheres com evidência clínica e/ou sorológica de infecção toxoplásmica aguda. Entretanto, em 19% das mulheres apresentando persistência de anticorpos IgM e alto índice de avidez dos anticorpos IgG, anticorpos IgA anti-T. gondii foram detectados em amostras de soros coletadas mais de 9 meses após o início da infecção, indicando que esses anticorpos não podem ser considerados marcadores confiáveis de infecção aguda primária. No segundo estudo, nós relatamos o diagnóstico de infecção toxoplásmica primária em um paciente com mieloma múltiplo submetido a transplante alogênico não-mieloablativo de células hematopoiéticas, provenientes de doador com sorologia negativa para toxoplasmose. A resposta primária contra o T. gondii foi baseada na soroconversão dos anticorpos IgM, IgG e IgA. O paciente foi prontamente tratado e nenhuma complicação relacionada à toxoplasmose foi observada nos meses subseqüentes. Esse caso ressalta a necessidade da detecção dos anticorpos anti-T. gondii no doador e no receptor antes do transplante e a importância do monitoramento sorológico do receptor durante o seguimento pós-transplante / Abstract: Toxoplasmosis, a cosmopolitan zoonotic disease caused by the intracellular parasite Toxoplasma gondii, is usually acquired through the ingestion of viable parasite cysts or oocysts, present, respectively, in raw or undercooked meat and in soil, food or water contaminated with feces of infected cats. Toxoplasmosis can be highly debilitating and occasionally fatal in children infected in utero and in transplant recipients. The diagnosis of acute primary infection in pregnant women is usually based on serology, because in the great majority of cases primary infection is not recognized clinically. The sustained persistence, in some persons, of specific IgM antibodies and the difficulty in demonstrating seroconversion or a significant increase in specific antibody concentrations, have complicated the interpretation of serological tests when acute infection is suspected. With regard to toxoplasmic infection in transplant patients, in many cases the serological status of the donor is not known and the periodic research of anti-T. gondii antibodies in the receptor is rarely performed. In the first study, we investigated the usefulness of detecting anti-T. gondii IgA for the diagnosis of an acute acquired Toxoplasma infection. Our results showed that anti-T. gondii IgA antibodies are detected with a high frequency in serum samples obtained from women with clinical and/or serologic evidence of acute acquired Toxoplasma infection. However, in 19% of the women presenting a sustained IgM antibody response and a high IgG avidity index, anti-T. gondii IgA antibodies were detected in serum samples collected more than nine months after the beginning of infection, indicating that IgA cannot be considered a dependable marker for acute primary infection. In the second study, we report the diagnosis of a primary toxoplasmic infection in a patient with multiple myeloma following a non-myeloablative allogeneic transplant with hematopoietic stem cells from a donor with negative serology for toxoplasmosis. The primary response to T. gondii was supported by IgM, IgG and IgA seroconversion. The patient was promptly treated and there were no complications related to toxoplasmosis in the subsequent months. This case stresses the importance of detecting anti-T. gondii antibodies in the donor and in the recipient before transplantation, and of serologically monitoring the recipient during long-term follow-u / Mestrado / Ciencias Biomedicas / Mestre em Ciências Médicas

Submitted by Ramon Santana (ramon.souza@ufpe.br) on 2015-03-10T14:49:55Z No. of bitstreams: 2 Dissertação Viviane Martha de Morais.pdf: 1592021 bytes, checksum: e29a3eec2c8393142f0c5e43344e1535 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-03-10T14:49:55Z (GMT). No. of bitstreams: 2 Dissertação Viviane Martha de Morais.pdf: 1592021 bytes, checksum: e29a3eec2c8393142f0c5e43344e1535 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2013-02-28 / Introdução: O diagnóstico rápido, simples e preciso para confirmar a infecção pelo vírus dengue (DENV) é uma necessidade real, uma vez que a doença pode se manifestar com um amplo espectro de sinais e sintomas, similares a outros quadros febris agudos. Durante a infecção por dengue se verifica também a produção da imunoglobulina A (IgA) específica, que aumenta ao mesmo tempo que a imunoglobulina M (IgM), permanece positiva por um período de tempo mais curto e se apresenta em níveis mais elevados na infecção secundária. Objetivo: O presente estudo teve como objetivo investigar a presença de IgA no soro durante a infecção primária e secundária (sequencial) pelo DENV. Metodologia: Foram avaliadas amostras de soro por meio do teste imunoenzimático de captura da IgA (AAC-ELISA) in house. Resultados: Avaliou-se um total de 445 amostras de soro, sendo 171 caracterizados como infecção primária e 194 secundária; 40 amostras de indivíduos saudáveis negativos para dengue e 40 de vacinados contra febre amarela. As amostras foram distribuídas em 13 grupos. A positividade da IgA foi de 42,2% (154/365), sendo 27,5% (47/171) na infecção primária e 55,2% (107/194) na secundária. Na infecção secundária, a IgA foi detectada do 2º ao 4º dias de sintomas (grupo 1), antes mesmo da IgM, assim como no grupo 11 no qual a IgM não havia sido detectada (infecção secundária). Na infecção primária o maior valor da sensibilidade foi de 60,0 (36,4 - 80,0) no grupo com 30-35 dias de sintomas e na secundária foi de 87,5% (60,4 – 97,8), grupo com 8 dias de sintomas. A especificidade foi de 100% nas duas infecções (94,3 – 100). Ao aplicar o teste em paralelo para ambas técnicas observou-se um aumento global de 6,6% na sensibilidade do diagnóstico; sendo 2,7% para a infecção primária e de 15,2% para a secundária. A IgA não foi detectada nas amostras dos indivíduos saudáveis, nem nas amostras dos indivíduos recentemente vacinados contra febre amarela. Conclusões: A detecção da IgA demonstrou ser útil como forma de diagnóstico sorológico e em conjunto com a detecção da IgM poderá auxiliar na confirmação de casos agudos de dengue e na interpretação dos resultados de casos inconclusivos, permitindo a adoção de medidas preventivas para evitar a ocorrência de epidemias e ocorrência de casos graves e óbitos.

High training loads or prolonged bouts of acute exercise can increase susceptibility to opportunistic infections. Such infections, although generally medically innocuous, can have profound negative implications for athletic performance. This thesis presents a series of studies investigating which factors influence infection risk in athletes, as well as exploring potential strategies to maintain immunocompetence during heavy training. In Chapters 2 and 3, a large cohort of elite winter endurance athletes were followed over a number of years to determine patterns and frequency of illness in this population, and to identify training- and competition-related predictors of infection. Incidence rates and seasonal patterns of illness were found to be broadly similar to elite athletes from summer sports, and to the general population. Competition, air travel, greater day-to-day fluctuations in training load and lower performance level were significant predictors of illness. When high training loads are combined with insufficient recovery, athletes may become overreached or overtrained. Previous studies suggest that increasing carbohydrate intake can be an effective means of preventing overreaching during periods of intense training. In Chapter 4 we therefore investigated the efficacy of carbohydrate supplementation in reducing immune disturbances and symptoms of overreaching. The lower carbohydrate does (20 g/h during exercise) was found to be equally effective in preserving immunity and power output as the higher dose (60 g/h), with modest immune and performance changes observed in both groups following eight days of intensified training. Many athletes fail to ingest sufficient fluid to maintain euhydration during exercise. However, Chapter 5 found that moderate hypohydration, elicited by a 24 h period of fluid restriction, had little effect on immune responses to prolonged exercise. Altitude training is an important component of the training process of most of today s elite endurance athletes. Chapters 6 and 7 explored the effects of acute and prolonged hypoxic training on immunity. Despite a somewhat augmented stress hormone response to exercise in hypoxia, altitude training was found to have little negative effect on host defence, providing relative exercise intensity at altitude and sea-level was matched.

IgA nephropathy is the world's most common primary glomerulonephropathy. Recent evidence in a rat model implicated excessive production of oxygen-free radicals in the pathogenesis and suggested that vitamin E-treatment ameliorated progression. We studied this antioxidant therapy on the glomerular filtration rate (GFR), proteinuria and hematuria in biopsy-proven IgA nephropathy in children. The duration of treatment or placebo was 2 years, with vitamin E treatment consisting of 400 IU/day in children weighing <30 kg, and twice that dose for those >30 kg. We measured GFR at entry, midpoint and exit. At baseline and at 4-month intervals after randomization, urinary protein/creatinine ratios and urinalysis were examined. The mixed model procedure with log transformation was used in data analysis to test treatment difference as well as the potential time effect. Fifty-five patients were randomized and 38 completed at least 1 year of follow-up. At entry, the clinical characteristics were not different between the treatment and placebo groups. There was a trend toward better preservation of GFR in vitamin E-treated versus placebo patients, 127±50 vs. 112±31 ml/min/1.73 m2, respectively (P=0.09). The urinary protein/creatinine ratio was significantly lower in the vitamin E-treated group vs. placebo; 0.24±0.38 vs. 0.61±1.37 (P<0.013). However, there was no difference in the prevalence of hematuria between the groups. Vitamin E treatment in our study patients was associated with significantly lower proteinuria, but no effect on hematuria. While there was a trend toward stabilization of GFR in the vitamin E-treated patients, longterm treatment and follow-up are needed to determine whether antioxidant therapy is associated with preservation of renal function in IgA nephropathy.

Background Glomerulonephritis is a major cause of end-stage kidney disease (ESRD) in Africa. There is scanty data on the clinico-pathological characteristics and outcome of the mesangioproliferative glomerulonephritides in Africa, despite the non-IgA subtype being reported as a common cause of nephrotic syndrome. This study will assess the outcome of patients with biopsy proven mesangioproliferative glomerulonephritis (MesPGN) from a single centre in Cape Town, South Africa. Methods The study is designed as 10-year retrospective analysis of patients with biopsy proven MesPGN. The MesPGN patterns were divided into non-IgA MesPGN and IgA nephropathy (IgAN), depending on the predominant type of immune deposit. Univariate cox regression analysis was used to determine factors associated with ESRD. Results Data of 109 patients with renal biopsy-proven MesPGN were included for the period between 2005-2014. The mean age at biopsy was 33.8 ±14.9 years, 53.2% were males, and 39.4% were black Africans. Clinically, 58.7% presented with nephrotic syndrome. On histology 79.8% had non-IgA MesPGN, and 20.2% had IgAN. Compared to the non-IgA group, most patients with IgAN were not treated with immunosuppression (72.7% vs. 40.2%; p=0.006). At the last visit, 10.1% reached ESRD (40.9% vs. 2.3%; p<0.0001) and 30.2% achieved complete remission (9.1% vs. 35.7%; p=0.015) for IgAN and non-IgA MesPGN respectively. The 5-year renal survival for IgAN and non-IgA MesPGN respectively, were: 63.3% vs. 97.6%, log rank p=0.001. Overall, hypertension (p=0.019), not receiving immunosuppression (p=0.046) and having IgAN (p=0.007) were independent predictors of progression to ESRD. Conclusion There is a significantly higher ESRD-free survival of patients with biopsy proven non-IgA MesPGN than IgAN. Whether this is related to the limited use of immunosuppressive therapy in IgAN patients or represents a true nature of the disease still requires further research.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第19021号 / 農博第2099号 / 新制||農||1030(附属図書館) / 学位論文||H27||N4903(農学部図書室) / 31972 / 京都大学大学院農学研究科応用生物科学専攻 / (主査)教授 久米 新一, 教授 祝前 博明, 教授 廣岡 博之 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

The use of the finite element method within an optimization workflow is fraught with challenges that limit the automation of such workflows. These challenges are inherent to the traditional finite element formulations which are heavily dependent on a manual meshing process that introduces variability that is challenging to account for within an automated workflow. The recently developed flex representation method (FRM) provides a salient solution to the manual meshing process without sacrificing solution accuracy. In response to the development of FRM a global automotive company requested a study to explore the applicability of FRM to one of their sizing-optimization problems: the constrained optimization of a wheel undergoing a rigidity test. In this study we develop an optimization framework based on the DAKOTA optimization framework, the open-source FreeCAD computer-aided design software, and an implementation of FRM within the Coreform IGA solver. Within this framework we demonstrate in the affirmative that FRM enables a highly robust sizing-optimization workflow while requiring minimal effort to prepare the FRM model.

Perturbing gut microbiota early in life can lead to the development of autoimmunity. We are just beginning to unravel how early immune programming by microbiota may have long-term effects on noncommunicable diseases. In this thesis, we lay groundwork for programming of the immune system by commensal bacteria early in life through our studies on the induction of early endogenous neonatal IgA, and we evaluate Limosilactobacillus reuteri's characteristics as an inducer. Garnering attention for use a probiotic, L. reuteri has many proven health promoting benefits, such as IgA induction, but emerging evidence also links specific strains to autoimmune disease. "Super-induction" of neonatal IgA can be achieved through cross-fostering immunocompetent pups on immunocompromised dams. We found that this phenomenon was categorically due to transferal of microbes from dam to offspring. By comparing strain CF48-3A to the non-gastric-related organism L. oris, we discovered that L. reuteri is a microorganism that can enhance early neonatal IgA induction. Further investigations revealed that the ability to induce neonatal IgA is not ubiquitous in all L. reuteri strains, as ATCC PTA 6475 did not significantly elevate IgA. We discovered that 6475 has the antigenic ability to stimulate B cell differentiation and IgA production, but it is suppressed by a mechanism related to differences in surface architecture of this strain. L. reuteri strains also vary in their potency of aryl hydrocarbon receptor (AhR) stimulation. In mice, activation of AhR during gestation by a potent prototypical ligand, TCDD, leads to development of autoimmunity offspring. We found that TCDD exacerbated autoimmunity in adult mice using a strain of mice with similar AhR affinity to humans. Further investigations can clarify whether differential AhR ligand expression between L. reuteri strains contributes to the relationship between L. reuteri and autoimmunity. Overall, we conclude that differences between strains of L. reuteri have profoundly different immunological consequences that contribute to our understanding of the linkage between strains and autoimmunity. / Doctor of Philosophy / Differences in microbes transferred to infants through maternal routes shapes the early development of the immune system. In general, transferred microbes are healthy for the infant, and studies suggest that disruption of healthy microbes in the infant gut is linked to long-term health consequences, like autoimmune diseases. We found that a particular difference in maternally transferred microbes increases the early appearance of immunoglobulin A (IgA, a gut-related antibody) in neonatal mice, which is an antibody important for protecting against gut-related infections. We were able to link this early IgA production to a probiotic species Limosilactobacillus reuteri. Within the species classification as L. reuteri, several genetically different strains are health-promoting and broadly marketed over-the-counter for use in probiotic supplements for infants, children, and pregnant and nursing mothers. Emerging scientific evidence also points to a potential connection between other L. reuteri strains and autoimmune disease. Secreted products of genetically different L. reuteri strains have been discovered to activate aryl hydrocarbon receptor (AhR) with various potency. We used a prototypical AhR ligand and found exacerbation of autoimmune disease in adult mice. Thus, we have concluded that different strains of L. reuteri have broadly different effects on immune system development, and strain variability may explain the different effects on autoimmunity that have been observed.

Expressão aumentada do receptor Fc de IgA (CD89) e da cadeia g associada, avaliadas respectivamente por citometria de fluxo e por “immunobloting", foram encontradas em fagócitos do sangue de pacientes com bacteremia por germes gram-negativos, em comparação com controles e pacientes com bacteremia por gram-positivos. A Mr do CD89 avaliada por SDS-PAGE estava diminuída, com núcleo protéico de 32kDa, sugerindo alteração de glicosilação. O aumento da expressão do CD89 correlacionou-se com aumento dos níveis séricos de IL-6. A cadeia g estava fosforilada nos neutrófilos, sugerindo participação do CD89 na sepsis por gram-negativos. / The expression and function of FcaRI (CD89) were analyzed on blood monocytes and neutrophils of patients with gram-positive and gram-negative bacteremia. Eighteen patients with gram-positive bacteremia, sixteen patients with gram-negative bacteremia and twenty healthy individuals were studied. CD89 expression were analyzed by flow cytometry using specific stained antibodies. Analysis of the surface iodinated CD89 molecules by SDS-PAGE and of the CD89 g-associated chain by immunoblotting also were performed. A marked increase in expression of the a and g subunits of the FcaRI were found on both types of cells in patients with gram-negative bacteremia, but not in patients with gram-positive bacteremia. This increase was independent of serum IgA levels. FcaRI Mr was lower on cells from gram negative patients than on cells from controls (50-65 kDa vs 55-75 kDa) despite of similar 32 kDa backbone, indicating altered glycosylation. Increased levels of FcaRI on blood phagocytes correlated with enhanced serum IL-6 levels, but not with IFN-g or TNF-a levels. The CD89 g-associated chain was phosphorylated on neutrophils, suggesting an engagement of CD89 during gram negative sepsis.

Η ΙgΑ νεφροπάθεια αποτελεί την πιο συχνή μορφή πρωτοπαθούς σπειραματονεφρίτιδας η οποία αντιμετωπίζεται με την εφαρμογή διαφόρων θεραπευτικών σχημάτων. Στο πρώτο σκέλος της παρούσας διδακτορικής διατριβής, αξιολογήθηκε η αποτελεσματικότητα θεραπευτικών σχημάτων που χορηγήθηκαν με βάση τα κλινικά και ιστολογικά χαρακτηριστικά των ασθενών με ΙgΑ νεφροπάθεια για περίοδο παρακολούθησης 5 ετών. Στη μελέτη συμπεριελήφθησαν 50 ασθενείς, οι οποίοι χωρίστηκαν σε 4 ομάδες. Ασθενείς με φυσιολογική νεφρική λειτουργία και πρωτεϊνουρία <1,0 g/24h δεν έλαβαν ειδική αγωγή (Ομάδα Α, n=6). Ασθενείς με φυσιολογική νεφρική λειτουργία, πρωτεϊνουρία >1,0 g/24h και με ήπια έως μέτρια μεσαγγειακή υπερπλασία και διαμεσοσωληναριακή συμμετοχή αντιμετωπίστηκαν με αγωγή που περιελάμβανε α-ΜΕΑ και κορτικοστεροειδή (Ομάδα Β, n=23). Ασθενείς με αρχική κρεατινίνη ορού Scr<2,5 mg/dL, πρωτεϊνουρία >3,5 g/24h και/ή μέτριες έως σοβαρές ιστολογικές αλλοιώσεις αντιμετωπίστηκαν με συνδυασμό α-ΜΕΑ, κορτικοστεροειδών και άλλων ανοσοκατασταλτικών φαρμάκων (Ομάδα Γ, n=18). Ασθενείς με αρχική τιμή Scr >2,5 mg/dL, με παρουσία σοβαρής σπειραματοσκλήρυνσης και διαμεσοσωληναριακής βλάβης αντιμετωπίστηκαν μόνο με α-ΜΕΑ και ιχθυέλαια (Ομάδα Δ, n=3). Από τους 50 ασθενείς, οι 9 (18%) παρουσίασαν διπλασιασμό της αρχικής κρεατινίνης ορού, 2 από την ομάδα Β (8,7%), 5 από την ομάδα Γ (27,7%) και 2 από την ομάδα Δ (66,7%). Από τους 7 ασθενείς που εμφάνισαν νεφρική ανεπάρκεια τελικού σταδίου, ένας ήταν από την ομάδα Β (4,3%), 4 από την ομάδα Γ (22%) και 2 από την ομάδα Δ (66,7%). Ύφεση της πρωτεϊνουρίας παρατηρήθηκε σε όλους τους ασθενείς της ομάδας Β και σε 15 ασθενείς της ομάδας Γ (83,3%). Παρενέργειες παρατηρήθηκαν σε 3 (7,3%) ασθενείς που έλαβαν ανοσοκατασταλτική αγωγή. Σε ασθενείς με IgA νεφροπάθεια η χορήγηση εξατομικευμένου θεραπευτικού σχήματος που βασίζεται στα κλινικά και ιστολογικά χαρακτηριστικά ενός εκάστου των ασθενών φαίνεται ότι μπορεί να οδηγήσει στην επίτευξη του μέγιστου θεραπευτικού αποτελέσματος με τον ελάχιστο κίνδυνο ανεπιθύμητων ενεργειών. Στο δεύτερο σκέλος μελετήθηκαν δυνητικά πρώιμοι δείκτες εξέλιξης της νεφρικής βλάβης τόσο στον νεφρικό ιστό, όσο και σε ούρα ασθενών. Μελέτες στο γονιδίωμα και το πρωτέωμα υποδηλώνουν ότι η τρανσγελίνη πιθανόν εμπλέκεται στην νεφρική βλάβη μέσω ενεργοποίησης των μυοϊνοβλαστών. Σε τομές νεφρικού ιστού 67 ασθενών έγινε ανίχνευση με ανοσοϊστοχημική μέθοδο και ανοσοφθορισμό και ποσοτική εκτίμηση της παρουσίας αφενός μεν της τρανσγελίνης αφετέρου της α-ακτίνης των λείων μυϊκών ινών (α-SMA), γνωστού δείκτη ενεργοποίησης των μυοϊνοβλαστών στο νεφρικό ιστό. Έκφραση της τρανσγελίνης και της α-SMA εντοπίστηκαν στα σπειράματα και στο διάμεσο χώρο. Σε ασθενείς με IgA νεφροπάθεια και εστιακή τμηματική σπειραματοσκλήρυνση η έκφραση της τρανσγελίνης ήταν εντονότερη από αυτήν της α-SMA. Η ανοσοϊστοχημική έκφραση της τρανσγελίνης σχετιζόταν με το βαθμό σπειραματικής σκλήρυνσης (p=0,035) και ίνωσης του διάμεσου χώρου (p=0,047), με το βαθμό μεσαγγειακής υπερπλασίας (p=0,034), με την ύφεση της λευκωματουρίας (p=0,041) και την έκβαση της νεφρικής λειτουργίας (p=0,009). Η μελέτη συνεντοπισμού των τρανσγελίνης και α-SMA στο νεφρικό ιστό έδειξε ότι σε κάποιες περιοχές οι δύο πρωτεΐνες εκφράζονταν ταυτόχρονα και σε άλλες περιοχές κάθε πρωτεΐνη εκφραζόταν χωριστά. Έντονη παρουσία της τρανσγελίνης παρατηρήθηκε στο νεφρικό ιστό ασθενών με διάφορους τύπους σπειραματονεφριτίδων. Η παρουσία μυοϊνοβλαστών που παράγουν τρανσγελίνη ή α-SMA ή και τις δύο πρωτεΐνες μαζί, υποδηλώνει την πιθανή παρουσία διαφορετικών υποπληθυσμών μυοϊνοβλαστών στο νεφρικό ιστό ασθενών με διάφορους τύπους σπειραματονεφριτίδων. Επιπλέον, προσδιορίστηκαν τα επίπεδα απέκκρισης προφλεγμονωδών (IL-2, IL-17, TNF-α, INF-γ, IL-6) και αντιφλεγμονωδών (IL-4, IL-10, TGF-β1) κυτταροκινών καθώς και της χημειοκίνης MCP-1 σε ούρα ασθενών με IgA νεφροπάθεια και συγκρίθηκαν με αυτά υγειών εθελοντών δοτών καθώς και ασθενών με διαφόρους τύπους σπειραματο-νεφρίτιδας που βρίσκονταν σε κλινική ύφεση της νόσου. Μελετήθηκαν 97 ασθενείς με πρωτοπαθή σπειραματονεφρίτιδα και όλοι ήταν σε κλινική ύφεση μετά από ανοσοκατασταλτική αγωγή. Από τους 97 ασθενείς, οι 31 είχαν IgA νεφροπάθεια (IgAΝ), οι 36 μεμβρανώδη (MN) και οι 30 νόσο ελαχίστων αλλοιώσεων ή εστιακή τμηματική σπειραματοσκληρυνση (MC/FSGS). Τα επίπεδα των Th-κυτταροκινών και της MCP-1 μετρήθηκαν σε τυχαίο δείγμα ούρων και συγκρίθηκαν με τα επίπεδα 17 υγειών εθελοντών δοτών. Οι ασθενείς με σπειραματονεφρίτιδα, σε σύγκριση με τους υγιείς, είχαν σημαντικά υψηλότερα επίπεδα απέκκρισης σε όλες τις κυτταροκίνες που μελετήθηκαν, εκτός από την IL-4 και τον TNF-α. Στην ομάδα των ασθενών διαπιστώθηκε ισχυρή θετική συσχέτιση μεταξύ των επιπέδων του TGF- β1 και της MCP-1 (p=0,000, r=0,721). Σε ασθενείς με IgAΝ εντοπίστηκαν σημαντικές διαφορές στην απέκκριση των IL-2 (p=0,0345), IL-17A (p=0,005), TNF-α (p=0,0098), TGF-β1 (p=0,0092) και MCP-1 (p=0,0301) σε σύγκριση με ασθενείς με MN και στην απέκκριση της IL-2 σε σχέση με ασθενείς με MC/FSGS (p=0,0018). Η παρατήρηση αυτή υποδηλώνει τη συνεχιζόμενη παρουσία φλεγμονώδους διαδικασίας στο νεφρικό ιστό των ασθενών με διαφόρους τύπους σπειραματονεφριτίδων ακόμα και μετά την επίτευξη ύφεσης. Η διαφορετική έκφραση των Th κυτταροκινών μεταξύ των διαφόρων τύπων σπειραματονεφριτίδων μπορεί να αντανακλά ειδικούς δείκτες εξέλιξης της νεφρικής βλάβης αλλά αυτό απαιτεί περαιτέρω μελέτη. Η συνύπαρξη τόσο της Th1/Th17 όσο και της Th2 απόκρισης ενδεχομένως υποδηλώνει την πιθανή ύπαρξη ενός ανοσορυθμιστικού μηχανισμού που στοχεύει στην ανοσολογική ομοιόσταση του νεφρικού ιστού, ο οποίος χρήζει περαιτέρω διερεύνησης. / IgA nephropathy represents a common glomerular disease treated by various therapeutic regimens. In the first part of the current thesis, the effect of different therapeutic regimens based on the severity of clinical and histological involvement, in the clinical outcome of patients with IgA nephropathy over a follow-up period of 5 years was estimated. Fifty patients were included in the study and were divided in four groups. Patients with normal renal function and proteinuria <1g/24h received no treatment (Group A, n=6). Patients with normal renal function, proteinuria >1g/24h and mild to moderate histological lesions received angiotensin converting enzyme inhibitors (ACEi) and corticosteroids (Group B, n=23). Patients with baseline serum creatinine (Scr) <2.5mg/dl, proteinuria >3.5g/24h and severe histological lesions received ACEi, corticosteroids and other immunosuppressive drugs (Group C, n=18). Patients with Scr >2.5mg/dl, glomerulosclerosis and tubulointerstitial fibrosis received ACEi and fish oil (Group D, n=3). Doubling of baseline Scr was observed in 9 of 50 patients (18%), 2 from group B (8.7%), 5 from group C (27.7%) and 2 from group D (66.7%). Out of 7 (14%) who reached ESRD, 1was from group B (4.3%), 4 from group C (22%) and 2 from group D (66.7%). Reduction of proteinuria was observed in all patients from group B and in 15 from group C (83.3%). Adverse reactions occurred in 3 (7.3%) patients treated with immunosupressive drugs. The choice of therapeutic regimen used in the treatment of patients with IgA nephropathy could be based on the severity of clinical and histological involvement in order to achieve the maximun effect with less adverse reactions. In the second part potential early markers for progression of renal injury in both kidney tissue and urine of patients were studied. Genomic and proteomic studies suggest that transgelin represents a protein that may be involved in renal injury. Transgelin was identified in biopsy sections of 67 patients by immunohistochemistry and immunofluorescence. Its distribution was compared to that of α-smooth muscle actin (α-SMA), a marker of myofibroblast activation in the kidney. Transgelin and α-SMA expression was identified within glomeruli and interstitium. In patients with IgA nephropathy and focal segmental glomerulosclerosis, glomerular expression of transgelin was higher than that of α-SMA. The extent of transgelin immunostaining was related to mesangial proliferation (p=0.034), glomerular sclerosis (p=0.035), interstitial fibrosis (p=0.047) and to the clinical course (p=0.009). Colocalization studies showed that in some areas of kidney tissue both proteins were expressed with comparable intensity, whereas in other areas expression of either transgelin or α-SMA was predominant. Strong transgelin expression was observed in renal tissue of patients with glomerulonephritis. The observed differences in the pattern of transgelin and α-SMA expression suggest that either different subpopulation of myofibroblasts exist, or that these proteins are activated at different stages of renal injury/scarring. Moreover, the levels of pro-inflammatory (IL-2, IL-17, TNF-α, INF-γ, IL-6) and anti-inflammatory (IL-4, IL-10, TGF-β1) cytokines as well as chemokine MCP-1 excretion in the urine of patients with IgA nephropathy were measured and compared with those of healthy individuals and patients with various types of glomerulonephritis after clinical remission of the disease. Ninety seven patients with primary glomerulonephritis were studied. All patients were in clinical remission following immunosuppressive treatment. The original diagnoses were IgA nephropathy (IgAN) in 31, membranous nephropathy (MN) in 36, and minimal changes disease or focal segmental glomerulosclerosis (MC/FSGS) in 30 out of 97 patients. Th- cytokine and MCP-1 levels were measured in a random urine sample and compared to those of 17 healthy individuals. Subgroup analysis of various types of GN was also performed. Patients with glomerulonephritides had significantly higher urinary levels of all tested cytokines, apart from IL-4 and TNF-α, in comparison to healthy individuals. A strong positive correlation of TGF- β1 concentration in the urine with that of MCP-1 was noted in patients with various glomerulonephritides (p=0.000, r=0.721). Subgroup analysis showed statistically significant differences in the concentration of IL-2 (p=0.0345), IL-17A (p=0.005), TNF-α (p=0.0098), TGF-β1 (p=0.0092) and MCP-1 (p=0.0301) between patients with IgAN and MN. Furthermore, a significant difference was observed in the urinary levels of IL-2 between patients with IgAN and those with MC/FSGS (p=0.0018). Th-cytokines and MCP-1 urinary levels of IgAN patients in clinical remission showed an ongoing inflammation of renal tissue. The different concentration of Th cytokines in various types of GN, may represent specific markers of disease activity but this needs to be further investigated. The coexistence of Th1/Th17 and Th2 immune responses may suggest the presence of an immunoregulatory mechanism that triggers renal immune homeostasis.