Progressive Multifocal Leukoencephalopathy (PML) is caused by lytic infection of oligodendrocytes by JC Virus (JCV). JCV Encephalopathy (JCVE) is a newly identified disease characterized by JCV infection of cortical pyramidal neurons. JCVCPN was isolated from the brain of a JCVE patient. JCVCPN contains a unique 143 base pair deletion in the agnogene and has an archetype-like regulatory region (RR), of the type typically found in the kidneys. In this dissertation, we studied the JCVCPN virus to better understand the role of the agnogene and the RR in JCV replication. We used kidney, glial and neuronal cell lines to compare the replication of JCVCPN to the prototype virus JCVMad-1. JCVCPN was able to replicate viral DNA in all cell lines tested, but was unable to establish the high level of infection seen with JCVMad-1. Levels of VP1 capsid protein were undetectable in JCVCPN transfected cells, and few infectious virions were produced. JCVCPN did not have a replication advantage in the neuronal cell line tested. To determine if the agnogene deletion or the archetype-like RR was responsible for the observed phenotype of JCVCPN, we generated a series of chimeric viruses between JCVCPN and JCVMad-1. We found that the phenotype of JCVCPN was due predominantly to the deletion in the agnogene, in particular the loss of the DNA and not the lack of a full length agnoprotein. To further study the role of the agnogene DNA in JCV replication, we introduced a series of small agnogene deletions into a virus with a start codon mutation which prevents agnoprotein expression. We characterized the replication of these additional mutants and found that nucleotides 376-396 are crucial for the expression of VP1 capsid protein. Previous studies have provided evidence for the binding of host cell proteins to the agnogene DNA. We used DNA-Immunoprecipitations with the agnogene to identify candidate binding proteins, but were unable to confirm any candidate proteins as binding specifically to the JCV agnogene. Studying this naturally occurring pathogenic variant of JCV provided a valuable tool for understanding the functions of the agnogene and RR form in JCV replication.

Das humanes WU Polyomavirus wurde im Jahr 2007 als ein neues Virus in Proben des Respirationstraktes beschrieben und gehört zur Familie der Polpymaviridae. Das Ziel der Arbeit war es, eine WUPyV-rea-time-PCR zu etablieren und zu evaluieren und mit dieser neuen Methode WUPyV-DNA in Nasenrachenskreten (NRS) zu detektieren und zu quantifizieren. Insgesamt wurden 1232 NRS von Patientin mit akuten respiratoischen Erkrankungen, die an der Universitätskinderklinik Würzburg im Zeitraum von Januar 2002 bis September 2005 und Januar 2007 bis July 2007 stationär behandelt worden waren, auf WUPyV-DNA getestet. Zusätzlich wurden 14 Serum- und 14 Stuhlproben von Kindern mit WUPyV-DNA-pos. NRS getestet. Mit der real-time PCR wurde WUPyV-DNA in 5,2 % der 1232 NRS detektiert. Der Viruslastmedian aller WUPyV-positiven NRS betrug 950 Kopien/m. Neben einigen sehr hohen Viruslasten (4,7 % > E9 Kopien/ml) wurden vor allem niedirge Viruslaten (51,6 % < 1000 Kopien/ml) mit der WUPyV-real-time PCR nachgewiesen. Es ergaben sich keine statistisch signifikanten Zusammenhänge zwischen der Viruslast und der Koinfektionen mit anderen respiratorischen Viren, mit klinischer Diagnose, mit dem Alter der infizierten Kinder und mit dem jahreszeitlichen Auftreten. In 3 der 14 Serum und 2 der 14 Stuhlproben konnte WUPyV-DNA detektiert werden. Virämische Kinder hatten tendenzen zu höhrer Viruslast im NRS.Weitere Studien sind notwendig um die pathogenetische Relevanz des WUPyV für den Menschen zu untersuchen. Die in dieser Arbeit etablierte real-time PCR zur WUPyV-Quantifizierung kann dabei zur Anwendung kommen. / The human WU polyomavirus (WUPyV) has been recently described as a novel virus in respiratory tract samples. To investigate the viral load of WUPyV in nasopharyngeal aspirates (NPA´s), stool samples, and serum samples of pediatric patients with acute respiratory tract diseases, obtained between 2002 and 2007, we etablished a real-time PCR for WUPyV DNA. WUPyV was found in 5,2 % of 1232 NPA. The median viral load in the NPA was 950 copies/ml (maximun: 3.4 E10 copies/ml). The WUPyV load in NPA was neither associated wtih the coinfection status nor with the clinical diagnoses. WUPyV was found in 3 of 14 serum samples and 2 of 14 stool samples. The WUPyV load in NPA tended to be hihger in viremic children. Further stuies are necessary to determine whether WUPyV is a human pathogen.

Glioblastomas continue to carry poor prognoses for patients despite advances in surgical, chemotherapeutic, and radiation regimens. One feature of glioblastoma associated with poor prognosis is the degree of hypoxia and elevated expression levels of hypoxia-inducible factor-1 á (HIF-1á). HIF-1á expression allows metabolic adaptation to low oxygen availability, partly through upregulation of vascular endothelial growth factor (VEGF) and increased tumor angiogenesis as well as induction of anaerobic glycolysis. In this study, we demonstrate an induced level of astrocyte-elevated gene-1 (AEG-1) by hypoxia in glioblastoma cells. AEG-1 has the capacity to promote anchorage-independent growth and cooperates with Ha-ras in malignant transformation. In addition, AEG-1 was recently demonstrated to serve as an oncogene and can induce angiogenesis and autophagy in glioblastoma. Results from in vitro studies show that hypoxic induction of AEG-1 is dependent on HIF-1á stabilization during hypoxia and that phosphatidylinositol 3-kinase (PI3K) inhibition abrogates AEG-1 induction during hypoxia through loss of HIF-1á stability. Furthermore, we show that AEG-1 is induced by glucose deprivation and that prevention of intracellular reactive oxygen species (ROS) production prevents this induction. Additionally, AEG-1 knockdown results in increased ROS production and increased glucose deprivation-induced cytotoxicity, whereas AEG-1 overexpression prevents ROS production and decreases glucose deprivation-induced cytotoxicity, indicating that AEG-1 induction is necessary for cells to survive this type of cell stress. From studies examining the expression of enzymes involved in glucose metabolism, we demonstrate that AEG-1 alters the tumor metabolic profile in a partially 5'-adenosine monophosphate (AMP)-activated protein kinase (AMPK)-dependent manner. Moreover, glycolytic inhibition modulates the metabolic effects induced by AEG-1, and AEG-1 knockdown reduces the growth and alters the metabolic phenotype of glioblastoma subcutaneous xenografts. These observations link AEG-1 overexpression observed in glioblastoma with hypoxia and glucose metabolic signaling, and targeting these physiological pathways may lead to therapeutic advances in the treatment of glioblastoma in the future. Recent studies have reported the detection of the human neurotropic virus, JC Virus (JCV), in a significant population of brain tumors, including medulloblastomas. Accordingly, expression of the JCV early protein, T-antigen, which has transforming activity in cell culture and in transgenic mice, results in the development of a broad range of tumors of neural crest and glial origin. Evidently, the association of T-antigen with a range of tumor-suppressor proteins, including p53 and pRb, and signaling molecules, such as â-catenin and IRS-1, play a role in the oncogenic function of JCV T-antigen. We demonstrate that T-antigen expression is suppressed by glucose deprivation in medulloblastoma cells that endogenously express T-antigen. Mechanistic studies indicate that glucose deprivation-mediated suppression of T-antigen is partly influenced by AMPK, a critical sensor of the AMP/ATP ratio in cells. We have found that AMPK activation inhibits T-antigen expression, whereas AMPK inhibition prevents glucose deprivation-mediated T-antigen suppression. In addition, glucose deprivation-induced cell cycle arrest in the G1 phase is blocked with AMPK inhibition, which also prevents T-antigen downregulation. Furthermore, T-antigen-expressing medulloblastoma cells, as compared to those which do not express T-antigen, exhibit less G1 arrest and an increased percentage of cells in the G2 phase of the cell cycle during glucose deprivation. On a functional level, T-antigen downregulation is partially dependent on ROS production during glucose deprivation. Additionally, studies indicate that T-antigen prevents ROS induction, loss of ATP production, and cytotoxicity induced by glucose deprivation. We have also found that T-antigen is downregulated by the glycolytic inhibitor, 2-deoxy-D-glucose (2-DG), and the pentose phosphate inhibitors, 6-aminonicotinamde (6-AN) and oxythiamine (OT). Enzyme expression studies also indicate that T-antigen upregulates the expression of the pentose phosphate enzyme, transaldolase-1 (TALDO1), demonstrating a potential link between T-antigen and glucose metabolic regulation. These studies highlight the potential involvement of JCV T-antigen in the proliferation and metabolic phenotype of medulloblastoma and may enhance our understanding of the role of viral proteins in tumor glycolytic metabolism, thus implicating these proteins as potential targets for the treatment of virus-associated tumors. / Biomedical Neuroscience

A relação entre os diferentes genótipos do VJC e a patogênese da LEMP permanece ainda uma questão controversa. O presente estudo visou a caracterização genotipica do vírus JC (VJC) e sua associação com a leucoencefalopatia multifocal progressiva (LEMP) em pacientes com aids em São Paulo, no período de 2000 a 2008. Foram avaliadas 51 amostras de líquor de pacientes com LEMP e 47 amostras de pacientes com aids sem LEMP positivas para os genes T e VP1 pela reação em cadeia por polimerase (PCR). Foram identificados, através do sequenciamento genômico os genótipos 1, 2, 3, 4 e 6, sendo que os mais prevalentes foram os genótipos 2 (33%), 1 (27%), seguidos pelo genótipo 3 (23%). O genótipo 1 e mostrou associação positiva com a LEMP (p<0,05), enquanto que o genótipo 3 mostrou associação inversa à LEMP. Durante a internação em que foi diagnosticada LEMP observamos evolução para óbito em 27 (59%) pacientes e alta em 19 (41%). Não houve associação entre os genótipos do VJC ou carga viral liquórica e a evolução para óbito, mas houve associação significante entre o número de linfócitos TCD4+ e o óbito. No presente estudo, uma mutação no sítio 91 do gene VP1 com substituição da leucina para isoleucina ou valina, não descrita anteriormente, foi encontrada exclusivamente em pacientes com LEMP. Substituições nos sítios 123, 128 e 134 foram observadas mais frequentemente nos casos de LEMP. Não foram encontradas diferenças filogenéticas entre as sequências obtidas dos casos e dos controles. A PCR em tempo real padronizada mostrou sensibilidade e especificidade diagnósticas de 89% e 100%, respectivamente. / The relationship between the different genotypes of VJC and the pathogenesis of progressive multifocal leukoencephalopathy remains a controversial issue. This study aimed to genotypic characterization of JC virus (JCV) and its association with progressive multifocal leukoencephalopathy (PML) in AIDS patients in Sao Paulo, from 2000 to 2008. We analyzed 51 CSF samples from patients with PML and 47 samples of AIDS patients without PML positives for the T and VP1 genes by polymerase chain reaction (PCR). Were identified by genomic sequencing genotypes 1, 2, 3, 4 and 6, and were the most prevalent genotypes 2 (33%), 1 (27%), followed by genotype 3 (23%). Genotype 1 showed a positive association with PML (p <0.05), while genotype 3 was inversely associated with PML. Overall in-hospital mortality was 59% (27 patients). No association between the genotypes of VJC or CSF viral load and progression to death, but there was a significant association between number of CD4 + and death. In this study, a mutation in site 91 of VP1 gene with substitution of leucine for isoleucine or valine, not described previously, was found exclusively in patients with PML. Substitutions at sites 123, 128 and 134 were observed more frequently in cases of PML. No differences were found between phylogenetic sequences obtained from cases and controls. The real-time PCR showed standardized diagnostic sensitivity and specificity of 89% and 100%, respectively.

Os Poliomavírus JC e BK são os mais importantes integrantes da família Polyomaviridae, gênero Orthopolyomavirus, devido ao seu grau de patogenicidade no homem. O poliomavírus humano JC (JCV) foi isolado a partir de fragmento do cérebro de um paciente com linfoma de Hodgkin e leucoencefalopatia multifocal progressiva por Padgett e colaboradores. Já o poliomavírus humano BK foi isolado em 1971 por Gardner e colaboradores a partir da semeadura da urina de um paciente, submetido a transplante renal, em cultura de células da linhagem VERO Poucos são os dados sobre os poliomavírus humanos JC e BK em indivíduos sadios no mundo e no Brasil. Além disso, as formas de excreção e transmissão destes vírus ainda não estão completamente elucidadas. Este estudo teve como objetivos principais determinar a incidência, a dinâmica de excreção e a caracterização genotípica dos poliomavírus JC e BK em amostras sequenciais de urina de indivíduos sadios. Como objetivo secundário, foram analisados filogeneticamente os subtipos dos vírus encontrados. Foram incluídos 71 indivíduos de ambos os sexos, com idades variando entre 21 e 65 anos e destes foram coletadas amostras mensais de urina durante 6 meses. Todas as amostras do estudo foram submetidas a um teste de PCR em tempo real, que amplifica um fragmento do gene que codifica o antígeno T, e a um PCR convencional que amplifica um fragmento do gene da proteína VP1 do vírus. Ao final do período de 6 meses de acompanhamento a incidência de excreção urinária dos poliomavírus JCV e BKV na população estudada foi de 53,52% e 64,79% respectivamente. O perfil de excreção do JCV mostrou-se continuo em 42% dos casos , intermitente em 8% e esporádico em 50% dos casos. Analisando o perfil de excreção do BKV, em 80% dos casos mostrou-se esporádico, em 17% intermitente e em 3% dos casos contínuo. Posteriormente, as amostras positivas foram sequenciadas e analisadas filogeneticamente observando-se que os genótipos mais prevalentes do JCV foram o 1, subtipo 1B e 3, seguidos dos genótipos 1, subtipo 1A e 4. Com relação ao BKV, o genótipo 1, subtipo 1A foi o mais prevalente, seguido do 4 e do 1, subtipo 1B. / The Polyomavirus JC and BK are the most important members of Polyomaviridae family, genus Orthopolyomavirus, due to their degree of pathogenicity in humans. The human polyomavirus JC (JCV) was isolated from a fragment of the brain of a patient with Hodgkin\'s lymphoma and progressive multifocal leukoencephalopathy by Padgett et al. On the other hand the human polyomavirus BK was also isolated in 1971 by Gardner et al from the urine of a patient who underwent renal transplantation in VERO cell line. There are few data on human polyomavirus JC and BK in healthy individuals on the world and in Brazil. Moreover the forms of excretion and transmission of these viruses are not yet fully elucidated. This study aimed to determine the incidence, the dynamics of excretion, and the molecular characterization of polyomavirus JC and BK in serial samples of urine of healthy individuals. A secondary objective was to analyze phylogenetically the subtypes of the viruses found during the study. Were included 71 patients of both genders, aged from 21 to 65 yearsold. Urine samples were collected every month for six months. All samples in the study were screened by a real time PCR which amplifies a fragment of the T antigen gene, and to a conventional PCR that amplifies a fragment of the gene of VP1 protein of the virus. At the end of 6 months of follow-up, the incidence of urinary excretion of polyomaviruses BKV and JCV in the study population was 53.52% and 64.79% respectively The profile of excretion of JCV was continuous in 42% of cases, intermittent in 8% and sporadic in 50% of cases. The profile of excretion of BKV was shown to be sporadic in 80% of cases, intermittent in 17% and continuous in only 3% of cases. Subsequently, the positive samples were sequenced and analyzed phylogenetically showing that the more prevalent genotypes were JCV 1, subtype 1b and 3, followed by genotype 1, subtypes 1a and 4. Regarding the BKV genotype 1 subtype 1a was the most prevalent, followed by 4 and 1, subtype 1b.

A Esclerose Múltipla (EM) é uma doença autoimune caracterizada por um processo neuroinflamatório com degeneração axonal progressiva. O medicamento Natalizumab (Biogen Idec, NC, USA) representa hoje um dos tratamentos mais promissores para EM. Entretanto, pacientes sob esse tratamento possuem maiores chances de desenvolver Leucoencefalopatia Multifocal Progressiva (LEMP), em decorrência de uma possível reativação do poliomavírus JC (VJC). Além do VJC, o poliomavírus BK (VBK) pode representar uma preocupação adicional para tais pacientes, uma vez que também apresenta capacidade de causar encefalopatias. Apesar do Natalizumab ser uma ótima ferramenta contra a EM, o fato de interagir de alguma maneira com os poliomavírus, em especial o VJC, impede que seja utilizado em larga escala. Dessa forma,o objetivo desse trabalho foi investigar os padrões de excreção e reativação dos VJC e VBK em pacientes com EM durante o tratamento com Natalizumab e comparar aos padrões observados em pacientes que se encontram sob outros tratamentos. Amostras seriadas de sangue e urina foram coletadas e submetidas a testes de biologia molecular para detecção do vírus e caracterização molecular. Foram analisados 97 pacientes em diferentes tempos de acompanhamento. Não foi observada presença de poliomavírus no sangue de nenhum dos indivíduos analisados. Entretanto, 36% excretavam poliomavírus na urina em pelo menos uma das coletas, sendo que 21,7% excretavam VJC, 9,3% excretavam VBK e 5,1% excretavam ambos os poliomavírus. Não foi observada diferença entre as taxas de excreção urinária de poliomavírus entre pacientes que tratavam com Natalizumab (38,9%) e pacientes que sob outros tratamentos (34,5%), sendo que para o Grupo Controle (GC); 21,3%, 8,2% e 4,9% excretavam VJC, VBK e ambos os vírus, respectivamente e para o grupo Grupo Natalizumab (GN) 22,2%, 11,1% e 5,6% excretavam VJC, VBK e ambos os vírus, respectivamente. As análises moleculares da Região Regulatória do VJC revelaram sequências de característica arquetípica. A reconstrução filogenética de sequências do gene VP1 do VJC revelou predominância do genótipo 3 e do genótipo 1 para o VBK. Não foi observada diferença estatística da carga viral do VJC e do VBK entre os dois grupos. Foi detectada uma mutação (E29G) na VP1 de uma paciente que apresentou alta carga viral do VJC. Do grupo GN, 14 apresentaram anticorpos para VJC, sendo que desses 58% apresentou excreção de VJC, 42% não apresentou excreção urinária, interessantemente uma paciente não apresentou anticorpos contra o VJC, mas apresentou excreção de VJC. Pode-se concluir principalmente que a detecção de anticorpos, concomitantemente com a investigação molecular do VJC poderá contribuir para uma melhor determinação da estratificação do risco de desenvolvimento de LEMP em indivíduos com EM sob tratamento com Natalizumab. / Multiple sclerosis (MS) is an autoimmune disease characterized by neuronal inflamatory process with progressive axonal degeneration. The drug Natalizumab (Biogen Idec, NC, USA) is today one of the most promising treatments for MS. However, patients undergoing this treatment have higher chances of developing progressive multifocal leukoencephalopathy (PML), due to a possible reactivation of the polyomavirus JC (VJC). Besides VJC, the BK polyomavirus (VBK) may represent an additional concern for such patients, since it also has ability to cause encephalopathies. Despite Natalizumab be a great tool against MS, the fact that drug some way may interact with polyomavirus, especially VJC, prevents it from being used on a large scale. Thus, aim of this study was to investigate the patterns of excretion and reactivation of VJC and VBK in MS patients during treatment with Natalizumab and compare the patterns observed in patients who are under other treatments. Serial blood samples and urine were collected and submitted to molecular biology tests for virus detection and molecular characterization. Ninety seven patients were analyzed at different follow-up times. There was no polyomavirus DNA in the blood of none subjects analyzed. However, 36% of patients excreted polyomavirus in the urine in at least one of the samples, of those 21.7%, 9.3% and 5.1% excreted VJC, VBK and both polyomavirus, respectively. No difference was observed between the rates of urinary excretion of polyomavirus patients treated with Natalizumab (38.9%) and patients treated with other drugs (34.5%), for the Control Group (GC); 21,3%, 8,2% and 4,9 shed VJC, VBK and both viruses, respectevely and for the Natalizumab Group (GN) 22,2%, 11,1% and 5,6% shed VJC, VBK and both viruses, respectvely. Molecular analysis of the Regulatory Region of VJC revealed sequences similar to the archetype form of VJC. A phylogenetic reconstruction of the VP1 gene sequences revealed VJC predominance of genotype 3 and genotype 1 for VBK. There was no statistical difference in the viral load VJC and VBK between the two groups. It was detected a mutation (E29G) in VP1 of a patient who had a high viral load VJC, however the mutation disappeared after a few months of monitoring. Fourteen patients of GN had antibodies to VJC, and of these 58% had excretion VJC, 42% showed no urinary excretion, interestingly one patient had no antibodies against VJC but showed excretion of VJC. It can be concluded that mostly anti-VJC antibodies detection, concurrently with the VJC molecular research may contribute to a better determination of risk stratification for development of PML in patients with MS undergoing treatment with Natalizumab.

Introdução: Cistite Hemorrágica (CH) é uma grave complicação do Transplante de Células-Tronco Hematopoiéticas (TCTH) Alogênico. Sua incidência varia de 12 a 25,5%. A forma precoce desenvolve-se devido aos efeitos tóxicos de certos quimioterápicos usados no regime de condicionamento, especialmente Ciclofosfamida. Já a CH tardia ocorre a partir do terceiro dia após o TCTH e sua etiologia é multifatorial. Vários fatores de risco para o desenvolvimento de CH tardia foram descritos, incluindo Doença do Enxerto Contra o Hospedeiro (DECH) aguda, doador não relacionado, infecções por vírus urotrópicos, sexo masculino e condicionamento mieloablativo. Materiais e Métodos: O presente estudo tem como objetivos descrever a incidência de CH em pacientes adultos e pediátricos submetidos a TCTH alogênico e identificar fatores de risco associados ao desenvolvimento de CH nesta população. Foram analisados dados de prontuário de 347 pacientes submetidos a TCTH Alogênico no Hospital de Clínicas de Porto Alegre no período de Janeiro de 2001 a Dezembro de 2014. Resultados: CH ocorreu em 42 pacientes (12,1%, IC: 8,9 - 16%), em uma média de 53.4 dias após o procedimento (desvio padrão: 28.1 dias). Apenas 1 paciente (2,4%) desenvolveu CH precoce, com início dos sintomas no D+1. Entre os 41 pacientes que desenvolveram CH tardia, BK vírus foi o principal agente viral identificado. CH ocorreu em 12.8% dos pacientes que receberam condicionamento mieloablativo e em 10.5% dos restantes (P = 0,704). Dos 197 pacientes que apresentaram DECH aguda, 35 (17,8%) desenvolveram CH e somente 7 (4,9%) apresentaram CH na ausência de DECH aguda (P < 0,001). CH foi mais frequente também em pacientes do sexo masculino (P = 0,027). Conclusão: A incidência de CH em nossa amostra foi semelhante à encontrada em outros trabalhos. DECH aguda e sexo masculino estiveram associados a um maior risco de desenvolvimento de CH. / Introduction: Hemorrhagic cystitis (HC) is a serious complication of Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) afecting 12 to 25.5% of the patients. The early-onset form of HC develops during or until 72 hours after the conditioning regimen containing high doses of certain chemotherapy drugs such as Busulfan and especially Cyclophosphamide. Late-onset HC occurs from the third day on after HSCT and its etiology is multifactorial. Several risk factors for the late-onset form have been reported including graft-versus-host disease (GVHD), unrelated donor, urotropic infections, male gender and myeloblative conditioning regimen. Methods: This study aims to evaluate the incidence of HC in adult and pediatric patients undergoing Allogeneic HSCT and to identify risk factors associated with the development of HC in this population. Medical records of 347 patients who underwent Allogeneic HSCT at Hospital de Clínicas, Porto Alegre, Brazil, from January 2001 to December 2014 were analyzed. Results: HC occurred in 42 patients (12.1% CI: 8.9 - 16%) at an average of 53.4 days after the procedure (standard deviation: 28.1 days). Only one of them developed early-onset HC, with onset of symptoms on D+1. Among the 41 patients who developed late-onset HC, BKV was the main identified viral agent. HC developed in 12.8% of the patients treated with myeloablative conditioning and in 10.5% of the remaining patients (P = 0.704). Of the 197 patients with acute GVHD, 35 (17.8%) developed HC and only 7 (4.9%) showed HC in the absence of GVHD (P<0.001). HC was also more frequent in males than females (P = 0.027). Conclusion: The incidence of HC in our sample was similar to that found in other studies. In our cohort of patients being male and having acute GVHD increased the risk of developing HC.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010.

Introdução: Cistite Hemorrágica (CH) é uma grave complicação do Transplante de Células-Tronco Hematopoiéticas (TCTH) Alogênico. Sua incidência varia de 12 a 25,5%. A forma precoce desenvolve-se devido aos efeitos tóxicos de certos quimioterápicos usados no regime de condicionamento, especialmente Ciclofosfamida. Já a CH tardia ocorre a partir do terceiro dia após o TCTH e sua etiologia é multifatorial. Vários fatores de risco para o desenvolvimento de CH tardia foram descritos, incluindo Doença do Enxerto Contra o Hospedeiro (DECH) aguda, doador não relacionado, infecções por vírus urotrópicos, sexo masculino e condicionamento mieloablativo. Materiais e Métodos: O presente estudo tem como objetivos descrever a incidência de CH em pacientes adultos e pediátricos submetidos a TCTH alogênico e identificar fatores de risco associados ao desenvolvimento de CH nesta população. Foram analisados dados de prontuário de 347 pacientes submetidos a TCTH Alogênico no Hospital de Clínicas de Porto Alegre no período de Janeiro de 2001 a Dezembro de 2014. Resultados: CH ocorreu em 42 pacientes (12,1%, IC: 8,9 - 16%), em uma média de 53.4 dias após o procedimento (desvio padrão: 28.1 dias). Apenas 1 paciente (2,4%) desenvolveu CH precoce, com início dos sintomas no D+1. Entre os 41 pacientes que desenvolveram CH tardia, BK vírus foi o principal agente viral identificado. CH ocorreu em 12.8% dos pacientes que receberam condicionamento mieloablativo e em 10.5% dos restantes (P = 0,704). Dos 197 pacientes que apresentaram DECH aguda, 35 (17,8%) desenvolveram CH e somente 7 (4,9%) apresentaram CH na ausência de DECH aguda (P < 0,001). CH foi mais frequente também em pacientes do sexo masculino (P = 0,027). Conclusão: A incidência de CH em nossa amostra foi semelhante à encontrada em outros trabalhos. DECH aguda e sexo masculino estiveram associados a um maior risco de desenvolvimento de CH. / Introduction: Hemorrhagic cystitis (HC) is a serious complication of Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) afecting 12 to 25.5% of the patients. The early-onset form of HC develops during or until 72 hours after the conditioning regimen containing high doses of certain chemotherapy drugs such as Busulfan and especially Cyclophosphamide. Late-onset HC occurs from the third day on after HSCT and its etiology is multifactorial. Several risk factors for the late-onset form have been reported including graft-versus-host disease (GVHD), unrelated donor, urotropic infections, male gender and myeloblative conditioning regimen. Methods: This study aims to evaluate the incidence of HC in adult and pediatric patients undergoing Allogeneic HSCT and to identify risk factors associated with the development of HC in this population. Medical records of 347 patients who underwent Allogeneic HSCT at Hospital de Clínicas, Porto Alegre, Brazil, from January 2001 to December 2014 were analyzed. Results: HC occurred in 42 patients (12.1% CI: 8.9 - 16%) at an average of 53.4 days after the procedure (standard deviation: 28.1 days). Only one of them developed early-onset HC, with onset of symptoms on D+1. Among the 41 patients who developed late-onset HC, BKV was the main identified viral agent. HC developed in 12.8% of the patients treated with myeloablative conditioning and in 10.5% of the remaining patients (P = 0.704). Of the 197 patients with acute GVHD, 35 (17.8%) developed HC and only 7 (4.9%) showed HC in the absence of GVHD (P<0.001). HC was also more frequent in males than females (P = 0.027). Conclusion: The incidence of HC in our sample was similar to that found in other studies. In our cohort of patients being male and having acute GVHD increased the risk of developing HC.

Introdução: Cistite Hemorrágica (CH) é uma grave complicação do Transplante de Células-Tronco Hematopoiéticas (TCTH) Alogênico. Sua incidência varia de 12 a 25,5%. A forma precoce desenvolve-se devido aos efeitos tóxicos de certos quimioterápicos usados no regime de condicionamento, especialmente Ciclofosfamida. Já a CH tardia ocorre a partir do terceiro dia após o TCTH e sua etiologia é multifatorial. Vários fatores de risco para o desenvolvimento de CH tardia foram descritos, incluindo Doença do Enxerto Contra o Hospedeiro (DECH) aguda, doador não relacionado, infecções por vírus urotrópicos, sexo masculino e condicionamento mieloablativo. Materiais e Métodos: O presente estudo tem como objetivos descrever a incidência de CH em pacientes adultos e pediátricos submetidos a TCTH alogênico e identificar fatores de risco associados ao desenvolvimento de CH nesta população. Foram analisados dados de prontuário de 347 pacientes submetidos a TCTH Alogênico no Hospital de Clínicas de Porto Alegre no período de Janeiro de 2001 a Dezembro de 2014. Resultados: CH ocorreu em 42 pacientes (12,1%, IC: 8,9 - 16%), em uma média de 53.4 dias após o procedimento (desvio padrão: 28.1 dias). Apenas 1 paciente (2,4%) desenvolveu CH precoce, com início dos sintomas no D+1. Entre os 41 pacientes que desenvolveram CH tardia, BK vírus foi o principal agente viral identificado. CH ocorreu em 12.8% dos pacientes que receberam condicionamento mieloablativo e em 10.5% dos restantes (P = 0,704). Dos 197 pacientes que apresentaram DECH aguda, 35 (17,8%) desenvolveram CH e somente 7 (4,9%) apresentaram CH na ausência de DECH aguda (P < 0,001). CH foi mais frequente também em pacientes do sexo masculino (P = 0,027). Conclusão: A incidência de CH em nossa amostra foi semelhante à encontrada em outros trabalhos. DECH aguda e sexo masculino estiveram associados a um maior risco de desenvolvimento de CH. / Introduction: Hemorrhagic cystitis (HC) is a serious complication of Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) afecting 12 to 25.5% of the patients. The early-onset form of HC develops during or until 72 hours after the conditioning regimen containing high doses of certain chemotherapy drugs such as Busulfan and especially Cyclophosphamide. Late-onset HC occurs from the third day on after HSCT and its etiology is multifactorial. Several risk factors for the late-onset form have been reported including graft-versus-host disease (GVHD), unrelated donor, urotropic infections, male gender and myeloblative conditioning regimen. Methods: This study aims to evaluate the incidence of HC in adult and pediatric patients undergoing Allogeneic HSCT and to identify risk factors associated with the development of HC in this population. Medical records of 347 patients who underwent Allogeneic HSCT at Hospital de Clínicas, Porto Alegre, Brazil, from January 2001 to December 2014 were analyzed. Results: HC occurred in 42 patients (12.1% CI: 8.9 - 16%) at an average of 53.4 days after the procedure (standard deviation: 28.1 days). Only one of them developed early-onset HC, with onset of symptoms on D+1. Among the 41 patients who developed late-onset HC, BKV was the main identified viral agent. HC developed in 12.8% of the patients treated with myeloablative conditioning and in 10.5% of the remaining patients (P = 0.704). Of the 197 patients with acute GVHD, 35 (17.8%) developed HC and only 7 (4.9%) showed HC in the absence of GVHD (P<0.001). HC was also more frequent in males than females (P = 0.027). Conclusion: The incidence of HC in our sample was similar to that found in other studies. In our cohort of patients being male and having acute GVHD increased the risk of developing HC.