La mammite bovine à S. aureus se caractérise, dune part, par la persistance dun nombre élevé de cellules inflammatoires (principalement des neutrophiles) dans le lait, et, dautre part, par la persistance de la bactérie dans la glande mammaire. La guérison dune mammite à S. aureus repose donc sur deux facteurs étroitement liés : le retour à une concentration normale de cellules dans le lait et léradication de lorganisme pathogène. Or, laccumulation et lactivation des neutrophiles au site de linfection requiert lexpression de cytokines pro-inflammatoires, telles que lIL-8 et le GM-CSF, et la croissance intra- et extracellulaire de S. aureus est augmentée en présence de cytokines inflammatoires, telles que lIL-1b et le TNF-a. La persistance de cytokines pro-inflammatoires semble donc être défavorable à la guérison dune mammite à S. aureus. Lexpression de la plupart des cytokines pro-inflammatoires est sous la dépendance du facteur nucléaire kB (NF kB). Par conséquent, NF kB pourrait être une cible thérapeutique intéressante pour améliorer ou soigner la mammite à S. aureus. Au début de ce travail aucune étude navait été dédiée au rôle de NF kB dans la mammite. Nous avons par conséquent tenté de clarifier ce rôle, ainsi que le potentiel de NF-kB en tant que cible thérapeutique dans la mammite bovine.

A tuberculose (TB) é um grave problema de saúde pública. Somente no ano de 2004, cerca de 9 milhões de pessoas desenvolveram TB ativa e mais de 2 milhões de pessoas morreram da doença. O desenvolvimento de novos modelos experimentais de TB seriam de grande utilidade para para elucidar mecanismos fisiopatológicos da doença e testar esquemas terapêuticos para a prevenção e contenção da doença. Além disso, o desenvolvimento de novas vacinas torna-se indispensável como ferramenta de prevenção e controle da TB. A TB no sistema nervoso central (SNC), assim como em outros tecidos do organismo, promove a ativação de células inflamatórias. No SNC a micróglia desempenha este papel, sendo capaz de produzir ou ser influenciada por mediadores solúveis. Vários mediadores estão envolvidos nos mecanismos moleculares decorrentes da infecção e inflamação causados pela TB, entre eles: NFB, iNOS e VEGF. A ativação do NFB, um fator de transcrição citoplasmático que sob estímulo migra para o núcleo celular, tem íntima relação com a indução da iNOS e de VEGF. A resistência intracelular a patógenos, inclusive ao Mycobacterium tuberculosis, parece estar associada a expressão de iNOS em macrófagos. O óxido nítrico (NO) tem papel importante na comunicação intercelular, estimulando a síntese de mediadores inflamatórios, como as citocinas, e regulando sua própria produção endógena. Estas citocinas por sua vez também podem induzir a atividade do NFB e a expressão da iNOS e VEGF. O VEGF é um potente ativador de permeabilidade vascular e de angiogênese, envolvido na ruptura da barreira hemato-encefálica. Neste estudo, mostramos a caracterização morfológica e imuno-histoquímica de um modelo murino de TB no SNC, com a indução da doença pela inoculação de BCG. Com este modelo experimental obtivemos importantes resultados que podem esclarecer mecanismos envolvidos na fisiopatologia da neuro-TB humana. A indução de meningite e tuberculomas foi possível através da inoculação de 104 cfu de BCG no cerebelo de camundongos, por estereotaxia, e esta indução foi dependente do tempo. A confirmação do diagnóstico foi feita pela detecção de bacilos álcool-ácido resistentes (BAAR), nas lesões tuberculosas. Observamos, ao longo do tempo (1 a 6 dias; 1, 2, 4 e 8 semanas) o recrutamento de diferentes populações gliais (micróglia e astrócitos) no sítio de injeção. Houve aumento de produção e ativação NFB nas lesões tuberculosas, caracterizada pela translocação da molécula do citoplasma para o núcleo celular. Houve expressão de iNOS restrita às lesões tuberculosas, além do aumento de expressão de VEGF nestas lesões. Além disso, camundongos imunizados com a vacina gênica hsp65, contra a TB, não expressam VEGF em suas lesões. Esta vacina parece conferir um efeito protetor em nosso modelo experimental, reduzindo a expressão de VEGF, e consequentemente reduzindo seu efeito angiogênico decorrente do processo inflamatório. O recrutamento glial, e a produção de mediadores solúveis (NFB, iNOS e VEGF) pelo hospedeiro, em resposta à invasão do patógeno no SNC, parecem estar envolvidos na fisiopatologia da neurotuberculose, como demonstrado neste modelo experimental. Nosso modelo permitirá investigar fatores possivelmente responsáveis pelo desenvolvimento e manutenção de lesões tuberculosas no SNC. O objetivo final seria elucidar a fisiopatologia desta grave doença e compreender eventos moleculares envolvidos na produção de lesões. O conhecimento gerado poderá permitir o delineamento de terapias específicas e efetivas. / Tuberculosis (TB) is a serious public health problem; in 2004, 9 million people developed active TB and the disease killed 2 million patients. Development of experimental models and new vaccines are essential both to elucidate physiopathological mechanisms and to control the disease. This infection in the central nervous system (CNS), as in other tissues of the organism, activates inflammatory cells. In CNS, this role is performed by the microglia, which is capable of producing or be influenced by soluble mediators. Several mediators are involved in the molecular mechanisms of the infection and inflammation by mycobacteria , such as NFB, iNOS and VEGF. NFB activation, a cytoplasmic transcriptional factor that migrates to the cellular nucleus under stimuli, is involved with the iNOS and VEGF induction of expression. The intracellular resistance to Mycobacterium tuberculosis has been associated with iNOS expression in macrophage cells. Nitric oxide (NO) is crucial in intercellular communication, modulating the synthesis of mediators of inflammation, such as cytokines, and modulation itself. These cytokines induces NFB activity, and induces iNOS and VEGF expression. VEGF is a potent activator of vascular permeability and of angiogenesis and it is a factor involved in the breakdown of the blood brain-barrier in tuberculous meningitis. In this study, we showed the morphologic and immunohistochemistry characterization of an experimental model of TB in the CNS, with inoculation of BCG in mice. In this model we elicited important outcome that can elucidate mechanisms involved in the physiopathology of human neuron-TB. Induction of meningitis and tuberculomas were possible with stereotaxic inoculation of 104 cfu of BCG in mice cerebellum, in a time-dependent way. Diagnostic was confirmed by detection of alcohol-acid resistant bacilli (BAAR), in tuberculous lesions. We observed, the time-course (1 to 6 days; 1, 2, 4 e 8 weeks) of the recruitment of different glial populations (microglia and astrocytes) in the injection site. There was increased production and activation of NFB in the tuberculous lesions, it was characterized by its nuclear translocation from cytoplasm. There was iNOS expression only in the tuberculous lesions, and expression increased of VEGF in these lesions. Furthermore, mice immunizated with vaccine DNA-hsp65 there was no expression of VEGF in its lesions. This vaccine seems confer a protector effect in our experimental model, reducing the expression of VEGF, and then reducing its angiogenic effect derived from inflammatory process. Glial recruitment, and the soluble mediators production (NFB, iNOS e VEGF) by the host, producing in response to invasion of the pathogen in the CNS, has been involved in the pathophysiology of the neuro-TB, such as demonstrated in this experimental model. Our model will allow investigate possible factors responsible for the development and maintenance of tuberculous lesions in the CNS. The final aim is to elucidate the physiopathology of this serious illness and understand the molecular events involved in the production of the lesions. The knowledge created may permit to pave the way to delineate specific and effective therapies.

Japanese encephalitis virus (JEV) is one of the major causes of encephalitis in Asia. JEV causes serious inflammation of the brain, which may lead to permanent brain damage and has a high mortality rate. Almost 3 billion people live in JE endemic areas and JEV causes an estimated 20,000 cases of disease and 6000 deaths per year. JEV is a positive stranded RNA virus belonging to the Flavivirus genus of the family Flaviviridae. The genome of JEV is about 11 kb long and codes for a polyprotein which is cleaved by both host and viral encoded proteases to form 3 structural and 7 non-structural proteins. JEV transmission occurs through a zoonotic cycle involving mosquitoes and vertebrate amplifying hosts, chiefly pigs and ardeid birds. Humans are infected when bitten by an infected mosquito and are dead end hosts. The role of humoral and cell mediated immune responses during JEV infection have been studied by several groups. While the humoral responses play a central role in protection against JEV, the cell mediated immune responses contributing to this end are not fully understood. The MHC molecules have been known to play predominant roles in host responses to viral infections and the consequences of virus infection on the expression of MHC molecules are varied. The expression of MHC-I molecules is known to decrease upon infection with many viruses such as HIV, MCMV, HCMV, Adv, and EBV. In contrast, infection with flavivirus such as West Nile Virus (WNV) has been shown to increase the cell surface expression of both MHC-I and MHC-II molecules. It has been reported previously that WNV infection increases the cell surface expression of adhesion molecules such as ICAM-1, VCAM-1 as well as E-Selectin and these changes were mediated directly by WNV and not by soluble cytokines. In contrast to classical MHC-I molecules, the nonclassical MHC-I molecules do not belong to a single group of structurally and functionally homologous proteins and normally have lower cell surface expression. Earlier studies have shown that the expression of nonclassical MHC-I molecules were induced during infection with JHM strain of mouse hepatitis virus (MHV). However, the functional significance of this induction is unclear. Expression of nonclassical MHC-I molecules upon flaviviral infection is not very well understood. In this thesis, evidence is presented that JEV infection induces the expression of both classical and nonclassical MHC-I molecules on primary mouse brain astrocytes, mouse embryonic fibroblasts (MEFs) and H6 (hepatoma cell). The levels of adhesion molecules as well as molecules involved in antigen processing and presentation were also analyzed and our results clearly demonstrate that JEV infection induces their expression on astrocytes, MEFs and H6. The role of NF-κB and type-I IFNs in the induction of classical and nonclassical MHC-I molecules as well as molecules involved in antigen processing and presentation were also analyzed and our results demonstrated that type-I IFN mediated signaling is responsible for the induction of these molecules during JEV infection. Chapter 1 discusses the innate and adaptive immune system, the role of classical and nonclassical MHC molecules in the initiation of immune response and diverse strategies adapted by different viruses to evade the immune response. It also includes a detailed discussion about the IFN and NF-κB signaling pathways and their modulation by viral infection. Finally, the genome organization, epidemiology, transmission cycle, pathogenesis and pathology, clinical features, humoral as well as cell mediated immune response to JEV infection and the current vaccine status to JEV infection are briefly discussed. Chapter 2 describes the general materials and methods used in this study. It includes the details of the reagents and cell lines used in the experiments. It also discusses the various techniques such as RT-PCR, FACS analysis, EMSA and ELISA. Chapter 3 focusses on the validation of different knockout MEFs used in the study as well as confirming the purity of primary astrocyte cultures established from pub brains. The susceptibility of various cells to JEV infection has also been investigated. Our results confirmed the authenticity of all the cells and the purity of primary astrocyte cultures used in the study. Our results also indicated that all the cells used in the study are susceptible to JEV infection. Chapter 4 discusses the expression of MHC and related genes involved in immune response upon JEV infection of primary mouse brain astrocytes, MEFs and H6. Chapter 4 demonstrates for the first time that JEV infection induces the expression of nonclassical MHC-I or class Ib molecules namely Qa-1, Qb1 and T10 in addition to the induction of classical MHC-I molecules. In contrast to WNV, there was no increase in the cell surface expression of MHC-II molecules upon JEV infection of primary mouse brain astrocytes. JEV infection also induces the expression of adhesion molecules as well as molecules involved in antigen processing and presentation namely Tap1, Tap2, Tapasin, Lmp2, Lmp7 and Lmp10. Chapter 5 demonstrates that JEV infection induces NF-κB activation in astrocytes and MEFs. Studies using MEFs deficient in classical and alternate pathways of NF-κB activation indicate that JEV activates the classical pathway of NF-κB activation and is dependent on canonical lKKβ/IKK2 activity. JEV infection of astrocytes, MEFs and H6 induces the production of type-I IFNs. To determine the mechanism of type-I IFN induction during JEV infection, MEFs deficient in NF-κB signaling and IFN signaling were used. Results indicate that type-I IFN production in MEFs occurs by both NF-κB dependent and independent mechanisms. In contrast, the production of IFN-α was completely abrogated in IFNAR-\- MEFs whereas IFN-β production was greatly reduced. Production of type-I IFNs in IFNGR-\- MEFs is also reduced upon JEV infection but the reason for this is unclear. Chapter 6 demonstrates that JEV induced expression of classical MHC-I molecules occurs by type-I IFN mediated signaling. This result is in contrast to WNV infection, in which both NF-κB and type-I IFNs are involved in the induction of classical MHC-I molecules. Type-I IFNs were also shown to be involved in the induction of nonclassical MHC molecules namely, Qa-1 and Qb1 during JEV infection. In contrast, the expression of T10, another nonclassical MHC molecule occurs independent of type-I IFN signaling. The expression of molecules involved in antigen processing and presentation namely, Tap1, Tap2, Lmp2 and Lmp7 was type-I IFN-mediated, whereas the expression of Tapasin and Lmp10 was mediated by both type-I IFN dependent and independent mechanisms. The expression of VCAM-1 was dependent on NF-κB mediated signaling. Chapter 7 precisely describes the underlying mechanism of induction of MHC and various other related molecules and their significance during JEV infection. In addition, it also includes a working model for the induction of these molecules during JEV infection. In summary, this is the first study in which the mechanism of JEV mediated induction of classical as well as nonclassical MHC molecules has been studied in detail. This study clearly demonstrated that type-I IFNs are involved in the induction of classical and nonclassical MHC-I molecules during JEV infection. The functional significance of this JEV mediated induction of classical MHC-I molecules is unclear, but it has been proposed that this is to escape from the action of NK cells. The absence of MHC-II induction during JEV infection could be important because it may lead to the initiation of an immune response which is different from that induced during other viral infections which induce the expression of MHC-II molecules. In contrast to classical MHC-I molecules, the functional and biological significance of nonclassical MHC-I molecules are poorly studied. Nonclassical MHC-I molecules play an important role in bridging adaptive and innate immune response. So the nonclassical MHC molecules induced during JEV infection may play an important role in the initiation of immune response during JEV infection. The role these nonclassical MHC-I molecules in antigen presentation during JEV infection is not known. These nonclassical antigens are also recognized by NK and γδT cells, thus the expression of nonclassical MHC-I molecules during JEV infection might also confer a protective role.

Orientador: Eduardo Melani Rocha / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-13T17:43:11Z (GMT). No. of bitstreams: 1 Alves_MonicadeCassia_D.pdf: 5666644 bytes, checksum: 2d8f41985c6b80374f8fd6d042fc9ff0 (MD5) Previous issue date: 2006 / Resumo: Este estudo avaliou as possíveis vias comuns na fisiopatogênese da síndrome do olho seco no Diabetes Mellitus (DM) e no envelhecimento, envolvendo o acúmulo dos produtos de glicosilação ("Advanced Glycation End-products" - AGEs), seu receptor RAGE e a ativação do Fator Nuclear-?B (NF-?B) na glândula lacrimal (GL) e alterações do filme lacrimal nessas condições. Modelos animais de DM induzido com estreptozotocina e animais senis (24 meses de vida) foram comparados a animais controle tratados com tampão citrato e adultos jovens (2 meses de vida). Foram avaliadas vias de sinalização, envolvendo AGEs, RAGE e a ativação do NF-?B na GL e alterações no filme lacrimal em ratos Wistar de todos os grupos. A análise do filme lacrimal foi realizada através de medidas de volume de secreção basal e dosagem de citocinas como a Interleucina-1 ß (IL-1 ß) e Fator de Necrose Tumoral - a (TNF- a). A capacidade secretória da GL foi avaliada através de medidas da atividade de peroxidase. Técnicas de "western blot" foram utilizadas para avaliar a expressão e ativação do NF-?B na GL. A expressão de AGE, RAGE e NF-?B na GL e córnea nos grupos estudados, foi avaliada através de microscopia confocal com imunofluorescência. O volume lacrimal foi significativamente menor nos animais diabéticos e senis (P=0,02 e 0,016, respectivamente). Concentrações de IL-1ß e TNF-a na lágrima foram mais altas nos ratos diabéticos e senis do que nos controles e adultos jovens (P=0,007 e 0,02 nos diabéticos e P= 0,007 e P= 0,05 nos senis). A atividade de peroxidase foi significativamente mais baixa no grupo senil (P=0,016), mas não nos experimentos, comparando animais diabéticos e controles (P=0,34). A expressão de AGE, RAGE e NF-?B na GL foi aumentada nos animais diabéticos e senis. O DM e a senilidade induzem alterações estruturais e secretórias significativas na GL e nos tecidos da superfície ocular e associa-se a maior incidência de olho seco. A expressão aumentada de AGE, RAGE e NF-?B na GL sugere a participação desses fatores nas vias de sinalização e subseqüentes alterações inflamatórias relacionadas ao olho seco nessas condições. / Abstract: The present study evaluates the dry eye syndrome related to diabetes and aging through the involvement of the Advanced Glycation End-product (AGEs), the Advanced Glycation End-product Receptor (RAGE) axis and Nuclear Factor-?B (NF-?B) activation in lacrimal gland (LG) and tear film dysfunction in these conditions. To evaluate whether AGE, RAGE and NF-?B signaling in LG are altered in diabetes and aging, streptozotocin-induced diabetic, normoglicaemic, aging (24 month-old) and young adults (2 month-old) male Wistar rats were compared. Tear film alterations and the expression of AGE, RAGE and NF-?B in ocular tissues were evaluated in all considered groups. Tear secretion parameters as basal tear secretion volume, Interleukin-1 ß (IL-1 ß) and Tumor Necrosis Factor- a (TNF- a) levels and LG and peroxidase activity in LG tissue were measured. NF-?B expression and activation was evaluated in LG by western blot. Immunohistochemistry with confocal microscopy was used to assess AGE, RAGE and NF-?B expression in LG of all groups. Tear volume was significantly lower in diabetic and aging rats (P=0.02 and 0.016, respectively). IL-1ß and TNF-a concentrations in tears were higher in diabetic and aging than in control and young rats (P=0.007 and 0.02 in diabetic and P= 0.007 and P= 0.05 in aging). Peroxidase activity was significantly lower in the aging group (P=0.016) but not the assays with diabetic rats (P=0.34). AGE, RAGE and NF-?B expressions were increased in LG of diabetic and aging rats. Diabetes and aging induce significant alterations in rat LG structure and secretion. The higher expression of AGE, RAGE and NF-?B in LG may suggest that these factors are involved in signalling and in subsequent inflammatory alterations related to dry eye related to these conditions. / Doutorado / Clinica Medica / Doutor em Clínica Médica

A tuberculose (TB) é um grave problema de saúde pública. Somente no ano de 2004, cerca de 9 milhões de pessoas desenvolveram TB ativa e mais de 2 milhões de pessoas morreram da doença. O desenvolvimento de novos modelos experimentais de TB seriam de grande utilidade para para elucidar mecanismos fisiopatológicos da doença e testar esquemas terapêuticos para a prevenção e contenção da doença. Além disso, o desenvolvimento de novas vacinas torna-se indispensável como ferramenta de prevenção e controle da TB. A TB no sistema nervoso central (SNC), assim como em outros tecidos do organismo, promove a ativação de células inflamatórias. No SNC a micróglia desempenha este papel, sendo capaz de produzir ou ser influenciada por mediadores solúveis. Vários mediadores estão envolvidos nos mecanismos moleculares decorrentes da infecção e inflamação causados pela TB, entre eles: NFB, iNOS e VEGF. A ativação do NFB, um fator de transcrição citoplasmático que sob estímulo migra para o núcleo celular, tem íntima relação com a indução da iNOS e de VEGF. A resistência intracelular a patógenos, inclusive ao Mycobacterium tuberculosis, parece estar associada a expressão de iNOS em macrófagos. O óxido nítrico (NO) tem papel importante na comunicação intercelular, estimulando a síntese de mediadores inflamatórios, como as citocinas, e regulando sua própria produção endógena. Estas citocinas por sua vez também podem induzir a atividade do NFB e a expressão da iNOS e VEGF. O VEGF é um potente ativador de permeabilidade vascular e de angiogênese, envolvido na ruptura da barreira hemato-encefálica. Neste estudo, mostramos a caracterização morfológica e imuno-histoquímica de um modelo murino de TB no SNC, com a indução da doença pela inoculação de BCG. Com este modelo experimental obtivemos importantes resultados que podem esclarecer mecanismos envolvidos na fisiopatologia da neuro-TB humana. A indução de meningite e tuberculomas foi possível através da inoculação de 104 cfu de BCG no cerebelo de camundongos, por estereotaxia, e esta indução foi dependente do tempo. A confirmação do diagnóstico foi feita pela detecção de bacilos álcool-ácido resistentes (BAAR), nas lesões tuberculosas. Observamos, ao longo do tempo (1 a 6 dias; 1, 2, 4 e 8 semanas) o recrutamento de diferentes populações gliais (micróglia e astrócitos) no sítio de injeção. Houve aumento de produção e ativação NFB nas lesões tuberculosas, caracterizada pela translocação da molécula do citoplasma para o núcleo celular. Houve expressão de iNOS restrita às lesões tuberculosas, além do aumento de expressão de VEGF nestas lesões. Além disso, camundongos imunizados com a vacina gênica hsp65, contra a TB, não expressam VEGF em suas lesões. Esta vacina parece conferir um efeito protetor em nosso modelo experimental, reduzindo a expressão de VEGF, e consequentemente reduzindo seu efeito angiogênico decorrente do processo inflamatório. O recrutamento glial, e a produção de mediadores solúveis (NFB, iNOS e VEGF) pelo hospedeiro, em resposta à invasão do patógeno no SNC, parecem estar envolvidos na fisiopatologia da neurotuberculose, como demonstrado neste modelo experimental. Nosso modelo permitirá investigar fatores possivelmente responsáveis pelo desenvolvimento e manutenção de lesões tuberculosas no SNC. O objetivo final seria elucidar a fisiopatologia desta grave doença e compreender eventos moleculares envolvidos na produção de lesões. O conhecimento gerado poderá permitir o delineamento de terapias específicas e efetivas. / Tuberculosis (TB) is a serious public health problem; in 2004, 9 million people developed active TB and the disease killed 2 million patients. Development of experimental models and new vaccines are essential both to elucidate physiopathological mechanisms and to control the disease. This infection in the central nervous system (CNS), as in other tissues of the organism, activates inflammatory cells. In CNS, this role is performed by the microglia, which is capable of producing or be influenced by soluble mediators. Several mediators are involved in the molecular mechanisms of the infection and inflammation by mycobacteria , such as NFB, iNOS and VEGF. NFB activation, a cytoplasmic transcriptional factor that migrates to the cellular nucleus under stimuli, is involved with the iNOS and VEGF induction of expression. The intracellular resistance to Mycobacterium tuberculosis has been associated with iNOS expression in macrophage cells. Nitric oxide (NO) is crucial in intercellular communication, modulating the synthesis of mediators of inflammation, such as cytokines, and modulation itself. These cytokines induces NFB activity, and induces iNOS and VEGF expression. VEGF is a potent activator of vascular permeability and of angiogenesis and it is a factor involved in the breakdown of the blood brain-barrier in tuberculous meningitis. In this study, we showed the morphologic and immunohistochemistry characterization of an experimental model of TB in the CNS, with inoculation of BCG in mice. In this model we elicited important outcome that can elucidate mechanisms involved in the physiopathology of human neuron-TB. Induction of meningitis and tuberculomas were possible with stereotaxic inoculation of 104 cfu of BCG in mice cerebellum, in a time-dependent way. Diagnostic was confirmed by detection of alcohol-acid resistant bacilli (BAAR), in tuberculous lesions. We observed, the time-course (1 to 6 days; 1, 2, 4 e 8 weeks) of the recruitment of different glial populations (microglia and astrocytes) in the injection site. There was increased production and activation of NFB in the tuberculous lesions, it was characterized by its nuclear translocation from cytoplasm. There was iNOS expression only in the tuberculous lesions, and expression increased of VEGF in these lesions. Furthermore, mice immunizated with vaccine DNA-hsp65 there was no expression of VEGF in its lesions. This vaccine seems confer a protector effect in our experimental model, reducing the expression of VEGF, and then reducing its angiogenic effect derived from inflammatory process. Glial recruitment, and the soluble mediators production (NFB, iNOS e VEGF) by the host, producing in response to invasion of the pathogen in the CNS, has been involved in the pathophysiology of the neuro-TB, such as demonstrated in this experimental model. Our model will allow investigate possible factors responsible for the development and maintenance of tuberculous lesions in the CNS. The final aim is to elucidate the physiopathology of this serious illness and understand the molecular events involved in the production of the lesions. The knowledge created may permit to pave the way to delineate specific and effective therapies.