A Fashi lymphoproliferative phenotype reveals non-apoptotic Fas signalling in HTLV-1-associated neuroinflammation.

Human T-cell lymphotropic virus (HTLV)-1 was the first human retrovirus to be associated to cancer, namely Adult T-cell Leukemia (ATL), but its pathogenesis remains enigmatic, since only a minority of infected individuals develops either ATL or the neuroinflammatory disorder HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). A functional FAS -670 polymorphism in an interferon (IFN)-regulated STAT1-binding site has been associated to both ATL and HAM/TSP susceptibility. Fashi T stem cell memory (Tscm) cells have been identified as the hierarchical apex of ATL, but have not been investigated in HAM/TSP. In addition, both FAS and STAT1 have been identified in an IFN-inducible HAM/TSP gene signature, but its pathobiological significance remains unclear. We comprehensively explored Fas expression (protein/mRNA) and function in lymphocyte activation, apoptosis, proliferation and transcriptome, in PBMC from a total of 47 HAM/TSP patients, 40 asymptomatic HTLV-1-infected individuals (AC) and 58 HTLV-1 -uninfected healthy controls. Fas surface expression followed a two-step increase from HC to AC and from AC to HAM/TSP. In HAM/TSP, Fas levels correlated positively to lymphocyte activation markers, but negatively to age of onset, linking Fashi cells to earlier, more aggressive disease. Surprisingly, increased lymphocyte Fas expression in HAM/TSP was linked to decreased apoptosis and increased lymphoproliferation upon in vitro culture, but not to proviral load. This Fashi phenotype is HAM/TSP-specific, since both ex vivo and in vitro Fas expression was increased as compared to multiple sclerosis another neuroinflammatory disorder. To elucidate the molecular mechanism underlying non-apoptotic Fas signalling in HAM/TSP, we combined transcriptome analysis with functional assays, i.e. blocking vs. triggering Fas receptor in vitro with antagonist and agonist- anti-Fas mAb, respectively. Treatment with agonist anti-Fas mAb restored apoptosis, indicating biased but not defective Fas signalling in HAM/TSP. In silico analysis revealed biased Fas signalling towards proliferation and inflammation, driven by RelA/NF-kB. Correlation of Fas transcript levels with proliferation (but not apoptosis) was confirmed in HAM/TSP ex vivo transcriptomes. In conclusion, we demonstrated a two-step increase in Fas expression, revealing a unique Fashi lymphocyte phenotype in HAM/TSP, distinguishable from multiple sclerosis. Non-apoptotic Fas signalling might fuel HAM/TSP pathogenesis, through increased lymphoproliferation, inflammation and early age of onset

Estudo da regulação molecular da apoptose Ex vivo e In vitro em pacientes com mielopatia associada ao HTLV-I

Submitted by Repositório Arca ([email protected]) on 2019-07-08T18:54:12Z No. of bitstreams: 1 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2019-07-12T12:11:50Z (GMT) No. of bitstreams: 2 Daniele Decanine Estudo da regulação...2001.pdf: 34890638 bytes, checksum: 02d5e047c190e1660cee8abcdb8afe37 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Made available in DSpace on 2019-07-12T12:11:50Z (GMT). No. of bitstreams: 2 Daniele Decanine Estudo da regulação...2001.pdf: 34890638 bytes, checksum: 02d5e047c190e1660cee8abcdb8afe37 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2001Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil.O vírus linfotrópico de células T humano - tipo I (HTLV-I) é o agente etiológico da Leucemia/Linfoma de células T no adulto (ATL), da Mielopatia Associada ao HTLV-I/Paraparesia Espástica Tropical (HAM/TSP), e de outras patologias. Os IFN-α e IFN-β foram recentemente introduzidos no tratamento de ATL e HAM/TSP, embora seus mecanismos de ação ainda estejam pouco esclarecidos. A presença da iNOS e seu recém-descoberto papel anti-apoptótico na ATL iniciou novas perspectivas terapêuticas. Considerando a correlação entre a linfoproliferação induzida pelo HTLV-I e as patologias associadas, buscamos estudar a regulação molecular da linfoproliferação e da apoptose ex vivo e in vitro em indivíduos soropositivos assintomáticos e pacientes com HAM/TSP. A expressão ex vivo do mRNA das moléculas pró-apoptóticas Fas, FasL e pró-caspase-3 foi maior nos indivíduos assintomáticos quando comparados aos pacientes com HAM/TSP, sugerindo que uma diminuição da morte celular programada poderia ter influência no processo patológico. O aumento do mRNA da iNOS nos assintomáticos foi correlacionado a inibição da linfoproliferação in vitro pelo L-NMMA (inibidor da iNOS). Por outro lado, apenas o IFN-β mostrou atividade anti-proliferativa significante, mas não pró-apoptótica in vitro em células mononucleares de pacientes com HAM/TSP. Porém, as células mostraram-se sensíveis ao estímulo com anti-CD3 na indução da apoptose. A estimulação com o IFN-β ou anti-CD3 não diminuiu a expressão do mRNA da proteína viral Tax, sugerindo que os efeitos antiproliferativos e pró-apoptóticos ocorrem independente da transcrição viral. De acordo com nossos resultados, o uso combinado de IFN-β_e outras drogas antivirais ou pró-apoptóticas poderia ser considerado em futuros ensaios terapêuticos.Human T cell Lymphotropic Virus Type I (HTLV-I) is the ethiologic factor for ATL (Adult T cell Lymphoma), HTLV-I-associated myelopathy (HAM), as well as other pathologies. Interferon (IFN)-a e -ß have been shown to be effective in ATL e HAM, but their mechanism of action remains unclear. Recently, the presence of iNOS and its anti-apoptotic effect has been demonstrated in ATL, initiating new perspectives for therapy. Considerating the correlation between HTLV-I induced lymphoproliferation and associated pathologies, we investigated the molecular regulation of lymphoproliferation and apoptosis ex vivo and in vitro in seropositive asymptomatic individuals and HAM/TSP patients. We found that ex vivo expression of Fas, FasL and pro-caspase-3 mRNA was higher in asymptomatic individuals compared to HAM/TSP patients, suggesting that programmed cell death reduced influence the pathologic process. iNOS mRNA expression was higher in asymptomatic individuals and correlated to the inhibitory effect of L-NMMA (iNOS inhibitor) on in vitro lymphoproliferation. On the other hand, only IFN-ß displayed a profound anti-proliferative, but not pro-apoptotic effect in HAM/TSP patients. However, cells were susceptible to apoptosis induction upon anti-CD3 stimulation. IFN-ß and anti-CD3 stimulation did not modulate Tax mRNA levels, suggesting that anti-proliferative and pro- apoptotic effects occurred independent of viral transcription. In accordance to our results, the association of IFN-ß and others antivirais or pro-apoptotic drugs might possibly be considered in future clinical trials

Ativação celular, proliferação e apoptose nas patologias associadas ao HTLV-I

Submitted by Repositório Arca ([email protected]) on 2019-09-04T17:23:29Z No. of bitstreams: 1 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2019-09-10T14:29:22Z (GMT) No. of bitstreams: 2 Daniele Decanine Ativação...2006.pdf: 45081071 bytes, checksum: 2132180d82c158a76471b434ccd43ecf (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Made available in DSpace on 2019-09-10T14:29:22Z (GMT). No. of bitstreams: 2 Daniele Decanine Ativação...2006.pdf: 45081071 bytes, checksum: 2132180d82c158a76471b434ccd43ecf (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2006CNPq.Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil.O HTLV-I é o agente etiológico da Leucemia/Linfoma de células T no adulto (ATL), da Mielopatia Associada ao HTLV- l/Paraparesia Espástica Tropical (HAM/TSP), e de outras patologias. Os IFN-a e IFN-p foram recentemente introduzidos no tratamento de ATL e HAM/TSP, embora seus mecanismos de ação ainda estejam pouco esclarecidos. A presença da iNOS e seu recém-descoberto papel anti-apoptótico na ATL inciou novas perspectivas terapêuticas. Considerando o provável papel da ativação e proliferação linfocitária induzida pelo HTLV-I nas patologias associadas, buscamos estudar a regulação molecular e celular da ativação celular, linfoproliferação e apoptose ex vivo e in vitro em indivíduos soropositivos assintomáticos, pacientes com HAM/TSP e pacientes com ATL. Observamos uma correlação significante do estágio clínico na HAM/TSP com a linfoproliferação in vitro e com a expressão de CDSO em células B ex vivo. O efeito marcante do IFN-p no aumento da expressão in vitro de Fas e CD86, sugere a utilização destas moléculas como possíveis marcadores biológicos do uso clinico dos IFNs. Observamos que, apenas o IFN-p, mas não o IFN-a, mostrou atividade anti-proliferativa significante, mas não pró-apoptótica in vitro nos pacientes com HAM/TSP, ATL e em doadores normais. Porém, as células mostraram-se sensíveis ao estímulo com anti-CD3 na indução da apoptose, sugerindo que os IFNs, principalmente o IFN-p possui efeitos anti-proliferativos e o anti-CD3 pró-apoptóticos. Em relação ao inibidor da iNOS, não observamos nenhum efeito anti-proliferativo e/ou pró-apoptótico nos indivíduos infectados por HTLV-I. De acordo com nossos resultados, o conhecimento de marcadores biológicos e o uso combinado de IFN-p e outras drogas pró-apoptóticas, como o anti-CD3, poderia ser considerado em futuros ensaios terapêuticos em HAM/TSP ATL.HTLV-I is the ethiologic factor for ATL (Adult T ceil Lymphoma), HTLV-l-associated myelopathy (HAM), as well as other pathologies. Interferon (IFN)-a e -|3 have been shown to be effective in ATL e HAM, but their mechanism of action remains unclear. Recently, the presence of iNOS and its anti-apoptotic effect has been demonstrated in ATL, initiating new perspectives for therapy. Considering the correlation between HTLV-I induced lymphoproliferation and associated pathologies, we investigated the cellular activation, lymphoproliferation and apoptosis ex vivo and in vitro in seropositive asymptomatic individuals, ATL and HAM/TSP patients. We found that ex vivo expression of CDSO in B cells and in vitro lymphoproliferation significantly correlacionates with clinical stage in HAM/TSP. The marked effect of IFN-p upon in vitro expression of Fas and CDS6 suggests their possible use as biological markers in clinical evaluation. IFN-p displayed significantly increased anti-proliferative activity, as compared to IFN-a, in HAM/TSP, ATL and health donors. However, only anti-CD3 stimulation was able to exert a significant pro-apoptotic effect in vitro, in all groups studied. Inhibition of iNOS had no significant anti-proliferative or pro-apoptotic effect in HTLV-l-infected individuals. In summary, this study points at the possible use of IFN-p, in combination with pro-apoptotic drugs such anti-CD3, as a therapeutic strategy in HAM/TSP and ATL

IFN-β induces greater antiproliferative and proapoptotic effects and increased p53 signaling compared with IFN-α in PBMCs of Adult T-cell Leukemia/Lymphoma patients

Current first-line treatment for Adult T-cell leukemia (ATL) includes combination therapy with interferon alpha (IFN-α) and zidovudine (AZT). The use of IFN-α in this treatment is mostly empirical in origin, whereas the therapeutic potential of interferon beta (IFN-β), has not yet been thoroughly explored in this context. Here we compare the effects of IFN-α and IFN-β treatment in short term ex vivo peripheral blood mononuclear cell (PBMC) cultures from 22 ATL patients. Using proliferation, apoptosis and antiviral bioassays, complemented with microarray and gene set analysis, we demonstrate that in this setting IFN-β has superior antiproliferative and pro‑apoptotic effects than IFN-α. Increased p53 signaling is observed under the IFN-β treatment, while the antiviral effects are equivalent to those of IFN-α treatment. Notably, the genes in a published in vivo AZT/IFN-α response profile are affected more strongly by IFN-β than by IFN-α stimulus in these ex vivo PBMCs. In conclusion, this first comprehensive analysis comparing the effects of IFN-α and IFN-β on ex vivo PBMCs of ATL patients demonstrates that IFN-β has a greater impact than IFN-α on biological processes which have been shown to be crucial in the treatment of ATL, making IFN-β an intriguing candidate for further in vivo testing.status: publishe

Avaliação da função sexual e qualidade de vida de mulheres infectadas pelo o vírus HTLV-I / Evaluation of sexual function and quality of life in HTLV-I infected women

Introdução:O presente estudo objetivou avaliar a função sexual de mulheres com diagnóstico soropositivo de HTLV-I, com idade acima de 18 anos. Metodologia: Foram avaliadas 46 mulheres, sendo 22 com HTLV-I e 24 mulheres saudáveis utilizando o questionário Female Sexual Function Index (FSFI) e SF-36. As análises estatísticas foram realizadas por meio do programa Graphpad Prism 6.0 e os resultados expressos pela diferença entre as médias. Resultados:As mulheres portadoras de HTLV-I tiveram um escore muito abaixo do valor mínimo aceitável como uma boa função sexual (escore 17,9), e quando as comparamos com o grupo controle (escore 26) pode-se observar uma diferença estatisticamente significante entre os dois grupos (p=0,001). Quanto à qualidade de vida, as mulheres do grupo controle foram as que mais se aproximam da normalidade (escore 88,84), apresentando uma qualidade de vida superior a observada nas mulheres portadoras do vírus HTLV-I (escore 43,25) (p<0,0001).Conclusão: As pacientes infectadas com HTLV-I demonstraram disfunção sexual acentuada e uma redução da qualidade de vida. Entretanto, são necessários mais trabalhos comparativos que demonstrem as causas de disfunção sexual nestas pacientes e seu impacto na qualidade de vida das mesmas