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Investigation of varicella zoster virus glycoprotein-specific T cell responsesMalavige, Gathsaurie Neelika January 2007 (has links)
T cells are believed to be important in the control of varicella zoster virus (VZV) replication but little is known of T cell epitopes and the relationships between T cell responses, viral load and clinical disease during primary infection. I initially set to investigate the immune responses to two of the main VZV glycoproteins (gE and gI) using ex vivo and cultured IFNγ ELISpot assays. I identified several novel CD4+ T cell epitopes within gE and gI and characterized the phenotype of gE DRB1*1501 tetramer specific responses in healthy immune donors. I then set out to investigate the function and phenotype of VZV specific T cells in primary infection and their relationship to viral loads and clinical disease severity by using glycoprotein E/DRB1*1501 specific MHC class II tetramers, ex vivo IFNγ ELISpot assays and quantitative real time PCR assays. I compared the frequency and phenotype of specific T cells with virological and clinical outcomes in 32 adult individuals with primary VZV infection. In healthy immune donors, the gE specific T cells showed a early intermediate stage of differentiation with evidence of recent activation. Patients with acute primary infection had higher VZV/DRB1*1501 tetramer specific T cell responses and expressed markers of activation and effector differentiation. Viral loads were found to be significantly higher in patients with moderate to severe infection compared to those with mild infection (p<0.001). A significant inverse correlation was seen between the viral loads and the ex vivo IFNγ ELISpot responses of the patients (p<0.05, r=-0.64). These data would be compatible with a role for gE and gl-specific T cells in the control of viral replication during both primary infection and re-activation.
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Estudo imuno-histoquímico de β2-glicoproteína I em fígado e intestino de ratos submetidos à sepse pela via endovenosa ou associada à translocação bacteriana / β2-glycoprotein expression in sepsis: an immunohistochemical study of rat liver and intestine following endovenous inoculation or bacterial translocationAugustinis, Sheila Vieira da Cruz 23 September 2005 (has links)
β2-glicoproteina I (β2GPI), uma proteína circulante produzida em fígado e intestino, foi estudada no fígado e íleo de ratos durante sepse controlada (S) ou translocação bacteriana (TB) com E. coli R-6. A sepse foi induzida por inoculação endovenosa de 109 UFC/mL/100g. A TB, pelo confinamento de 10 mL 1010 UFC/mL, no intestino. Grupos controle receberam veículo. Após 2h, os animais foram sacrificados. β2GPI foi espressa nos tecidos de todos animais, em cortes fixados em metacarn e impregnados com parafina, após reação com anti-β2GPI amplificada com Envision-AP. O fígado mostrou dois padrões citoplasmáticos distintos. Um padrão difuso, atribuído à síntese protéica, aumentado somente no grupo S e um padrão focal, invariável. No íleo, a mucosa foi negativa; a submucosa, o endotélio sangüíneo e linfático, positivos. Esta marcação aumentou somente no grupo TB. Os resultados sugerem a participação da β2GPI na regulação local da resposta hepática e intestinal de fase aguda. / β2-glycoprotein I is a circulating protein (β2GPI) produced in liver and intestines. β2GPI was studied in liver and ileum in rats under controlled sepsis (S) or bacterial translocation (BT) with E. coli R-6. Sepsis was induced by endovenous inoculation with 109 CFU/mU100g. BT was induced by confining 10 mL 1010 CFU/mL, to the intestine. Control groups received vehicle. After 2h, animals were sacrificed. Metacarn fixed, paraffin embedded tissue slices were reacted with monoclonal anti-β2GPI, revealed with Envision-AP and hematoxylin counterstained. Ali animals stained for β2GPI in the liver and ileum. Liver presented two distinct cytoplasm staining patterns. The diffuse pattern, ascribed to protein synthesis, increased in group S animals only. The spotted pattern was invariant. Ileum mucosa was negative, while submucosa, blood and Iymphatic endothelium were positive. The ileum staining increased after TB, only. Results underline the hypothesis for locally regulated liver and intestine contribution to acute phase response modulation.
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Estudo imuno-histoquímico de β2-glicoproteína I em fígado e intestino de ratos submetidos à sepse pela via endovenosa ou associada à translocação bacteriana / β2-glycoprotein expression in sepsis: an immunohistochemical study of rat liver and intestine following endovenous inoculation or bacterial translocationSheila Vieira da Cruz Augustinis 23 September 2005 (has links)
β2-glicoproteina I (β2GPI), uma proteína circulante produzida em fígado e intestino, foi estudada no fígado e íleo de ratos durante sepse controlada (S) ou translocação bacteriana (TB) com E. coli R-6. A sepse foi induzida por inoculação endovenosa de 109 UFC/mL/100g. A TB, pelo confinamento de 10 mL 1010 UFC/mL, no intestino. Grupos controle receberam veículo. Após 2h, os animais foram sacrificados. β2GPI foi espressa nos tecidos de todos animais, em cortes fixados em metacarn e impregnados com parafina, após reação com anti-β2GPI amplificada com Envision-AP. O fígado mostrou dois padrões citoplasmáticos distintos. Um padrão difuso, atribuído à síntese protéica, aumentado somente no grupo S e um padrão focal, invariável. No íleo, a mucosa foi negativa; a submucosa, o endotélio sangüíneo e linfático, positivos. Esta marcação aumentou somente no grupo TB. Os resultados sugerem a participação da β2GPI na regulação local da resposta hepática e intestinal de fase aguda. / β2-glycoprotein I is a circulating protein (β2GPI) produced in liver and intestines. β2GPI was studied in liver and ileum in rats under controlled sepsis (S) or bacterial translocation (BT) with E. coli R-6. Sepsis was induced by endovenous inoculation with 109 CFU/mU100g. BT was induced by confining 10 mL 1010 CFU/mL, to the intestine. Control groups received vehicle. After 2h, animals were sacrificed. Metacarn fixed, paraffin embedded tissue slices were reacted with monoclonal anti-β2GPI, revealed with Envision-AP and hematoxylin counterstained. Ali animals stained for β2GPI in the liver and ileum. Liver presented two distinct cytoplasm staining patterns. The diffuse pattern, ascribed to protein synthesis, increased in group S animals only. The spotted pattern was invariant. Ileum mucosa was negative, while submucosa, blood and Iymphatic endothelium were positive. The ileum staining increased after TB, only. Results underline the hypothesis for locally regulated liver and intestine contribution to acute phase response modulation.
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