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Background/Purpose: Glucocorticoids (GC) remain the frontline treatment in Systemic Lupus Erythematosus (SLE) despite their predictable metabolic adverse effects. Type I interferons (IFN), produced by TLR7 and TLR9 activated plasmacytoid dendritic cells (pDC), play a critical role in SLE pathogenesis, but are not suppressed by GC. Glucocorticoid- Induced Leucine Zipper (GILZ) is an endogenous anti-inflammatory protein, which regulates T and B cell activation and is suppressed in SLE; whether GILZ regulates type 1 IFN production in SLE is not known. We tested the hypothesis that GILZ inhibits Type I IFN production in SLE. Method(s): We performed in vitro analysis on pDC and bone marrow-derived DC (BMDC), and in vivo studies, of WTand GILZ-/- mice using stimuli of TLR4 (LPS), TLR7 (Imiquimod), TLR7/8 (Resiquimod) and TLR9 (CpG). IFN was measured using a IFN luciferase assay, ELISA and Luminex. IFN-stimulated gene signatures (ISG) were measured by qPCR. To determine whether GILZ regulates IFN in human SLE, we mined a public gene expression dataset GSE61635. We examined IRF7 and NF-kappaB involvement in GILZ effects using ChIP and reporter assays. Result(s): GILZ deficiency resulted in increased pDC and BMDC secretion of IFN in response to TLR7, TLR7/8 and TLR9 stimulation. GILZ deficiency was also associated with increased ISG in naive spleen cells, and TLR-stimulated pDC (Fig 1A) and BMDC of GILZ-/- mice. Correspondingly, increased IFN was seen in GILZ-/- mice in response to TLR7/8 stimulation in vivo. In human SLE whole blood, GILZ mRNA was negatively correlated with ISG (IFI44, IFI44L, RSAD2, IFI27) (Fig 1B). In BMDC, TLR activation suppressed GILZ, but increased IRF7 expression. CHIP-qPCR showed GILZ enrichment at the IRF7 locus in unstimulated cells, which decreased following TLR7/8/9 activation, indicating direct binding of GILZ to the IRF7 locus as the mechanism through which it regulates IFNa production. In addition, TLR7/8/9- induced NF-kappaB activation was s

Background/Purpose: Glucocorticoids (GC) remain the frontline treatment in Systemic Lupus Erythematosus (SLE) despite their predictable metabolic adverse effects. Type I interferons (IFN), produced by TLR7 and TLR9 activated plasmacytoid dendritic cells (pDC), play a critical role in SLE pathogenesis, but are not suppressed by GC. Glucocorticoid- Induced Leucine Zipper (GILZ) is an endogenous anti-inflammatory protein, which regulates T and B cell activation and is suppressed in SLE; whether GILZ regulates type 1 IFN production in SLE is not known. We tested the hypothesis that GILZ inhibits Type I IFN production in SLE. Method(s): We performed in vitro analysis on pDC and bone marrow-derived DC (BMDC), and in vivo studies, of WTand GILZ-/- mice using stimuli of TLR4 (LPS), TLR7 (Imiquimod), TLR7/8 (Resiquimod) and TLR9 (CpG). IFN was measured using a IFN luciferase assay, ELISA and Luminex. IFN-stimulated gene signatures (ISG) were measured by qPCR. To determine whether GILZ regulates IFN in human SLE, we mined a public gene expression dataset GSE61635. We examined IRF7 and NF-kappaB involvement in GILZ effects using ChIP and reporter assays. Result(s): GILZ deficiency resulted in increased pDC and BMDC secretion of IFN in response to TLR7, TLR7/8 and TLR9 stimulation. GILZ deficiency was also associated with increased ISG in naive spleen cells, and TLR-stimulated pDC (Fig 1A) and BMDC of GILZ-/- mice. Correspondingly, increased IFN was seen in GILZ-/- mice in response to TLR7/8 stimulation in vivo. In human SLE whole blood, GILZ mRNA was negatively correlated with ISG (IFI44, IFI44L, RSAD2, IFI27) (Fig 1B). In BMDC, TLR activation suppressed GILZ, but increased IRF7 expression. CHIP-qPCR showed GILZ enrichment at the IRF7 locus in unstimulated cells, which decreased following TLR7/8/9 activation, indicating direct binding of GILZ to the IRF7 locus as the mechanism through which it regulates IFNa production. In addition, TLR7/8/9- induced NF-kappaB activation was s

Le développement de vaccins sécuritaires capables de générer une réponse immunitaire cellulaire, essentielle pour protéger contre la plupart des infections persistantes et des cancers, est encore aujourd’hui un défi. Une des méthodes utilisées pour augmenter l’efficacité de ces vaccins est de les combiner avec un adjuvant. Cependant, encore peu d’adjuvants sont autorisés pour l’utilisation chez l’humain et ceux qui le sont ont un mécanisme d’action parfois mal défini et engendrent principalement une réponse immunitaire de type humoral. Ainsi, l’élaboration de nouveaux adjuvants et de nouvelles méthodes de vaccination ainsi que la compréhension de leur mécanisme d’action est cruciale pour réussir à prévenir ou traiter plusieurs maladies infectieuses ou encore le cancer. Parmi les nouveaux adjuvants et systèmes de vaccination en développement, un intérêt grandissant a récemment été observé pour les pseudoparticules virales (VLP) et les molécules immunomodulatrices comme les ligands de récepteur de motifs moléculaires associés aux pathogènes (PRR). Notre équipe a démontré précédemment que les pseudoparticules du virus de la mosaïque de la papaye (PapMV), composées des protéines de capside d’un virus de plante et d’un acide ribonucléique simple brin (ARNsb) non codant, sont hautement immunogéniques chez la souris. De plus, PapMV peut être utilisé comme plateforme vaccinale, en fusionnant à sa surface des épitopes d’un agent pathogène, ou comme adjuvant, en l’ajoutant à un vaccin existant. Dans ces contextes, une augmentation de la réponse immunitaire humorale spécifique aux antigènes vaccinaux a été obtenue. Dans cette étude, nous avons caractérisé le mécanisme d’action de PapMV et évaluer son potentiel dans la lutte contre les infections et le cancer. En utilisant un modèle d’immunisation chez la souris, nous avons démontré que le PapMV activait les cellules du système immunitaire via la liaison du récepteur de type Toll 7 (TLR7) par son ARNsb et la production d’interf

Aim: To evaluate the efficacy and safety of bevacizumab in the adjuvant cancer therapy setting within different subset of patients. Methods & Design/Results: PubMed, EMBASE, Cochrane and Clinical trials.gov databases were searched for English language studies of randomized controlled trials comparing bevacizumab and adjuvant therapy with adjuvant therapy alone published from January 1966 to 7thof May 2014. Progression free survival, overall survival, overall response rate, safety and quality of life were analyzed using random- or fixed-effects models according to the PRISMA guidelines. We obtained data from 44 randomized controlled trials (30,828 patients). Combining bevacizumab with different adjuvant therapies resulted in significant improvement of progression free survival (log hazard ratio, 0.87; 95% confidence interval (CI), 0.84-0.89), overall survival (log hazard ratio, 0.96; 95% CI, 0.94-0.98) and overall response rate (relative risk, 1.46; 95% CI: 1.33-1.59) compared to adjuvant therapy alone in all studied tumor types. In subgroup analyses, there were no interactions of bevacizumab with baseline characteristics on progression free survival and overall survival, while overall response rate was influenced by tumor type and bevacizumab dose (p-value: 0.02). Although bevacizumab use resulted in additional expected adverse drug reactions except anemia and fatigue, it was not associated with a significant decline in quality of life. There was a trend towards a higher risk of several side effects in patients treated by high-dose bevacizumab compared to the lowdose e.g. all grade proteinuria (9.24; 95% CI: 6.60-12.94 vs. 2.64; 95% CI: 1.29-5.40). Conclusions: Combining bevacizumab with different adjuvant therapies provides a survival benefit across all major subsets of patients, including by tumor type, type of adjuvant therapy, and duration and dose of bevacizumab therapy. Though bevacizumab was associated with increased risks of some adverse drug reactions s

The functional capacity of NK cells is dynamically tuned by integrated signals from inhibitory and activating cell surface receptors in a process termed NK cell education. However, the understanding of the cellular and molecular mechanisms behind this functional tuning is limited. In this study, we show that the expression of the adhesion molecule and activation receptor DNAX accessory molecule 1 (DNAM-1) correlates with the quantity and quality of the inhibitory input by HLA class I-specific killer cell Ig-like receptors and CD94/NKG2A as well as with the magnitude of functional responses. Upon target cell recognition, the conformational state of LFA-1 changed in educated NK cells, associated with rapid colocalization of both active LFA-1 and DNAM-1 at the immune synapse. Thus, the coordinated expression of LFA-1 and DNAM-1 is a central component of NK cell education and provides a potential mechanism for controlling cytotoxicity by functionally mature NK cells., QC 20150526

Background and Aims: CXCL-8 is a pro-inflammatory chemokine that induces neutrophil chemotaxis. Low serum CXCL-8 levels correlate with virological response to IFNalfa in HCV patients. We examined the correlation of CXCL-8 and other markers with HBeAg seroconversion and ALT flares in CHB patients treated with IFNalfa. Method(s): Treatment naive CHB patients received either Peg-IFN alone or Peg-IFN and Tenofovir for 48 weeks. Plasma was collected prior to, during and post treatment. CXCL-8, CXCL-10, MCP-1, MIG and IP-10 were measured with cytometric bead array. HBV load, quantitative HBeAg and HBsAg were measured at 9 time points. Result(s): 17 HBeAg positive patients were recruited and 119 longitudinal plasma specimens collected. Serial ALT levels correlated with IL-10 (Spearman's rank correlation coefficient (rho) = 0.399, p < 0.001), MIG (rho = 0.492, p < 0.001) and IP-10 (rho = 0.534, p < 0.001). HBeAg seroconversion occurred in 31.3% of patients (no difference between treatment groups). Serum CXCL-8 >10,000 fg/ml at baseline predicted failure to achieve HBeAg seroconversion (PPV: 100%, NPV: 45%). 50% experienced a post-treatment ALT flare (median 87 days, IQR: 64-150 days post treatment). CXCL-8 >10,000 fg/ml one week post IFNalfa completion predicted a HBV flare within 6 months (PPV: 83%, NPV: 60%). Among patients who flared, an increase in CXCL-8 was detected 4-12 weeks before an ALT rise. Other markers rose in tandem with ALT but did not herald the occurrence of flares. Conclusion(s): CXCL-8 is a potential biomarker for predicting pre and post treatment response to IFNalfa therapy in CHB. CXCL-8 may affect the virological response to IFN-alfa therapy and requires further investigation.

Background and Aims: OPTIMIZE confirmed that telaprevir twice daily (bid) plus pegylated interferon-alfa (P) and ribavirin (R) was noninferior to every 8 hours (q8h) for SVR12 in treatmentnaive patients with genotype 1 chronic HCV infection. Decreased glomerular filtration rate (GFR) may occur with PI-based triple therapy. We examined the effect of telaprevir and comorbidities on GFR during and post treatment. Method(s): Patients (screening creatinine clearance >=50 mL/min) received telaprevir bid or q8h for 12 weeks, then PR for 12/36 weeks. GFR (Cockcroft-Gault) was measured at baseline, and regularly during Weeks 1-12 and post-telaprevir treatment (Weeks 12-48) in the overall treatment phase. Each patient's lowest GFR recorded during each phase of the study was used. Result(s): At baseline, mean(SD) GFR was 110.5(31.5) mL/min (n = 739). GFR decreased in both treatment groups during telaprevir treatment (bid: -15 mL/min; q8h: -14 mL/min; P = 0.40), but returned to baseline levels post telaprevir (bid: 0 mL/min; q8h: -1.1 mL/min) by Week 13. Similar numbers of patients had GFR <60 mL/min (bid: 41/365 versus q8h: 40/368; P = 0.91), mostly during the telaprevir treatment phase (bid: 40/41; q8h: 38/40). GFR reductions were slightly greater with advanced fibrosis stage during and post telaprevir (Table 1). Greater GFR reductions from baseline during telaprevir treatment occurred in patients with diabetes (P = 0.026), hypertension (P <0.001) or those receiving ACE inhibitors (P = 0.002) versus those without. Conclusion(s): GFR decreased during telaprevir treatment and was <60 mL/min in 11% of patients. GFR returned to baseline levels following telaprevir completion. Advanced fibrosis and comorbidities contributed to decreases in GFR in the presence of telaprevir. (Table Presented).

Background and Aims: CXCL-8 is a pro-inflammatory chemokine that induces neutrophil chemotaxis. Low serum CXCL-8 levels correlate with virological response to IFNalfa in HCV patients. We examined the correlation of CXCL-8 and other markers with HBeAg seroconversion and ALT flares in CHB patients treated with IFNalfa. Method(s): Treatment naive CHB patients received either Peg-IFN alone or Peg-IFN and Tenofovir for 48 weeks. Plasma was collected prior to, during and post treatment. CXCL-8, CXCL-10, MCP-1, MIG and IP-10 were measured with cytometric bead array. HBV load, quantitative HBeAg and HBsAg were measured at 9 time points. Result(s): 17 HBeAg positive patients were recruited and 119 longitudinal plasma specimens collected. Serial ALT levels correlated with IL-10 (Spearman's rank correlation coefficient (rho) = 0.399, p < 0.001), MIG (rho = 0.492, p < 0.001) and IP-10 (rho = 0.534, p < 0.001). HBeAg seroconversion occurred in 31.3% of patients (no difference between treatment groups). Serum CXCL-8 >10,000 fg/ml at baseline predicted failure to achieve HBeAg seroconversion (PPV: 100%, NPV: 45%). 50% experienced a post-treatment ALT flare (median 87 days, IQR: 64-150 days post treatment). CXCL-8 >10,000 fg/ml one week post IFNalfa completion predicted a HBV flare within 6 months (PPV: 83%, NPV: 60%). Among patients who flared, an increase in CXCL-8 was detected 4-12 weeks before an ALT rise. Other markers rose in tandem with ALT but did not herald the occurrence of flares. Conclusion(s): CXCL-8 is a potential biomarker for predicting pre and post treatment response to IFNalfa therapy in CHB. CXCL-8 may affect the virological response to IFN-alfa therapy and requires further investigation.

Background and Aims: OPTIMIZE confirmed that telaprevir twice daily (bid) plus pegylated interferon-alfa (P) and ribavirin (R) was noninferior to every 8 hours (q8h) for SVR12 in treatmentnaive patients with genotype 1 chronic HCV infection. Decreased glomerular filtration rate (GFR) may occur with PI-based triple therapy. We examined the effect of telaprevir and comorbidities on GFR during and post treatment. Method(s): Patients (screening creatinine clearance >=50 mL/min) received telaprevir bid or q8h for 12 weeks, then PR for 12/36 weeks. GFR (Cockcroft-Gault) was measured at baseline, and regularly during Weeks 1-12 and post-telaprevir treatment (Weeks 12-48) in the overall treatment phase. Each patient's lowest GFR recorded during each phase of the study was used. Result(s): At baseline, mean(SD) GFR was 110.5(31.5) mL/min (n = 739). GFR decreased in both treatment groups during telaprevir treatment (bid: -15 mL/min; q8h: -14 mL/min; P = 0.40), but returned to baseline levels post telaprevir (bid: 0 mL/min; q8h: -1.1 mL/min) by Week 13. Similar numbers of patients had GFR <60 mL/min (bid: 41/365 versus q8h: 40/368; P = 0.91), mostly during the telaprevir treatment phase (bid: 40/41; q8h: 38/40). GFR reductions were slightly greater with advanced fibrosis stage during and post telaprevir (Table 1). Greater GFR reductions from baseline during telaprevir treatment occurred in patients with diabetes (P = 0.026), hypertension (P <0.001) or those receiving ACE inhibitors (P = 0.002) versus those without. Conclusion(s): GFR decreased during telaprevir treatment and was <60 mL/min in 11% of patients. GFR returned to baseline levels following telaprevir completion. Advanced fibrosis and comorbidities contributed to decreases in GFR in the presence of telaprevir. (Table Presented).

Background: Vitamin D is believed to play an important role outside the endocrine system in the regulation of the immune system, and in cellular proliferation and differentiation. The aim of the study was to investigate the impact of vitamin D levels on innate immunity. Method(s): Participants for this prospective, longitudinal study were recruited amongst otherwise healthy staff of a large hospital in Victoria, Australia. Those fulfilling the inclusion criteria, including a vitamin D level of <50 nmol/L, were supplemented. Using flow cytometry, expression of the innate immune receptors TLR2, TLR4 and CD86 was measured on peripheral blood mononuclear cells (PBMCs) collected prior to vitamin D treatment and then at 1 and 3 months. Additonally, PBMCs at each timepoint were stimulated with specific TLR ligands and resultant supernatants were assayed for the cytokines TNFalpha, IL-6, IFN-alpha and IP-10. Result(s): In participants whose vitamin D level was >100 nmol/L post supplementation (n=11), TLR2 expression on PBMCs increased significantly, with no change noted in TLR4 or CD86 expression. Stimulation of vitamin D deficient samples with TLR ligands produced a number of proinflammatory cytokines, which were significantly reduced upon vitamin D normalisation. In patients whose levels returned to a deficient level at 3 months despite ongoing low-level supplementation, an increase in the pro-inflamamtory state returned. This suggests that vitamin D may play an important role in ensuring an appropriate baseline pro-inflammatory state. Conclusion(s): This ex-vivo pilot study adds clinical evidence supporting a possibly important role for vitamin D in innate immunity. If confirmed, this unique clinical study has potentially significant implications for the treatment of a variety of inflammatory conditions, where achieving optimal vitamin D levels may help reduce inflammation. © 2013 Ojaimi et al.; licensee BioMed Central Ltd.

Toll like receptors (TLR) are crucial mediators of innate immune responses, but their influence on natural killer (NK) cell function in chronic hepatitis B infection (CHB) is not well understood. Here we evaluated the responses of peripheral NK cells from CHB patients to multiple TLR agonists. CHB was associated with an impaired NK cell IFN-gamma response to TLR9 stimulation compared to controls. This deficiency corrected with recombinant IFN-alpha, while anti-IFN-alpha neutralizing antibody diminished NK IFN-gamma production in controls. NK cell CD69 upregulation in response to TLR9 was maintained in CHB. No differences were noted in responses to the other TLR ligands. Our results demonstrate a dichotomous NK cell response to TLR9 that is mediated by IFN-alpha and reflect the multiple mechanisms involved with NK activation. Consequently, it is possible that when activated these cells are unable to contribute to viral clearance while still having the potential to mediate tissue injury. © 2012 Elsevier Inc.

Background: Vitamin D is believed to play an important role outside the endocrine system in the regulation of the immune system, and in cellular proliferation and differentiation. The aim of the study was to investigate the impact of vitamin D levels on innate immunity. Method(s): Participants for this prospective, longitudinal study were recruited amongst otherwise healthy staff of a large hospital in Victoria, Australia. Those fulfilling the inclusion criteria, including a vitamin D level of <50 nmol/L, were supplemented. Using flow cytometry, expression of the innate immune receptors TLR2, TLR4 and CD86 was measured on peripheral blood mononuclear cells (PBMCs) collected prior to vitamin D treatment and then at 1 and 3 months. Additonally, PBMCs at each timepoint were stimulated with specific TLR ligands and resultant supernatants were assayed for the cytokines TNFalpha, IL-6, IFN-alpha and IP-10. Result(s): In participants whose vitamin D level was >100 nmol/L post supplementation (n=11), TLR2 expression on PBMCs increased significantly, with no change noted in TLR4 or CD86 expression. Stimulation of vitamin D deficient samples with TLR ligands produced a number of proinflammatory cytokines, which were significantly reduced upon vitamin D normalisation. In patients whose levels returned to a deficient level at 3 months despite ongoing low-level supplementation, an increase in the pro-inflamamtory state returned. This suggests that vitamin D may play an important role in ensuring an appropriate baseline pro-inflammatory state. Conclusion(s): This ex-vivo pilot study adds clinical evidence supporting a possibly important role for vitamin D in innate immunity. If confirmed, this unique clinical study has potentially significant implications for the treatment of a variety of inflammatory conditions, where achieving optimal vitamin D levels may help reduce inflammation. © 2013 Ojaimi et al.; licensee BioMed Central Ltd.

Toll like receptors (TLR) are crucial mediators of innate immune responses, but their influence on natural killer (NK) cell function in chronic hepatitis B infection (CHB) is not well understood. Here we evaluated the responses of peripheral NK cells from CHB patients to multiple TLR agonists. CHB was associated with an impaired NK cell IFN-gamma response to TLR9 stimulation compared to controls. This deficiency corrected with recombinant IFN-alpha, while anti-IFN-alpha neutralizing antibody diminished NK IFN-gamma production in controls. NK cell CD69 upregulation in response to TLR9 was maintained in CHB. No differences were noted in responses to the other TLR ligands. Our results demonstrate a dichotomous NK cell response to TLR9 that is mediated by IFN-alpha and reflect the multiple mechanisms involved with NK activation. Consequently, it is possible that when activated these cells are unable to contribute to viral clearance while still having the potential to mediate tissue injury. © 2012 Elsevier Inc.

Background Hepatitis C (HCV) recurrence post liver transplant follows a more aggressive course than pre-transplantation and the mechanisms remain poorly understood. Toll-like receptors (TLRs) are critical to innate immune antiviral responses. The interplay between TLR function and immunosuppressive drugs and how this may affect recurrence of diseases such as HCV infection post transplant is unknown. Aims To determine the relationship between serum calcineurin inhibitor levels and TLR function post liver transplantation. Methods Peripheral blood mononuclear cells (PBMCs) from 83 HCV post liver transplant patients, 53 non-HCV post transplant controls (matched for sex, race and time post transplant) and 10 healthy controls were stimulated with TLR specific ligands LPS (TLR4), P3C (TLR2), PIC (TLR3), R848 (TLR7/8) and CpG (TLR9) for 24 hrs. Production of interleukin 6 (IL-6), tumour necrosis factor (TNF) and interferon alpha (IFNalpha) was then measured using ELISA. Flow cytometry was used to determine intracellular cytokine production by monocytes, NK, NKT and T cells. Results were compared between groups using Mann-Whitney and Spearman correlation. Results Cyclosporine levels correlated with reduced TLR3-induced IFNgamma production by NK cells (NK cells p = 0.011, CD56bright NK cells p = 0.017) and T cells (p = 0.049). Cyclosporine levels also correlated with reduced TLR3-induced monocyte IL-6 (p = 0.028) and TNF (p = 0.070) production. Cyclosporine level correlated positively with T cell frequency (p = 0.040). In contrast, tacrolimus levels correlated inversely with TLR4-induced PBMC IL-6 production (p = 0.030), TLR3-induced TNF production by PBMCs (p = 0.027) and TLR7/8-induced NK cell TNF (NK cells p = 0.009, CD56dim NK cells p = 0.013). Tacrolimus levels correlated inversely with NKT cell frequency (p = 0.032). Conclusion NK cells and monocytes have impaired TLR3-induced cytokine production that inversely correlated with increasing cyclosporine level. In contrast

Background: Hepatitis C (HCV) recurrence post liver transplant follows a more aggressive course than pre-transplantation and the mechanisms remain poorly understood. Toll-like receptors (TLRs) are critical to innate immune antiviral responses. The interplay between TLR function and immunosuppressive drugs and how this may affect HCV recurrence post transplant is unknown. Method(s): Peripheral blood mononuclear cells (PBMCs) from 83 HCV post liver transplant patients, 53 non-HCV post transplant controls (matched for sex, race and time post transplant) and 10 healthy controls were stimulated with TLR specific ligands LPS (TLR4), P3C (TLR2), PIC (TLR3), R848 (TLR7/8) and CpG (TLR9) for 24 hrs. Production of interleukin 6 (IL-6), tumour necrosis factor (TNF) and interferon alpha (IFNa) was then measured using ELISA. Flow cytometry was used to determine intracellular cytokine production by monocytes, NK, NKT and T cells. Results were compared between groups using Mann-Whitney and Spearman correlation. Result(s): Cyclosporine levels correlated with reduced TLR3-induced IFNg production by NK cells (NK cells p = 0.011, CD56bright NK cells p = 0.017) and T cells (p = 0.049). Cyclosporine levels also correlated with reduced TLR3-induced monocyte IL-6 (p = 0.028) and TNF (p = 0.070) production. Cyclosporine level correlated positively with T cell frequency (p = 0.040). In contrast, tacrolimus levels correlated inversely with TLR4- induced PBMC IL-6 production (p = 0.030), TLR3-induced TNF production by PBMCs (p = 0.027) and TLR7/8-induced NK cell TNF (NK cells p = 0.009, CD56dim NK cells p = 0.013). Tacrolimus levels correlated inversely with NKT cell frequency (p = 0.032). Conclusion(s): NK cells and monocytes have impaired TLR3-induced cytokine production that varied with increasing cyclosporine level. In contrast, tacrolimus levels correlated inversely with TLR4- induced IL-6 by PBMCs, TLR3-induced TNF from monocytes, and TLR7/8 induced TNF from NK cells. These data demonst

Background In patients with chronic hepatitis B (CHB), hepatitis B surface antigen levels (QHBsAg) have recently been associated with predicting response to peg-interferon-alfa, including HBsAg seroconversion (SC) and loss or decline. Long-term nucleos(t)ide analogues (NA) suppresses HBV replication in the majority of patients, however very few actually undergo HBsAg SC. The utility of HBsAg quantitative analysis in these patients is poorly understood, and the impact of multi-drug resistant (MDR) variants has not been explored. Aims To evaluate the QHBsAg decline in a well-characterized cohort of patients with MDR (rtA181T/V) HBV who were treated with >7 years with multiple NA, including at least 3 years of tenofovir (TDF). Methods TDF-109 was an investigator-initiated, prospective, multicentre, open-label study of 60 patients treated with TDF salvage/rescue therapy. All patients had previously failed LAM and subsequent ADVadd- on or switch therapy. The median age was 48.5 years (range 21-80), 77% were males, 67% were HBeAg positive, 67% were of Asian ethnicity and 52% were infected with HBV genotype C. Median follow-up is now 4 years; all patients have achieved an adequate virological response, with 75% (38/51) having undetectable HBV DNA levels. All patients have been maintained on TDF in long-term follow-up, now out to 4 years. QHBsAg titres were retrospectively quantified at baseline and every 6 months and expressed as IU/ml (Roche Elecsys). Results At baseline, median QHBsAg titre was 5226 IU/ml. Baseline QHBsAg titre were positively correlated with HBV DNA levels at baseline, R = 0.33, p = 0.0168). There was a trend for higher baseline QHBsAg levels in HBeAg positive patients (p = 0.1257). The median HBsAg titres were 2772 IU/ml at year 1, 2284 IU/ml at year 2 and 1819 IU/ml at year 3. There was a trend for patients with detectable rtA181T/V variants to have a lower QHBsAg at baseline. There was a trend for patients with no detectable resistance changes or lam

Hepatitis B virus (HBV) infection in immunocompetent adults usually results in a self-limited, transient liver disease and viral clearance, with only a small percentage (5 to 10%) developing chronic hepatitis associated with viral persistence. In contrast, when neonates are infected, more than 90% become persistently infected, suffering differing degrees of chronic liver disease. Activation of immunity plays a central role in host-virus interactions, greatly influencing viral replication and the clinical outcome of infection. Although all of the specific mechanisms and consequences of this interaction have not been elucidated, the purpose of this article is to describe the basic arms of the immune system as they interact with the HBV and describe the present state of knowledge in this area. These arms may be divided broadly into innate and specific immune responses, and they have different roles and responses in acute and chronic infection. Copyright © 2006 by Thieme Medical Publishers, Inc.

Synthetic small interfering RNAs (siRNAs) can trigger a strong innate immune response in mammalian cells. This nonspecific side effect may hinder the application of siRNAs as tools in gene silencing. Chemically synthesized siRNAs, including traditional 19-mers with 2-nt 3' overhangs, longer duplexes with blunt or 3' overhangs, and asymmetric duplexes with a blunt end and a 2-nt 3' overhang, can evoke strong dose-dependent interferon-alpha (IFN-alpha) and tumor necrosis factor-alpha (TNF-alpha) release in human peripheral blood mononuclear cells (PBMCs). This response is independent of retinoic acid-inducible gene I but may involve endosomal toll-like receptors (TLRs). The immunostimulatory effect of the siRNAs is directly related to either or both of the strands of the duplex in a sequence-dependent manner. However, although some single-stranded RNAs and siRNAs potently evoked both IFN-alpha and TNF-alpha induction, these responses were not always coupled. In accordance with this, specific chemical modifications differentially altered cytokine production, suggesting recruitment of different TLRs in a sequence-dependent manner. © 2008 Mary Ann Liebert, Inc.

Hepatitis B virus (HBV) infection in immunocompetent adults usually results in a self-limited, transient liver disease and viral clearance, with only a small percentage (5 to 10%) developing chronic hepatitis associated with viral persistence. In contrast, when neonates are infected, more than 90% become persistently infected, suffering differing degrees of chronic liver disease. Activation of immunity plays a central role in host-virus interactions, greatly influencing viral replication and the clinical outcome of infection. Although all of the specific mechanisms and consequences of this interaction have not been elucidated, the purpose of this article is to describe the basic arms of the immune system as they interact with the HBV and describe the present state of knowledge in this area. These arms may be divided broadly into innate and specific immune responses, and they have different roles and responses in acute and chronic infection. Copyright © 2006 by Thieme Medical Publishers, Inc.

Background Hepatitis C (HCV) recurrence post liver transplant follows a more aggressive course than pre-transplantation and the mechanisms remain poorly understood. Toll-like receptors (TLRs) are critical to innate immune antiviral responses. The interplay between TLR function and immunosuppressive drugs and how this may affect recurrence of diseases such as HCV infection post transplant is unknown. Aims To determine the relationship between serum calcineurin inhibitor levels and TLR function post liver transplantation. Methods Peripheral blood mononuclear cells (PBMCs) from 83 HCV post liver transplant patients, 53 non-HCV post transplant controls (matched for sex, race and time post transplant) and 10 healthy controls were stimulated with TLR specific ligands LPS (TLR4), P3C (TLR2), PIC (TLR3), R848 (TLR7/8) and CpG (TLR9) for 24 hrs. Production of interleukin 6 (IL-6), tumour necrosis factor (TNF) and interferon alpha (IFNalpha) was then measured using ELISA. Flow cytometry was used to determine intracellular cytokine production by monocytes, NK, NKT and T cells. Results were compared between groups using Mann-Whitney and Spearman correlation. Results Cyclosporine levels correlated with reduced TLR3-induced IFNgamma production by NK cells (NK cells p = 0.011, CD56bright NK cells p = 0.017) and T cells (p = 0.049). Cyclosporine levels also correlated with reduced TLR3-induced monocyte IL-6 (p = 0.028) and TNF (p = 0.070) production. Cyclosporine level correlated positively with T cell frequency (p = 0.040). In contrast, tacrolimus levels correlated inversely with TLR4-induced PBMC IL-6 production (p = 0.030), TLR3-induced TNF production by PBMCs (p = 0.027) and TLR7/8-induced NK cell TNF (NK cells p = 0.009, CD56dim NK cells p = 0.013). Tacrolimus levels correlated inversely with NKT cell frequency (p = 0.032). Conclusion NK cells and monocytes have impaired TLR3-induced cytokine production that inversely correlated with increasing cyclosporine level. In contrast

Background: Hepatitis C (HCV) recurrence post liver transplant follows a more aggressive course than pre-transplantation and the mechanisms remain poorly understood. Toll-like receptors (TLRs) are critical to innate immune antiviral responses. The interplay between TLR function and immunosuppressive drugs and how this may affect HCV recurrence post transplant is unknown. Method(s): Peripheral blood mononuclear cells (PBMCs) from 83 HCV post liver transplant patients, 53 non-HCV post transplant controls (matched for sex, race and time post transplant) and 10 healthy controls were stimulated with TLR specific ligands LPS (TLR4), P3C (TLR2), PIC (TLR3), R848 (TLR7/8) and CpG (TLR9) for 24 hrs. Production of interleukin 6 (IL-6), tumour necrosis factor (TNF) and interferon alpha (IFNa) was then measured using ELISA. Flow cytometry was used to determine intracellular cytokine production by monocytes, NK, NKT and T cells. Results were compared between groups using Mann-Whitney and Spearman correlation. Result(s): Cyclosporine levels correlated with reduced TLR3-induced IFNg production by NK cells (NK cells p = 0.011, CD56bright NK cells p = 0.017) and T cells (p = 0.049). Cyclosporine levels also correlated with reduced TLR3-induced monocyte IL-6 (p = 0.028) and TNF (p = 0.070) production. Cyclosporine level correlated positively with T cell frequency (p = 0.040). In contrast, tacrolimus levels correlated inversely with TLR4- induced PBMC IL-6 production (p = 0.030), TLR3-induced TNF production by PBMCs (p = 0.027) and TLR7/8-induced NK cell TNF (NK cells p = 0.009, CD56dim NK cells p = 0.013). Tacrolimus levels correlated inversely with NKT cell frequency (p = 0.032). Conclusion(s): NK cells and monocytes have impaired TLR3-induced cytokine production that varied with increasing cyclosporine level. In contrast, tacrolimus levels correlated inversely with TLR4- induced IL-6 by PBMCs, TLR3-induced TNF from monocytes, and TLR7/8 induced TNF from NK cells. These data demonst

Background In patients with chronic hepatitis B (CHB), hepatitis B surface antigen levels (QHBsAg) have recently been associated with predicting response to peg-interferon-alfa, including HBsAg seroconversion (SC) and loss or decline. Long-term nucleos(t)ide analogues (NA) suppresses HBV replication in the majority of patients, however very few actually undergo HBsAg SC. The utility of HBsAg quantitative analysis in these patients is poorly understood, and the impact of multi-drug resistant (MDR) variants has not been explored. Aims To evaluate the QHBsAg decline in a well-characterized cohort of patients with MDR (rtA181T/V) HBV who were treated with >7 years with multiple NA, including at least 3 years of tenofovir (TDF). Methods TDF-109 was an investigator-initiated, prospective, multicentre, open-label study of 60 patients treated with TDF salvage/rescue therapy. All patients had previously failed LAM and subsequent ADVadd- on or switch therapy. The median age was 48.5 years (range 21-80), 77% were males, 67% were HBeAg positive, 67% were of Asian ethnicity and 52% were infected with HBV genotype C. Median follow-up is now 4 years; all patients have achieved an adequate virological response, with 75% (38/51) having undetectable HBV DNA levels. All patients have been maintained on TDF in long-term follow-up, now out to 4 years. QHBsAg titres were retrospectively quantified at baseline and every 6 months and expressed as IU/ml (Roche Elecsys). Results At baseline, median QHBsAg titre was 5226 IU/ml. Baseline QHBsAg titre were positively correlated with HBV DNA levels at baseline, R = 0.33, p = 0.0168). There was a trend for higher baseline QHBsAg levels in HBeAg positive patients (p = 0.1257). The median HBsAg titres were 2772 IU/ml at year 1, 2284 IU/ml at year 2 and 1819 IU/ml at year 3. There was a trend for patients with detectable rtA181T/V variants to have a lower QHBsAg at baseline. There was a trend for patients with no detectable resistance changes or lam

Synthetic small interfering RNAs (siRNAs) can trigger a strong innate immune response in mammalian cells. This nonspecific side effect may hinder the application of siRNAs as tools in gene silencing. Chemically synthesized siRNAs, including traditional 19-mers with 2-nt 3' overhangs, longer duplexes with blunt or 3' overhangs, and asymmetric duplexes with a blunt end and a 2-nt 3' overhang, can evoke strong dose-dependent interferon-alpha (IFN-alpha) and tumor necrosis factor-alpha (TNF-alpha) release in human peripheral blood mononuclear cells (PBMCs). This response is independent of retinoic acid-inducible gene I but may involve endosomal toll-like receptors (TLRs). The immunostimulatory effect of the siRNAs is directly related to either or both of the strands of the duplex in a sequence-dependent manner. However, although some single-stranded RNAs and siRNAs potently evoked both IFN-alpha and TNF-alpha induction, these responses were not always coupled. In accordance with this, specific chemical modifications differentially altered cytokine production, suggesting recruitment of different TLRs in a sequence-dependent manner. © 2008 Mary Ann Liebert, Inc.

Background: In CHB patients, quantitative hepatitis B surface antigen (QHBsAg) have recently been associated with predicting response to peg-interferon-alfa, including HBsAg seroconversion (SC) & loss or decline. Long-term nucleos(t)ide analogues (NA) suppresses HBV replication in the majority of patients, however very few actually undergo HBsAg SC. The utility of QHBsAg analysis in these patients is poorly understood & the impact of multi-drug resistant (MDR) variants has not been explored. Aim(s): To evaluate the QHBsAg decline in a wellcharacterized cohort of patients with MDR (rtA181T/V) HBV who were treated with >7years with multiple NA, including at least 3 years of tenofovir (TDF). Method(s): TDF109 was an investigator-initiated, prospective, multi-centre, open-label study of the efficacy of TDF rescue therapy in 60 patients who had previously failed lamivudine (LAM) & subsequent adefovir (ADV) add-on or switch therapy. We have previously presented virological outcome at 2 years (Patterson, Gut 2011). All patients have been maintained on TDF in long-term follow-up, now out to 4 years. QHBsAg titers were retrospectively quantified at baseline & every 6 months, expressed as IU/ml (Roche Elecsys). Sub-analysis according to baseline viral load & the type of HBV resistance-associated substitutions present at baseline was then performed. Result(s): At baseline, median QHBsAg titer was 5226IU/ml. Baseline QHBsAg positively correlated with HBV DNA levels at baseline, (R = 0.33, p=0.0168). There was a trend for higher baseline QHBsAg levels in HBeAg positive patients (p=0.13). The median HBsAg titers were 2772IU/ml, 2284IU/ml &1819IU/ml at year 1, 2&3 respectively (P<0.0001 for comparison of baseline to year 3). Patients with detectable rtA181T/V variants have a lower QHBsAg at baseline, but more gradual decline in QHBsAg (p=0.002). There was a trend for patients with no detectable resistance changes or LAM resistance HBV Pol at baseline, to have higher baseline QHBsA

Small intestinal neuroendocrine tumors (carcinoids) are the most common neuroendocrine tumor (NET) within the gastrointestinal tract (15–20%). The incidence and prevalence is steadily increasing during the last decade and the estimated incidence is now about 2.5/100 000/year. The majority of the tumors produce peptides and amines giving rise to clinical symptoms related to the hormone production such as the carcinoid syndrome with flushing, diarrhea, bronchial constriction, and carcinoid heart disease. The majority of patients present with metastatic disease (60%) and therefore a surgical cure is not obtainable. The medical treatment consists of somatostatin analogs, alpha interferons, and targeted treatment with mTOR- and tyrosine kinase-inhibitors. Somatostatin analogs are still the gold standard for treatment of carcinoid tumors with hormone-related clinical symptoms. A possible antitumor effect has been debated, but a recent study (PROMID) has demonstrated an antitumor effect of octreotide LAR, 30 mg, every 4 weeks versus placebo. Tyrosine kinase inhibitors have been applied mostly for antiangiogenic purposes with objective responses in 10–20% of the patients and PFS of 10–15 months. The mTOR-inhibitor everolimus have demonstrated an antitumor effect in patients with carcinoid tumors in the RADIANT-2 trial. Cytotoxic treatment has demonstrated a limited efficacy in classical low proliferation well-differentiated neuroendocrine gastrointestinal tumors and should not be considered first-line treatment. The future treatment will be based on tumor biology and molecular genetics taking into account the new classification systems. The treatment will be personalized.

Infection with hepatitis C virus (HCV) is a major global health issue. Only a small proportion of patients clear the virus spontaneously and the majority develop chronic hepatitis C infection. Chronic hepatitis C is one of the most common causes of advanced liver disease in the western world and is now the leading indication for liver transplantation. Unfortunately, the standard treatment, consisting of pegylated-interferon and ribavirin, is suboptimal. Less than 50% of patients infected with HCV genotype 1 are cured, treatment is costly and is associated with significant toxicity. Therefore, there has been a need to identify accurate predictors of treatment outcome to facilitate treatment decision-making. Four independent genome-wide association studies have recently confirmed an association between genetic variation in the region of the IL28B gene and treatment outcome in HCV-1 patients. Patients who carry the good response variant are two-to three-fold more likely to be cured. The difference in the frequency of the good response variant between patients of different ethnic background explains much of the recognized ethnic disparity in treatment response rates. The IL28B variants are also associated with likelihood of spontaneous clearance of HCV infection. This discovery represents a significant advance in our ability to personalize HCV therapy, as well as suggesting novel avenues for research into viral pathogenesis and therapeutic development. © 2011 Future Medicine Ltd.

Background: Toll like receptors (TLRs) are critical to innate immune antiviral responses. Hepatitis C (HCV) alters TLR function to evade immune clearance. Whether TLRs play a role in rapid HCV recurrence and fibrosis progression post liver transplant is unknown. Method(s): Peripheral blood mononuclear cells (PBMCs) from 73 HCV post liver transplant patients and 53 non-HCV post transplant controls (matched for sex, race and time post transplant) were stimulated with TLR subclass-specific ligands LPS (TLR4), P3C (TLR2), PIC (TLR3), R848 (TLR7/8) and CpG (TLR9) for 24 hrs. Production of interleukin 6 (IL-6), tumour necrosis factor (TNFalpha) and interferon alpha (IFNalpha) was then measured by analyzing cell culture supernatants using enzyme-linked immunosorbent assay and compared between groups using Mann Whitney test. Rate of fibrosis progression was calculated using post transplant liver biopsies graded by Metavir scoring (F0-4; R = fibrosis stage/ year post transplant). Rapid fibrosis was defined as >0.4 units/yr, dichotomized about the median observed rate (0.4). Result(s): 28 of 73 (38%) HCV post liver transplant patients had rapid fibrosis progression. PBMCs from HCV subjects produced greater amounts of IL-6 at baseline (p = 0.03) and in response to TLR3 stimulation (p = 0.02) compared with controls. In contrast, PBMCs from HCV rapid-fibrosers produced less IL-6 in response to TLR3 (p = 0.007) and TLR9 stimulation (p = 0.02), and less IFNalpha in response to TLR7/8 stimulation compared with HCV slow-fibrosers (p = 0.02). We confirmed these findings in a prospective cohort of 16 patients who had PBMCs collected and liver biopsy 1 yr post transplant. 8 (50%) had fibrosis progression (F1 or greater). Rapid fibrosers produced less IL-6 in response to TLR3 (p = 0.05) and TLR9 stimulation (p = 0.06) and less IFNalpha in response to TLR7/8 stimulation (p = 0.01). Additionally, rapid fibrosers produced less TNFalpha in response to TLR3 stimulation (p = 0.009). Conclusio

Infection with hepatitis C virus (HCV) is a major global health issue. Only a small proportion of patients clear the virus spontaneously and the majority develop chronic hepatitis C infection. Chronic hepatitis C is one of the most common causes of advanced liver disease in the western world and is now the leading indication for liver transplantation. Unfortunately, the standard treatment, consisting of pegylated-interferon and ribavirin, is suboptimal. Less than 50% of patients infected with HCV genotype 1 are cured, treatment is costly and is associated with significant toxicity. Therefore, there has been a need to identify accurate predictors of treatment outcome to facilitate treatment decision-making. Four independent genome-wide association studies have recently confirmed an association between genetic variation in the region of the IL28B gene and treatment outcome in HCV-1 patients. Patients who carry the good response variant are two-to three-fold more likely to be cured. The difference in the frequency of the good response variant between patients of different ethnic background explains much of the recognized ethnic disparity in treatment response rates. The IL28B variants are also associated with likelihood of spontaneous clearance of HCV infection. This discovery represents a significant advance in our ability to personalize HCV therapy, as well as suggesting novel avenues for research into viral pathogenesis and therapeutic development. © 2011 Future Medicine Ltd.

Background: Toll like receptors (TLRs) are critical to innate immune antiviral responses. Hepatitis C (HCV) alters TLR function to evade immune clearance. Whether TLRs play a role in rapid HCV recurrence and fibrosis progression post liver transplant is unknown. Method(s): Peripheral blood mononuclear cells (PBMCs) from 73 HCV post liver transplant patients and 53 non-HCV post transplant controls (matched for sex, race and time post transplant) were stimulated with TLR subclass-specific ligands LPS (TLR4), P3C (TLR2), PIC (TLR3), R848 (TLR7/8) and CpG (TLR9) for 24 hrs. Production of interleukin 6 (IL-6), tumour necrosis factor (TNFalpha) and interferon alpha (IFNalpha) was then measured by analyzing cell culture supernatants using enzyme-linked immunosorbent assay and compared between groups using Mann Whitney test. Rate of fibrosis progression was calculated using post transplant liver biopsies graded by Metavir scoring (F0-4; R = fibrosis stage/ year post transplant). Rapid fibrosis was defined as >0.4 units/yr, dichotomized about the median observed rate (0.4). Result(s): 28 of 73 (38%) HCV post liver transplant patients had rapid fibrosis progression. PBMCs from HCV subjects produced greater amounts of IL-6 at baseline (p = 0.03) and in response to TLR3 stimulation (p = 0.02) compared with controls. In contrast, PBMCs from HCV rapid-fibrosers produced less IL-6 in response to TLR3 (p = 0.007) and TLR9 stimulation (p = 0.02), and less IFNalpha in response to TLR7/8 stimulation compared with HCV slow-fibrosers (p = 0.02). We confirmed these findings in a prospective cohort of 16 patients who had PBMCs collected and liver biopsy 1 yr post transplant. 8 (50%) had fibrosis progression (F1 or greater). Rapid fibrosers produced less IL-6 in response to TLR3 (p = 0.05) and TLR9 stimulation (p = 0.06) and less IFNalpha in response to TLR7/8 stimulation (p = 0.01). Additionally, rapid fibrosers produced less TNFalpha in response to TLR3 stimulation (p = 0.009). Conclusio

Background: Toll like receptors (TLRs) are critical to innate immune antiviral responses and hepatitis C (HCV) alters TLR function to evade immune clearance. The role of TLRs in HCV recurrence post liver transplantation has not been characterised and whether rapid progression of HCV fibrosis is related to altered TLR responses is unknown. Method(s): Peripheral blood mononuclear cells (PBMCs) from 55 HCV post liver transplant patients and 30 non-HCV post transplant controls (matched for sex, race and time post transplant) were stimulated with TLR subclass-specific ligands LPS (TLR4), P3C (TLR2), PIC (TLR3), R848 (TLR7/8), CpGA (monocyte TLR9) and CpGB (plasmacytoid dendritic cell TLR9) for 24 h. Production of interleukin 6 (IL-6), tumour necrosis factor (TNF) and interferon alpha (IFNalpha) was then measured by analyzing cell culture supernatants using enzyme-linked immunosorbent assay and compared between groups using Mann Whitney and ANOVA tests. Rate of fibrosis progression was calculated using post transplant protocol liver biopsies graded by Metavir scoring (F0-4; R = fibrosis stage/year post transplant). Rapid fibrosis was defined as >0.5 units/year, dichotomized about the median observed rate (0.4). Results 11 of 55 HCV post liver transplant patients had rapid fibrosis progression (20%). PBMCs from HCV subjects produced greater amounts of IL-6 in response to TLR2 (p = 0.009), TLR3 (p = 0.02) and monocyte TLR9 stimulation (p = 0.04) compared with controls. HCV subjects also produced greater levels of TNF in response to TLR3 (p = 0.009) and TLR7/8 stimulation (p = 0.02). Baseline levels of both IL-6 (p = 0.014) and TNF (p = 0.05, NS) were also higher in HCV subjects. IFNalpha production was not different between HCV subjects and controls. In contrast, PBMCs from HCV rapid- fibrosers produced less IFNalpha in response to TLR7/8 stimulation compared with HCV slow-fibrosers (p = 0.014) and controls (p = 0.02). However, there was no significant difference in IL-6 an

Background: The effect of serum ALT changes in the outcome of combination pegylated interferon alpha (PEG IFNalpha) and ribavirin therapy for chronic hepatitis C virus (HCV) infection is not well known. We evaluated changes in ALT during standard therapy for HCV to determine correlations with sustained virological response (SVR), non-response (NR) and relapse (R). Method(s): 179 patients with HCV infection who completed standard combination therapy in our institution from 2004 to 2008 were identified. Age, sex, pre-treatment ALT and viral load, fibrosis score, type of PEG IFNalpha, on treatment viral loads and ALT (weeks 2, 4, 8, 12, 16, 20, 24, 36, 48) and during follow-up at 6 months were retrospectively collected and evaluated. Patients were divided into SVR, NR and R. ALT and baseline characteristics were compared using Student's T-test and Chi squared analysis to determine correlations between ALT level and virological response. Result(s): Overall SVR (all genotypes) was 72.07%, with NR 16.76% and R 11.17%. There was no significant ALT rise after week 12 in any of the outcome groups, however the absolute ALT level was significantly higher in NR compared to SVR at weeks 2 to 24 (p < 0.001). The percentage decrease in ALT was also significantly reduced at weeks 8, 12, 36, 48 in NR compared to SVR (p < 0.03). ALT levels in the R group were significantly higher than the SVR group from weeks 12 to 36 (p < 0.01), and had a reduced percentage change in ALT at weeks 8 to 24 (p < 0.02). Comparing baseline demographics, clinical and virological factors, only males were significantly more likely to relapse after completing treatment (p = 0.027). Of patients with initial viral suppression continuing to week 24 (n = 170), we found that ALT <= 25 U/L was strongly associated with SVR compared to NR and R (p < 0.05). Conclusion(s): Higher ALT levels between weeks 2 to 24 and a smaller percentage drop in ALT compared to pre-treatment were associated with NR. Higher ALT levels a

Despite the introduction of new nucleos(t)ide analogues in recent years, peginterferon is still recommended as a potential first-line treatment option by current practice guidelines for the management of chronic hepatitis B. Peginterferon offers the advantage of higher sustained off-treatment response rates compared to nucleos(t)ide analogues because of its immunomodulatory effects. Sustained transition to the inactive hepatitis B surface antigen (HBsAg) carrier state can be achieved in about 30% of hepatitis B e antigen (HBeAg)-positive patients and 20% of HBeAg-negative patients. Recent studies have focused on identification of pretreatment and on-treatment factors that allow the selection of patients who are likely to achieve a sustained response to peginterferon therapy in order to avoid the side-effects and costs associated with unnecessary treatment. Future studies need to address whether specific virologic benchmarks can guide individualized decisions concerning therapy continuation and whether peginterferon combined with new potent nucleos(t)ide analogues improves treatment outcomes.

Background: The effect of serum ALT changes in the outcome of combination pegylated interferon alpha (PEG IFNalpha) and ribavirin therapy for chronic hepatitis C virus (HCV) infection is not well known. We evaluated changes in ALT during standard therapy for HCV to determine correlations with sustained virological response (SVR), non-response (NR) and relapse (R). Method(s): 179 patients with HCV infection who completed standard combination therapy in our institution from 2004 to 2008 were identified. Age, sex, pre-treatment ALT and viral load, fibrosis score, type of PEG IFNalpha, on treatment viral loads and ALT (weeks 2, 4, 8, 12, 16, 20, 24, 36, 48) and during follow-up at 6 months were retrospectively collected and evaluated. Patients were divided into SVR, NR and R. ALT and baseline characteristics were compared using Student's T-test and Chi squared analysis to determine correlations between ALT level and virological response. Result(s): Overall SVR (all genotypes) was 72.07%, with NR 16.76% and R 11.17%. There was no significant ALT rise after week 12 in any of the outcome groups, however the absolute ALT level was significantly higher in NR compared to SVR at weeks 2 to 24 (p < 0.001). The percentage decrease in ALT was also significantly reduced at weeks 8, 12, 36, 48 in NR compared to SVR (p < 0.03). ALT levels in the R group were significantly higher than the SVR group from weeks 12 to 36 (p < 0.01), and had a reduced percentage change in ALT at weeks 8 to 24 (p < 0.02). Comparing baseline demographics, clinical and virological factors, only males were significantly more likely to relapse after completing treatment (p = 0.027). Of patients with initial viral suppression continuing to week 24 (n = 170), we found that ALT <= 25 U/L was strongly associated with SVR compared to NR and R (p < 0.05). Conclusion(s): Higher ALT levels between weeks 2 to 24 and a smaller percentage drop in ALT compared to pre-treatment were associated with NR. Higher ALT levels a

Introduction : While consumption of omega-3 long-chain polyunsaturated fatty acids (n-3 LCPUFA) has been recommended for those at risk of inflammatory disease such as rheumatoid arthritis, the mechanism of their anti-inflammatory effect remains to be clearly defined, particularly in relation to the dose and type of n-3 LCPUFA. The objective of this study was to determine whether varying the levels of n-3 LCPUFA in erythrocyte membrane lipids, following dietary supplementation, is associated with altered numbers and function of circulating leukocytes conducive to protection against inflammation. Methods : In a double-blind and placebo-controlled study, 44 healthy subjects aged 23 to 63 years consumed either standard or n-3 LCPUFA-enriched versions of typical processed foods, the latter allowing a target daily consumption of 1 gram n-3 LCPUFA. After six months, peripheral blood leukocyte and subpopulation proportions and numbers were assessed by flow cytometry. Leukocytes were also examined for lymphoproliferation and cytokine production, neutrophil chemotaxis, chemokinesis, bactericidal, adherence and iodination activity. Erythrocytes were analyzed for fatty-acid content. Results : Erythrocyte n-3 LCPUFA levels were higher and absolute leukocyte and lymphocyte numbers were lower in subjects consuming n-3 enriched foods than in controls. There were no changes in the number of neutrophils, monocytes, T cells (CD3+), T-cell subsets (CD4+, CD8+) and B cells (CD19+). However, natural killer (NK) (CD3-CD16+CD56+) cell numbers were lower in n-3 supplemented subjects than in controls and were inversely related to the amount of eicosapentaenoic acid or docosahexaenoic acid in erythrocytes. No significant correlations were found with respect to lymphocyte lymphoproliferation and production of IFN-γ and IL-2, but lymphotoxin production was higher with greater n-3 LCPUFA membrane content. Similarly, neutrophil chemotaxis, chemokinesis, bactericidal activity and adherence d

Adult soft tissue sarcomas are relatively rare tumours which are curable with radical surgery. Approximately 50% of patients will develop inoperable disease or metastases for which chemotherapy may be inappropriate. Only two cytotoxic agents - doxorubicin and ifosfamide - have activity in > 20% of patients. For both these agents there is evidence of a dose-response relationship. There is currently no good evidence that combination chemotherapy confers a clinical benefit compared with single agents. Outside a clinical trial, standard first-line therapy should be with single agent doxorubicin at a dose intensity ≥ 70 mg2 every 3 weeks. Approximately 25% of patients may be expected to respond to this regimen. There is the suggestion that responses may occur to ifosfamide in patients who progress on doxorubicin. The role of chemotherapy in the adjuvant setting remains uncertain. Several trials have suggested a modest relapse-free and overall survival benefit for the use of post-operative chemotherapy and a recent overview of 14 randomised trials confirms a small though significant benefit. These benefits have to be weighed against the toxicity of chemotherapy. The importance of treating all patients with soft tissue sarcomas in clinical trials is stressed. There is an urgent need to define new active agents to treat this disease.

Several immunomodulators were effective against infection of adult mice with herpes simplex virus type 2 (HSV-2), Venezualean equine encepalitis alphavirus (VEE), Banzi flavivirus or Caraparu bunyavirus. Similar effectiveness was found among the various immunodulators, although the viruses did differ in sensitivity. Infection with Banzi virus was the most sensitive, while infection with Caraparu virus was least sensitive to treatment with immunomodulators. Greatest antiviral protection was observed with C. parvum, alpha interferon, and the synthetic immunomodulators MVE-2, CL246,738 and amplign. Moderate protection was produced with gamma interferon and various microbially derived cell wall materials. Greatest effects with the immunomodulators were observed with prophylactic treatment, but early or repeated therapeutic treatment with such drugs as CL246,738, ampligen and alpha or gamma interferon was also partially effective. Thus, a variety of chemically diverse agents can produce broad spectrum antiviral protection. Immunomodulator induced alterations in nonspecific immunity may be involved in the protection. Many of the compounds induced NK cell activity and activated macrophages. A common mechanism for the antiviral action, however, has yet to be established. Keywords: Antiviral agents.

[No abstract available]