Human platelet antigen-3 polymorphism as a risk factor for rheumatological manifestations in hepatitis C

Abstract INTRODUCTION: Hepatitis C virus (HCV) infection is involved in the pathogenesis of autoimmune and rheumatic disorders. Although the human platelet antigens (HPA) polymorphism are associated with HCV persistence, they have not been investigated in rheumatological manifestations (RM). This study focused on verifying associations between allele and genotype HPA and RM in patients with chronic hepatitis C. METHODS: Patients (159) with chronic hepatitis C of both genders were analyzed. RESULTS: Women showed association between HPA-3 polymorphisms and RM. CONCLUSIONS: An unprecedented strong association between rheumatological manifestations and HPA-3 polymorphism, possibly predisposing women to complications during the disease course, was observed.

IgG1 and IgG4 antibodies against core and NS3 antigens of hepatitis C virus

Abstract INTRODUCTION: IgG subclasses involved in the immune response to hepatitis C virus (HCV) antigens have been rarely studied. We investigated the immune response mediated by IgG1 and IgG4 antibodies against the recombinant core and NS3 antigens in patients with chronic hepatitis C. METHODS: Sixty patients infected with HCV genotype 1 without antiviral treatment and 60 healthy subjects participated in the study. Serum levels of alanine aminotransferase, HCV viremia, and the presence of cryoglobulinemia and liver fibrosis were determined. We investigated the serum IgG1 and IgG4 antibodies against recombinant HCV core and NS3 non-structural protein antigens using amplified indirect ELISA. RESULTS: Anti-core and anti-NS3 IgG1 antibodies were detected in 33/60 (55%) and 46/60 (77%) patients, respectively, whereas only two healthy control samples reacted with an antigen (NS3). Anti-core IgG4 antibodies were not detected in either group, while 30/60 (50%) patients had anti-NS3 IgG4 antibodies. Even though there were higher levels of anti-NS3 IgG4 antibodies in patients with low viremia (< 8 × 105 IU/mL), IgG1 and IgG4 antibody levels did not correlate with ALT levels, the presence of cryoglobulinemia, or degree of hepatic fibrosis. High production of anti-core and anti-NS3 IgG1 antibodies was observed in chronic hepatitis C patients. In contrast, IgG4 antibodies seemed to only be produced against the NS3 non-structural antigen and appeared to be involved in viremia control. CONCLUSIONS: IgG1 antibodies against structural and non-structural antigens can be detected in chronic hepatitis C, while IgG4 antibodies seem to be selectively stimulated by non-structural HCV proteins, such as the NS3 antigen.

Association of serum levels of C-reactive protein with CRP-717 T/C polymorphism and viremia in HCV and HBV carriers

Abstract INTRODUCTION: The present study investigated the association of the rs2794521 polymorphism in the CRP gene in individuals with chronic hepatitis B and C, correlating it with markers of hepatic inflammation, fibrosis scores, viral load, and plasma protein levels. METHODS: The study analyzed 185 blood samples obtained from patients with hepatitis B (n=74) and hepatitis C (n=111) and 300 samples from healthy donors. Genotyping was performed by real-time polymerase chain reaction, and protein levels were quantified using the automated immunoturbidimetric method. RESULTS: The TT genotype was the most frequent in all studied groups and was associated with higher plasma levels of the protein but not with the progression of liver disease. Low levels of C-reactive protein were associated with increased viremia and scores indicative of severe fibrosis and cirrhosis. CONCLUSIONS: The present results demonstrated a close relationship between the ability of the virus to replicate and cause liver damage and low serum concentrations of C-reactive protein. Future research may determine if these results can be interpreted as a possible form of escape for the virus by decreasing its action as an opsonin and decreasing phagocytosis, which are functions of C-reactive protein in the immune response.

Influence of the inflammatory response on treatment of hepatitis C with triple therapy

Abstract INTRODUCTION: Chronic hepatitis C is a leading cause of liver disease. Infection triggers an immediate immune response in the host that is mediated by humoral/cellular mechanisms. T cells respond to infection via secretion of cytokines, which inhibit or stimulate one another, leading to cytokine imbalance and ultimately affecting treatment. Studies using interferon (IFN) and ribavirin (RBV) showed that TCD8+ cells and cytokine levels are associated with sustainable virological response (SVR). However, studies that investigated the effects of triple therapy (TT) are limited. METHODS: The study included hepatitis C virus (HCV)+ RNA, naives, genotype 1, ≥18 years, and advanced fibrosis (F≥3) patients. Samples were collected at baseline and after 12 weeks (W12) of TT. Six cytokines were analyzed by flow cytometry. RESULTS: Of 31 patients, four were excluded (two deaths, one interrupted TT, and one F2 patient). Of the 27 remaining patients, 21 (78%) were cirrhotic. SVR was achieved in 63% of the patients. The patients had a mean age of 55.11 ± 10.03 years. Analyses at baseline showed that the chemokine CCL5/Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES) (p=0.04) and interleukin (IL)-6 (p=0.02), which was associated with SVR. RANTES (p=0.04) and IL-8 (p=0.01) levels were associated with SVR at W12. CONCLUSIONS Similar to patterns observed during double therapy, IL-6, IL-8, and RANTES levels were associated with SVR in TT, indicating the potential role of interferon in immune response to hepatitis C virus.

El polimorfismo rs12979860 C> T en el gen Interferón lambda 4 no está asociado a riesgo de fibrosis hepática en pacientes chilenos con hepatitis crónica por virus C / Interferon Lambda 4 RS12979860 C> T polymorphism is not associated with liver fibrosis in patients with hepatitis C

Background. Host genetic predispositions may be important determinants of liver fibrosis in patients with chronic hepatitis C (CHC). The association between Interferon-L 4 (IFNL4) rs12979860 C>T polymorphism and risk of liver fibrosis in CHC is contradictory. Aim: To evaluate the impact of IFNL4 rs12979860 polymorphism on the risk of fibrosis in patients with CHC. Material and Methods: One hundred fifty patients with CHC aged 50 ± 11 years (89 females) were genotyped for IFNL4 rs12979860 using real time PCR. Fibrosis present in liver biopsies was assessed using the METAVIR score, comparing patients with either no fibrosis, mild fibrosis, or intermediate fibrosis (F0+F1+F2, n = 96), with patients with severe fibrosis or cirrhosis (F3+F4, n = 54). Results: In F0-F2 patients the distribution of rs12979860 genotypes was 22 CC, 57 CT and 17 TT, whereas in patients F3-F4 the distribution was 10, 29 and 15, respectively. No association between IFNL4 rs12979860 genotype and risk of fibrosis was observed in uni or multivariate analyses. Conclusions: IFNL4 rs12979860 C>T polymorphism is not associated with risk of liver fibrosis in this group of patients with CHC.

Is serum angiotensin converting enzyme level a useful non-invasive marker for liver fibrosis in patients with chronic hepatitis C?

Summary Objective: Chronic hepatitis C (CHC) continues to be a critical problem. The liver fibrosis score is the most valuable tool in determining treatment and prognosis. Liver biopsy is still considered a gold method but, due to unmet needs, new non-invasive markers are required. The aim of this study was to investigate any possible relationship between serum angiotensin-converting enzyme (ACE) levels and the stages of liver fibrosis in patients with CHC. Method: A total 100 CHC and 100 healthy subjects were enrolled in this study. The relationship between serum ACE level and the stages liver fibrosis was investigated using three different formats, as follows: (group [G]-I, classic Ishak's Score from F1 to F6; G-II, mild [F1-2], moderate [F3-4] and severe [F5-6]; G-III, mild [≤ F2] and advanced [F > 2]). The clinical usability of serum ACE level for both groups was also investigated. Results: Median serum ACE levels were higher in the healthy group than in CHC (42.5 [7-119] vs. 36 [7-91] U/I, p=0.002). There was no statistical difference among the three different fibrosis groups (G-I, G-II, G-III, p=0.797, p=0.986, and p=0.874) and no correlation between serum ACE level and the stages of liver fibrosis (r=0.026, p=0.923). The usability of serum ACE for evaluated patients with CHC and healthy subjects were calculated as 47% and 64%, respectively. Conclusion: Our study indicated that there is no relationship or correlation between serum ACE levels and stages of liver fibrosis in patients with CHC. The assessment of serum ACE level using genetically corrected reference values may provide more accurate results.

High prevalence of insulin resistance among Brazilian chronic hepatitis C patients

ABSTRACT Objective: This study aims to estimate the prevalence of insulin resistance (IR) among chronic hepatitis C (CHC) patients and their related laboratory and demographic data. Subjects and methods: In this study, non-diabetic CHC patients referred to Viral Hepatitis Ambulatories from Rio de Janeiro (Brazil) donated blood samples. Insulin was measured using a chemiluminescence immunoassay. IR was determined by HOMA-IR, where HOMA-IR > 2 was defined as IR. Results: A total of 214 CHC patients were recruited (123 females aged 53.6 years ± 10.9 years). IR was present in 133 patients (62.1%) and was associated in bivariate analysis to higher mean values of age (p = 0.040), triglycerides (p = 0.032), glucose (p = 0.000), insulin (p = 0.000), waist circumference (p = 0.001), and body mass index (p = 0.007); however, none of these variables were significant in the multivariate analysis. Conclusions: The high prevalence of IR was observed among CHC patients, and there was no difference in clinical or laboratory parameters when both groups were compared in the multivariate analysis. This high IR prevalence could lead to a high risk for development of cardiovascular disease and metabolic disorders.

Accuracy of transient elastography-FibroScan®, acoustic radiation force impulse (ARFI) imaging, the enhanced liver fibrosis (ELF) test, APRI, and the FIB-4 index compared with liver biopsy in patients with chronic hepatitis C

OBJECTIVES: Although liver biopsy is the gold standard for determining the degree of liver fibrosis, issues regarding its invasiveness and the small amount of liver tissue evaluated can limit its applicability and interpretation in clinical practice. Non-invasive evaluation methods for liver fibrosis can address some of these limitations. The aim of this study was to evaluate the accuracy of transient elastography-FibroScan®, acoustic radiation force impulse (ARFI), enhanced liver fibrosis (ELF), the aspartate aminotransferase-to-platelet ratio index (APRI), and the FIB-4 index compared with liver biopsy in hepatitis C. METHODS: We evaluated chronic hepatitis C patients who were followed at the Division of Clinical Gastroenterology and Hepatology, Hospital das Clínicas, Department of Gastroenterology of University of São Paulo School of Medicine, São Paulo, Brazil, and who underwent liver biopsy. The accuracy of each method was determined by a receiver operating characteristic (ROC) curve analysis, and fibrosis was classified as significant fibrosis (≥F2), advanced fibrosis (≥F3), or cirrhosis (F4). The Obuchowski method was also used to determine the diagnostic accuracy of each method at the various stages of fibrosis. In total, 107 FibroScan®, 51 ARFI, 68 ELF, 106 APRI, and 106 FIB-4 analyses were performed. RESULTS: A total of 107 patients were included in the study. The areas under the ROC curve (AUROCs) according to fibrosis degree were as follows: significant fibrosis (≥F2): FibroScan®: 0.83, FIB-4: 0.76, ELF: 0.70, APRI: 0.69, and ARFI: 0.67; advanced fibrosis (≥F3): FibroScan®: 0.85, ELF: 0.82, FIB-4: 0.77, ARFI: 0.74, and APRI: 0.71; and cirrhosis (F4): APRI: 1, FIB-4: 1, FibroScan®: 0.99, ARFI: 0.96, and ELF: 0.94. The accuracies of transient elastography, ARFI, ELF, APRI and FIB-4 determined by the Obuchowski method were F0-F1: 0.81, 0.78, 0.44, 0.72 and 0.67, respectively; F1-F2: 0.73, 0.53, 0.62, 0.60, and 0.68, respectively; F2-F3: 0.70, 0.64, 0.77, 0.60, and 0.67, respectively; and F3-F4: 0.98, 0.96, 0.82, 1, and 1, respectively. CONCLUSION: Transient elastography remained the most effective method for evaluating all degrees of fibrosis. The accuracy of all methodologies was best at F4.

Is there an association between vitamin D and liver fibrosis in patients with chronic hepatitis C? / Existe associação entre vitamina D e fibrose hepática em pacientes com hepatite C crônica?

ABSTRACT BACKGROUND Vitamin D is known for its immunomodulatory, anti-inflammatory and antifibrotic properties, which are quite relevant in the pathogenesis and treatment of many causes of chronic liver disease. OBJECTIVE This study aimed to evaluate the association between serum vitamin D levels and the histopathological findings in patients with chronic hepatitis C virus infection. METHODS Cross-sectional study composed of patients with chronic hepatitis C. All patients underwent vitamin D 25 dosage and anthropometric data analysis. Liver biopsy was performed in a maximum 36-month period before inclusion in the study. RESULTS Of the 74 patients included in the study, 45 (60.8%) were women, mean age was 57.03±9.24 years, and 63 (85.1%) were white. No association was observed between the serum levels of vitamin D and inflammatory activity (P=0.699) nor with the degree of liver fibrosis (P=0.269). CONCLUSION In this study, no association was observed between vitamin D and inflammatory activity, as well as the degree of liver fibrosis, in patients with chronic hepatitis C.

RESUMO CONTEXTO A vitamina D é conhecida por possuir propriedades imunomoduladoras, anti-inflamatórias e antifibróticas, relevantes na patogênese e tratamento de muitas causas de doença hepática crônica. OBJETIVO Este estudo tem como objetivo avaliar a associação entre os níveis séricos de vitamina D e os achados histopatológicos em pacientes com infecção crônica do vírus da hepatite C. MÉTODOS Estudo transversal, composto por pacientes com hepatite C crônica. Todos os pacientes foram submetidos à dosagem de vitamina D 25 e análise de dados antropométricos. A biópsia hepática foi realizada em um período máximo de 36 meses antes da inclusão no estudo. RESULTADOS Dos 74 pacientes incluídos no estudo, 45 (60,8%) eram mulheres, média de idade de 57,03±9,24 anos e 63 (85,1%) eram brancos. Não foi observada associação entre os níveis séricos de vitamina D e atividade inflamatória (P=0,699), nem com o grau de fibrose hepática (P=0,269). CONCLUSÃO No presente estudo, não foi observada associação entre a vitamina D e a atividade inflamatória, bem como com o grau de fibrose hepática, em pacientes com hepatite C crônica.

Platelet-derived growth factor A mRNA in platelets is associated with the degree of hepatic fibrosis in chronic hepatitis C

Abstract: INTRODUCTION: Transforming growth factor beta 1 (TGFB1) and platelet-derived growth factor (PDGF) are the main cytokines related to hepatic fibrogenesis. METHODS: RNA isolated from the platelets and hepatic tissue of 43 HCV carriers was used for quantitative polymerase chain reaction to determine TGFB1, PDGFA, and PDGFB RNA expression. RESULTS: The mRNA expression of PDGFA in platelets was significantly lower in the group with advanced fibrosis than in the group with early-stage fibrosis. TGFB1 was more frequently expressed in platelets than in hepatic tissue, which was different from PDGFB. CONCLUSIONS: A pathway mediated by overexpression of TGFB1 via PDGFA in megakaryocytes could be involved in the development of fibrosis.

GAMMA GLUTAMYLTRANSFERASE IMPACT IN THERAPEUTIC RESPONSE OF CHRONIC HEPATITIS C: a systematic review of the literature / Impacto da gama glutamiltransferase na resposta terapêutica da hepatite C crônica: revisão sistemática da literatura

BackgroundThe standard treatment of chronic hepatitis C is the administration of pegylated interferon α2a or α2b in combination with ribavirin, but adverse effects can be observed, as well as the high cost of this therapy. Therefore, there is interest in understanding the predictors of sustained virologic response, as the gamma glutamyltransferase.ObjectiveTo evaluate the serum levels of gamma glutamyltransferase as a predictor of response to treatment with pegylated interferon α and ribavirin in chronic hepatitis C.MethodsThis is a systematic review of literature, conducted by consulting PUBMED, LILACS, MEDLINE, SCOPUS, Cochrane electronic databases, and active search of articles selected between January 2000 and April 2013.ResultsA total of 4,785 titles were iden tified. Out of those material, following inclusion and exclusion criteria, 273 abstracts were selected, by two independent researchers. After reading those texts, the reviewers consensually included ten studies for systematization and classification, according to the criteria of the Oxford Scale. 1B studies are predominant (prospective cohort study - six studies). Rapid virologic response and early virological response were considered as estimates for the sustained virological response. The frequency of virologic response was identified in three studies and early virological response in two, with a total of 392 and 413 patients, respectively; sustained virologic response was reported in nine articles corresponding to 3,787 patients (76.5 %).ConclusionGamma glutamyltransferase is a predictor of sustained virologic response in the treatment of chronic hepatitis C with pegylated interferon α2a or α2b associated with ribavirin.

ContextoO tratamento padrão da hepatite C crônica consiste na administração de interferon peguilado α2a ou α2b associado à ribavirina. Contudo, podem ser observados efeitos adversos além do alto custo desta terapêutica. Por isso há interesse no conhecimento dos fatores preditivos de resposta virológica sustentada como a gama glutamiltransferase.ObjetivoAvaliar os níveis séricos da gama glutamiltransferase como fator preditivo de resposta terapêutica com interferon peguilado α e ribavirina na hepatite C crônica.MétodosTrata-se de uma revisão sistemática da literatura, conduzida através de consulta às bases eletrônicas de dados PUBMED, LILACS, MEDLINE, SCOPUS e COCHRANE, e busca ativa das referências dos artigos selecionados, no período de janeiro de 2000 a abril de 2013.ResultadosForam identificados 4.785 títulos. Destes, seguindo os critérios de inclusão e exclusão, foram selecionados 273 resumos para a leitura por dois pesquisadores independentes. Após a leitura dos artigos, na íntegra e em consenso, os revisores incluíram dez estudos, para sistematização e qualificação, segundo os critérios da Escala de Oxford. Predominaram os estudos 1B A (Coorte prospectivo - seis estudos), publicações da Alemanha. A frequência de resposta virológica rápida foi identificada em três estudos e resposta virológica precoce em dois estudos, com um total de 392 e 413 pacientes respectivamente; a resposta virológica sustentada foi registrada em nove artigos correspondendo a 3.787 pacientes (76,5%).ConclusãoO nível sérico da gama glutamiltransferase é um fator preditivo de resposta virológica sustentada no tratamento da hepatite C crônica com interferon peguilado α2a ou α2b associado à ribavirina.

Serum levels of immunoglobulin free light chains in patients with chronic hepatitis C presenting cryoglobulinemia

Hepatitis C virus (HCV) infects B-lymphocytes, provokes cellular dysfunction and causes lymphoproliferative diseases such as cryoglobulinemia and non-Hodgkin's B-cell lymphoma. In the present study, we investigated the serum levels of kappa and lambda free light chains (FLC) of immunoglobulins and the kappa/lambda FLC ratio in Brazilian patients with chronic HCV infection and cryoglobulinemia. We also analyzed the immunochemical composition of the cryoglobulins in these patients. Twenty-eight cryoglobulinemic HCV patients composed the target group, while 37 HCV patients without cryoglobulinemia were included as controls. The median levels of kappa and lambda FLC were higher in patients with cryoglobulinemia compared to controls (p = 0.001 and p = 0.003, respectively), but the kappa/lambda FLC ratio was similar in patients with and without cryoglobulinemia (p > 0.05). The median FLC ratio was higher in HCV patients presenting with advanced fibrosis of the liver compared to HCV patients without fibrosis (p = 0.004). Kappa and lambda FLC levels were strongly correlated with the IgA, IgG and IgM levels in the patients with cryoglobulinemia. In patients without cryoglobulinemia, the kappa FLC level was only correlated with the IgG level, whereas the lambda FLC were weakly correlated with the IgA, IgG and IgM levels. An immunochemical pattern of mixed cryoglobulins (MC), predominantly IgM, IgG, IgA and kappa light chain, was verified in these immune complexes. We concluded that HCV-infected patients presenting cryoglobulinemia have vigorous polyclonal B-lymphocyte activation due to chronic HCV infection and persistent immune stimulation.

Epidemiological, serological and molecular aspects of hepatitis B and C in children and teenagers of municipal daycare facilities schools and schools in the city of Santos / Aspectos epidemiológicos, sorológicos e moleculares das hepatites B e C em crianças e adolescentes de creches e escolas municipais na cidade de Santos

Objective: To estimate the prevalence of the serological markers anti-HBc, HBsAg and anti-HBs of hepatitis B and anti-HCV of hepatitis C among children and teenagers enrolled at daycare facilities, kindergartens and municipal elementary education network in the city of Santos, São Paulo, Brazil. Methods: A cross-sectional study was carried out from June 28 to December 14, 2007, in which 4,680 finger-prick blood samples were collected from children and teenagers. A survey questionnaire was applied to their family members. The sample was dimensioned using the software Epi Info version 6 with expected frequency of 1%, acceptable error of 0.5% and confidence interval of 95%. The serological tests were performed using the ELISA technique. The molecular analysis was performed using the technique of polymerase chain reaction in House. Results: Age of the studied population ranged from 7 months to 18 years and 1 month. The general prevalence of anti-HBc reagent was 0.1%, HBsAg was 0.02% and anti-HCV was 0.02%. Conclusions: In children, the general prevalence of serological markers for hepatitis B and C in the city of Santos was low when compared with literature data. .

Objetivo: Estimar a prevalência de marcadores sorológicos anti-HBc, AgHBs e anti-HBs da hepatites B e anti-HCV da hepatite C em crianças e adolescentes matriculados em creches e escolas de ensino infantil e fundamental da rede municipal na cidade de Santos, São Paulo. Métodos: Estudo transversal realizado no período de 28 de junho a 14 de dezembro de 2007, no qual foram coletadas 4.680 amostras de sangue colhidas através de punção capilar. Foi aplicado um questionário nos familiares das crianças e adolescentes. Para o cálculo da amostra, foi utilizado o programa Epi Info versão 6 com frequência esperada de 1%, erro aceitável de 0,5% e nível de confiança de 95%. Os exames sorológicos foram realizados utilizando a técnica de ELISA. O estudo molecular foi realizado pela técnica de reação em cadeia de polimerase in House. Resultados: A idade da população estudada variou de 7 meses a 18 anos e 1 mês. A prevalência geral do anti-HBc reagente foi de 0,1%, do AgHBs foi de 0,02% e do anti-HCV foi de 0,02%. Conclusão: A prevalência geral em crianças dos marcadores sorológicos para hepatites B e C na cidade de Santos foi baixa, quando comparada com os dados de literatura. .

Perfil imunofenotípico e nível de citocinas plásmáticas em portadores de hepatite C crônica (genótipo 1) com diferentes graus de comprometimento hepático / Immunophenotypic profile and level of plasmatic cytokines in patients with chronic hepatitis C with varying degrees of hepatic impairment

O vírus da hepatite C crônica (VHC) é causa de doença hepática crônica afetando cerca de 3% da população mundial, aproximadamente 170 milhões de pessoas. Investigações sobre a resposta imune em indivíduos infectados com HCV é justificável pelo fato desta constituir doença infecciosa cujas alterações imunológicas têm relação direta com o desenvolvimento e a manutenção da infecção. Este projeto visou avaliar aspectos que compõem a resposta imune de pacientes com hepatite C crônica com diferentes graus de lesão hepática. Foram incluídos no estudo pacientes VHC+ (n=83), genótipo 1, pré-tratamento, os quais foram estratificados conforme o grau de fibrose hepática determinada por biópsia (classificação METAVIR), compondo: G1 (n = 15) pacientes nos estágios F0 (nenhuma fibrose) + F1 (fibrose em expansão dos espaços porta); G2 (n = 21) pacientes no estágio F2 (poucas fontes de fibrose nos espaços porta); G3 (n = 15) pacientes em estágio F3 (início da formação de nódulos); G4 (n = 32) pacientes em estágio F4 (fibrose severa); G5 (n = 16) doadores de sangue voluntários (grupo controle). Foram avaliadas características imunofenotípicas de subpopulações de leucócitos do sangue periférico (células NK totais, NKdim, NKbright, células dendríticas mielóides e plasmocitóides, monócitos clássicos e pro-inflamatórios, linfócitos T CD4+/CD8+), níveis de citocinas e quimiocinas plasmáticas (TGF-β, TNF-α, CCL2, CCL3, CCL4, CCL5, CXCL8, CXCL9, CXCL10), analisando a relação destes com a progressão da fibrose. Todas as frequências das subpopulações leucocitárias avaliadas estão alteradas em portadores de hepatite C crônica comparados com indivíduos saudáveis. Além disso, há correlação entre o grau de fibrose e a quantidade de linfócitos T CD4, T CD8, monócitos pro-inflamatórios e células NKbright circulantes...

The chronic hepatitis C virus (HCV) is the cause of chronic liver disease affecting about 3% of world population, approximately 170 million people. Investigations of the immune response in individuals infected with HCV is justifiable because this form infectious disease whose immunological changes are directly related to the development and maintenance of infection. This project aimed to evaluate aspects that make up the immune chronic hepatitis C patients with different degrees of liver damage response. The study included patients HCV+ (n = 83), genotype 1, pretreatment, which were classified according to the rate of hepatic fibrosis determined by biopsy (METAVIR classification), composed of: G1 (n = 15) patients in stages F0 (no fibrosis) + F1 (fibrosis expansion of port spaces), G2 (n = 21) patients in the F2 stage (few sources of fibrosis in portal ), G3 ( n = 15 ) patients with stage F3 (early nodule formation), G4 (n = 32) patients with stage F4 (severe fibrosis), G5 (n = 16) volunteer blood donors (control group). Immunophenotypic characteristics of subpopulations of peripheral blood leukocytes (total NK cells, NKdim , NKbright , myeloid dendritic cells and plasmacytoid , classical monocytes, pro- inflammatory CD4 + / CD8 + cells), cytokine levels and plasma chemokines (were evaluated TGF- β , TNF - α , CCL2 , CCL3 , CCL4 , CCL5 , CXCL8 , CXCL9, CXCL10), analyzing their relationship with the progression of fibrosis. All frequencies of evaluated leukocyte subpopulations are altered in patients with chronic hepatitis C compared with healthy subjects. In addition, no correlation between the degree of fibrosis and the amount of CD4, CD8, pro-inflammatory monocytes and circulating NKbright cells. Increased levels of TNF-α were observed in cirrhotic patients (G4) and TGF - β in HCV + groups with moderate hepatic fibrosis (G2 and G3)...

Perfil imunofenotípico e nível de citocinas plásmáticas em portadores de hepatite C crônica (genótipo 1) com diferentes graus de comprometimento hepático / Immunophenotypic profile and level of plasmatic cytokines in patients with chronic hepatitis C with varying degrees of hepatic impairment

O vírus da hepatite C crônica (VHC) é causa de doença hepática crônica afetando cerca de 3% da população mundial, aproximadamente 170 milhões de pessoas. Investigações sobre a resposta imune em indivíduos infectados com HCV é justificável pelo fato desta constituir doença infecciosa cujas alterações imunológicas têm relação direta com o desenvolvimento e a manutenção da infecção. Este projeto visou avaliar aspectos que compõem a resposta imune de pacientes com hepatite C crônica com diferentes graus de lesão hepática. Foram incluídos no estudo pacientes VHC+ (n=83), genótipo 1, pré-tratamento, os quais foram estratificados conforme o grau de fibrose hepática determinada por biópsia (classificação METAVIR), compondo: G1 (n = 15) pacientes nos estágios F0 (nenhuma fibrose) + F1 (fibrose em expansão dos espaços porta); G2 (n = 21) pacientes no estágio F2 (poucas fontes de fibrose nos espaços porta); G3 (n = 15) pacientes em estágio F3 (início da formação de nódulos); G4 (n = 32) pacientes em estágio F4 (fibrose severa); G5 (n = 16) doadores de sangue voluntários (grupo controle). Foram avaliadas características imunofenotípicas de subpopulações de leucócitos do sangue periférico (células NK totais, NKdim, NKbright, células dendríticas mielóides e plasmocitóides, monócitos clássicos e pro-inflamatórios, linfócitos T CD4+/CD8+), níveis de citocinas e quimiocinas plasmáticas (TGF-β, TNF-α, CCL2, CCL3, CCL4, CCL5, CXCL8, CXCL9, CXCL10), analisando a relação destes com a progressão da fibrose. Todas as frequências das subpopulações leucocitárias avaliadas estão alteradas em portadores de hepatite C crônica comparados com indivíduos saudáveis. Além disso, há correlação entre o grau de fibrose e a quantidade de linfócitos T CD4, T CD8, monócitos pro-inflamatórios e células NKbright circulantes...

The chronic hepatitis C virus (HCV) is the cause of chronic liver disease affecting about 3% of world population, approximately 170 million people. Investigations of the immune response in individuals infected with HCV is justifiable because this form infectious disease whose immunological changes are directly related to the development and maintenance of infection. This project aimed to evaluate aspects that make up the immune chronic hepatitis C patients with different degrees of liver damage response. The study included patients HCV+ (n = 83), genotype 1, pretreatment, which were classified according to the rate of hepatic fibrosis determined by biopsy (METAVIR classification), composed of: G1 (n = 15) patients in stages F0 (no fibrosis) + F1 (fibrosis expansion of port spaces), G2 (n = 21) patients in the F2 stage (few sources of fibrosis in portal ), G3 ( n = 15 ) patients with stage F3 (early nodule formation), G4 (n = 32) patients with stage F4 (severe fibrosis), G5 (n = 16) volunteer blood donors (control group). Immunophenotypic characteristics of subpopulations of peripheral blood leukocytes (total NK cells, NKdim , NKbright , myeloid dendritic cells and plasmacytoid , classical monocytes, pro- inflammatory CD4 + / CD8 + cells), cytokine levels and plasma chemokines (were evaluated TGF- β , TNF - α , CCL2 , CCL3 , CCL4 , CCL5 , CXCL8 , CXCL9, CXCL10), analyzing their relationship with the progression of fibrosis. All frequencies of evaluated leukocyte subpopulations are altered in patients with chronic hepatitis C compared with healthy subjects. In addition, no correlation between the degree of fibrosis and the amount of CD4, CD8, pro-inflammatory monocytes and circulating NKbright cells. Increased levels of TNF-α were observed in cirrhotic patients (G4) and TGF - β in HCV + groups with moderate hepatic fibrosis (G2 and G3)...

Performance of diagnostic biomarkers in predicting liver fibrosis among hepatitis C virus-infected Egyptian children

The aim of the present study was to identify specific markers that mirror liver fibrosis progression as an alternative to biopsy when biopsy is contraindicated, especially in children. After liver biopsies were performed, serum samples from 30 hepatitis C virus (HCV) paediatric patients (8-14 years) were analysed and compared with samples from 30 healthy subjects. All subjects were tested for the presence of serum anti-HCV antibodies. Direct biomarkers for liver fibrosis, including transforming growth factor-β1, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), hyaluronic acid (HA), procollagen type III amino-terminal peptide (PIIINP) and osteopontin (OPN), were measured. The indirect biomarkers aspartate and alanine aminotransferases, albumin and bilirubin were also tested. The results revealed a significant increase in the serum marker levels in HCV-infected children compared with the healthy group, whereas albumin levels exhibited a significant decrease. Significantly higher levels of PIIINP, TIMP-1, OPN and HA were detected in HCV-infected children with moderate to severe fibrosis compared with children with mild fibrosis (p < 0.05). The diagnostic accuracy of these direct biomarkers, represented by sensitivity, specificity and positive predictive value, emphasises the utility of PIIINP, TIMP-1, OPN and HA as indicators of liver fibrosis among HCV-infected children.

Autoantibody profile in individuals with chronic hepatitis C

Introduction Autoantibodies are often produced during infection with chronic hepatitis C virus (HCV), but it remains controversial whether they influence the biochemical profile and histological features of this disease. Therefore, this current study sought to describe these autoantibodies and evaluate their impact on the clinical and histological presentation of hepatitis C. Methods This cross-sectional analytical study assessed patients with HCV (RNA+) from October 2011 to July 2012. Results This study included 66 patients, with a mean age of 53.2±10.5 years. Of these patients, 60.6% were male, and 54.3% presented with genotype 1. Non-organ-specific autoantibodies (NOSA) were detected in 24% of the patients; of these, 7.6% were anti-mitochondrial antibodies (AMA+), 26.7% were anti-smooth muscle antibodies (SMA+) and 6.8% were liver kidney microsomal type 1 antibodies (LKM1+). With respect to the thyroid autoantibodies, 7.4% were anti-peroxidase (ATPO+) antibodies, and none were anti-thyroglobulin (ATG+) antibodies. Regarding celiac disease autoantibodies, 5.8% were endomysial antibodies (EMA+), and no transglutaminase (TTG+) antibodies were detected. Cryoglobulins were found in 2.1% of patients. When NOSA+ individuals were compared to patients without the presence of NOSAs, they exhibited higher median alkaline phosphatase (0.7 vs. 0.6 xULN; p=0.041), lower median platelet counts (141,500.0 vs. 180,500.0/mm 3 ; p=0.036), lower mean prothrombin activity (72.6±11.5% vs. 82.2±16.0%; p=0.012) and an increased prevalence of significant fibrosis (E≥2) (45.5% vs. 18.2%; p=0.012). There was also a tendency for a greater proportion of NOSA+ cases to have marked periportal activity (APP≥3) (44.5% vs. 15.6%; p=0.087). Conclusions In addition to the high prevalence of autoantibodies associated with HCV infection, it was observed that NOSA ...

Hepatitis C virus quantification in serum and saliva of HCV-infected patients

The hepatitis C virus (HCV) can be detected in blood and other bodily fluids, such as saliva, semen and gastric juices. The aim of this study was to compare the HCV viral loads in the serum and saliva of infected patients. Twenty-nine patients with detectable HCV RNA in their serum and saliva were included in this study. The HCV viral loads were determined through quantitative real-time polymerase chain reactions. The median viral RNA levels were 5.78 log10 copies in the serum and 3.32 log10 copies in the saliva. We observed that the salivary HCV viral load was significantly lower than the viral load in the serum. Further studies are required to understand the role of saliva in the diagnosis, management and potential transmission of HCV.

Effect of HFE gene polymorphism on sustained virological response in patients with chronic hepatitis C and elevated serum ferritin / Efeito do polimorfismo do gene HFE sobre a resposta viral sustentada em portadores de hepatite crônica C com ferritina sérica elevada

CONTEXT: Abnormal serum ferritin levels are found in approximately 20%-30% of the patients with chronic hepatitis C and are associated with a lower response rate to interferon therapy. OBJECTIVE: To determine if the presence of HFE gene mutations had any effect on the sustained virological response rate to interferon based therapy in chronic hepatitis C patients with elevated serum ferritin. METHODS: A total of 44 treatment naÏve patients with histologically demonstrated chronic hepatitis C, all infected with hepatitis C virus genotype non-1 (38 genotype 3; 6 genotype 2) and serum ferritin above 500 ng/mL were treated with interferon (3 MU, 3 times a week) and ribavirin (1.000 mg, daily) for 24 weeks. RESULTS: Sustained virological response was defined as negative qualitative HCV-RNA more than 24 weeks after the end of treatment. Serum HCV-RNA was measured by qualitative in house polymerase chain reaction with a limit of detection of 200 IU/mL. HFE gene mutation was detected using restriction-enzyme digestion with RsaI (C282Y mutation analysis) and BclI (H63D mutation analysis) in 16 (37%) patients, all heterozygous (11 H63D, 2 C282Y and 3 both). Sustained virological response was achieved in 0 of 16 patients with HFE gene mutations and 11 (41%) of 27 patients without HFE gene mutations (P = 0.002; exact Fisher test). CONCLUSION: Heterozigozity for H63D and/or C282Y HFE gene mutation predicts absence of sustained virological response to combination treatment with interferon and ribavirin in patients with chronic hepatitis C, non-1 genotype and serum ferritin levels above 500 ng/mL.

CONTEXTO: Níveis séricos anormais de ferritina são encontrados em aproximadamente 20%-30% dos pacientes com hepatite crônica C e estão associadas a uma baixa taxa de resposta à terapia com interferon. OBJETIVO: Avaliar a associação entre a presença de mutações do gene HFE e a taxa de resposta virológica sustentada ao interferon em pacientes portadores de hepatite crônica C com ferritina sérica elevada. MÉTODOS: Um total de 44 pacientes, virgem de tratamento, infectado pelo vírus da hepatite C de genótipos não-1 (38 genótipo 3; 6 genótipo 2) e ferritina sérica acima de 500 ng/mL foi tratado com interferon (3 MU, três vezes por semana) e ribavirina (1000 mg/dia) por 24 semanas. Resposta virológica sustentada foi definida como HCV-RNA indetectável 24 semanas após o fim do tratamento. Foi utilizado técnica de reação em cadeia da polimerase em tempo-real com limite de detecção de 200 UI /mL. RESULTADOS: Mutações do gene HFE foram detectadas por "restriction-enzyme digestion" com RsaI (análise de mutação C282Y) e BclI (análise de mutação H63D) em 16 pacientes (37%), todos heterozigotos (11 H63D, 2 C282Y e 3 ambos). Resposta virológica sustentada foi alcançada em 0 de 16 pacientes com mutações do gene HFE e 11 (41%) dos 27 pacientes sem mutações do gene HFE (P = 0,002; teste exato de Fisher). CONCLUSÃO: A heterozigose para os genes H63D e/ou C282Y HFE está associada à redução significativa da taxa de resposta virológica sustentada ao tratamento com interferon e ribavirina em pacientes com hepatite crônica C, genótipo não-1 e com níveis séricos de ferritina acima de 500 ng/mL.

Soluble inflammatory markers as predictors of virological response in patients with chronic hepatitis C virus infection treated with interferon-α plus ribavirin

The host immune response plays an important role in viral clearance in patients who are chronically infected with hepatitis C virus (HCV) and are treated with interferon and ribavirin. Activation of the immune system involves the release of pro and anti-inflammatory molecules that can be measured in plasma samples. The present study aimed to evaluate the association between pretreatment plasma levels of chemokines and soluble tumor necrosis factor receptors (sTNF-R) and the virological response in treated patients with chronic hepatitis C infection. Forty-one chronically-infected HCV patients that were being treated with interferon-α (IFN-α) plus ribavirin were included in the study. Socio-demographic, clinical and laboratory data were collected and pretreatment plasma levels of chemokine CCL2, CCL3, CCL11, CCL24, chemokine CXCL9, CXCL10, sTNF-R1 and sTNF-R2 were measured. The virological response was assessed at treatment week 12, at the end of treatment and 24 weeks after treatment. Pretreatment CXCL10 levels were significantly higher in patients without an early virological response (EVR) or sustained virological response (SVR) compared to responders [512.9 pg/mL vs. 179.1 pg/mL (p = 0.011) and 289.9 pg/mL vs. 142.7 pg/mL (p = 0.045), respectively]. The accuracy of CXCL10 as a predictor of the absence of EVR and SVR was 0.79 [confidence interval (CI) 95 percent: 0.59-0.99] and 0.69 (CI 95 percent: 0.51-0.87), respectively. Pretreatment plasma levels of the other soluble inflammatory markers evaluated were not associated with a treatment response. Pretreatment CXCL10 levels were predictive of both EVR and SVR to IFN-α and ribavirin and may be useful in the evaluation of candidates for therapy.