Substance P enhances cytokine-induced vascular cell adhesion molecule-1 (VCAM-1) expression on cultured rheumatoid fibroblast-like synoviocytes

There is evidence that TNF-α contributes to the pathogenesis of chronic viral hepatitis. The cellular effects of this cytokine are regulated by two specific receptors, and membranous shedding of these receptors reflects activation of the TNF system. We performed a study of TNF-α and functionally active soluble TNF-receptors (TNFR-p55 and -p75) in 105 patients with chronic HCV infection. In HCV RNA-positive patients a significant enhancement of TNF-α and both receptor types was observed compared with controls (TNF-α 83.8 ± 91.7 pg/ml versus 18.8 ± 8.4 pg/ml, P < 0.001; TNFR-p55 1.4 ± 0.4 ng/ml versus 0.9 ± 0.2 ng/ml, P < 0.0001; TNFR-p75 6.4 ± 2.4 ng/ml versus 2.9 ± 0.6 ng/ml, P < 0.0001, respectively). The enhanced serum levels of TNF-α and TNFRs were reflected by a significant expression of TNFR-specific mRNA in peripheral mononuclear cells of HCV-infected patients (P < 0.001). Serum aminotransferases correlated with soluble TNFR-p75 (P < 0.001) but not with TNFR-p55 and TNF-α. We demonstrated an association of the degree of histological inflammation with both TNFRs (P < 0.01). Furthermore, enhanced hepatocellular expression of TNF-α and TNFRs could be demonstrated by immunohistochemical staining in HCV-infected patients. Sixty-eight out of 105 patients were treated with interferon-alpha (IFN-α) (3 × 10⁶ U × 3/week). Pretreatment levels of TNF-α and TNFRs did not differ between responders and non-responders. Our results demonstrate that TNF-α and TNFRs are enhanced in chronic HCV infection and reflect histological activity of the disease. This up-regulation of TNFRs might modify host response and potentially contribute to liver damage in chronic HCV infection.     

Assessment of early virological response to antiviral therapy by comparing four assays for HCV RNA quantitation using the international unit standard: implications for clinical management of patients with chronic hepatitis C virus infection

WHO International Standards for nucleic acid tests are used widely to compare the different assays used in HCV RNA quantitation. The aim of the study was to assess the impact of the international unit standard for measuring HCV RNA in the management of patients with chronic hepatitis C virus (HCV) infection. Twenty‐seven naïve patients infected chronically by HCV were treated with ribavirin plus PEG‐interferon‐alfa‐2b for 48 weeks. SVR was obtained for 16 patients (the other were non‐responders). For HCV RNA quantitation, four assays were undertaken: Versant HCV RNA 3.0 (Bayer), Real time PCR (TaqMan, Roche), LCx HCV RNA (Abbott), and Cobas Amplicor‐Monitor v2 (Roche). Considering a 2‐log decline at Week 12 after the beginning of therapy, discordant results were found with the four HCV RNA methods in predicting SVR or non‐response. At Week 4 and Week 12, significant differences were observed between Versant HCV RNA 3.0 versus PCR HCV Taqman, Versant HCV RNA 3.0 versus LCx HCV RNA, Cobas Monitor Amplicor HCV 2.0 versus LCx HCV RNA, and Cobas Monitor Amplicor HCV 2.0 versus PCR HCV Taqman (P < 0.001). The HCV RNA cutoff, given a 100% negative predictive value at Week 4 and Week 12, differed with the assays used to quantify HCV RNA, despite the use of the IU/ml units. Eighty‐nine percent of serum values for HCV RNA were concordant by the IU standard. All assays, however, failed to detect HCV RNA in some cases. Despite the use of the IU standard HCV‐infected patients might be monitored with only one assay. J. Med. Virol. 78:208–215, 2006. © 2005 Wiley‐Liss, Inc.     

Hepatitis C (HCV) genotype and viral titer distribution among Argentinean hemophilic patients in the presence or absence of human immunodeficiency virus (HIV) co-infection

Hepatits C (HCV) infection is frequent among hemophilic patients treated with non-inactivated factor-concentrates. Both HCV genotype and viral load have been suggested to be important prognostic markers of disease progression and treatment outcome. In addition, co-infection with the human immunodeficiency virus (HIV) has been associated with increased level of HCV replication and higher risk of developing liver failure. Thus, HCV genotype, viral load, and HIV co-infection are important factors in HCV infection. Using restriction fragment length polymorphism analysis (RFLP) and the branched-DNA (bDNA) assay, we retrospectively investigated the HCV genotypes and viral loads present in 59 Argentinean hemophiliacs, in the presence or absence of HIV infection. HCV genotype 1 was the predominant viral variant detected among HIV-negative (HIV⁻) (76%) and HIV-positive (HIV⁺) (82.5%) patients, followed by genotypes 3 (10.4%), 2(2%) and a small proportion of multiply co-infected patients including genotypes 4 and 5 (6.25%). HIV⁺ patients had higher plasma HCV RNA levels than HIV⁻ patients (88.4 ± 16.5 × 10⁵ Eq/ml vs. 24.7 ± 5.8 × 10⁵ Eq/ml) (P < 0.001); however, no correlation between HCV replication and level of immune suppression, evaluated by CD4⁺ T-cell measurement, was observed among HIV⁺ patients (r = 0.017). Abnormal and higher ALT levels were more frequently detected among HIV⁺ (93%; 123.6 ± 15.7 U/liter) than HIV⁻ (41%; 70.2 ± 24.2 U/liter) patients (P < 0.001; P < 0.05). Although we were able to confirm previous reports suggesting the existence of increased HCV replication in HIV/HCV co-infected hemophiliacs, our data did not support the conclusion that HIV-induced immune suppression is directly responsible for this phenomena. It is possible that other factors induced by HIV are responsible for the increased levels in HCV replication observed. J. Med. Virol. 52:219–225, 1997. © 1997 Wiley-Liss, Inc.     

Immunodetection of a Hepatitis C Virus (HCV) Antigen and Th1/Th2 Cytokines in Cerebrospinal Fluid of Meningitis Patients

Abstract

Infection with hepatitis C virus (HCV) has become the most important public health problem in Egypt. HCV infection has been implicated in diseases of the central nervous system. Cerebrospinal fluid (CSF) and serum samples from 91 patients with meningitis (62 males and 29 females, mean age of 37 years) were investigated. Anti‐HCV antibodies and HCV antigen were evaluated in patients CSF and serum using enzyme linked immunosorbent assay. The levels (mean?±?SD?pg/ml) of Th1 cytokines (IFN‐γ and TNF‐α) and Th2 interleukines (IL‐10 and IL‐4) were also determined. The anti‐HCV antibodies were detected in high percentages both in CSF samples (71%) and in sera (90%). Also, the HCV antigen was detected in about 60% of tested CSF and serum samples. The levels of IFN‐γ and IL‐10 cytokines were significantly higher (P?<?0.05) in both serum and CSF of patients positive for HCV antigen than those negative. HCV antigen was detected in the CSF of meningitis patients with a significant upregulation of Th1 and Th2 responses. The high incidence of HCV infection may draw light on the etiological role of HCV in the pathogensis of meningitis diseases in our study group.     

Hepatitis C virus screening and clinical monitoring of biomarkers in patients undergoing hemodialysis

In this study, 395 volunteers were enrolled to investigate the seroprevalence of hepatitis C virus, the immunological and the alanine aminotransferase (ALT) biomarkers amongst hemodialysis patients, living in Manaus, Brazil. An overall seroprevalence of 13.9% was found in the hemodialysis patients. Analysis of seroconversion patterns demonstrated that most patients with HCV seroconverted up to 10 years following the first hemodialysis session. Anti‐NS5 antibody was detectable in 60.4% of patients with HCV. A lower percentage of circulating CD3⁺ and CD4⁺ T‐cells was found in patients seronegative for HCV, whereas a higher frequency of CD8⁺ T‐cells was the hallmark of patients with HCV. An overall low activation state of monocytes and eosinophils were observed in hemodialysis patients. In contrast, a higher frequency of activated neutrophils was observed in patients with HCV, selectively in the NS5+ subgroup. All hemodialysis patients had a higher percentage of activated lymphocytes, with the higher activation state in patients with NS5? reactivity. Higher ALT levels were observed in patients with HCV, especially in the NS5+ subgroup. Interestingly, the ALT levels were correlated negatively with the lymphocyte activation state, selectively in the NS5? subgroup, suggesting a protective role of these activated lymphocytes in patients with HCV. These findings reinforce the importance of the transmission of HCV among hemodialysis patients, suggesting that apart from the HCV screening, the serological and ALT biomarkers may represent important predictors of morbidity and/or mortality among patients undergoing hemodialysis. J. Med. Virol. 81:1220–1231, 2009. © 2009 Wiley‐Liss, Inc.     

Efficacy and safety of peginterferon alpha‐2a/ribavirin in treatment‐naive Cameroonian patients with chronic hepatitis C

Data were examined from a day‐to‐day clinical practice in Yaounde, Cameroon to evaluate the efficacy and safety of peginterferon alfa‐2a and ribavirin in treatment‐naive Cameroonian patients with chronic hepatitis C. Ninety adults with chronic hepatitis C (mean age, 53 ± 8 years; 79% males; 37.8% genotype 1; 23.3% genotype 2; and 38.9% genotype 4) were given at least 12 weeks of combination therapy between February 2003 and August 2007. Of these, 54 completed the treatment and the 24‐week follow up. Subsequently, 18 continued treatment and 18 (20%) discontinued the treatment, 6 (6.7%) due to adverse effects. An intention‐to‐treat analysis showed that 38 (52.8%) had an end‐of‐treatment virologic response and 34 (47.2%) had a sustained virologic response. Sustained virologic response were significantly higher among patients with hepatitis C virus (HCV) genotype 2 (83.4%) than in those with genotype 1 (31%) or genotype 4 (42.3%) (P < 0.05). Non HCV‐2 genotype, pretreatment fibrosis score >2, HCV RNA level >8.0 × 10⁵ IU/ml and a non‐virologic response at 12 weeks of treatment were associated with poor sustained virologic response (P < 0.05). Thus, HCV can be treated in a Sub‐Saharan African country. It indicates that Cameroonian HCV‐1 and ‐4 patients have a poorer sustained virologic response than the published results for Western and Middle‐East countries. Virus subtype may influence the treatment outcome, since there is a great genetic diversity within Cameroonian HCV‐1 and ‐4 genotypes. J. Med. Virol. 80:2079–2085, 2008. © 2008 Wiley‐Liss, Inc.     

Serum hepatitis C virus RNA level as a predictor of subsequent response to interferon-α therapy in Japanese patients with chronic hepatitis C

Serum hepatitis C virus (HCV) RNA level has been shown to be a good predictor of subsequent response to interferon-α (IFN) therapy in US patients in whom genotype 1a/1b are both predominant. To determine whether serum HCV RNA level is a predictor of subsequent response to IFN in Japanese patients or not, appropriately collected pre-IFN therapy serum samples from 35 Japanese patients with chronic HCV infection were studied. Serum HCV RNA level and HCV genotype were determined and correlated with the subsequent response to IFN. Response to IFN was defined by serum alanine transaminase level: complete and sustained response (n = 15), complete response followed by relapse (n = 10), and no response (n = 10). Patients with complete and sustained response had lower pre-IFN serum HCV RNA level (median: RT-PCR+, bDNA-) compared to the complete response to relapse group (median: 5.25 × 10⁶ genome equivalent/ml [eq/ml], P < 0.001) and the no response group (median: 10.63 × 10⁶ eq/ml, P < 0.001). Seven (46.7%) of the complete and sustained response patients had HCV genotype 2a and three patients had a mixture of genotypes 1b and 2a. In contrast, all 10 patients in the complete response to relapse group had genotype 1b whereas 8 of 10 patients in the non-response group had genotype 1b and 2 had genotype 2b. The patients with HCV genotype 2a had lower serum HCV RNA level than those with 1b (P = 0.002). When the HCV viremia were controlled by stratifying them into < and > 10⁶ eq/ml, patients with genotype 1 had a similar complete and sustained response rate compared to genotype 2. These data indicated that pre-IFN serum HCV RNA is also a good predictor of subsequent complete and sustained response in Japanese patients with chronic HCV infection. © 1994 Wiley-Liss, Inc.     

Viral replication in patients with concomitant hepatitis B and C virus infections

The aim of this study was to assess the implications of dual infection with hepatitis B virus (HBV) and hepatitis C virus (HCV). The HBV and HCV status in 100 patients with chronic hepatitis was analysed. HBV DNA was studied using liquid hybridization and the polymerase chain reaction (PCR). HCV viremia was measured using qualitative and quantitative PCR. The HCV genotype was determined by PCR. Patients were divided into three groups according to their HCV-RNA and HBsAg status: group I consisted of 40 patients with chronic hepatitis caused by HBV; group II, 40 patients with chronic hepatitis caused by HCV; and group III, 20 patients infected with both viruses. The HBV-DNA level was higher in group I than in group III (66.4 vs. 11.5 pg/ml; p<0.05). Quantification of HCV viremia revealed mean values of 36.9 copies × 10⁵/ml in group II and 5.5 copies/ml × 10⁵ in group III (p<0.05). The mean aminotransferase level and histological activity were higher in group III. HCV genotype 1b was the predominant type. The data suggest that there is reciprocal inhibition of viral replication in patients with dual HBV and HCV infection. Liver disease appears to be more severe in patients with chronic hepatitis B and C.     

A European multicentre study on the comparison of HCV viral loads between VERIS HCV assay and COBAS® TaqMan® HCV Test and RealTime HCV Assay

BackgroundBeckman Coulter has developed the VERIS HCV Assay for use on the new fully automated DxN VERIS Molecular Diagnostic System^¥ for HCV viral load monitoring.ObjectivesEvaluate the clinical performance of the new quantitative VERIS HCV Assay.Study designComparison was performed on 279 plasma specimens from HCV infected patients tested with the VERIS HCV Assay and COBAS^® Ampliprep/COBAS^® Taqman^® HCV Test and 369 specimens tested with the VERIS HCV Assay and RealTime HCV Assay. Patient monitoring sample results from four time points were also compared.ResultsThe average bias between the VERIS HCV Assay and the COBAS^® Ampliprep/COBAS^® Taqman^® HCV Test was 0.04 log₁₀ IU/mL, while between the VERIS HCV Assay and the RealTime HCV Assay average bias was 0.21 log₁₀ IU/mL. Bias, however, was not consistent across the measuring range. Analysis at the lower end of quantification levels 50, 100, and 1000 IU/mL showed a predicted bias for VERIS HCV Assay versus COBAS^® Ampliprep/COBAS^® Taqman^® HCV Test between −0.42 and −0.22 log₁₀ IU/mL and for VERIS HCV Assay versus RealTime HCV Assay between 0.00 and 0.13 log₁₀ IU/mL. Patient monitoring of HCV viral load over time demonstrated similar levels between VERIS HCV Assay results and COBAS^® Ampliprep/COBAS^® Taqman^® HCV Test (52 samples from 13 patients) and RealTime HCV Assay (112 samples from 28 patients).ConclusionsVERIS HCV Assay for use on the DxN VERIS Molecular Diagnostic System represents a reliable new tool for easy sample to result HCV RNA viral load monitoring.     

Reduction of hepatitis C virus by dialysis membrane

Recently, it has been reported that hepatitis C virus (HCV) RNA levels in blood are reduced after a hemodialysis procedure and that HCV RNA levels in blood are significantly lower in hemodialysis patients than in nonuremic subjects. The present study was designed to examine whether there were differences in the reduction of HCV RNA levels in blood between different dialysis membranes used in a hemodialysis procedure in maintenance hemodialysis patients with HCV. Dialyzers made of polysulfone, ethylenevinylalcohol, polyacrylonitril, polymethylmethacrylate, cellulose diacetate, and cellulose triacetate were used. We took an aliquot of blood from blood tubing at the inlet and the outlet of each dialyzer to measure serum HCV RNA levels. Furthermore, the filtrate was concurrently collected to measure its HCV RNA level at the dialysis outlet. We found that serum HCV RNA levels were reduced when dialyzers made of polysulfone, polyacrylonitril, polymethylmethacrylate, cellulose diacetate, and cellulose triacetate were used. Particularly, serum HCV RNA levels were reduced significantly when dialyzers made of polymethylmethacrylate (P<0.05) and cellulose diacetate (P<0.1) were used. No HCV RNA was detected in the filtrate of any dialysis membrane. These results suggest that the reduction of hepatitic C viral particles in blood is different in each dialysis membrane used in a hemodialysis procedure, and that viral particles may be adsorbed onto the inner surface of the dialysis membranes. This work was presented at the Annual Meeting of the Japanese Society of Dialysis Therapy in Fukuoka, Japan, in 2000     

Serum cytokine and chemokine profiles in patients with myasthenia gravis

Myasthenia gravis (MG) is an autoimmune‐mediated inflammatory disease of the neuromuscular junction. Previous studies of animal MG models have suggested important roles of cytokines in MG pathogenesis, but adequate studies on cytokines in human MG are lacking. Using a multiplex suspension array system, we measured the serum levels of 27 cytokines/chemokines in 47 anti‐acetylcholine receptor antibody‐positive patients with MG and 20 normal controls (NC) to investigate the contribution of cytokines/chemokines toward MG pathogenesis. Correlations between clinical parameters and cytokine/chemokine levels in patients with MG were also examined. The serum levels of interleukin (IL)‐15 (mean ± standard deviation: 6·85 ± 6·97 pg/ml) and vascular endothelial growth factor (VEGF) (96·21 ± 71·60 pg/ml) significantly increased, whereas IL‐4 levels (3·57 ± 0·86 pg/ml) decreased in patients with MG compared with NC (IL‐15: 4·42 ± 1·55 pg/ml; VEGF: 63·51 ± 32·95 pg/ml; IL‐4: 4·15 ± 0·81 pg/ml, P < 0·05). In addition, eight cytokines (IL‐4, IL‐8, IL‐15, eotaxin, macrophage inflammatory protein‐1α, macrophage inflammatory protein‐1β, VEGF and IL‐1b) were significantly changed among MG patients with thymoma, MG patients without thymoma and NC (P < 0·05). Some cytokines, such as IL‐4, IL‐15, and VEGF, may play roles in the pathogenesis of MG.     

Infection of B cells with hepatitis C virus for the development of lymphoproliferative disorders in patients with chronic hepatitis C

Infection with hepatitis C virus (HCV) is associated with lymphoproliferative disorders, represented by essential mixed cryoglobulinemia and B‐cell non‐Hodgkin's lymphoma, but the pathogenic mechanism remains obscure. HCV may infect B cells or interact with their cell surface receptors, and induce lymphoproliferation. The influence of HCV infection of B cells on the development of lymphoproliferative disorders was evaluated in 75 patients with persistent HCV infection. HCV infection was more prevalent (63% vs. 16%, 14%, or 17% P < 0.05 for each), and HCV RNA levels were higher (3.35 ± 3.85 vs. 1.75 ± 2.52, 2.15 ± 2.94 or 2.10 ± 2.90 log copies/100 ng, P < 0.01 for each) in B cells than CD4⁺, CD8⁺ T cells or other cells. Negative‐strand HCV RNA, as a marker of viral replication, was detected in B cells from four of the 75 (5%) patients. Markers for lymphoproliferative disorders were more frequent in the 50 patients with chronic hepatitis C than the 32 with chronic hepatitis B, including cryoglobulinemia (26% vs. 0%, P < 0.001), low CH₅₀ levels (48% vs. 3%, P = 0.012), and the clonality of B cells (12% vs. 0%, P < 0.01). By multivariate analysis, HCV RNA in B cells was an independent factor associated with the presence of at least one marker for lymphoproliferation (odds ratio: 1.98 [95% confidence interval: 1.36–7.24], P = 0.027). Based on the results obtained, the infection of B cells with HCV would play an important role in the development of lymphoproliferative disorders. J. Med. Virol. 81:619–627, 2009 © 2009 Wiley‐Liss, Inc.     

T helper type 2-like cells and therapeutic effects of interferon-γ in combined immunodeficiency with hypereosinophilia (Omenn's syndrome)

We characterized the defects of CD4+ cells in a 17-month-old girl suffering from combined immunodeficiency with hypereosinophilia (Omenn's syndrome). Because the vast majority of peripheral blood CD4⁺ cells expressed the CD45R0 isoform, we purified circulating CD4⁺ CD45R0⁺ cells from the patient and healthy individuals in order to compare their production of cytokines. The patient's CD4⁺ CD45R0⁺ cells spontaneously produced high levels of interleukin-5 (IL-5) in vitro (1600 pg/ml after 24 h of culture) and this was associated with the presence of IL-5 in serum (323 pg/ml). After stimulation with phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187, they produced higher levels of IL-4 (306 vs. 55 ± 4 pg/ml) and IL-5 (2900 vs. 213 ± 72 pg/ml) and lower levels of IL-2 (17 vs. 63 ± 17 IU/ml) and interferon-γ (IFN-γ) (16 vs. 299 ± 70 IU/ml) than controls CD4⁺ CD45R0⁺ cells. This T helper type 2 (Th2) pattern was confirmed by the detection using reverse polymerase chain reaction of IL-4, IL-5 and IL-10 mRNA within peripheral blood mononuclear cells. During a therapeutic trial with human IFN-γ (40 μg/day) which ameliorated the clinical status of the patient, we observed a down-regulation of the in vivo expression of IL-5 and IL-10, a normalization of the eosinophil count and an improvement of the Tcell response to phytohemagglutinin. This observation indicates for the first time that Th2-like cells might be involved in certain forms of congenital immunodeficiency and that IFN-γ might down-regulate their activities in vivo.     

Vitamin D deficiency affects the immunity against Mycobacterium tuberculosis infection in mice

The aim of the study is to investigate the immunological changes after stimulation with bacillus Calmette–Guerin (BCG) in mice with vitamin D deficiency. After weaning, mice were divided into the vitamin D-deficient group (?D group), the normal group (N group), and the vitamin D-supplemented group (+D group). Twelve-week-old mice were intraperitoneally injected with 0.5 mg/ml BCG (≥1.0 × 10⁶ CFU/mg) and maintained for 6 weeks. Spleen lymphocytes were isolated, and the percentages of CD4⁺ and CD8⁺ lymphocytes were determined by flow cytometry. IFN-γ levels, IL-10 levels, and the TB-PPD-specific antibody titer were determined by ELISA. The inter-group difference was analyzed using one-way ANOVA, and multiple comparisons were analyzed using the LSD test. The percentage of CD4⁺ cells was 27.1 ± 0.6 in the ?D group, 23.62 ± 0.42 in the N group, and 19.46 ± 0.32 in the +D group (P < 0.05). The percentage of CD8⁺ lymphocytes was 12.15 ± 0.61 in the ?D group, 8.7 ± 0.64 in the N group, and 7.12 ± 0.48 in the +D group (P < 0.05). The CD4⁺/CD8⁺ ratio was 2.23 ± 0.15 in the ?D group, 2.71 ± 0.21 in the N group, and 2.73 ± 0.31 in the +D group (P < 0.05). The plasma IFN-γ levels were 416.42 ± 16.42 pg/ml in the ?D group, 325.41 ± 11.16 pg/ml in the N group, and 276.26 ± 25.32 pg/ml in the +D group (P < 0.005). The plasma IL-10 levels were 16.45 ± 1.58 pg/ml in the ?D group, 24.31 ± 2.16 pg/ml in the N group, and 26.28 ± 0.42 pg/ml in the +D group (P < 0.005). The serum TB-PPD-specific antibody level was significantly higher in the ?D group than in the N and +D groups. Vitamin D deficiency affects the immunity against Mycobacterium tuberculosis infection in mice.     

Serum hepatitis B surface antigen levels correlate with high serum HBV DNA levels in patients with chronic hepatitis B: a cross-sectional study

Purpose: The hallmark of chronic hepatitis B (CHB) infection is the presence of hepatitis B surface antigen (HBsAg) positivity for at least 6 months. Recently, serum levels of HBsAg have been compared with serum HBV DNA as a surrogate marker to monitor CHB patients. However, data correlating these two markers are scarce. Hence, the present study was done to correlate HBV DNA with HBsAg in CHB patients. Materials and Methods: Consecutive patients of CHB were included. HBV DNA was measured by real-time polymerase chain reaction (PCR). Serum HBsAg was measured by Architect HBsAg. Results: Of the 198 patients enrolled, 166 fulfilled the inclusion criteria (mean age 43 ± 14 years, 87% males) and the median HBV DNA was 1.7 × 10³ (range 6.0–1.1 × 10⁸) IU/ml. Median HBsAg was 8.7 × 10³ (range 5.0–3.2 × 10⁵) IU/ml. Overall correlation between HBV DNA and HBsAg was weak but significant (Spearman ρ = 0.443, P < 0.01). Correlation in HBe antigen-positive group was better (ρ = 0.402, P < 0.01) in comparison to HBe antigen-negative group (ρ = 0.193 P = 0.05). Good correlation existed in treatment-naïve group (ρ = 0.538, P < 0.01). Correlation was regardless of normal or raised alanine transaminase (ALT). Eighty (48%) patients had high HBV DNA (≥2000 IU/ml). Correlation in high DNA group was significant (P < 0.01). The best cut-off of HBsAg for diagnosing high DNA is 3.36 ×10³ IU/ml. Conclusions: Serum HBsAg correlates with HBV DNA in CHB patients, especially in high serum HBV DNA, HBe antigen-positive and treatment-naïve group. HBsAg levels can be used for predicting high serum HBV DNA levels.     

Emergence of lamivudine resistance hepatitis B virus mutations in pregnant women infected with HBV and HIV receiving antiretroviral prophylaxis for the prevention of mother-to-infant transmission in Malawi

HIV/HBV co‐infection is highly prevalent in sub‐Saharan Africa. The aim of this study was to determine if the use of triple combination lamivudine‐containing prophylaxis for the prevention of mother‐to‐infant HIV transmission was associated with the emergence of lamivudine HBV mutations. The study included 21 pregnant co‐infected women in Malawi who received either zidovudine or stavudine plus lamivudine and nevirapine from week 25 of gestation until 6 months after delivery or indefinitely if they met the criteria for treatment (CD4+ <350/mm³). HBV‐DNA was determined using the Roche COBAS assay. Resistance mutations were assessed by the Trugene assay (Siemens Diagnostics). At baseline 33% of the women were HBeAg positive and had HBV‐DNA > 10⁴ IU/ml. Median CD4 count was 237 cells/mm³ and median HIV‐RNA was 3.8 log₁₀ copies/ml. After a median of 259 days of treatment, HBV‐DNA was detectable in 9 out of 21 patients (42.8%). In three cases the HBV‐DNA level was >10⁴ IU/ml. Resistance mutations (M204I in five cases and L180M + M204I/V in one case) were present in 6 (28.6%) patients. Women with a resistant virus had significantly higher baseline HBV‐DNA levels than those not developing resistance (1.1 × 10⁷ IU/ml vs. 20.8 IU/ml, P = 0.022). Levels of ALT and AST were higher in women with resistant viruses compared to those retaining a wild‐type virus. A high rate of lamivudine resistance was seen in this cohort of pregnant women. Follow‐up of these patients will clarify if the presence of resistance has a significant impact on liver disease. J. Med. Virol. 84:1553–1557, 2012. © 2012 Wiley Periodicals, Inc.     

Monocyte implication in renal allograft dysfunction

Macrophages are involved in the development and progression of kidney fibrosis. The aim of this study was to analyse the phenotype of circulating monocytes and their ability to predict kidney allograft dysfunction in living kidney transplant recipients. Whole blood samples from 25 kidney recipients and 17 donors were collected at five time‐points. Monocyte phenotype was analysed by flow cytometry, and interleukin (IL)‐10 and soluble CD163 by enzyme‐linked immunosorbent assay. One week after transplantation, surface CD163 and IL‐10 levels increased significantly from baseline [2·99 ± 1·38 mean fluorescence intensity (MFI) to 5·18 ± 2·42 MFI for CD163; 4·5 ± 1·46 pg/ml to 6·7 ± 2·5 pg/ml for IL‐10]. This CD163 increase correlated with 4‐month creatinine levels (r = 0·4394, P = 0·04). However, soluble CD163 decreased significantly from baseline at 1 week (797·11 ± 340·45 ng/ml to 576·50 ± 293·60 ng/ml). CD14⁺CD16^– monocytes increased at 4 months and correlated positively with creatinine levels at 12 and 24 months (r = 0·6348, P = 0·002 and r = 0·467, P = 0·028, respectively) and negatively with Modification of Diet in Renal Disease (MDRD) at 12 months (r = 0·6056, P = 0·003). At 4 months, IL‐10 decreased significantly (P = 0·008) and correlated positively with creatinine at 2 years (r = 0·68, P = 0·010) and with CD14⁺CD16^– monocytes at 4 months (r = 0·732, P = 0·004). At 24 h, levels of human leucocyte antigen D‐related declined from 12·12 ± 5·99 to 5·21 ± 3·84 and CD86 expression decreased from 2·76 ± 1·08 to 1·87 ± 0·95. Both markers recovered progressively until 12 months, when they decreased again. These results indicate that monitoring monocytes could be a promising new prognostic tool of graft dysfunction in renal transplant patients.     

Comparison of serum hepatitis C virus RNA and core antigen concentrations and determination of whether levels are associated with liver histology or affected by specimen storage time

An enzyme immunoassay has recently been developed for the hepatitis C virus (HCV) core antigen. To evaluate the possible association between core antigen and HCV RNA levels with regards to the change in liver histology over time as well as study the effect of duration of storage on viral load results, sequential sera were analyzed from 45 patients with chronic HCV infection who had undergone two or more liver biopsies. A relatively strong association was found between the core antigen and HCV RNA concentrations (r(s) = 0.8), with a core antigen level of 1 pg/ml corresponding to approximately 1,000 IU/ml. All 42 sera with detectable HCV RNA at the time of the second biopsy had core antigen concentrations above 1 pg/ml, and the three sera without detectable HCV RNA had concentrations below 1 pg/ml. No association was found between HCV RNA or core antigen levels and the stage of fibrosis in biopsy samples, progression of fibrosis, necro-inflammatory grade, steatosis, genotype, alanine aminotransferase level, or alcohol consumption. A significant association was demonstrated between the storage time of the samples and both the HCV RNA and core antigen concentrations. The median log HCV RNA concentrations (international units/milliliter) were 3.92 for the sera obtained at the time of the first biopsy (median storage time, 13.0 years) and 4.41 for the sera obtained at the time of the second biopsy (median storage time, 6.6 years) compared to 5.96, the median for 102 different routine clinical patient samples.