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This study aimed to evaluate the prognostic value of baseline macrophage inflammatory protein (MIP)-1β/IL12p40 ratio for antiviral treatment outcome in HCV genotype 4 patients., Methods: Sera of 450 treatment-naïve chronic HCV patients and 50 healthy individuals were collected. Liver transaminases, total bilirubin and albumin were biochemically tested, viral RNA was quantified, and circulating MIP-1β and IL-12p40 were estimated using human anti-MIP-1β and IL-12p40 antibodies in Sandwich ELISA., Results: No difference was observed in the baseline chemokines levels between responders and relapsers, but the later had a significantly higher MIP-1β/IL-12p40 ratio (P< 0.0001). Multivariate regression analysis of baseline characteristics showed that gender, age, viral load, albumin level and chemokine ratios can significantly predict treatment outcome (P= 0.0114, 0.0095, 0.042, 0.0004 and < 0.0001; respectively). Accordingly, a predictive threshold of baseline chemokine ratio was calculated and it showed an AUC of 0.6917 (P= 0.0108; 95% CI: 0.5566 to 0.8268). The calculated threshold for predicting virologic response was 8.245, with positive and negative predictive values of 92.98% and 100%; respectively. The chemokine ratios had significant correlations with liver transaminases in treated groups whether pre or post-treatment., Conclusion: Baseline MIP-1β/IL-12p40 ratio represents a non-invasive prognostic biomarker that would provide shorter treatment duration and minimizes the emergence of drug-resistant variants in HCV genotype 4-patients.

Background and Aims: Assessment of the potential predictive value of serum inducible protein-10 chemokine (IP-10) in the clearance of HCV in Egyptian patients with and without treatment., Materials and Methods: Ninety Egyptian individuals were involved in the current study where, 20 patients (23%) were chronic HCV (positive HCV antibodies and positive HCV RNA without treatment, 20 (22%) were healthy individuals (negative for both HCV antibodies and HCV RNA, 20 (22%) were natural clearance (positive HCV antibodies and negative for HCV RNA without treatment), 20 (22%) were achieved SVR after treatment (responders group, HCV positive and negative for HCV RNA after treatment) and 10 (11%) were non responders (positive HCV antibodies and still positive HCV RNA after treatment). HCV RNA was quantitated by real time PCR and serum IP10 level was measured by commercial ELISA kit. All biochemical and hematological examinations included liver function, CBC and alphefeto protein were assessed., Results: The mean serum levels of IP-10 were significantly higher (p< 0.001) in CHC patients (345.4 ± 100) pg/ml compared with healthy control group (101.5 ± 31.4) and natural clearance group (103.2 ± 40.7). Also serum levels of IP-10 was significantly elevated in non-responders group (257.4 ± 52.5) compared with each of SVR group (103.5 ± 43.5) (p< 0.001) and healthy group (101.5 ± 31.4), (p< 0.001). Prediction of a clinical response based on a IP10 chemokine revealed high sensitivity (93%), specificity (96%), negative predictive value (96%), and area under curve is (1.00). Moreover, there is no correlation ((R= 0.05), P value p< 0.795) between serum level of IP-10 and HCV viral load., Conclusion: IP10 is a useful non-invasive biomarker for viral clearance and might be used to apply patients according to the predictable treatment outcome. Accordingly, patients who are unlikely to respond to treatment would avoid unnecessary exposure to medication that is related with high morbidity.

Background and Aims: In this study, the safety and tolerability of new candidate HCV vaccine named Cenv6 were screened in mice. Cenv6 peptide is composed of 6 synthetic HCV peptides (3 structural and 3 nonstructural peptides)., Methods: Forty eight mice were enrolled in this study, 12 controls and 36 mice (the thirty-six mice were categorized into 3 groups according to administered doses (3 monthly doses of 800 ng, 1600 ng, and 16 μg/25 gm mouse body weight (bw))). Hematological, biochemical and histopathological changes were appraised., Results: Our data indicated that the doses of 800 ng and 1600 ng of Cenv6 per 25 gm mouse body weight were safe as compared to the dose 16 μg/25 gm bw (10 times more than the potential therapeutic dose) for all examined tissues while the 16 μg Cenv6 dose provoked histopathological changes in kidneys, liver and lungs., Conclusions: The extravagant histopathological changes in different organs have exiled the 16 μg dose out of acceptable range and validated that Cenv6 is safe and tolerable at the two lower doses (800 and 1600 ng/25 gm bw).

Aim: Assessment of the neutralizing activity of human monoclonal antibodies against HCV and also study their safety in experimental small animals (Swiss mice)., Materials and Methods: Assessment of neutralizing activity of human monoclonal antibodies against HCV envelope regions (E1, E2) by two methods: by HCV cc infectious system 1) and by using positive HCV positive serum as source of HCV particles genotype 4a (neutralizing assay 2). Dot ELISA was used to study the activity of the generated antibodies. Safety and toxicity of the generated human antibodies were tested by assessing the changes in the biochemistry of liver function and kidney function tests, Complete blood counts (CBC) and studying the pathological changes with different concentrations of purified human antibodies were carried out.., Results: Human Abs # 5 & 11 showed neutralizing activity by (neutralizing assay 2) but were not neutralizing by HCV cc assay. Human Abs # 12 & 15 showed neutralizing activity by the two methods i.e our generated human antibodies Abs# 5 &11 & 12 & 15 were neutralizing for HCV genotype 4a and Abs # 12 & 15 were neutralizing for HCV genotypes 4a and 2a. Liver and kidney functions and CBC results indicated that doses of 10 μg, 100 μg were safe. The histopathological results indicated that the dose of 10 μg of purified human monoclonal antibodies per mouse body weight was safe., Conclusion: The generated human monoclonal antibodies can be used to develop potent immunotherapy agents that can be administrated for the post-transplantation patients to prevent the recurrence of HCV infection. Also, the monoclonal antibodies can be used to develop a candidate vaccine against HCV.

We aimed to establish Human cell lines producing human monoclonal antibodies to the envelope E1/E2 protein of hepatitis C virus (HCV). Two protocols for EBV immortalization of CD22⁺ cells separated from HCV positive patients were used; 1) Immortalization with 100% EBV only, 2) immortalization by 30% EBV and CPG2006. Immortalization was checked microscopically and verified by screening the culture supernatant for antibody production using dot blot and ELISA analysis. ELISA plates were coated by HCV E1/E2 derived from cell lysate transfected by plasmid expressed HCV E1/E2. Also we tested the reactivity of human antibodies based on ELISA plates coating with one linear peptides derived from HCV E1 (a.a 315-319) and two peptides derived from HCV E2 (a.a 412-419) and (a.a 517-530). Neutralization activity was measured using H77C HCV retroviral pseudoparticles (HCVpp). Fifteen clones secreting human immunoglobulin G against HCV E1/E2 protein were isolated. Results of ELISA plates coated with HCV peptides showed that one antibody was binding to E2 peptide (a.a 517-530), and two antibodies binding to HCV E2 peptide (a.a 412-419). The three generated antibodies showed extremely neutralization activity against HCV pp. The three human antibodies were IgG3 and IgG2. These antibodies may be useful for passive immunotherapy of HCV infection.