Mona Rafik

The ABCs of viral hepatitis that define biomarker signatures of acute viral hepatitis

Viral hepatitis is the leading cause of liver disease worldwide and can be caused by several agents, including hepatitis A (HAV), B (HBV), and C (HCV) virus. We employed multiplexed protein immune assays to identify biomarker signatures of viral hepatitis in order to define unique and common responses for three different acute viral infections of the liver. We performed multianalyte profiling, measuring the concentrations of 182 serum proteins obtained from acute HAV‐ (18), HBV‐ (18), and HCV‐infected (28) individuals, recruited as part of a hospital‐based surveillance program in Cairo, Egypt. Virus‐specific biomarker signatures were identified and validation was performed using a unique patient population. A core signature of 46 plasma proteins was commonly modulated in all three infections, as compared to healthy controls. Principle component analysis (PCA) revealed a host response based upon 34 proteins, which could distinguish HCV patients from HAV‐ and HBV‐infected individuals or healthy controls. When HAV and HBV groups were compared directly, 34 differentially expressed serum proteins allowed the separation of these two patient groups. A validation study was performed on an additional 111 patients, confirming the relevance of our initial findings, and defining the 17 analytes that reproducibly segregated the patient populations. Conclusions: This combined discovery and biomarker validation approach revealed a previously unrecognized virus‐specific induction of host proteins. The identification of hepatitis virus specific signatures provides a foundation for functional studies and the identification of potential correlates of viral clearance. (Hepatology 2014;59:1273‐1282)

CXCL10 antagonism and plasma sDPPIV correlate with increasing liver disease in chronic HCV genotype 4 infected patients

Egypt has the highest prevalence of hepatitis C virus infection worldwide. CXCL10 is a potent chemoattractant that directs effector lymphocytes to sites of inflammation. It has been reported that plasma CXCL10 is processed by dipeptidylpeptidase IV (DPPIV) thus leading to the generation of an antagonist form. Using Luminex-based immunoassays we determined the concentration of different forms of CXCL10 (total, agonist, and antagonist). We also evaluated plasma soluble DPPIV (sDPPIV) concentration and plasma dipeptidylpeptidase (DPP) activity. Using flow cytometry and immunohistochemistry, we analyzed the distribution of lymphocyte subsets. Plasma CXCL10 was elevated in chronic HCV patients, however the agonist form was undetectable. Increased sDPPIV concentration and DPP activity supported the NH2-truncation of CXCL10. Finally, we demonstrated an increased frequency of CXCR3(+) cells in the peripheral blood, and low numbers of CXCR3(+) cells within the lobular regions of the liver. These findings generalize the observation of chemokine antagonism as a mechanism of immune modulation in chronic HCV patients and may help guide the use of new therapeutic immune modulators.

Detection of occult hepatitis C virus among healthy spouses of patients with HCV infection

The criterion standard for the diagnosis of occult hepatitis C virus (HCV) infection is detection of HCV‐RNA in liver cells. However, because of the invasive nature of liver biopsy, other methods have been studied. The present study aimed to identify subjects with occult HCV‐4 infection among healthy sexual partners of patients with chronic HCV‐4 infection by detecting HCV‐RNA in peripheral blood mononuclear cells (PBMCs) using real‐time polymerase chain reaction (PCR). Fifty healthy Egyptian spouses of patients with chronic HCV‐4 infection were included in this study. Real‐time PCR was used to detect HCV‐RNA in PBMCs in all the study subjects. The prevalence of occult HCV‐4 infection was 4%, and a statistically significant higher prevalence was found among patients with a history of sexually transmitted infection. The results of the present study indicate the importance of intra‐spousal transmission of HCV‐4 infection, especially in subjects with a history of sexually transmitted infection. J. Med. Virol. 87:424–427, 2015.

Frequent Transient Hepatitis C viremia without Seroconversion among Healthcare Workers in Cairo, Egypt

Backgrounds With 10% of the general population aged 15–59 years chronically infected with hepatitis C virus (HCV), Egypt is the country with the highest HCV prevalence worldwide. Healthcare workers (HCWs) are therefore at particularly high risk of HCV infection. Our aim was to study HCV infection risk after occupational blood exposure among HCWs in Cairo. Methodology/Principal Findings The study was conducted in 2008–2010 at Ain Shams University Hospital, Cairo. HCWs reporting an occupational blood exposure at screening, having neither anti-HCV antibodies (anti-HCV) nor HCV RNA, and exposed to a HCV RNA positive patient, were enrolled in a 6-month prospective cohort with follow-up visits at weeks 2, 4, 8, 12 and 24. During follow-up, anti-HCV, HCV RNA and ALT were tested. Among 597 HCWs who reported a blood exposure, anti-HCV prevalence at screening was 7.2%, not different from that of the general population of Cairo after age-standardization (11.6% and 10.4% respectively, p = 0.62). The proportion of HCV viremia among index patients was 37%. Of 73 HCWs exposed to HCV RNA from index patients, nine (12.3%; 95%CI, 5.8–22.1%) presented transient viremia, the majority of which occurred within the first two weeks after exposure. None of the workers presented seroconversion or elevation of ALT. Conclusions/Significance HCWs of a general University hospital in Cairo were exposed to a highly viremic patient population. They experienced frequent occupational blood exposures, particularly in early stages of training. These exposures resulted in transient viremic episodes without established infection. These findings call for further investigation of potential immune protection against HCV persistence in this high risk group.

Studies on Infertility in Males

One hundred and fifteen infertile men were examined for circulating spermagglutinating antibodies by the Kibrick spermagglutination test; thirty-three (28%) were found to have positive agglutination titers--1:32 or more in thirteen samples. This high figure may be explained by the high incidence of genital tract infection and of urinary schistosomiasis in our study group. Of the 33 men who demonstrated autoantibodies in their sera, 21 had microscopic agglutination of more than 10%. There was a positive correlation between the serum autoantibody titer and spermagglutination. Eight cases (6.9%) of sperm-immobilizing antibodies were found.

Apolipoprotein H expression is associated with IL28B genotype and viral clearance in hepatitis C virus infection

BACKGROUND & AIMS HCV requires host lipid metabolism for replication, and apolipoproteins have been implicated in the response to treatment. METHODS We examined plasma apolipoprotein concentrations in three cohorts of patients: mono-infected patients with symptomatic acute hepatitis C (aHCV); those undergoing treatment for chronic hepatitis C (cHCV); and HIV/HCV co-infected patients being treated for their chronic hepatitis C. We also evaluated associations between apolipoproteins and IL28B polymorphisms, a defined genetic determinant of viral clearance. RESULTS Plasma apolipoprotein H (ApoH) levels were significantly higher in patients who achieved spontaneous clearance or responded to pegylated-interferon/ribavirin therapy. Strikingly, patients carrying the IL28B rs12979860 CC SNP correlated with the plasma concentration of ApoH in all three cohorts. Both ApoH and IL28B CC SNP were associated with HCV clearance in univariate analysis. Additional multivariate analysis revealed that the association between IL28B and HCV clearance was closely linked to that of Apo H and HCV clearance, suggesting that both belong to the same biological pathway to clearance. The association between IL28B CC SNP and ApoH was not observed in healthy individuals, suggesting that early post-infection events trigger differential ApoH expression in an IL28B allele dependent manner. CONCLUSIONS This relationship identifies ApoH as the first induced protein quantitative trait associated with IL28B, and characterises a novel host factor implicated in HCV clearance.

Circulating Plasmacytoid Dendritic Cells in Acutely Infected Patients with Hepatitis C Virus Genotype 4 are Normal in Number and Phenotype

The incidence of hepatitis C virus (HCV) genotype 4 infection in Egypt provides a unique opportunity to study the innate immune response to symptomatic acute HCV infection. We investigated whether plasmacytoid dendritic cells (pDCs) are activated as a result of HCV infection. We demonstrate that, even during symptomatic acute infection, circulating pDCs maintained a similar precursor frequency and resting phenotype, compared with pDCs in healthy individuals. Moreover, stimulation with a Toll-like receptor 9 agonist resulted in an intact inflammatory response. These data support the growing consensus that pDCs are not directly activated by HCV and therefore are viable targets for immunotherapy throughout HCV infection.

TH1 cytokine response to HCV peptides in Egyptian health care workers: a pilot study

Our objective was to elucidate the effects of different HCV peptides on TH1 cytokine synthesis (interleukin 2(IL2), gamma interferon (INFγ) and tumor necrosis factor α (TNF α)), in a proliferative response in a high risk population of HCV seronegative aviremic Egyptian healthcare workers (HCW). We studied the TH1 cytokine response to different HCV peptides among 47 HCW with and without evidence of HCV infection. Participants were classified according to the proliferation index (PI) in a CFSE proliferation assay as an indicator of previous exposure to HCV. Cytokines were analyzed using Luminex xMAP technology. Results showed that positive PI HCW produced a higher IL2 in response to all HCV peptides except NS4, a higher IFNγ response to NS3 and NS4 and no difference in TNFα response when compared to the negative PI HCWs. When compared to chronic HCV HCW, positive PI HCW showed no difference in the IL2 response, a higher IFNγ response to NS4 and NS5 HCV peptides and a higher TNFα response to all peptides. In conclusion the magnitude and type of cytokines produced in HCV infection is critical in determining the outcome of infection. NS4 & NS5 HCV peptides induce a protective TH1 response in positive PI HCW

Sexual Transmission of HCV in Heterologous Monogamous Spouses

We screened for evidence of HCV infection in healthy heterologous monogamous spouses of chronic HCV patients and studied the relation with various risk factors. A cross-sectional study of fifty healthy monogamous heterosexual spouses of HCV-positive index cases was carried out. All participants were HBV and HIV negative. The association with various risk factors was studied. Five spouses (10%) showed evidence of HCV infection. Two partners were positive for HCV antibody alone (4%) and 3 for antibody and HCV PCR (6%). No association was found between HCV infection and various sociodemographic parameters with the exception of older age categories. Intraspousal transmission of HCV may be an important source of spread of HCV infection. The reservoir of HCV-infected individuals in Egypt is sizable, and sexual transmission of HCV may contribute to the total burden of infection in Egypt.

455 APOLIPOPROTEIN H IS STRONGLY ASSOCIATED WITH IL28B SNP GENOTYPE AND PREDICTS VIRAL CLEARANCE IN BOTH ACUTE AND CHRONIC HEPATITIS C VIRUS INFECTION

455 APOLIPOPROTEIN H IS STRONGLY ASSOCIATED WITH IL28B SNP GENOTYPE AND PREDICTS VIRAL CLEARANCE IN BOTH ACUTE AND CHRONIC HEPATITIS C VIRUS INFECTION M.E. Laird, A. Mohsen, R. Saleh, D. Duffy, L. Le Fouler, M. El-Daly, M. Rafik, M. Abdel-Hamid, M.K. Mohamed, P. Bonnard, V. Mallet, S. Pol, M. Albert, A. Fontanet, The ANRS HC 20 ETOC Study Group. Immunology, Institut Pasteur, Paris, France; Community Medicine Department, National Research Center, Faculty of Medicine, Ain Shams University, Cairo, Egypt; Emerging Disease Epidemiology, Institut Pasteur, Paris, France; Liver Disease Research, National Hepatology and Tropical Medicine Research Institute, Cairo, Faculty of Medicine, Minia University, Minia, Egypt; Maladies Infectieuses et Tropicales, Hopital Tenon (APHP), INSERM U 707, UPMC, Universite Paris Descartes, Institut Cochin, INSERM (IMR-S 1016), CNRS (UMR 8104), Unite d’Hepatologie, Assistance Publique, Hopitaux de Paris (APHP), Groupe Hospitalier Cochin Saint-Vincent de Paul, INSERM U 818, Conservatoire National des Arts et Metiers, Paris, France E-mail: fontanet@pasteur.fr