HCV-specific Immune Responses Research Articles

Immune Responses to Anti-Hepatitis C Virus Antibodies during Pre-Liver Transplantation Direct-Acting Antiviral Therapy in Hepatitis C Virus-Infected Recipients Associated with Post-Liver Transplantation Allograft Injury.

The impact of antibody responses following direct-acting antiviral (DAA) therapy in hepatitis C virus (HCV)-infected recipients before and after liver transplantation (LT) is still undetermined. In this observational cohort study, we aimed to explore the association between changes in anti-HCV antibody titers following pre-LT DAA therapy and allograft injury, including biliary complications (BCs) and acute cellular rejection (ACR). A total of 153 cases were enrolled from January 2015 to February 2021. Serum anti-HCV antibody titers were assessed before and after (day 30) LT. Among all recipients, 31/153 (20.3%) had pre-LT DAA therapy (the DAA group) and 122/153 (79.7%) did not undergo pre-LT DAA therapy (the DAA-naïve group). A higher incidence of post-LT BCs was observed in the DAA group (p = 0.028). Compared with the DAA-naïve group, the DAA group had a significantly higher mean level of anti-HCV titer upregulation (p = 0.0024); furthermore, among the recipients with BCs (n = 28) and ACR (n = 41), those in the DAA group exhibited significantly higher mean levels of anti-HCV antibody titer upregulation (p < 0.005). In conclusion, we speculate that the upregulation of anti-HCV antibody titers, which might have been induced via the restoration of HCV-specific immune responses through pre-LT DAA therapy, was associated with post-LT allograft injury.

Hepatitis-C during Pregnancy: Antenatal period challenges, management and the way forward.

Pregnant women and the general public are both directly impacted by cirrhosis which is a chronic liver disease. It is also widely known that women who have a history of injectable drug use and cirrhosis are more prone to experience unfavorable consequences that have a negative impact on the health of both the mother and the unborn child. Higher maternal Hepatitis-C Virus (HCV) in pregnancy viral load, length of labor, use of amniocentesis or fetal scalp monitoring and protracted membrane rupture are all the risk of perinatal transmission of HCV in newborns. Globally, a large number of childbearing-age women become affected by HCV every year and vertical transmission of HCV is still a serious public health concern. Pregnancy-related immune alterations have a significant impact on the course of HCV infection throughout the third trimester and provide favorable circumstances for the spread of the virus. The exacerbation of hepatic damage during pregnancy and the postpartum period is mostly responsible for HCV-specific cell-mediated immune responses. An extensive literature search done via electronic search engine including Cochrane library databases, PubMed, Google Scholar, Science Direct and HCV in pregnancy articles were included.

Restoration of HCV-Specific Immune Responses with Antiviral Therapy: A Case for DAA Treatment in Acute HCV Infection.

Worldwide, 71 million individuals are chronically infected with Hepatitis C Virus (HCV). Chronic HCV infection can lead to potentially fatal outcomes including liver cirrhosis and hepatocellular carcinoma. HCV-specific immune responses play a major role in viral control and may explain why approximately 20% of infections are spontaneously cleared before the establishment of chronicity. Chronic infection, associated with prolonged antigen exposure, leads to immune exhaustion of HCV-specific T cells. These exhausted T cells are unable to control the viral infection. Before the introduction of direct acting antivirals (DAAs), interferon (IFN)-based therapies demonstrated successful clearance of viral infection in approximately 50% of treated patients. New effective and well-tolerated DAAs lead to a sustained virological response (SVR) in more than 95% of patients regardless of viral genotype. Researchers have investigated whether treatment, and the subsequent elimination of HCV antigen, can reverse this HCV-induced exhausted phenotype. Here we review literature exploring the restoration of HCV-specific immune responses following antiviral therapy, both IFN and DAA-based regimens. IFN treatment during acute HCV infection results in greater immune restoration than IFN treatment of chronically infected patients. Immune restoration data following DAA treatment in chronically HCV infected patients shows varied results but suggests that DAA treatment may lead to partial restoration that could be improved with earlier administration. Future research should investigate immune restoration following DAA therapies administered during acute HCV infection.

Alphavirus-based hepatitis C virus therapeutic vaccines: can universal helper epitopes enhance HCV-specific cytotoxic T lymphocyte responses?

Background:Antigen-specific T cell immune responses play a pivotal role in resolving acute and chronic hepatitis C virus (HCV) infections. Currently, no prophylactic or therapeutic vaccines against HCV are available. We previously demonstrated the preclinical potency of therapeutic HCV vaccines based on recombinant Semliki Forest virus (SFV) replicon particles. However, clinical trials do not always meet the high expectations of preclinical studies, thus, optimization of vaccine strategies is crucial. In efforts to further increase the frequency of HCV-specific immune responses in the candidate SFV-based vaccines, the authors assessed whether inclusion of three strong, so-called universal helper T cell epitopes, and an endoplasmic reticulum localization, and retention signal (collectively termed sigHELP-KDEL cassette) could enhance HCV-specific immune responses.Methods:We included the sigHELP-KDEL cassette in two of the candidate SFV-based HCV vaccines, targeting NS3/4A and NS5A/B proteins. We characterized the new constructs in vitro for the expression and stability of the transgene-encoded proteins. Their immune efficacy with respect to HCV-specific immune responses in vivo was compared with the parental SFV vaccine expressing the corresponding HCV antigen. Further characterization of the functionality of the HCV-specific CD8+ T cells was assessed by surface and intracellular cytokine staining and flow cytometry analysis.Results:Moderate, but significantly, enhanced frequencies of antigen-specific immune responses were achieved upon lower/suboptimal dosage immunization. In optimal dosage immunization, the inclusion of the cassette did not further increase the frequencies of HCV-specific CD8+ T cells when compared with the parental vaccines and the frequencies of effector and memory populations were identical.Conclusion:We hypothesize that the additional effect of the sigHELP-KDEL cassette in SFV-based vaccines depends on the immunogenicity, nature, and stability of the target antigen expressed by the vaccine.

Reversing immune dysfunction and liver damage after direct-acting antiviral treatment for hepatitis C.

The introduction of small molecules targeting viral functions has caused a paradigm shift in hepatitis C virus (HCV) treatment. Administration of these direct-acting antivirals (DAAs) achieves a complete cure in almost all treated patients with short-duration therapy and minimal side effects. Although this is a major improvement over the previous pegylated interferon plus ribavirin (PEG-IFNα/RBV) standard-of-care treatment for HCV, remaining questions address several aspects of the long-term benefits of DAA therapy. Interferon (IFN)-based treatment with successful outcome was associated with substantial reduction in liver disease-related mortality. However, emerging data suggest a complex picture and several confounding factors that influence the effect of both IFN-based and DAA therapies on immune restoration and limiting liver disease progression. We review current knowledge of restoration of innate and HCV-specific immune responses in DAA-mediated viral elimination in chronic HCV infection, and we identify future research directions to achieve long-term benefits in all cured patients and reduce HCV-related liver disease morbidity and mortality.

Development of a recombinant murine tumour model using hepatoma cells expressing hepatitis C virus nonstructural antigens.

Hepatitis C virus (HCV) chronically infects 2%-3% of the world's population, causing liver disease and cancer with prolonged infection. The narrow host range of the virus, being restricted largely to human hepatocytes, has made the development of relevant models to evaluate the efficacy of vaccines a challenge. We have developed a novel approach to accomplish this by generating a murine hepatoma cell line stably expressing nonstructural HCV antigens which can be used in vitro or in vivo to test HCV vaccine efficacies. These HCV-recombinant hepatoma cells formed large solid-mass tumours when implanted into syngeneic mice, allowing us to test candidate HCV vaccines to demonstrate the development of an HCV-specific immune response that limited tumour growth. Using this model, we tested the therapeutic potential of recombinant anti-HCV-specific vaccines based on two fundamentally different attenuated pathogen vaccine systems-attenuated Salmonella and recombinant adenoviral vector based vaccine. While attenuated Salmonella that secreted HCV antigens limited growth of the HCV-recombinant tumours when used in a therapeutic vaccination trial, replication-competent but noninfectious adenovirus expressing nonstructural HCV antigens showed overall greater survival and reduced weight loss compared to non-replicating nondisseminating adenovirus. Our results demonstrate a model with anti-tumour responses to HCV nonstructural (NS) protein antigens and suggest that recombinant vaccine vectors should be explored as a therapeutic strategy for controlling HCV and HCV-associated cancers.

Neonatal Exposure to Hepatitis C Virus Antigens in Uninfected Children Born to Infected Mothers.

Vertical transmission of hepatitis C virus (HCV) infection is uncommon and occurs in approximately 5% of births from HCV-infected mothers. The reason for the low transmission rate is unclear. We aimed to investigate whether there is evidence of HCV exposure also in the noninfected children born to HCV-infected mothers by the presence of a detectable immune response. Serum and peripheral blood mononuclear cells from 9 HCV vertically infected children, 32 uninfected children born to HCV infected mothers, and 15 HCV chronically infected mothers, were analyzed. HCV-RNA-negative adults and children were used as controls. HCV-specific T cell responses were analyzed by interferon gamma using an enzyme-linked immunospot assay and 3H-thymidine incorporation assay. HCV antibodies were also analyzed. An HCV-specific T cell response was detected in 73% (11/15) of the HCV-infected mothers, 67% (6/9) of the vertically infected children, 56% (18/32) of the exposed but uninfected children and in 10% and 20% of the control groups, respectively. The 2 groups of HCV-exposed children both had a significantly higher proportion of HCV-specific T cell responders compared to pediatric controls (P = 0.01 and P = 0.02). HCV-specific immune responses were more common in children born to HCV-infected mothers, regardless of the presence of HCV RNA. We conclude that noninfected children born to HCV-infected mothers may have been exposed to HCV antigens.

Immune-mediated effects targeting hepatitis C virus in a syngeneic replicon cell transplantation mouse model

ObjectiveHCV is characterised by its ability to establish chronic infection in hepatocytes and to replicate in the presence of an inflammation. We mimicked this situation in vivo in immune-competent mice...

Dendritic Cells in Hepatitis C Virus Infection

Background: The global prevalence of chronic hepatitis C is estimated at 2.8%. There is markedly higher prevalence in the Middle East about 14.7% in Egypt. Dendritic cells (DCs) are one of the major Antigen presenting cells in the body. They bridge innate and adaptive immunity and impact priming of HCV-specific immune responses. The current study was aimed to investigate the DC activation status, and their role in interaction with natural killer (NK) cells utilizing different setups with healthy NK and HCV+ DC, HCV+ NK and healthy DC, healthy DC and healthy NK and finally HCV+ NK and HCV+ DC in the presence of HCV peptides and a ratio of 5 NK: 1DC. Results: DC-NK interaction in chronic HCV infection is mainly affected by the affection of DCs by HCV leading to a maturation defect (decreased expression of HLA DR, CD 86 and CD 83). Healthy NK cells were able to stimulate the maturation of DCs particularly with core peptide whereas NS3-4 had no effect. When DCs were healthy, all peptides were able to produce significant maturation of DCs even when co-cultured with HCV+ NK cells. Co-cultured HCV+ NK cells and HCV+ DCs showed significantly higher apoptosis of both cells. This could be attributed to the immature moDCs more with chronic HCV infection due to the fact that immature DCs typically under express HLA-class I molecules that would protect from NK-mediated lysis. Conclusion: Cross-talk between DCs and NK cells plays an important role in the induction of both the innate and adaptive immune systems. HCV infection was found to impair the maturation of DCs. Thus consequently affecting its antigen presentation and T cell allostimulatory capacity and rendering them more liable to NK mediated lysis which could explain the persistence of infection and chronicity.

Cellular immune response to hepatitis-C-virus in subjects without viremia or seroconversion: is it important?

Hepatitis C Virus (HCV) causes chronic infection and represents a global health burden. To date, there is no licensed vaccine for HCV. The high viral replication rate and the existence of several HCV genotypes and quasispecies hamper the development of an effective universal vaccine. In this regard, the current HCV vaccine candidates show genotype-specific protection or narrow cross reactivity against other genotypes. Importantly, HCV spontaneous clearance occurs in 15–50 % of infected subjects, indicating that natural resistance to chronic infection exists. This phenomenon was demonstrated among humans and chimpanzees and continues to motivate researchers attempting to develop an effective HCV vaccine. However, what constitutes a protective immune response or correlate of protection against HCV infection is still vague. Additionally, the mechanisms behind successful HCV clearance suggest the coordination of several arms of the immune system, with cell-mediated immunity (CMI) playing a crucial role in this process. By contrast, although neutralizing antibodies have been identified, they are isolate-specific and poorly correlate with viral clearance. Antigen-specific CD4 T cells, instead, correlate with transient decline in HCV viremia and long-lasting control of the infection. Unfortunately, HCV has been very successful in evading host immune mechanisms, leading to complications such as liver fibrosis, cirrhosis and hepatocellular carcinoma. Interestingly, CMI to HCV antigens were shown among exposed individuals without viremia or seroconversion, suggesting the clearance of prior HCV infection(s). These individuals include family members living with HCV-infected subjects, healthcare workers, IV drug users, and sexual contacts. The correlates of protection could be closely monitored among these individuals. This review provides a summary of HCV-specific immune responses in general and of CMI in particular in these cohorts. The importance of these CMI responses are discussed.

Methods to Evaluate Novel Hepatitis C Virus Vaccines.

The hepatitis C virus (HCV) is a major cause of severe liver disease worldwide. It is estimated that around 130-170 million individuals are chronic carriers of the infection and they are over time at an increased risk of developing severe liver disease. HCV is often referred to as a silent epidemic because the majority of infected individuals do not develop any symptoms. Hence, many individuals are diagnosed at a late stage and thus in need of immediate treatment. Today we have very effective direct-acting antivirals (DAAs), which cure more than 90-95 % of all treated patients. However, this treatment is associated with high-costs and the use is limited to the patients with most advanced liver disease in high-income countries. Notably, a majority of the chronic HCV carriers live in resource-poor countries and do not have access to the new effective DAAs. We therefore need to develop alternative treatments for chronic HCV infection such as therapeutic vaccines. The idea with therapeutic vaccines is to reactivate the infected patient's own immune system. It is well known that patients with chronic HCV infection have dysfunctional immune responses to the virus. Hence, the vaccine should activate HCV-specific T cells that will home to the liver and eradicate the HCV infected hepatocytes. Importantly, one should also consider the combination of a therapeutic vaccine and DAAs as a treatment strategy to equip the resolving patients with post-cure HCV-specific immune responses. This would provide patients with a better protection against reinfection. Numerous genetic vaccine candidates for HCV have been developed and tested in clinical trials with limited effects on viral load and in general inefficient activation of HCV-specific immune responses. In this chapter we describe the rational of developing highly immunogenic vaccines for HCV. Different strategies to improve vaccine immunogenicity and methods to evaluate vaccine efficacy are described. Detailed description of vaccine delivery by intramuscular immunization in combination with in vivo electroporation/electrotransfer (EP/ET) is covered, as well as immunological analysis of primed immune responses by determination of interferon-γ (IFN-γ) production by ELISpot assay and direct ex vivo quantification of HCV NS3/4A-specific CD8+ T cells by pentamer staining. To analyze the in vivo functionality of primed NS3/4A-specific T cells we utilized the in vivo bioluminescence imaging technology. In conclusion, this chapter describes a method to design HCV vaccines and also a protocol to assess their efficacy.

HCV-specific immune responses under direct-acting antiviral therapy of chronic HCV infection

HCV-specific immune responses under direct-acting antiviral therapy of chronic HCV infection

Virus-Specific Cellular Response in Hepatitis C Virus Infection.

Studies performed on chimpanzees and humans have revealed that strong, multispecific and sustained CD4(+) and CD8(+) T cell immune responses is a major determinant of hepatitis C virus (HCV) clearance. However, spontaneous elimination of the virus occurs in minority of infected individuals and cellular response directed against HCV antigens is not persistent in individuals with chronic infection. This review presents characteristics of the HCV-specific T cell response in patients with different clinical course of infection, including acute and chronic infection, persons who spontaneously eliminated HCV and non-infected subjects exposed to HCV. Detection of HCV-specific response, especially in non-infected subjects exposed to HCV, may be indicative of HCV prevalence in population and rate of spontaneous viral clearance. Understanding the mechanisms and role of HCV-specific cellular immune response would contribute to better understanding of HCV epidemiology, immunopathogenesis and may help to design an effective vaccine.

Characterisation of liver pathogenesis, human immune responses and drug testing in a humanised mouse model of HCV infection

ObjectiveHCV infection affects millions of people worldwide, and many patients develop chronic infection leading to liver cancers. For decades, the lack of a small animal model that can recapitulate HCV...

Differential distribution of IL28B.rs12979860 single-nucleotide polymorphism among Egyptian healthcare workers with and without a hepatitis C virus-specific cellular immune response.

The CC genotype of the interleukin (IL)-28B.rs12979860 gene has been associated with spontaneous hepatitis C virus (HCV) clearance and treatment response. The distribution and correlation of an IL28B.rs12979860 single-nucleotide polymorphism (SNP) with HCV-specific cell-mediated immune (CMI) responses among Egyptian healthcare workers (HCWs) is not known. We determined this relationship in 402 HCWs who serve a patient cohort with ~85% HCV prevalence. We enrolled 402 HCWs in four groups: group 1 (n = 258), seronegative aviremic subjects; group 2 (n = 25), seronegative viremic subjects; group 3 (n = 41), subjects with spontaneously resolved HCV infection; and group 4 (n = 78), chronic HCV patients. All subjects were tested for an HCV-specific CMI response using an ex-vivo interferon-gamma (IFNγ) ELISpot assay with nine HCV genotype-4a overlapping 15-mer peptide pools corresponding to all of the HCV proteins. All subjects were tested for IL28B.rs12979860 SNP by real-time PCR. An HCV-specific CMI was demonstrated in ~27% of the seronegative aviremic HCWs (group 1), suggesting clearance of infection after low-level exposure to HCV. The frequency of IL28B.rs12979860 C allele homozygosity in the four groups was 49%, 48%, 49%, and 23%, while that of the T allele was 14%, 16%, 12 and 19%, respectively, suggesting differential distributions among subjects with different HCV status. As reported, IL28B.rs12979860 predicted the outcome of HCV infection (p < 0.05), but we did not find any relationship between the IL28B genotypes and the outcome of HCV-specific CMI responses in the four groups (p > 0.05). The data show differential IL28B.rs12979860 genotype distribution among Egyptian HCWs with different HCV status and could not predict the outcome of HCV-specific CMI responses.

Increased frequency of late-senescent T cells lacking CD127 in chronic hepatitis C disease.

Hepatitis C virus (HCV) causes persistent disease in ~85% of infected individuals, where the viral replication appears to be tightly controlled by HCV-specific CD8+ T cells. Accumulation of senescent T cells during infection results in considerable loss of functional HCV-specific immune responses. We characterized the distinct T-cell phenotypes based on the expression of costimulatory molecules CD28 and CD27, senescence markers PD-1 and CD57, chronic immune activation markers CD38 and HLA-DR, and survival marker CD127 (IL-7R) by flow cytometry following activation of T cells using HCV peptides and phytohemagglutinin. HCV-specific CD4+ and CD8+ T cells from chronic HCV (CHC) patients showed increased expression of PD-1. Furthermore, virus-specific CD4+ T cells of CHC-infected subjects displayed relatively increased expression of HLA-DR and CD38 relative to HCV-specific CD8+ T cells. The CD4+ and CD8+ T cells from HCV-infected individuals showed significant increase of late-differentiated T cells suggestive of immunosenescence. In addition, we found that the plasma viral loads positively correlated with the levels of CD57 and PD-1 expressed on T cells. Chronic HCV infection results in increased turnover of late-senescent T cells that lack survival potentials, possibly contributing to viral persistence. Our findings challenge the prominence of senescent T-cell phenotypes in clinical hepatitis C infection.

Prediction of T-cell epitopes of hepatitis C virus genotype 5a.

BackgroundHepatitis C virus (HCV) is a public health problem with almost 185 million people estimated to be infected worldwide and is one of the leading causes of hepatocellular carcinoma. Currently, there is no vaccine for HCV infection and the current treatment does not clear the infection in all patients. Because of the high diversity of HCV, protective vaccines will have to overcome significant viral antigenic diversities. The objective of this study was to predict T-cell epitopes from HCV genotype 5a sequences.MethodsHCV near full-length protein sequences were analyzed to predict T-cell epitopes that bind human leukocyte antigen (HLA) class I and HLA class II in HCV genotype 5a using Propred I and Propred, respectively. The Antigenicity score of all the predicted epitopes were analysed using VaxiJen v2.0. All antigenic predicted epitopes were analysed for conservation using the IEDB database in comparison with 406, 221, 98, 33, 45, 45 randomly selected sequences from each of the HCV genotypes 1a, 1b, 2, 3, 4 and 6 respectively, downloaded from the GenBank. For epitope prediction binding to common HLA alleles found in South Africa, the IEDB epitope analysis tool was used.ResultsA total of 24 and 77 antigenic epitopes that bind HLA class I and HLA class II respectively were predicted. The highest number of HLA class I binding epitopes were predicted within the NS3 (63%), followed by NS5B (21%). For the HLA class II, the highest number of epitopes were predicted in the NS3 (30%) followed by the NS4B (23%) proteins. For conservation analysis, 8 and 31 predicted epitopes were conserved in different genotypes for HLA class I and HLA class II alleles respectively. Several epitopes bind with high affinity for both HLA class I alleles and HLA class II common in South Africa.ConclusionThe predicted conserved T-cell epitopes analysed in this study will contribute towards the future design of HCV vaccine candidates which will avoid variation in genotypes, which in turn will be capable of inducing broad HCV specific immune responses.

Do the epidemiology, physiological mechanisms and characteristics of hepatocellular carcinoma in HIV-infected patients justify specific screening policies?

Reducing the incidence of hepatocellular carcinoma (HCC) in HIV-infected patients has become a serious problem when managing these patients. There are many explanations for this disease evolution, which notably include their longer survival under effective antiviral therapy and also the more rapid evolution of chronic liver disease. Despite recent advances in the management of hepatitis B (HBV) and hepatitis C (HCV) viral diseases, which will probably increase the number of patients achieving a virological response, HIV-infected patients with cirrhosis are still at risk of the onset of HCC. This evolution to HCC is also correlated to other comorbidities such as excessive alcohol consumption and nonalcoholic steatohepatitis (NASH). HCC thus remains a public health issue in this population. The poor prognosis and aggressiveness of HCC have been fully demonstrated, but the mechanisms underlying this aggressiveness are not yet well defined. As well as underlying mechanisms that contribute to accelerating hepatocarcinogenesis in HIV-infected patients, there are other reasons why HIV-infected patients should be considered a higher risk population. This review discusses the principal epidemiological determinants; the mechanisms of pathogenesis; and the treatment of HCC in HIV/HBV and HIV/HCV coinfected patients. It also discusses the probable need to develop a specific screening policy for HCC in this population in order to prevent the rapid development and to make them more amenable to a curative treatment.

Frequency of regulatory T-cell and hepatitis C viral antigen-specific immune response in recurrent hepatitis C after liver transplantation

IntroductionRegulatory T (Treg) and type 1 regulatory T (Tr1) cells facilitate hepatitis C virus (HCV) recurrence after orthotopic liver transplantation (OLT). However, their frequencies and effects on HCV-specific immune responses have not been well investigated. MethodsWe determined Treg and Tr1 frequencies in OLT patients with hepatitis C and assessed their associations with HCV-specific T cell responses. These patients comprised the following groups: an early post-transplantation group (n=14); an OLT-chronic active hepatitis C group (n=14) with active hepatitis C (alanine aminotransferase of>upper limit of normal/positive for HCV-RNA); an OLT-persistently normal alanine aminotransferase group (n=12) without active hepatitis C (not interferon/positive for HCV-RNA); and an OLT-sustained viral response group (n=6) with sustained viral responses using interferon treatment (negative for HCV-RNA). The frequencies of HCV-specific CD4+ T cells that secreted interferon-γ were determined by enzyme-linked immunosorbent spot assay (except for the OLT early group). ResultsTreg and Tr1 frequencies were low during the early post-transplantation period. OLT patients with sustained viral responses had lower Treg frequencies than those with chronic hepatitis C, whereas Tr1 frequencies were significantly reduced in OLT patients with persistently normal alanine aminotransferase levels compared to those with chronic hepatitis C (p<0.05). Treg frequencies positively correlated with HCV NS3 antigen-specific interferon-γ responses, which corresponded to HCV clearance. ConclusionsIncreased Treg frequencies and reduced HCV-NS3 antigen-specific responses recovered after viral eradication in post-OLT chronic hepatitis C patients. Reduced Tr1 frequencies were associated with hepatitis activity control, which may facilitate controlling chronic hepatitis C in patients after OLT.

A polyantigenic genotype 1a/1b consensus hepatitis C virus DNA vaccine induces broadly reactive HCV-specific cellular immune responses in both mice and non-human primates (VAC7P.985)

Abstract Chronic HCV is a surreptitious disease that can lead to cirrhosis and cancer decades following infection. Each year, 3-4 million individuals are infected. However, there are currently no vaccines available to prevent chronic HCV infection. From patients who acutely resolve (A.R.) infection, a strong and broadly reactive cytotoxic T lymphocyte (CTL) response is associated with viral clearance. With in vivo electroporation (E.P.), DNA vaccines induce strong CTL responses, making them well suited for an HCV vaccine. Our vaccine strategy maximizes recipients' opportunity to induce a broad T cell response with genotype 1a/1b consensus sequences of multiple HCV antigens. Immunogenicity of the HCV vaccine was assessed in mice and non-human primates (NHP). Animals were immunized three times, two weeks apart for mice and six weeks apart for NHP as an intramuscular injection followed by E.P. In mice, interferon-γ (IFNγ) responses spanned all HCV antigens and with total values exceeding 6,000 spot-forming units per million mononuclear cells (SFU) within both the spleen and liver by IFNγ ELIspot. Similarly in NHP, immune responses up to 4,680 SFU per million PBMCs were detectable by IFNγ ELISpot. Flow cytometry revealed HCV-specific CD4+ and CD8+ T cell responses by cytokine staining and significant frequencies of cytolytic CTL compared to baseline. Here, we have demonstrated the magnitude and breadth of HCV-specific immune responses elicited by an HCV DNA vaccine in mice and NHP.