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U.S. Department of Health and Human Services

Vaccines, Blood & Biologics

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Developing New Methods to Measure the Efficacy, Safety and Potency of Preventive and Therapeutic Vaccines Against Hepatitis Viruses

Principal Investigator: Marian Major, PhD
Office / Division / Lab: OVRR / DVP / LHV


General Overview

Our research program is focused on learning more about the way hepatitis C virus (HCV) causes disease in humans and how the immune system responds to this virus.

This work addresses a serious public health threat: an estimated 3.2 million Americans have chronic HCV infection and the results of a variety of studies suggest that over 200 million people worldwide are infected (about 3.3% of the world's population). Moreover, 8,000 to 10,000 Americans die from this disease each year, according to the US Centers for Disease Control and Prevention. In addition, 85% of people infected with the virus develop persistent infections that can eventually cause severe liver problems, such as cirrhosis and liver cancer (hepatocellular carcinoma, or HCC). In fact, HCV infection is considered one of the major risk factors for primary liver cancer in the US, Europe and Japan. (A primary cancer is in the site where it originated rather than having spread from a different site). Furthermore, HCC is one of the few cancers that is increasing in frequency and rate of mortality in the US: studies show that about 50% of HCC cases arise from chronic HCV infection.

Despite these alarming statistics, there is as yet no vaccine to prevent HCV infection. Therapy for HCV has improved over the past five years. Yet for most of the world daily treatment with these drugs is not an option because they are expensive and must be administered carefully due to their toxicity. Two options that hold promise for reducing the rates of infection, disease, and death due to HCV are vaccines and immunotherapy. (Immunotherapy of HCV would involve manipulating the immune system to treat an existing HCV infection.)

In response to this need our laboratory is developing tools to help us study the immune response to HCV and how HCV causes disease. These studies include the development of a small-animal model for HCV infection, identifying the types of immune system activities that demonstrate that a treatment is successful, and the development of neutralization tests for the virus (i.e., tests that show whether the body has produced enough effective antibodies against the virus).

We are developing a system to deliver HCV proteins into specific cells in the body in order to efficiently stimulate the immune system. In addition, we are trying to develop vaccines that do not involve stimulating the production of antibodies, but rather trigger the action of T lymphocytes, which kill cells that are infected with HCV. Such vaccines could be either preventive (blocking infection) or therapeutic (eliminating HCV after it has already established a persistent infection).

These studies will be critical to our ability to guide manufacturers of HCV vaccines and therapies and to the ability of FDA to evaluate the safety of these products.


Scientific Overview

Our laboratory is focused on developing scientific tools to understand the immunobiology and pathogenesis of hepatitis C virus (HCV).

Studies include the identification of efficacy biomarkers (i.e., immunologic correlates of protection), development of neutralization tests for the virus, the use of nanotechnology for targeting DNA vaccines to antigen presenting cells, induction of protective immune responses, and studies on the safety of therapeutic vaccines for this virus. Many of these studies use reagents and data obtained from the chimpanzee model. We are now studying data from several chimpanzees to better understand the kinetics of viral infection and the immune responses that control the infection during the acute phase. Following our earlier studies of re-infections in recovered chimpanzees we began to develop a highly innovative method of vaccination using targeted liposomes for the delivery of DNA to antigen presenting cells. We are now extending this principle to influenza virus vaccines using these same nanolipoplexes to induce immune responses in the respiratory tract through intranasal administration.

HCV could not previously be propagated in a robust cell culture system, which made classical in vitro neutralization tests impossible. In 2005 other investigators showed that RNA from a genotype 2a strain of HCV replicates in cells and produces infectious virus (HCVcc). However, the most prevalent HCV in the US is the 1a genotype. Therefore we generated a 1a/2a and 1b/2a chimeric viruses carrying the respective genotype structural proteins (core, E1, E2, p7) in the 2a replicating backbone. We showed that these chimeric viruses replicate in cell culture as efficiently as the 2a genotype, and have used them to assess neutralizing antibody in a large set of samples.

We are now investigating why vaccines fail and are developing biomarkers to predict clearance of virus in vaccinees. Our approach is to qualitatively compare the recall T-cell response in recovered or re-challenged chimpanzees with the recall response in vaccinated chimpanzees by comparing T-cell markers and cytokine production in vitro, following stimulation with HCV-specific peptides.

Three previously recovered chimpanzees have been re-challenged with HCV. Three naïve chimpanzees have been immunized with our DNA prime-boost vaccination (a systemically delivered prime comprising targeted liposomes followed by a recombinant adenovirus boost) to induce T cell responses, and then challenged with HCV. We obtained serum, T-cells, and liver biopsies from all animals and will analyze HCV-specific T-cells using flow cytometry for surface markers (CD4+, CD8+, CD62L, CD27, CCR7, CD127) and intracellular cytokines (IFN-gamma, IL-2 and TNF-alpha). Based on this data, we will determine the phenotype and perform qualitative comparisons between T-cells induced by vaccination versus those induced by natural infection.


Publications

J Virol 2013 Apr;87(8):4772-7
The Frequency of CD127+ HCV-specific T cells but not the Expression of Exhaustion Markers Predict the Outcome of Acute Hepatitis C Virus Infection.
Shin EC, Park SH, Nascimbeni M, Major M, Caggiari L, de Re V, Feinstone SM, Rice CM, Rehermann B

J Virol 2012 Dec;86(23):12686-94
Amino Acid residue-specific neutralization and nonneutralization of hepatitis C virus by monoclonal antibodies to the e2 protein.
Duan H, Kachko A, Zhong L, Struble E, Pandey S, Yan H, Harman C, Virata-Theimer ML, Deng L, Zhao Z, Major M, Feinstone S, Zhang P

Clin Infect Dis 2012 Jul;55 Suppl 1:S25-32
Prospects for prophylactic and therapeutic vaccines against hepatitis C virus.
Feinstone SM, Hu DJ, Major ME

Vaccine 2011 Dec 9;30(1):69-77
New neutralizing antibody epitopes in hepatitis C virus envelope glycoproteins are revealed by dissecting peptide recognition profiles.
Kachko A, Kochneva G, Sivolobova G, Grazhdantseva A, Lupan T, Zubkova I, Wells F, Merchlinsky M, Williams O, Watanabe H, Ivanova A, Shvalov A, Loktev V, Netesov S, Major ME

Gastroenterology 2011 Aug;141(2):686-695
Delayed induction, not impaired recruitment, of specific CD8(+) T cells causes the late onset of acute hepatitis C.
Shin EC, Park SH, Demino M, Nascimbeni M, Mihalik K, Major M, Veerapu NS, Heller T, Feinstone SM, Rice CM, Rehermann B

Virus Res 2010 Oct;153(1):121-33
Genetic factors of Ebola virus virulence in guinea pigs.
Subbotina E, Dadaeva A, Kachko A, Chepurnov A

Gastroenterology 2010 Sep;139(3):965-74
Meta-analysis of hepatitis C virus vaccine efficacy in chimpanzees indicates an importance for structural proteins.
Dahari H, Feinstone SM, Major ME

Vaccine 2010 Jun 7;28(25):4138-44
Hepatitis C virus with a naturally occurring single amino-acid substitution in the E2 envelope protein escapes neutralization by naturally-induced and vaccine-induced antibodies.
Duan H, Struble E, Zhong L, Mihalik K, Major M, Zhang P, Feinstone S, Feigelstock D

J Viral Hepat 2010 Apr;17(4):245-53
Clearance of hepatitis C in chimpanzees is associated with intrahepatic T-cell perforin expression during the late acute phase.
Watanabe H, Wells F, Major ME

Proc Natl Acad Sci U S A 2009 May 5;106(18):7537-41
Depletion of interfering antibodies in chronic hepatitis C patients and vaccinated chimpanzees reveals broad cross-genotype neutralizing activity.
Zhang P, Zhong L, Struble EB, Watanabe H, Kachko A, Mihalik K, Virata-Theimer ML, Alter HJ, Feinstone S, Major M

Vaccine 2009 Apr 28;27(19):2594-602
T-cell vaccines that elicit effective immune responses against HCV in chimpanzees may create greater immune pressure for viral mutation.
Zubkova I, Choi YH, Chang E, Pirollo K, Uren T, Watanabe H, Wells F, Kachko A, Krawczynski K, Major ME

Hepatology 2006 Dec;44(6):1478-86
Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver.
Heo J, Factor VM, Uren T, Takahama Y, Lee JS, Major M, Feinstone SM, Thorgeirsson SS

Hepatology 2006 Aug 29;44(3):736-45
CD4+ immune escape and subsequent T-cell failure following chimpanzee immunization against hepatitis C virus.
Puig M, Mihalik K, Tilton JC, Williams O, Merchlinsky M, Connors M, Feinstone SM, Major ME

Proc Natl Acad Sci U S A 2006 Mar 7;103(10):3805-9
Cell culture-grown hepatitis C virus is infectious in vivo and can be recultured in vitro.
Lindenbach BD, Meuleman P, Ploss A, Vanwolleghem T, Syder AJ, McKeating JA, Lanford RE, Feinstone SM, Major ME, Leroux-Roels G, Rice CM

Gastroenterology 2005 Apr;128(4):1056-66
Mathematical modeling of primary hepatitis C infection: noncytolytic clearance and early blockage of virion production.
Dahari H, Major M, Zhang X, Mihalik K, Rice CM, Perelson AS, Feinstone SM, Neumann AU

Virology 2004 Nov 10;329(1):53-67
The NS3 protein of hepatitis C virus induces caspase-8-mediated apoptosis independent of its protease or helicase activities.
Prikhod'ko EA, Prikhod'ko GG, Siegel RM, Thompson P, Major ME, Cohen JI

J Virol 2004 Sep;78(18):9782-9
Long-term persistence of infection in chimpanzees inoculated with an infectious hepatitis C virus clone is associated with a decrease in the viral amino Acid substitution rate and low levels of heterogeneity.
Fernandez J, Taylor D, Morhardt DR, Mihalik K, Puig M, Rice CM, Feinstone SM, Major ME

Proc Natl Acad Sci U S A 2004 Jul 6;101(27):10149-54
Neutralizing antibody response during acute and chronic hepatitis C virus infection.
Logvinoff C, Major ME, Oldach D, Heyward S, Talal A, Balfe P, Feinstone SM, Alter H, Rice CM, McKeating JA

Hepatology 2004 Jun;39(6):1709-20
Hepatitis C virus kinetics and host responses associated with disease and outcome of infection in chimpanzees.
Major ME, Dahari H, Mihalik K, Puig M, Rice CM, Neumann AU, Feinstone SM

Vaccine 2004 Feb 25;22(8):991-1000
Immunization of chimpanzees with an envelope protein-based vaccine enhances specific humoral and cellular immune responses that delay hepatitis C virus infection.
Puig M, Major ME, Mihalik K, Feinstone SM

     
 

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