Principal Investigator: Marian Major, PhD
Office / Division / Lab: OVRR / DVP / LHV
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.
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 naÃ¯ve 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.
Vaccine 2019 May 1;37(19):2608-16
Modeling indicates efficient vaccine-based interventions for the elimination of hepatitis C virus among persons who inject drugs in metropolitan Chicago.
Echevarria D, Gutfraind A, Boodram B, Layden J, Ozik J, Page K, Cotler SJ, Major M, Dahari H
Methods Mol Biol 2019;1911:421-32
Detection of antibodies to HCV E1E2 by lectin-capture ELISA.
Major M, Law M
Sci Transl Med 2018 Jul 11;10(449):eaao4496
Modeling of patient virus titers suggests that availability of a vaccine could reduce hepatitis C virus transmission among injecting drug users.
Major M, Gutfraind A, Shekhtman L, Cui Q, Kachko A, Cotler SJ, Hajarizadeh B, Sacks-Davis R, Page K, Boodram B, Dahari H
J Virol 2018 Feb 26;92(6):e01742-17
Determinants in the IgV domain of human HAVCR1 (TIM-1) are required to enhance hepatitis C virus entry.
Kachko A, Costafreda MI, Zubkova I, Jacques J, Takeda K, Wells F, Kaplan G, Major ME
Antiviral Res 2017 Aug;144:281-5
Modeling HCV cure after an ultra-short duration of therapy with direct acting agents.
Goyal A, Lurie Y, Meissner EG, Major M, Sansone N, Uprichard SL, Cotler SJ, Dahari H
PLoS One 2017 Jul 21;12(7):e0181578
Qualitative differences in cellular immunogenicity elicited by hepatitis C virus T-Cell vaccines employing prime-boost regimens.
Tan WG, Zubkova I, Kachko A, Wells F, Adler H, Sutter G, Major ME
Gut 2016 Jan;65(1):4-5
Hepatitis C: new clues to better vaccines?
Hepatology 2015 Dec;62(6):1670-82
Antibodies to an interfering epitope in hepatitis C Virus E2 can mask vaccine-induced neutralizing activity.
Kachko A, Frey SE, Sirota L, Ray R, Wells F, Zubkova I, Zhang P, Major ME
PLoS One 2015 Sep 30;10(9):e0137993
Agent-based model forecasts aging of the population of people who inject drugs in metropolitan Chicago and changing prevalence of hepatitis C infections.
Gutfraind A, Boodram B, Prachand N, Hailegiorgis A, Dahari H, Major ME
PLoS One 2015 Aug 21;10(8):e0135901
Mathematical modeling of hepatitis C prevalence reduction with antiviral treatment scale-up in persons who inject drugs in metropolitan Chicago.
Echevarria D, Gutfraind A, Boodram B, Major M, Del Valle S, Cotler SJ, Dahari H
EBioMedicine 2015 Jun 30;2(8):857-65
Reverse engineering of vaccine antigens using high throughput sequencing-enhanced mRNA display.
Guo N, Duan H, Kachko A, Krause BW, Major ME, Krause PR