Determining the Safety and Efficacy of Vaccines to Protect Against Viruses that Infect the Central Nervous System
Principal Investigator: Steven Rubin, PhD
Office / Division / Lab: OVRR / DVP / LMD
FDA regulators at CBER are now reviewing several live, attenuated (weakened versions of viruses) vaccines that are derived from neurotropic wild-type viruses (viruses that infect the central nervous system).
There are as yet no reliable markers (biological evidence) that can be used to determine whether such viruses have been successfully attenuated--other than the failure of the vaccine to produce obvious symptoms of disease in recipients. This problem is based on the lack of knowledge of 1) virus virulence factors (molecules that help viruses infect cells and cause disease); 2) characteristics of cells targeted by such viruses; and 3) how these viruses spread in the host. In addition, for many of these vaccines there are as yet no known markers of efficacy (measurable responses of the body that accurately signify that the vaccine is working effectively). This lack of markers of efficacy, such as a specific level of antibody, makes it difficult to interpret immune response data collected during clinical trials of these vaccines.
Our laboratory uses the mumps virus as a model to identify markers of successful virus attenuation as well as to identify markers in the blood that signify that the vaccine is providing significant protection. The present lack of sufficient knowledge in areas of mumps vaccine safety and efficacy is highlighted by the licensure of some mumps vaccines that have caused a complication called aseptic meningitis (inflammation of the membranes covering the brain and spinal cord) and the occurrence of mumps outbreaks in highly-vaccinated populations.
Problems with vaccine safety can be linked to an inadequate understanding of the infection process. Therefore, our research efforts are focused on identifying 1) cells that the virus naturally infects; and 2) mechanisms the virus uses to facilitate its spread in the infected host.
To help identify markers of vaccine efficacy, our laboratory is studying the ability of vaccine-induced antibodies to inactivate a broad range of variations of the virus obtained from different patients. Our goal is to determine the level of antibody that signifies that the immune response to the vaccine is providing protection against the virus.
We chose mumps virus as the model to study because, for the first time in over 40 years, new live attenuated mumps vaccines are being submitted to FDA for approval. Therefore, FDA regulators must understand what to test for in vaccines based on attenuated mumps virus to demonstrate that they are safe and effective. In response to these challenges, we are trying to learn more about mumps virus vaccine safety and efficacy and to apply this to other viral vaccines.
Identification of markers of virus neuroattenuation
Understanding viral pathogenesis is key to successful development of attenuated virus vaccines. In these studies we are trying to identify cell types infected following a natural route of inoculation (intranasal or intra-tracheal) in an animal model and follow the subsequent dissemination of the virus to other sites in the body, including the central nervous system. We inoculate animals via the respiratory route with recombinant mumps viruses expressing enhanced green fluorescent protein (eGFP). The viruses used for these studies include a highly attenuated mumps virus strain, a highly neurovirulent mump virus strain, and chimeric viruses consisting of mixtures of genes from these two viruses.
Disease-relevant host cells identified from the animal studies will then be used for in vitro testing to identify functional differences in the gene products (proteins) of both virulent and attenuated viruses. Our goal is to identify biomarkers of mumps virus neurovirulence, e.g., specific cellular targets of infection or functional properties of specific viral proteins, and to apply this knowledge to efforts at attenuating other neurotropic viruses, in order to facilitate the development and use of safer vaccines.
Examination of vaccine-induced protective efficacy
Over the past decade numerous mumps outbreaks have been reported in highly vaccinated populations in several countries. Widespread use of only one of the two recommended doses of vaccine was believed to be largely responsible. In 2006 the US experienced its largest mumps outbreak in 20 years. Multiple independently performed outbreak investigations found that between 70% and 99% of cases had received the recommended 2 doses of mumps-containing vaccine, indicating lower vaccine efficacy than previously estimated. While mumps was historically a disease of childhood, now mumps primarily occurs among young adults. The most likely explanations of this epidemiological change are (1) the ability of certain mumps virus strains to escape vaccine-induced immune responses, or (2) waning immunity.
To address the virus escape mutant theory, serum samples from recent vaccinees will be assessed for neutralizing antibody titer against a panel of phylogenetically distinct mumps virus strains, including an isolate from the 2006 US mumps outbreaks. The ability of serum to effectively neutralize all virus strains would argue against the virus escape mutant theory.
To address the waning theory, serum samples from individuals at 1 month to 15 years post vaccination will be assessed for neutralizing antibody titer against the vaccine virus itself as well as an isolate from the 2006 US mumps outbreaks. The anti-viral activity in serum will be assessed as a function of time post vaccination.
Finally, to identify a protective titer of mumps antibody, serum samples acquired via the CDC from a Red Cross blood drive at a university prior to a mumps outbreak will be assessed for pre-exposure mumps virus neutralizing antibody titer. The pre-exposure mumps virus neutralizing antibody titer in subjects who later developed or did not develop mumps during the outbreak will inform us of non-protective and protective levels of antibody.
J Pathol 2015 Jan;235(2):242-52
Molecular biology, pathogenesis and pathology of mumps virus.
Rubin S, Eckhaus M, Rennick LJ, Bamford CG, Duprex WP
Mol Pharm 2013 Dec 2;10(12):4590-602
Inhibition of hepatitis C virus by the cyanobacterial protein Microcystis viridis lectin: mechanistic differences between the high-mannose specific lectins MVL, CV-N, and GNA.
Kachko A, Loesgen S, Shahzad-Ul-Hussan S, Tan W, Zubkova I, Takeda K, Wells F, Rubin S, Bewley CA, Major ME
Pediatr Infect Dis J 2013 Oct;32(10):1156-7
Mumps vaccines: do we need a new one?
Rubin S, Beeler J
J Virol 2012 Feb;86(3):1768-76
The V protein of mumps virus plays a critical role in pathogenesis.
Xu P, Luthra P, Li Z, Fuentes S, D'Andrea JA, Wu J, Rubin S, Rota PA, He B
J Virol 2012 Jan;86(1):615-20
Recent mumps outbreaks in vaccinated populations: no evidence of immune escape.
Rubin SA, Link MA, Sauder CJ, Zhang C, Ngo L, Rima BK, Duprex WP
Procedia Vaccinol 2011;5:261-5
Toward replacement of the monkey neurovirulence test in vaccine safety testing
J Infect Dis 2011 Nov;204(9):1413-22
Mumps antibody levels among students before a mumps outbreak: in search of a correlate of immunity.
Cortese MM, Barskey AE, Tegtmeier GE, Zhang C, Ngo L, Kyaw MH, Baughman AL, Menitove JE, Hickman CJ, Bellini WJ, Dayan GH, Hansen GR, Rubin S
J Virol 2011 Jul;85(14):7059-69
Gene-specific contributions to mumps virus neurovirulence and neuroattenuation.
Sauder CJ, Zhang CX, Ngo L, Werner K, Lemon K, Duprex WP, Malik T, Carbone K, Rubin SA
J Virol 2011 Jun;85(12):6082-5
Discrimination of mumps virus small hydrophobic gene deletion effects from gene translation effects on virus virulence.
Malik T, Shegogue CW, Werner K, Ngo L, Sauder C, Zhang C, Duprex WP, Rubin S
Vaccine 2011 Apr 5;29(16):2850-5
Neurovirulence safety testing of mumps vaccines-Historical perspective and current status.
Rubin SA, Afzal MA
Biologicals 2010 Mar;38(2):278-83
A mouse-based assay for the pre-clinical neurovirulence assessment of vaccinia virus-based smallpox vaccines.
Zhang CX, Sauder C, Malik T, Rubin SA
Vaccine 2009 Sep 25;27(42):5822-9
Presence of lysine at aa 335 of the hemagglutinin-neuraminidase protein of mumps virus vaccine strain Urabe AM9 is not a requirement for neurovirulence.
Sauder CJ, Zhang CX, Link MA, Duprex WP, Carbone KM, Rubin SA
J Virol Methods 2009 Aug;159(2):239-43
High-throughput real-time PCR for early detection and quantitation of arenavirus Tacaribe.
Grajkowska LT, Pedras-Vasconcelos JA, Sauder C, Verthelyi D, Puig M
J Gen Virol 2009 Jul;90(Pt 7):1741-7
Single amino acid changes in the mumps virus HN and L proteins are associated with neuroattenuation.
Malik TH, Wolbert C, Nerret L, Sauder C, Rubin S
J Med Virol 2009 Jan;81(1):130-8
Identification of genetic mutations associated with attenuation and changes in tropism of Urabe mumps virus.
Shah D, Vidal S, Link MA, Rubin SA, Wright KE
Clin Infect Dis 2008 Dec 1;47(11):1458-67
Mumps Outbreaks in Vaccinated Populations: Are Available Mumps Vaccines Effective Enough to Prevent Outbreaks?
Dayan GH, Rubin S
J Neuroinflammation 2008 Nov 11;5:50
Astrocytes play a key role in activation of microglia by persistent Borna disease virus infection.
Ovanesov MV, Ayhan Y, Wolbert C, Moldovan K, Sauder C, Pletnikov MV
J Infect Dis 2008 Aug 15;198(4):508-515
Antibody Induced by Immunization with the Jeryl Lynn Mumps Vaccine Strain Effectively Neutralizes a Heterologous Wild-Type Mumps Virus Associated with a Large Outbreak.
Rubin SA, Qi L, Audet SA, Sullivan B, Carbone KM, Bellini WJ, Rota PA, Sirota L, Beeler J
J Infect Dis 2008 Jun 15;197(12):1662-8
Long-term persistence of mumps antibody after receipt of 2 measles-mumps-rubella (MMR) vaccinations and antibody response after a third MMR vaccination among a university population.
Date AA, Kyaw MH, Rue AM, Klahn J, Obrecht L, Krohn T, Rowland J, Rubin S, Safranek TJ, Bellini WJ, Dayan GH
J Immunol 2008 Jun 15;180(12):8231-40
Immunotherapy with CpG oligonucleotides and antibodies to TNF-alpha rescues neonatal mice from lethal arenavirus-induced meningoencephalitis.
Pedras-Vasconcelos JA, Puig M, Sauder C, Wolbert C, Ovanesov M, Goucher D, Verthelyi D
J Neurovirol 2008 Apr;14(2):136-42
Genetic contributions to influenza virus attenuation in the rat brain.
Qi L, Carbone KM, Ye Z, Liu T, Ovanesov M, Pletnikov M, Sauder C, Rubin SA
Vaccine 2008 Apr 16;26(17):2111-8
Measles cases in highly vaccinated population of Novosibirsk, Russia, 2000-2005.
Atrasheuskaya AV, Kulak MV, Neverov AA, Rubin S, Ignatyev GM
Lancet 2008 Mar 15;371(9616):932-44
Hviid A, Rubin S, Mühlemann K
Cytokine 2008 Feb;41(2):182-6
The IP10 (CXCL10) specific cDNA probe of the mCK-5c multiprobe RNase protection assay kit carries two nucleotide insertions that complicate the interpretation of results.
Sauder C, Pedras-Vasconcelos J, Puig M, Verthelyi D
J Neurovirol 2007 Dec;13(6):513-21
A single nucleotide change in the mumps virus F gene affects virus fusogenicity in vitro and virulence in vivo.
Malik T, Sauder C, Wolbert C, Zhang C, Carbone KM, Rubin S
J Gen Virol 2007 Sep;88(Pt 9):2533-41
Functional consequences of attenuating mutations in the haemagglutinin neuraminidase, fusion and polymerase proteins of a wild-type mumps virus strain.
Malik T, Wolbert C, Mauldin J, Sauder C, Carbone KM, Rubin SA
Clin Microbiol Infect 2007 Jul;13(7):670-6
Mumps vaccine failure investigation in Novosibirsk, Russia, 2002-2004.
Atrasheuskaya AV, Kulak MV, Rubin S, Ignatyev GM
Vaccine 2007 Jun 11;25(24):4651-8
Investigation of mumps vaccine failures in Minsk, Belarus, 2001-2003.
Atrasheuskaya AV, Blatun EM, Kulak MV, Atrasheuskaya A, Karpov IA, Rubin S, Ignatyev GM
J Virol 2006 Dec;80(24):12141-8
Activation of microglia by Borna Disease Virus infection: an in vitro study.
Ovanesov MV, Sauder C, Rubin SA, Richt J, Nath A, Carbone KM, Pletnikov MV
Virology 2006 Jun 20;350(1):48-57
Changes in mumps virus neurovirulence phenotype associated with quasispecies heterogeneity.
Sauder CJ, Vandenburgh KM, Iskow RC, Malik T, Carbone KM, Rubin SA
Vaccine 2006 Mar 24;24(14):2662-8
Serological and phylogenetic evidence of monotypic immune responses to different mumps virus strains.
Rubin S, Mauldin J, Chumakov K, Vanderzanden J, Iskow R, Carbone K
Vaccine 2006 Mar 6;24(10):1530-6
Horizontal transmission of the Leningrad-3 live attenuated mumps vaccine virus.
Atrasheuskaya AV, Neverov AA, Rubin S, Ignatyev GM
J Clin Microbiol 2005 Sep;43(9):4847-51
Mumps virus-specific antibody titers from pre-vaccine era sera: comparison of the plaque reduction neutralization assay and enzyme immunoassays.
Mauldin J, Carbone K, Hsu H, Yolken R, Rubin S
Bioinformatics 2005 Aug 1;21(15):3248-54
A quantitative determination of multi-protein interactions by the analysis of confocal images using a pixel-by-pixel assessment algorithm.
Goucher DR, Wincovitch SM, Garfield SH, Carbone KM, Malik TH
J Infect Dis 2005 Apr 1;191(7):1123-8
The rat-based neurovirulence safety test for the assessment of mumps virus neurovirulence in humans: an international collaborative study.
Rubin SA, Afzal MA, Powell CL, Bentley ML, Auda GR, Taffs RE, Carbone KM
Psychiatry Res 2005 Mar 30;134(1):105
Detection of anti-Borna disease virus antibodies by Western blot analysis.
Ghosh M, Sauder C, Carbone KM, Malik TH
Arch Virol 2004 Nov;149(11):2171-86
Susceptibility of Borna disease virus to the antiviral action of gamma-interferon: evidence for species-specific differences.
Sauder C, Herpfer I, Hassler C, Staeheli P
J Neurovirol 2004 Oct;10(5):305-14
Wild-type and attenuated influenza virus infection of the neonatal rat brain.
Rubin S, Liu D, Pletnikov M, McCullers J, Ye Z, Levandowski R, Johannessen J, Carbone K
J Neurovirol 2004 Oct;10(5):267-77
Developmental alterations in serotoninergic neurotransmission in Borna disease virus (BDV)-infected rats: a multidisciplinary analysis.
Dietz D, Vogel M, Rubin S, Moran T, Carbone K, Pletnikov M
Vaccine 2004 Mar 29;22(11-12):1486-93
Mouse neurotoxicity test for vaccinia-based smallpox vaccines.
Li Z, Rubin SA, Taffs RE, Merchlinsky M, Ye Z, Carbone KM