Roselyn J. Eisenberg, Ph.D.
Professor of Microbiology, Dept of Pathobiology, School of Dental Medicine

Microbiology, Virology and Parasitology Program

Address

216 Levy Building
240 S. 40th Street
Philadelphia, PA 19104

Office tel.: 215 898-6552
Lab tel.: 215 898-6558
Fax: 215 898-8385
E-mail: roselyn@biochem.dental.upenn.edu

Dr. Eisenberg's Veterinary Medicine Profile

Dr. Cohen & Dr. Eisenberg's lab website

Bryn Mawr College: AB (Biology), 1960.

University of Pennsylvania: PhD (Microbiology), 1965.

Princeton University: Postdoctoral Research (Molecular Biology), 1968.

Research Interests

• Herpes simplex virus entry mechanisms; structure and function of HSV glycoproteins, Vaccinia virus envelope proteins as vaccine components and entry molecules.

Key words: herpes, HSV, virology, glycoproteins, poxvirus, vaccinia, pox vaccine

Description of Research

All of my research is a collaborative effort with my long-term colleague, Dr. Gary H. Cohen. School of Dental Medicine at Penn. Our joint laboratory is located in the Leon Levy Building of the School of Dental Medicine. Our goal is to understand molecular events that mediate virus entry into susceptible cells and promote the pathogenesis of the virus in its human host. We are studying two viruses: herpes simplex virus (HSV) and vaccinia virus (VACV).

HSV entry requires four envelope glycoproteins, gD, gB, and a heterodimer of gH and gL. Binding of gD to one of its receptors triggers virus-cell fusion involving the other three HSV glycoproteins. The two protein receptors on cells are HVEM, a member of the TNF receptor superfamily, and nectin-1, a cell adhesion molecule that is a member of the Ig superfamily. These receptors may play different roles for the virus in its host, since HVEM is mainly found on immune cells and nectin-1 is mainly found on epithelial and neuronal cells. Ms. Katie Stiles (CAMB graduate student) is examining how gD competes for binding to HVEM with a major attenuator of the immune response, called BTLA. Having solved the three-dimensional structure of glycoprotein D bound to HVEM, we are also able to carry out detailed structure based mutagenesis studies to examine how gD triggers the downstream events leading to entry. Our studies have shown that the binding sites for both receptors are hidden in the native protein and that a major conformational changed in gD accompanies receptor binding and triggers fusion. Eric Lazear (CAMB graduate student) has focused his research on how gD accomplishes its triggering function. In collaboration with Drs. Katya Heldwein and Steve Harrison, we recently solved the 3-dimensional structure of gB, a critical component of the fusion machinery. Dr. Brian Hannah (just awarded his PhD as a CAMB graduate student) carried out structure based mutagenesis of gB to understand how it functions in fusion. To gain more information about how gB and gH/gL cooperate to accomplish virus-cell fusion, we are using a new approach called bimolecular complementation (BiMC). This allows us to follow protein-protein interactions during fusion using confocal microscopy. Thus far, we found that gD triggers an interaction between gB and gH simultaneously with triggering of cell-cell fusion. Additional studies indicate that this interaction may be an important step leading to fusion. Mutants created by Mr. Lazear and Dr. Hannah have been studied in the BiMC assay.

We began work on vaccinia virus (VV) glycoproteins in 2001 following events of September 11. We felt strongly that we could contribute in a direct way to developing reagents against the threat of a smallpox scare or outbreak. A major goal has been to develop a subunit vaccine against smallpox using proteins from VVor from variola virus (smallpox). Such a vaccine could be used for immuno-compromised patients who cannot tolerate the current live virus vaccines and could also be used in a prime-boost strategy. This work is being carried out in a collaborative project on poxviruses that is part of the Midlantic Regional Center of Excellence. A second goal is to understand the mechanism of VV entry into cells, using techniques and approaches we have applied to studies of HSV entry. Mr. Chwan Huan Foo (CAMB graduate student) has focused his studies on discerning which VACV protein is the receptor binding protein for this virus.

Selected Publications

Stiles, KM, Milne, RSB, Cohen GH, Eisenberg, RJ and Krummenacher C. 2008. The herpes simplex virus receptor nectin-1 is down-regulated after trans-interaction with glycoprotein D. Virology. 373:98-111.

Atanasiu D, Whitbeck JC, Cairns TM, Reilly B, Cohen GH and Eisenberg RJ. 2007. Bimolecular complementation reveals that glycoproteins gB and gH/gL of herpes simplex virus interact with each other during cell fusion. Proc. Nat’l Acad Sci USA. 104:18718-18723.

Hannah BP, Heldwein EE, Bender FC, Cohen GH and Eisenberg RJ. 2007. Mutational evidence of internal fusion loops in HSV glycoprotein B . J. Virol.81:4858-4865.

Lazear, E , Carfi A, Whitbeck JC, Cairns TM, Krummenacher C, Cohen GH and Eisenberg RJ. 2008. Engineered disulfide bonds in herpes simplex virus type 1 gD separate receptor binding from fusion initiation and viral entry. J. Virol. 82(2):700-709.

Cairns TM, Friedman LS, Lou H, Whitbeck JC, Shaner MS, Cohen GH and Eisenberg RJ. 2007. N-terminal mutants of HSV-2 gH are transported without gL but 5 require gL for function. J Virol. 81:5102-5111.

Search PubMed for more articles

Lab

Rotation Projects

1. We use a bimolecular complementation assay to study transient protein-protein interactions that occur among HSV as well as vaccinia virus (VACV) proteins during entry and cell-cell fusion. The assay relies on constructing hybrid proteins with either the N- or C-terminal half of yellow fluorescent protein (YFP) hooked to one end of the protein of interest. An interaction between the proteins of interest restores the flouresence of YFP which is visualized by confocal microscopy. Our state of the art microscope allows one to do these studies either on fixed cells or on live cells as the reaction occurs. The student will construct a pair of hybrid proteins and study possible interactions using confocal microscopy. People in the lab can assist with both aspects of this project.
   
   
2. Our lab is expert in the use of the baculovirus expression system for over-expressing proteins of interest for various biochemical and functional studies. We have a variety of mutants that have already been cloned into expression vectors. The student will subclone one of these mutants into the baculovirus transfer vector, carry out recombination to place the gene into the baculovirus genome and then learn how to infect insect cells, and prepare and purify proteins. This project is designed to provide students with technical training in both biochemistry, virology and various functional assays that are established in our laboratory.
   
   
3. A project that might be of interest to a first year graduate student wanting to learn how to work with antibodies would involve characterization of monoclonal antibodies (we have many, and not all of them are fully characterized) using such techniques as western blotting, immunofluorescence and virus neutralization. The student would be introduced to the use of biosensor analysis using our BIAcore 3000 to determine whether antibodies recognize similar or distinct epitopes.

Lab personnel

Eric Lazear, B.S., CAMB graduate student (5th year)
Katie Stiles, B.S., CAMB graduate student (4th year)
Chwang Hong Foo, B.S. CAMB graduate student (4th year)
Tina Cairns, Ph.D., Research Associate
Doina Atanasiu, Ph.D., Research Associate
J.Charles Whitbeck, Ph.D. Senior Research Scientist
Huan Lou, B.S. Research Specialist
Manuel Ponce de Leon, M.S., Research Specialist

Former graduate students

Dr. J.T. Matthews, Microbiology Graduate Group, Ph.D. 1983., Director, Aventis Pasteur Group, External Research &. Development, Sanofi Pasteur Inc

Dr. V. Isola, Immunology Graduate Group, Ph.D. 1988. Currently working as a teacher.

Dr. C. Seidel-Dugan, Molecular Biology Graduate Group. U. Penn, Ph.D. May, 1990. Director, New Targets, Oncology Research, Schering-Plough Research Institute, Kennilworth, NJ

Dr. D. L. Sodora, Molecular Biology Graduate Group, U. Penn, Ph.D. May, 1991; Associate Member, Seattle Biomedical Research Institute and Affiliate appointment, University of Washington, Seattle WA

Dr. Shan-Ling Hung, Molecular Biology Graduate Group, U. Penn, Ph.D. May, 1992; Professor of Oral Biology, National Yang-Ming University, Taipei, Taiwan R.O.C.

Dr. Deborah Long, Molecular Biology Graduate Group, U. Penn, Ph.D. Aug., 1992. Director, Virology, Genocea Biosciences, Inc, Cambridge, MA

Dr. Hsien-Yuan Chiang, Molecular Biology Grad. Group, U. Penn, Ph.D. May, 1993; Professor, Dept. Microbiology and Immunology National Defense Medical Center Taipei, Taiwan

Dr. Ruth Tal-Singer, Molecular Biology Grad. Group, U. Penn, Ph.D. May, 1995. Director of Research, Smith-Kline Beecham Pharmaceuticals.

Dr. Christopher Handler, Molec. Biology Grad. Group, U. Penn, Ph.D. May, 1996. CRA, Quintiles, Rockville, MD

Dr. Anthony Nicola, Molecular Biology Grad. Group, U. Penn, Ph.D., December, 1996. Associate Professor of Microbiology, Virginia Commenwealth University, Richmond VA

Dr. Tao Peng, Biology Graduate Group, Ph.D. May, 1998. Professor, Investigator & Head, Laboratory of Natural Product Drugs; Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou,China.

Dr. Sarah Connolly, Molecular Biology Grad. Group, U. Penn, Ph.D. May 2003. Currently Post-doctoral fellow with Dr. Robert Lamb, Northwestern University, Evanston, IL

Dr. Brian Hannah, Cell and Molecular Biology Graduate Group, 2004-2008. PhD 2008.