Our group uses a very broad array of techniques including molecular and cellular biology, state-of-the-art imaging and flow cytometry, bioinformatics, and bioengineering approaches to attack the scientific problems that are of interest to us. Our philosophy is to work on major human pathogens - we study model systems only when the direct study of the primary pathogen in humans is not feasible at a definitive level. Current studies center on respiratory syncytial virus, human metapneumovirus, rotavirus, and vaccinia virus.
|James E. Crowe, Jr., M.D.
Genetic Origins and Structural Determinants of Virus Neutralizing Antibodies
To study this area we have developed unique single B cell sorting and antibody engineering techniques including phage display combinatorial libraries, novel methods for generation of human B cell hybridomas, direct cloning of antibody variable genes from virus-specific B cells, and full-length expression of recombinant antibodies. This work has led to discovery of the molecular basis for the poor quality of infant antiviral antibodies, namely the lack of somatic hypermutations in infant antibody genes.
Currently, we are studying the repertoire of B cells that are gut homing cells, or B cells isolated from human intestinal tissues. These studies are yielding new insights into the nature of human mucosal immune responses to viruses. We are studying the signaling capabilities of human cord blood cells, and studying the differences among naive and memory human B cells in their functional capacity. We are studying the role of TLRs in B and T cell responses. We are also expressing many of the antibody genes that we have recovered as recombinant human monoclonal antibodies. This work has allowed us to understand better structure/function characteristics of antiviral antibodies. Several of the recombinant human antibodies that we have generated are being commercially developed.
Human T cell Response to Virus Vaccination or Infection
We are using state of the art techniques including 10-color flow cytometry to investigate the breadth of the T cell repertoire in response to viral pathogens, the nature of functional subsets of CTL, and the duration of T cell memory in humans. This work has taken us into the biodefense arena, especially in studies of the human response to smallpox immunization.
Trafficking of Virus Proteins in Polarized Epithelial Cells
We have recently discovered that a unique cell compartment, the apical recycling endosome, controls directional budding of respiratory syncytial virus. We are studying the role of particular proteins, including Rabs, Rab-FIPs, and myosins on viral trafficking. This work requires careful biochemistry, confocal and electron microscopy, and molecular biology.