a. Career Development Plan: The candidate will spend 80% of her time performing research as outlined in the proposal. A formal course in Macromolecular and Cellular Structure and Chemistry is proposed. In addition, the candidate will attend seminars at Scripps, and weekly lab meetings where she will give about 4 presentations a year. b. Research Plan: Septic shock is a tragic complication of Gram-negative bacterial infections. Myeloid cells clear the body of hazardous invading Gram-negative bacteria and bacterial endotoxin (LPS). CD14, a 55 kD glycophosphatidylinositol (GPI)-anchored plasma membrane protein, found on all myeloid lineage cells is an important mediator of responses to LPS. Secreted LPS-binding protein (LBP) binds to LPS with high affinity and serves as a carrier protein that enables efficient binding of LPS to CD14. The long term goal of the proposal is to understand how CD14 participates in LPS uptake and the phagocytosis of Gram-negative bacteria by myeloid cells. Preliminary studies suggest that the GPI-anchoring of CD14 confers special properties to this receptor that determine uptake of LPS and phagocytosis by myeloid cells. One of the primary goals is to determine the contribution of the CD14/LBP-dependent pathway to LPS uptake and phagocytosis by myeloid cells using flow cytometry. In addition, the role, if any, of the GPI-anchor of CD14 in mediating the signal(s) necessary for LPS uptake and phagocytosis of LPS-bearing particles will be determined. Finally, the localization of GPI-anchored CD14 in the plasma membrane will be characterized by biochemical and immunological approaches. Information obtained from these studies will help answer important questions pertaining to CD14 and its role in host defense, and may contribute to the development of new strategies for the treatment of Gram-negative infections.
