Medicine Project Abstract
DEVELOPMENT OF A WHOLE BLOOD FLOW CYTOMETRY METHOD FOR FUNCTIONAL ASSESSMENT OF PNEUMOCOCCAL PHAGOCYTOSIS (2002)
Presenters:
Emilie Dahod, Illinois Mathematics and Science Academy, 1500 West Sullivan Road, Aurora, IL 60506; dahodo16@imsa.edu
Shravani Pasupneti, Illinois Mathematics and Science Academy, 1500 West Sullivan Road, Aurora, IL 60506; shravani@imsa.edu
Mentor:
William Kabat, Children's Memorial Hospital, Infectious Diseases, 2300 Children's Plaza, Chicago, IL 60614
Abstract:
A large population of patients suffers from infection by Streptococcus pneumoniae. The process of developing a vaccine to prevent infection has been hindered by the different capsule types of pneumococci. A vaccine that provokes the development of antibodies that recognize all of the over one hundred different capsule types has not yet been produced. The best assessment of newly developed vaccines is the evaluation of functional antibodies. These assays evaluate the presence as well as the effectiveness of the antibody. The goal of this research is to develop a single tube, whole blood procedure to simultaneously observe opsonisation and phagocytosis by monocytes and granulocytes of Streptococcus pneumoniae by flow cytometric evaluation. This research team has made considerable progress toward the development of such an assay using FITC (Fluorescein-isothiocyanate) stained bacteria of different capsule types and flow cytometric analysis. At present, the basic procedures and conditions for the assay as well as the gating parameters for the flow assessment have been established. Monoclonal antibodies, including PE-conjugated anti-human IgG and CD14-PC5 have been evaluated. These developments have allowed us to simultaneously measure monocyte and granulocyte uptake of labeled bacteria. A presentation of future plans will follow.