J. Christopher Love1, Qing Han1, Qing Song1, Elizabeth M. Bradshaw2, Sally C. Kent2, and David A. Hafler2. (1) Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Bldg. 66-456, Cambridge, MA 02139, (2) Neurology, Brigham and Women's Hospital, 77 Avenue Louis Pasteur, NRB, Room 641, Boston, MA 02115
Generating highly-resolved quantitative profiles of human immune responses remains a significant challenge because clinical samples are often limited in size, and cells of particular interest (e.g., autoreactive T cells) can be rare. This talk will describe an approach for analyzing large populations of single cells for both phenotypic, surface-expressed markers and secreted factors such as cytokines and antigen-specific antibodies. A soft lithographic technique called microengraving is employed to allow concurrent detection of cytokines and antibodies from peripheral blood mononuclear cells. Application of the approach to examine B cells from a recent-onset Type I diabetic subject showed the frequency of proinsulin-reactive B cells to be ~0.5% of the circulating B cells tested. Extensions for detection of multiple analytes simultaneously will be presented.