Multiparametric Profiles of Human Immune Responses Produced by Single-Cell Analysis

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.