In dermatomyositis, B cells infiltrate the muscle tissue. The focus of this proposal will allow for the first time, examination of the antigen specificity of infiltrating B cells derived directly from muscle tissue of patients with dermatomyositis without preconceived bias. This proposal outlines molecular methods for studying the immunobiology of dermatomyositis. Single B cells will be isolated from muscle biopsy specimens by laser capture microdissection followed by PCR amplification and sequencing of the heavy and light variable domains. The combination of these techniques will lead to detailed topographic and molecular characterization of the B cells in dermatomyositis. Once the B cells have been characterized, large amounts of recombinant immunoglobulin will be generated, and the natural variable heavy and light pairing will be preserved allowing faithful reconstruction of these antibodies for antigen screening. These antibodies will be used to identify autoantigens driving the humoral response in dermatomyositis. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32AI065100-02
Application #
7222778
Study Section
Special Emphasis Panel (ZRG1-F07-L (20))
Program Officer
Prograis, Lawrence J
Project Start
2006-04-01
Project End
2009-03-31
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
2
Fiscal Year
2007
Total Cost
$50,428
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
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Han, Qing; Bradshaw, Elizabeth M; Nilsson, Bjorn et al. (2010) Multidimensional analysis of the frequencies and rates of cytokine secretion from single cells by quantitative microengraving. Lab Chip 10:1391-400
Beriou, Gaƫlle; Bradshaw, Elizabeth M; Lozano, Ester et al. (2010) TGF-beta induces IL-9 production from human Th17 cells. J Immunol 185:46-54
Bradshaw, Elizabeth M; Raddassi, Khadir; Elyaman, Wassim et al. (2009) Monocytes from patients with type 1 diabetes spontaneously secrete proinflammatory cytokines inducing Th17 cells. J Immunol 183:4432-9
Bradshaw, Elizabeth M; Kent, Sally C; Tripuraneni, Vinay et al. (2008) Concurrent detection of secreted products from human lymphocytes by microengraving: cytokines and antigen-reactive antibodies. Clin Immunol 129:10-8
Bradshaw, Elizabeth M; Orihuela, Ana; McArdel, Shannon L et al. (2007) A local antigen-driven humoral response is present in the inflammatory myopathies. J Immunol 178:547-56
Elyaman, Wassim; Bradshaw, Elizabeth M; Wang, Yue et al. (2007) JAGGED1 and delta1 differentially regulate the outcome of experimental autoimmune encephalomyelitis. J Immunol 179:5990-8