Characteristics and pharmacodynamics of severe neuroinflammation in a child with neurolupus

Methods. Two lumbar punctures were performed for clinical diagnostic purposes. Spun leukocytes in CSF and blood were stained promptly with a panel of directly conjugated monoclonal antibodies to identify lymphocyte subsets. Data were acquired and analyzed on a 5-color single laser Becton-Coulter Cytomics FC500 flow cytometer (St. John's Hospital, Springfield, IL). Also, CSF, serum, and blood samples were obtained as otherwise discarded samples from the St. John's Hospital clinical laboratory under an IRB protocol approved by St. John's Hospital and Southern Illinois University School of Medicine (Springfield, IL) and kept frozen at-80° C until use. The concentrations of chemokines and other cytokines were measured in CSF and serum (plasma for CXCL12) by human-specific enzyme-linked immunosorbent assays (ELISA) (R&D Systems, Inc., Minneapolis, MN) in the Pranzatelli laboratory. Control samples were from children with various non-inflammatory neurological disorders, collected from the clinical laboratory under the same IRB protocol and stored frozen.