Medical Student Research Fellowship for Summer 2012

Mentor:                       Steven Vernino MD PhD     
Department:               Neurology & Neurotherapeutics
Room number:           J3.118
Mail Code:                 9036
Phone number:          214-648-8816
E-mail:                        steven.vernino@utsouthwestern.edu                     
Project title:                Developing a flow cytometry based antibody assay for membrane antigens

Human subjects IRB approved project number (where applicable):      

Animal subjects IRB approved project number (where applicable):       N/A

Project Type (patient-based research, animal-based research, or basic research; this characterization is only to permit a general classification for grouping similar types of projects)
Laboratory-based project utilizing patient material

Brief Description of Project:
Antibody testing has become an integral part of diagnosis in neurology.  Many neurological antibodies are also important for the pathophysiology of the disease.  Thus, effective serological testing and identification of novel autoantibodies is critical for ongoing improvements in the evaluation and treatment of neurological diseases.  Testing for antibodies against soluble antigens is straightforward, but detecting antibodies against native membrane proteins (such as neurotransmitter receptors) is challenging since the proteins may lose antigenicity under denaturing conditions such as gel electrophoresis.  Measurement of antibody binding to membrane proteins in cell culture is challenging because the antigens have low abundance and are hard to detect by immunohistochemistry.  Radioimmunoprecipitation is the gold standard method, but this technique requires the use of a high affinity specific radioligand which is not available for most potential antigens.
This project aims to develop a method using antigens bound to fluorescent beads to detect antibodies against cell membrane neuronal proteins.
The student will be responsible for raising cell cultures of transfected cells expressing the membrane proteins of interest and then processing the cells for binding to fluorescent beads.  We will then use this material to test stored serum samples using a flow cytometry based assay.  This project has a potential to produce a novel (radiation-free) and highly sensitive method to detect antibodies against neuronal membrane proteins.  It will be readily adaptable to test both known antigens and to help identify new relevant neuronal autoantibodies.

Previous Research Activities or Publications with Medical Students:
Emil Tajzoy – 2010 summer research project and some ongoing work on pupillometry and autonomic ganglia histology.

• Publication: Tajzoy E, Mukherjee S, Vernino S. Autonomic Ganglia Neuronal Density and Synaptic Structure in Chronic Experimental Autoimmune Autonomic Ganglionopathy. Arch Neurology. 2011. 68:540-541
• Submitted manuscript: Muppidi S, Adams-Huet B, Tajzoy E, Scribner M, Blazek P, Spaeth EB, Frohman E, Davis S, Vernino S.  Dynamic Pupillometry as an Autonomic testing Tool. Clin Auton Res 2012 (submitted)

•   Abstract: Muppidi S, Huet B, Tajzoy E, Scribner M, Blazek P, Frohman E, Davis S, Vernino S.  Use of Dynamic Pupillometry as an Autonomic Testing Tool.  Carrel-Krusen Neuromuscular Symposium.  J Clin Neuromuscular Dis. 2011; 12(3): 18.
•   Abstract: Muppidi S, Tajzoy E, Huet B, Biggs-Scribner M, Blazek P, Davis S, Frohman E, Vernino S. Pupillometery: A Simple Test to Assess Autonomic Dysfunction. American Academy of Neurology. Neurology. 2011; 76(supp 4): A576.

Elaine Boydston – 2011 summer research project and ongoing work on autonomic ganglionopathy

• Abstract: Boydston E, Muppidi S, Vernino S.Long-term outcomes in Autoimmune Autonomic Ganglionopathy.  American Academy of Neurology.