Medical Student Research Fellowship for Summer 2010

Mentor: Olaf Stuve
Department: Neurology
Room number: G 3.118
Mail Code: 0936
Phone number: 214 648 8816
E-mail: olaf.stuve@utsouthwestern.edu
Project title: Cellular immune responses against aquaporin-4 in a humanized mouse
model of neuromyelitis optica

Human subjects IRB approved project number (where applicable):

Animal subjects IRB approved project number (where applicable): 20090202

Project Type: animal-based research, basic research

Brief Description of Project:

We hypothesize that T cell responses against AQP-4 constitute an important event in NMO pathogenesis. We also hypothesize that B cell depletion with anti-CD20 antibody therapy alters clinical and paraclinical disease activity, and reduces the frequency of AQP-4-specific T cells.

Specific Aim 1: To characterize cellular immune responses against full-length hAQP-4 in an AQP-4-mediated model of NMO in HLA-DR3, humanized mice

In this specific aim, we will characterize cellular immune responses in an animal model of NMO. Specifically, we will immunize humanized HLA-DR3 mice, deficient in murine MHC II and on a C57BL/6 background (H-2b), with full-length AQP-4 and determine clinical disease activity, the pupillary reflex pathway by pupillometry, CNS histopathology, as well as the phenotype of CNS-infiltrating leukocytes by flow cytometry.

Specific Aim 2: To identify dominant T cell determinants of hAQP-4 in humanized HLA-DR3 mice

We will utilize overlapping peptides of AQP-4 to determine dominant determinants by ELISPOT assay in HLA-DR3. Peptide(s) identified with this approach will then be utilized in HLA-DR3 mice to determine their encephalitogenic potential, as well as to study their effect on paraclinical disease activity as described under Specific Aim 1.

Specific Aim 3: To determine the differential role of B lymphocytes as antigen presenting cells in an hAQP-4-mediated model of NMO in HLA DR3 humanized mice
In this Specific Aim, we will deplete CD20+ B cells in HLA-DR3 mice immunized with full length AQP4 or the dominant linear determinant(s) identified in Specific Aim 2. To specifically determine the role of B cells outside the CNS in an animal model of NMO, we will backcross CD19 4-integrin-/- mice we generated onto HLA-DR3 mice and immunize them with AQP-4 protein or peptide(s). Clinical and paraclinical disease activity will be assessed as described under Specific Aim 1.

Previous Research Activities or Publications with Medical Students:

Martin MdelP, Cravens PD, Winger R, Frohman EM, Racke MK, Eagar TN, Monson NL, Zamvil SS, Weber MS, Hemmer B, Karandikar NJ, Kleinschmidt-DeMasters BK, Stüve O. Natalizumab decreases the numbers of dendritic cells and CD4+T cells in cerebral perivascular spaces. Archives of Neurology. 2008;65:1596-1603.

Martin MdelP, Cravens PD, Winger R, Kieseier BC, Cepok S, Eagar TN, Zamvil SS, Weber MS, Frohman EM, Kleinschmidt-DeMasters BK, Montine TJ, Hemmer B, Marra CM, Stüve O. Rituximab depletes B lymphocytes from cerebral perivascular spaces. Archives of Neurology. 2009;66:1016-1020.