Medical Student Research Fellowship for Summer 2009

Mentor: Philip Thorpe/Xianming Huang
Department: Pharmacology
Room number: NC7.304
Mail Code: 9041
Phone number: 81268/81623
E-mail: Philip.thorpe@utsouthwestern.edu / xianming.huang@utsouthwestern.edu
Project title: The Effect of Anti-PS Antibodies on Maturation and Antigen Presentation of Dendritic Cells.

Human subjects IRB approved project number (where applicable):

Animal subjects IRB approved project number (where applicable):

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) Basic Research

Brief Description of Project:
Specific aims:
1. To evaluate the effect of anti-PS antibody on maturation of immature dendritic cell loaded with tumor whole-cell antigen.
2. To evaluate the effect of anti-PS antibody on antigen presentation of immature dendritic cell loaded with tumor whole-cell antigen.

Methods:

Induction of phenotypic and functional maturation and activation of dendritic cells loaded with tumor cell antigen in the presence of anti-PS antibody will be examined. Mouse bone marrow derived dendritic cells will be loaded with various lethally irradiated tumor cells in the presence antibody or control antibody, the cells will then be harvested and used for analysis of CD80, CD83, CD86, CD40, and HLA-DR expression. The levels of chemokines and cytokines, such as IL-12p40, TNF- , IL-1ß and MIP-1 , MIP-1ß, and RANTES in supernatants will be measured by ELISA. Functional dendritic cell maturation will be assessed by (mixed lymphocyte reaction) MLR. Dendritic cells pretreated as described above will be irradiated (30 Gy) and used as stimulators, and will be co-cultured with T cells of the same species as responders. T-cell proliferation will be quantified. Induction of tumor specific cytotoxic T-lymphocyte (CTL) responses will be examined by CTL assay and IFN-? ELISPOT. Mouse T-cells will be stimulated with dendritic cells activated as described above. T-cells will then be harvested and used as effector cells, the original tumor cell used to load dendritic cells will be used as target cell. The CTL cytotoxicity will be determined. For IFN? ELISPOT assay, stimulated T-cells will be incubated with tumor cells used for dendritic cell loading, after overnight incubation, IFN?-producing T-cells will be determined.

Previous Research Activities or Publications with Medical Students: Laurie worked for Xianming Huang last summer (2008)