Abstract
The dynamic association between the immune system and the skeletal system has recently been appreciated. It has been suggested that cells involved in the inflammatory cascade might modulate the bone fracture repair process. Interestingly a number of studies have demonstrated that ability of the T lymphocyte to affect bone remodelling and health profoundly. For example the presence of T lymphocytes has been shown to increase bone resorption during experimental induced arthritis. We wanted to investigate the role of specific T lymphocytes in fracture repair and required an in vivo model to deplete CD4 and CD8 T lymphocytes selectively.
Method The cell lines of Anti-L3T4 (CD4), Anti-Lyt-2 (CD8) and Anti-phytochrome were obtained from ECACC and produced by Edinburgh University group, Immunosolv. Anti-phytochrome (AFRC MAC 51) antibody was used as the control. To each group of 5 murine models 50ìL of CD4 or CD8 or control antibody was injected ip on days 0, 1, 2, 7, 8, and 9. The body weight and behaviour were measured. On day 20 the spleens were sampled and a single cell suspension was created for each murine model. PE CD8 antibody and FITC CD4 antibody were then added to each sample. Each spleen sample was then cell sorted using the FACS machine.
Results Compared to the control group the murine model injected with CD4 antibody had only 1.14 % of CD4 T cells remaining (mean 2.462 % +/− 0.270). Similarly, the murine models injected with CD8 antibody had only 1.8% of CD8 T cells remaining (mean 1.723 % +/− 1.036).
Conclusion Our results suggest that to investigate the role of T lymphocytes in fracture repair, specific T lymphocytes can be successfully depleted with the repeated use of antibodies.
Correspondence should be addressed to Mr Carlos A. Wigderowitz, Senior Lecturer, University Dept of Orthopaedic and Trauma Surgery, Ninewells Hospital and Medical School, Dundee DD1 9SY
