Prolongation of GFP-expressed skin graft after intrathymic injection of GFP positive splenocytes in adult rats

Yoji Hakamata; Yuka Igarashi; Takashi Murakami; Eiji Kobayashi

doi:10.1117/12.648014

Abstract

GFP is a fluorescent product of the jellyfish Aequorea victoria and has been used for a variety of biological experiments as a reporter molecule. While GFP possesses advantages for the non-invasive imaging of viable cells, GFP-positive cells are still considered potential xeno-antigens. It is difficult to observe the precise fate of transplanted cells/organs in recipients without immunological control. The aim of this study was to determine whether intrathymic injection of GFP to recipients and the depletion of peripheral lymphocytes could lead to donor-specific unresponsiveness to GFP-expressed cell. LEW rats were administered intraperitoneally with 0.2 ml of anti-rat lymphocyte serum (ALS) 1 day prior to intrathymic injection of donor splenocytes or adeno-GFP vector. Donor cells and vector were non-invasively inoculated into the thymus under high frequency ultrasound imaging using an echo-guide. All animals subsequently received a 7 days GFP-expressed skin graft from the same genetic background GFP LEW transgenic rat. Skin graft survival was greater in rats injected with donor splenocytes (23.6+/-9.1) compared with adeno-GFP (13.0+/-3.7) or untreated control rats (9.5+/-1.0). Intrathymic injection of donor antigen into adult rats can induce donor-specific unresponsiveness. Donor cells can be observed for a long-term in recipients with normal immunity using this strategy.

Citation Download Citation

Yoji Hakamata, Yoji Hakamata, Yuka Igarashi, Yuka Igarashi, Takashi Murakami, Takashi Murakami, Eiji Kobayashi, Eiji Kobayashi, "Prolongation of GFP-expressed skin graft after intrathymic injection of GFP positive splenocytes in adult rat", Proc. SPIE 6098, Genetically Engineered Probes for Biomedical Applications, 60980F (23 February 2006); doi: 10.1117/12.648014; https://doi.org/10.1117/12.648014

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