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Title: Adoptive immunotherapy for Epstein-Barr Virus-Associated Post-Transplant Lymphoproliferative Disease
Author: Johannessen, Ingólfur
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2007
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The aim of this study was to establish a pre-clinical SCID mouse model of PTLD in which to test CTL immunotherapy. EBV in vitro infected B lymphoblastoid cell lines (BLCLs) were used to give rise to PTLD-like lesions in SCID mice. A minimum dose of 2x106 BLCL cells consistently induced subcutaneous (sc) or Intraperitoneal (ip) human CD45+ve, CD20+ve B immunoblastic tumours in vivo within 5-7 weeks. Tumours expressed EBV-encoded small RNAs (EBERs) EBV nuclear antigen (EBNA)2 and latent membrane protein (LMP)1 suggestive of unrestricted viral gene expression. The phenotype of the lesions mirrored PTLD, and they were used for testing CTL immunotherapy. Intravenous inoculation of 2 doses of 4x106-8x106 autologous CTLs a week apart significantly delayed sc tumour progression in sc BLCL-injected SCID mice (p=0.01). Similarly, ip transfer of 1 dose of 40xs106-50x106 autologous CTLs or 3x106-50x106 CD8-enriched T cells, into ip BLCL-inoculated animals significantly delayed tumour development in vivo (p=0.001 for both), and prevented tumour formation in a significant proportion (40%) of mice inoculated with CD8-enriched T cells (p=0.001). Immunostaining showed human T cells in the SCID tumours following CTL therapy, and that the cytotoxic mechanisms involved perforin and granzyme B cytotoxic molecules. Whilst CD4-enriched T cells significantly facilitated tumour outgrowth in sc BLCL-injected mice (p=0.04), they did not have a significant impact on tumour development in ip BLCL-inoculated animals. IL7 and 15 conditioning of CTLs in vitro prior to ip transfer into SCID mice significantly delayed ip BLCL-derived tumour formation in vivo when compared to CTLs expanded in vitro using standard methods involving IL2 stimulation (p=0.04). We conclude that PTLD can be modelled in SCID mice. In the animals, autologous CTLs and CD8-enriched T cells have a significant capacity to hinder PTLD-like tumour development.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (M.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available