Retrospectively alignments of the investigated hUTY-peptides with those of canine-, murine- and rat-sequences (Table 3) revealed conservation of K1234 in all four species. For W248, the most immunogenic peptide in our setting (positive
in 3 dogs) changes only appeared in 0-2 AAs which had no influence on peptide-sequence/properties and their immunogenic potential. Furthermore, buy ICG-001 W248-data could already be shown in mice [55]. Highest divergence was determined for T368, with the human- and canine-peptides being similar at most. As determined in this work for UTY, substantial homologies and conservation of immune-reactivity, functionality, proteins, peptides (including MHC-presentation) and isoforms were already described for canines in comparison to
humans, cats, mice, rats, apes and cows by others in vitro and in vivo [30, 56-68]. Further in vitro-culture experiments of lymphocytes from in vivo immunized females with DLA-identical-male cells should be performed to strengthen our preliminary data of our first proof-of-principle experiments. Furthermore, higher response of in vivo T cell proliferations might be exhibited by peptide-loaded (single-peptides or peptide-mix/pool) check details male-DCs or male-PBMCs, as well as investigating human- and canine-UTY-peptides in parallel. Thereby, using human- and canine-UTX-homologue peptides, unspecific X-chromosomally derived reactivity can be excluded and the DLA-binding efficacy of the human/canine-UTY/UTX peptides will be verified as well. Non-hematopoietic-cells (fibroblasts, keratinocytes) should be examined with respect to their target cell function as well as their UTY-expression profiles. In further studies we want to transfer our setting in clinical settings, especially in a context of stem-cell transplantation or T cell transfer for treatment SB-3CT of human leukemia: Normally, UTY is not restricted to cells of hematopoietic origin, but the level of expression may differ in various tissues. Adoptive immunotherapy with Y-chromosome-encoded UTY would be feasible in certain circumstances. This first proof-of-principle experiment should demonstrate that hUTY-peptides are presented on male-canine cell-surfaces triggering
a male-specific immune-response. Interpretation of our experiments could be enhanced by cloning some canine-T cells via limiting-dilution-culture recognizing one of the three hUTY-derived-peptides, permitting more detailed examinations of the antigenic specificity and functional properties of CD8+ as well as CD4+cells [43]. Adoptive immunotherapy with DLT after SCT provides a potent strategy to curatively treat haematological malignancies [69]. However, the use of DLT is limited by occurrence of GvHD and sometimes by the poor-response of the patients [70]. Optimized sensitization of donor T cells against antigens presented by leukemic cells could improve DLT. Therefore, ex vivo CTL-generation against UTY for the treatment of recurrent leukemia is reasonable.