Publicação
Impact of immune blockade receptor PD-1 in p53wt tumor cells
| Resumo: | Cancer is one of the leading causes of death, both in humans and dogs. Immunotherapies that target immune blockade receptors have been of particular interest over the past few years. Programmed cell death-1 (PD-1) is an immune blockade receptor expressed in activated T cells, B cells, and myeloid derived cells. In healthy individuals, PD- 1 and its ligands, PD-L1 and PD-L2 play an important role in the maintenance of peripheral tolerance by inhibition of T cell function. However, some tumor types express PD-L1, that when bound to PD-1 on activated T cells shuts down the immune response. Recent findings of this group, along with studies of other groups revealed PD-1 expression in tumor cells. The main objective of this thesis was to define whether canine cancer cells can produce tumor intrinsic PD-1 using two novel monoclonal antibodies and to develop cell based assays to define the PD-1 expression, regulation, and function of canine tumor cells. Two monoclonal antibodies were generated previously through immunization of mice with canine extracellular antigen and fusion with two distinct hybridomas. The monoclonal antibodies specificity was tested using different assays, such as ELISA, FACS, and immunofluorescence. Two OSA PD-1 +/- stable cell lines were created to characterize PD-1 expression on canine osteosarcoma. The generated monoclonal antibodies were showed previously to recognize and bind to PD-1 not only in normal tissue, but also melanoma and osteosarcoma tissue samples. In this thesis, different assays revealed that although both monoclonal antibodies have similar specificity they recognize different epitopes. Clonogenic assay of OSA31 PD-1 +/- cells compared with the glioma cell lines J3T PD- 1 +/- revealed that PD-1 enhances cell tumor growth in canine osteosarcoma but suppresses growth in glioma. It was also shown that PD-1 enhanced cellular migration in the canine osteosarcoma cells. This work suggest that PD-1 has a role in tumor cell growth and migration in canine osteosarcoma. |
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| Autores principais: | Lima, Ana Cristina da Silva |
| Assunto: | Ciências Naturais::Ciências Biológicas |
| Ano: | 2017 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso restrito |
| Instituição associada: | Universidade do Minho |
| Idioma: | inglês |
| Origem: | RepositóriUM - Universidade do Minho |
| Resumo: | Cancer is one of the leading causes of death, both in humans and dogs. Immunotherapies that target immune blockade receptors have been of particular interest over the past few years. Programmed cell death-1 (PD-1) is an immune blockade receptor expressed in activated T cells, B cells, and myeloid derived cells. In healthy individuals, PD- 1 and its ligands, PD-L1 and PD-L2 play an important role in the maintenance of peripheral tolerance by inhibition of T cell function. However, some tumor types express PD-L1, that when bound to PD-1 on activated T cells shuts down the immune response. Recent findings of this group, along with studies of other groups revealed PD-1 expression in tumor cells. The main objective of this thesis was to define whether canine cancer cells can produce tumor intrinsic PD-1 using two novel monoclonal antibodies and to develop cell based assays to define the PD-1 expression, regulation, and function of canine tumor cells. Two monoclonal antibodies were generated previously through immunization of mice with canine extracellular antigen and fusion with two distinct hybridomas. The monoclonal antibodies specificity was tested using different assays, such as ELISA, FACS, and immunofluorescence. Two OSA PD-1 +/- stable cell lines were created to characterize PD-1 expression on canine osteosarcoma. The generated monoclonal antibodies were showed previously to recognize and bind to PD-1 not only in normal tissue, but also melanoma and osteosarcoma tissue samples. In this thesis, different assays revealed that although both monoclonal antibodies have similar specificity they recognize different epitopes. Clonogenic assay of OSA31 PD-1 +/- cells compared with the glioma cell lines J3T PD- 1 +/- revealed that PD-1 enhances cell tumor growth in canine osteosarcoma but suppresses growth in glioma. It was also shown that PD-1 enhanced cellular migration in the canine osteosarcoma cells. This work suggest that PD-1 has a role in tumor cell growth and migration in canine osteosarcoma. |
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