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Patterns of T cell co-stimulation and co-inhibition in tumor microenvironment conditions, in vitro

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Summary:T cells (helper and cytotoxic) are present within the tumor microenvironment and constitute viable targets for immunotherapeutic approaches against cancer, either by adoptive T cell transfer or by the use of monoclonal antibodies. The activation, differentiation, function and survival of those T cells comprehends a complex process and it is regulated by co-stimulatory and co-inhibitory receptors which modulate the initial T cell receptor (TCR) signal, increasing or hindering their anti-tumor function, respectively. Furthermore, various factors secreted by tumor cells or non-lymphoid stromal cells are believed to mostly downmodulate T cell responses. In this work the patterns of expression of co-signaling molecules on T cells were assessed using peripheral blood mononuclear cells (PBMCs) from healthy donors cultured with transforming growth factor (TGF-β), in order to mimic one of the major immunosuppressive stimuli of the tumor microenvironment. Noteworthy, all herein reported effects of TGF-β were entirely dependent on the subsequent T cell activation by an antibody against the CD3ε chain that is part of the TCR complex. While co-inhibitory receptors such as CTLA4 and PD1 were upregulated, PD-L1, TIGIT and TIM3 (also co-inhibitory) had their expression decreased. The transcription factor FOXP3, associated with the regulatory T cell phenotype, was upregulated by TGF-β. For co-activating receptors, the same condition upregulated 4-1BB, CD30 and CD28 expression, whereas it decreased OX40, ICOS and DNAM1 and did not it interfere with CD27 or LIGHT expression. Moreover, alongside TGF-β and anti-CD3, recombinant proteins, such as 4-1BB or OX40 ligand, were added, in order to simulate potential therapeutic strategies using agonistic 4-1BB or OX40 antibodies. It was shown that even though they could induce expression of co-stimulatory receptors such as OX40, CD27 and CD28, there was also CTLA4, PD-1 and TIGIT upregulation. It seems crucial to unravel the details of the dynamics of the expression of co-signaling molecules on T cells to improve existent immunotherapies and design new approaches able to circumvent the immunosuppressive environment, created not only by tumor or stromal cells, but also by immune cells themselves.
Main Authors:Guerra, Luana Frias, 1992-
Subject:Biologia molecular Cancro Células T Imunoterapia Teses de mestrado - 2015
Year:2015
Country:Portugal
Document type:master thesis
Access type:open access
Associated institution:Universidade de Lisboa
Language:English
Origin:Repositório da Universidade de Lisboa
Description
Summary:T cells (helper and cytotoxic) are present within the tumor microenvironment and constitute viable targets for immunotherapeutic approaches against cancer, either by adoptive T cell transfer or by the use of monoclonal antibodies. The activation, differentiation, function and survival of those T cells comprehends a complex process and it is regulated by co-stimulatory and co-inhibitory receptors which modulate the initial T cell receptor (TCR) signal, increasing or hindering their anti-tumor function, respectively. Furthermore, various factors secreted by tumor cells or non-lymphoid stromal cells are believed to mostly downmodulate T cell responses. In this work the patterns of expression of co-signaling molecules on T cells were assessed using peripheral blood mononuclear cells (PBMCs) from healthy donors cultured with transforming growth factor (TGF-β), in order to mimic one of the major immunosuppressive stimuli of the tumor microenvironment. Noteworthy, all herein reported effects of TGF-β were entirely dependent on the subsequent T cell activation by an antibody against the CD3ε chain that is part of the TCR complex. While co-inhibitory receptors such as CTLA4 and PD1 were upregulated, PD-L1, TIGIT and TIM3 (also co-inhibitory) had their expression decreased. The transcription factor FOXP3, associated with the regulatory T cell phenotype, was upregulated by TGF-β. For co-activating receptors, the same condition upregulated 4-1BB, CD30 and CD28 expression, whereas it decreased OX40, ICOS and DNAM1 and did not it interfere with CD27 or LIGHT expression. Moreover, alongside TGF-β and anti-CD3, recombinant proteins, such as 4-1BB or OX40 ligand, were added, in order to simulate potential therapeutic strategies using agonistic 4-1BB or OX40 antibodies. It was shown that even though they could induce expression of co-stimulatory receptors such as OX40, CD27 and CD28, there was also CTLA4, PD-1 and TIGIT upregulation. It seems crucial to unravel the details of the dynamics of the expression of co-signaling molecules on T cells to improve existent immunotherapies and design new approaches able to circumvent the immunosuppressive environment, created not only by tumor or stromal cells, but also by immune cells themselves.