PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i>
(1) Purpose: To assess the use of the chicken embryo (<i>in ovo</i>) model as an alternative <i>in vivo</i> model for immuno-oncology (IO) drug development, focusing on programmed cell death protein-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) immune checkpoint inhibitors....
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/14/13/3095 |
| _version_ | 1797480692199718912 |
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| author | Yan Wang Xavier Rousset Chloé Prunier Paul Garcia Emilien Dosda Estelle Leplus Jean Viallet |
| author_facet | Yan Wang Xavier Rousset Chloé Prunier Paul Garcia Emilien Dosda Estelle Leplus Jean Viallet |
| author_sort | Yan Wang |
| collection | DOAJ |
| description | (1) Purpose: To assess the use of the chicken embryo (<i>in ovo</i>) model as an alternative <i>in vivo</i> model for immuno-oncology (IO) drug development, focusing on programmed cell death protein-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) immune checkpoint inhibitors. (2) Methods: First, the presence of immune cells in the model was detected through the immunophenotyping of chicken peripheral blood mononuclear cells (PBMCs) based on fluorescence activated cell sorting (FACS) analysis and the immunohistochemistry (IHC) analysis of <i>in ovo</i> tumor-infiltrating lymphocytes. Second, the cross-reactivity between one anti-human PD-1 Ab, pembrolizumab (KEYTRUDA<sup>®</sup>), and chicken PD-1 was verified through the labelling of chicken splenocytes with pembrolizumab by FACS analysis. Third, the blockade effect of pembrolizumab on chicken PBMCs was assessed <i>in vitro</i> through cytotoxicity assay based on MTT. Fourth, the CAM assay was used to estimate the anti-tumor performance of pembrolizumab through the analyses of tumor growth and chicken immune cell infiltration in tumors. Finally, the efficacy of several PD-1 or PD-L1 inhibitors (nivolumab, atezolizumab and avelumab) on tumor growth was further assessed using the CAM assay. (3) Results: The presence of CD3<sup>+</sup>, CD4<sup>+</sup>, CD8<sup>+</sup> T lymphocytes and monocytes was confirmed by FACS and IHC analyses. During <i>in vitro</i> assays, pembrolizumab cross-reacted with chicken lymphocytes and induced PD-1/PD-L1 blockade, which permitted the restoration of chicken T-cell’s cytotoxicity against human lung cancer H460 tumor cells. All these <i>in vitro</i> results were correlated with <i>in ovo</i> findings based on the CAM assay: pembrolizumab inhibited H460 tumor growth and induced evident chicken immune cell infiltration (with significant chicken CD45, CD3, CD4, CD8 and CD56 markers) in tumors. Furthermore, the potency of the CAM assay was not limited to the application of pembrolizumab. Nivolumab, atezolizumab and avelumab also led to tumor growth inhibition <i>in ovo</i>, on different tumor models. (4) Conclusions: The chicken embryo affords a physiological, immune reactive, <i>in vivo</i> environment for IO research, which allows observation of how the immune system defense against tumor cells, as well as the different immune tolerance mechanisms leading to tumor immune escape. The encouraging results obtained with PD-1/PD-L1 inhibitors in this study reveal the potential use of the chicken embryo model as an alternative, fast, and reliable <i>in vivo</i> model in the different fields of IO drug discovery. |
| first_indexed | 2024-03-09T22:04:39Z |
| format | Article |
| id | doaj.art-9186c35a606e48e3a1087561697e3cc5 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-09T22:04:39Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-9186c35a606e48e3a1087561697e3cc52023-11-23T19:44:18ZengMDPI AGCancers2072-66942022-06-011413309510.3390/cancers14133095PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i>Yan Wang0Xavier Rousset1Chloé Prunier2Paul Garcia3Emilien Dosda4Estelle Leplus5Jean Viallet6R&D Department, Inovotion, 38700 La Tronche, FranceR&D Department, Inovotion, 38700 La Tronche, FranceR&D Department, Inovotion, 38700 La Tronche, FranceR&D Department, Inovotion, 38700 La Tronche, FranceR&D Department, Inovotion, 38700 La Tronche, FrancePDC*line Pharma, 38700 La Tronche, FranceR&D Department, Inovotion, 38700 La Tronche, France(1) Purpose: To assess the use of the chicken embryo (<i>in ovo</i>) model as an alternative <i>in vivo</i> model for immuno-oncology (IO) drug development, focusing on programmed cell death protein-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) immune checkpoint inhibitors. (2) Methods: First, the presence of immune cells in the model was detected through the immunophenotyping of chicken peripheral blood mononuclear cells (PBMCs) based on fluorescence activated cell sorting (FACS) analysis and the immunohistochemistry (IHC) analysis of <i>in ovo</i> tumor-infiltrating lymphocytes. Second, the cross-reactivity between one anti-human PD-1 Ab, pembrolizumab (KEYTRUDA<sup>®</sup>), and chicken PD-1 was verified through the labelling of chicken splenocytes with pembrolizumab by FACS analysis. Third, the blockade effect of pembrolizumab on chicken PBMCs was assessed <i>in vitro</i> through cytotoxicity assay based on MTT. Fourth, the CAM assay was used to estimate the anti-tumor performance of pembrolizumab through the analyses of tumor growth and chicken immune cell infiltration in tumors. Finally, the efficacy of several PD-1 or PD-L1 inhibitors (nivolumab, atezolizumab and avelumab) on tumor growth was further assessed using the CAM assay. (3) Results: The presence of CD3<sup>+</sup>, CD4<sup>+</sup>, CD8<sup>+</sup> T lymphocytes and monocytes was confirmed by FACS and IHC analyses. During <i>in vitro</i> assays, pembrolizumab cross-reacted with chicken lymphocytes and induced PD-1/PD-L1 blockade, which permitted the restoration of chicken T-cell’s cytotoxicity against human lung cancer H460 tumor cells. All these <i>in vitro</i> results were correlated with <i>in ovo</i> findings based on the CAM assay: pembrolizumab inhibited H460 tumor growth and induced evident chicken immune cell infiltration (with significant chicken CD45, CD3, CD4, CD8 and CD56 markers) in tumors. Furthermore, the potency of the CAM assay was not limited to the application of pembrolizumab. Nivolumab, atezolizumab and avelumab also led to tumor growth inhibition <i>in ovo</i>, on different tumor models. (4) Conclusions: The chicken embryo affords a physiological, immune reactive, <i>in vivo</i> environment for IO research, which allows observation of how the immune system defense against tumor cells, as well as the different immune tolerance mechanisms leading to tumor immune escape. The encouraging results obtained with PD-1/PD-L1 inhibitors in this study reveal the potential use of the chicken embryo model as an alternative, fast, and reliable <i>in vivo</i> model in the different fields of IO drug discovery.https://www.mdpi.com/2072-6694/14/13/3095chicken embryo<i>in ovo</i>CAM assayPD-1/PD-L1 inhibitorimmuno-oncology |
| spellingShingle | Yan Wang Xavier Rousset Chloé Prunier Paul Garcia Emilien Dosda Estelle Leplus Jean Viallet PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i> Cancers chicken embryo <i>in ovo</i> CAM assay PD-1/PD-L1 inhibitor immuno-oncology |
| title | PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i> |
| title_full | PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i> |
| title_fullStr | PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i> |
| title_full_unstemmed | PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i> |
| title_short | PD-1/PD-L1 Checkpoint Inhibitors Are Active in the Chicken Embryo Model and Show Antitumor Efficacy <i>In Ovo</i> |
| title_sort | pd 1 pd l1 checkpoint inhibitors are active in the chicken embryo model and show antitumor efficacy i in ovo i |
| topic | chicken embryo <i>in ovo</i> CAM assay PD-1/PD-L1 inhibitor immuno-oncology |
| url | https://www.mdpi.com/2072-6694/14/13/3095 |
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