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. 2021 Jul 22;11:14965. doi: 10.1038/s41598-021-94017-7

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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No γδ T cells are present in blood and spleen of TRDC-knockout pigs. (A) PBMC of TRDC-knockout and wild type pigs were stained with anti-CD3, anti-δ chain and anti-γδ T cell-specific CD3 mABs. CD3 positive cells were gated and δ chain vs. γδ T cell-specific CD3 dotblots are displayed. About 20% of peripheral blood T cells were γδ T cells in wild type pigs while no γδ T cells were detected in TRDC-knockout pigs. (B) PBMC and spleen cells were isolated and staind for CD2 and γδ T cell-specific CD3. Typically in wild type pigs the majority (80%) of γδ T cells in the blood belong to the CD2 negative phenotype (left dot blot), while in the spleen the majority (66%) of γδ T cells are CD2 positive (middle dot blot). No γδ T cells of either type were detected in the spleen of TRDC-knockout pigs. (C) Most γδ T cells are CD4 and CD8 negative. Accordingly TRDC-knockout pigs have minimal numbers of CD4/CD8 double negative T cells. (D) The percentage of lymphocyte subpopulations of all lymphocytes without γδ T cells were compared of wild type and TRDC-knockout pigs. No significant difference of any lymphocyte subpopulation was detected.