{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T15:54:50Z","timestamp":1792598090179,"version":"4.5.0"},"reference-count":27,"publisher":"MyJove Corporation","issue":"219","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoVE"],"DOI":"10.3791\/68291","type":"journal-article","created":{"date-parts":[[2025,5,30]],"date-time":"2025-05-30T19:01:40Z","timestamp":1748631700000},"source":"Crossref","is-referenced-by-count":0,"title":["<em>In Vitro<\/em> Culture for H5N1-Specific Duck T Cells and Detection of Immune Responses Using Intracellular Cytokine Staining Method"],"prefix":"10.3791","author":[{"given":"Xueqing","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zimin","family":"Xie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanlin","family":"Jiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ming","family":"Liao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manman","family":"Dai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2098","published-online":{"date-parts":[[2025,5,30]]},"reference":[{"issue":"5","key":"ref1","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1016\/j.smim.2016.10.009","type":"journal-article","article-title":"Metabolic pathways in T cell activation and lineage differentiation","volume":"28","author":"Almeida","year":"2016","unstructured":"Almeida, L., Lochner, M., Berod, L., Sparwasser, T. Metabolic pathways in T cell activation and lineage differentiation. Semin Immunol. 28 (5), 514-524 (2016).","journal-title":"Semin Immunol"},{"issue":"6","key":"ref2","first-page":"267","type":"journal-article","article-title":"TCR-MHC\/peptide interaction: prospects for new anti-tumoral agents","volume":"11","author":"Weidle","year":"2014","unstructured":"Weidle, U. H., Georges, G., Tiefenthaler, G. TCR-MHC\/peptide interaction: prospects for new anti-tumoral agents. Cancer Genom Proteom. 11 (6), 267-277 (2014).","journal-title":"Cancer Genom Proteom"},{"issue":"4","key":"ref3","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.smim.2007.03.003","type":"journal-article","article-title":"What guides MHC-restricted TCR recognition","volume":"19","author":"Mazza","year":"2007","unstructured":"Mazza, C., Malissen, B. What guides MHC-restricted TCR recognition? Semin Immunol. 19 (4), 225-235 (2007).","journal-title":"Semin Immunol"},{"issue":"5","key":"ref4","doi-asserted-by":"crossref","first-page":"758","DOI":"10.1067\/mai.2002.124259","type":"journal-article","article-title":"T-cell activation through the antigen receptor. Part 1: signaling components, signaling pathways, and signal integration at the T-cell antigen receptor synapse","volume":"109","author":"Nel","year":"2002","unstructured":"Nel, A. E. T-cell activation through the antigen receptor. Part 1: signaling components, signaling pathways, and signal integration at the T-cell antigen receptor synapse. J Allergy Clin Immunol. 109 (5), 758-770 (2002).","journal-title":"J Allergy Clin Immunol"},{"issue":"14","key":"ref5","doi-asserted-by":"crossref","first-page":"2779","DOI":"10.1007\/s00018-019-03117-1","type":"journal-article","article-title":"Progress on chicken T cell immunity to viruses","volume":"76","author":"Dai","year":"2019","unstructured":"Dai, M., Xu, C., Chen, W., Liao, M. Progress on chicken T cell immunity to viruses. Cell Mol Life Sci. 76 (14), 2779-2788 (2019).","journal-title":"Cell Mol Life Sci"},{"issue":"3","key":"ref6","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1111\/imm.13843","type":"journal-article","article-title":"The role and therapeutic strategies for tissue-resident memory T cells, central memory T cells, and effector memory T cells in psoriasis","volume":"173","author":"Deng","year":"2024","unstructured":"Deng, G. et al. The role and therapeutic strategies for tissue-resident memory T cells, central memory T cells, and effector memory T cells in psoriasis. Immunology. 173 (3), 470-480 (2024).","journal-title":"Immunology"},{"issue":"1","key":"ref7","doi-asserted-by":"crossref","first-page":"148546","DOI":"10.1172\/JCI148546","type":"journal-article","article-title":"Targeting memory T cell metabolism to improve immunity","volume":"132","author":"Corrado","year":"2022","unstructured":"Corrado, M., Pearce, E. L. Targeting memory T cell metabolism to improve immunity. J Clin Invest. 132 (1), 148546 (2022).","journal-title":"J Clin Invest"},{"key":"ref8","doi-asserted-by":"crossref","first-page":"1100741","DOI":"10.3389\/fimmu.2023.1100741","type":"journal-article","article-title":"Maintenance and recall of memory T cell populations against tuberculosis: Implications for vaccine design","volume":"14","author":"Liu","year":"2023","unstructured":"Liu, X., Li, H., Li, S., Yuan, J., Pang, Y. Maintenance and recall of memory T cell populations against tuberculosis: Implications for vaccine design. Front Immunol. 14, 1100741 (2023).","journal-title":"Front Immunol"},{"issue":"1-2","key":"ref9","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.jim.2012.10.012","type":"journal-article","article-title":"An optimized method for establishing high purity murine CD8+ T cell cultures","volume":"387","author":"Zanker","year":"2013","unstructured":"Zanker, D. et al. An optimized method for establishing high purity murine CD8+ T cell cultures. J Immunol Methods. 387 (1-2), 173-180 (2013).","journal-title":"J Immunol Methods"},{"key":"ref10","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1007\/978-1-0716-0266-9_8","type":"journal-article","article-title":"CD4(+) T-Cell Differentiation In Vitro","volume":"2111","author":"Yang","year":"2020","unstructured":"Yang, W., Chen, X., Hu, H. CD4(+) T-Cell Differentiation In Vitro. Methods Mol Biol. 2111, 91-99 (2020).","journal-title":"Methods Mol Biol"},{"issue":"1","key":"ref11","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1186\/s13567-024-01415-6","type":"journal-article","article-title":"Revealing novel CD8(+) T-cell epitopes from the H5N1 avian influenza virus in HBW\/B1 haplotype ducks","volume":"55","author":"Jiao","year":"2024","unstructured":"Jiao, W. et al. Revealing novel CD8(+) T-cell epitopes from the H5N1 avian influenza virus in HBW\/B1 haplotype ducks. Vet Res. 55 (1), 169 (2024).","journal-title":"Vet Res"},{"issue":"1","key":"ref12","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1093\/toxsci\/kfs080","type":"journal-article","article-title":"The development of in vitro culture methods to characterize primary T-cell responses to drugs","volume":"127","author":"Faulkner","year":"2012","unstructured":"Faulkner, L. et al. The development of in vitro culture methods to characterize primary T-cell responses to drugs. Toxicol Sci. 127 (1), 150-158 (2012).","journal-title":"Toxicol Sci"},{"issue":"3","key":"ref13","doi-asserted-by":"crossref","first-page":"100758","DOI":"10.1016\/j.xpro.2021.100758","type":"journal-article","article-title":"A T-cell-based immunogenicity protocol for evaluating human antigen-specific responses","volume":"2","author":"Cimen Bozkus","year":"2021","unstructured":"Cimen Bozkus, C., Blazquez, A. B., Enokida, T., Bhardwaj, N. A T-cell-based immunogenicity protocol for evaluating human antigen-specific responses. STAR Protoc. 2 (3), 100758 (2021).","journal-title":"STAR Protoc"},{"issue":"11","key":"ref14","doi-asserted-by":"crossref","first-page":"1381","DOI":"10.1292\/jvms.11-0517","type":"journal-article","article-title":"Biological effects of chicken type III interferon on expression of interferon-stimulated genes in chickens: comparison with type I and type II interferons","volume":"74","author":"Masuda","year":"2012","unstructured":"Masuda, Y. et al. Biological effects of chicken type III interferon on expression of interferon-stimulated genes in chickens: comparison with type I and type II interferons. J Vet Med Sci. 74 (11), 1381-1386 (2012).","journal-title":"J Vet Med Sci"},{"key":"ref15","doi-asserted-by":"crossref","first-page":"113683","DOI":"10.1016\/j.biopha.2022.113683","type":"journal-article","article-title":"Role of interferon-gamma (IFN-\u03b3) and IFN-\u03b3 receptor 1\/2 (IFN\u03b3R1\/2) in regulation of immunity, infection, and cancer development: IFN-\u03b3-dependent or independent pathway","volume":"155","author":"Ding","year":"2022","unstructured":"Ding, H., Wang, G., Yu, Z., Sun, H., Wang, L. Role of interferon-gamma (IFN-\u03b3) and IFN-\u03b3 receptor 1\/2 (IFN\u03b3R1\/2) in regulation of immunity, infection, and cancer development: IFN-\u03b3-dependent or independent pathway. Biomed Pharmacother. 155, 113683 (2022).","journal-title":"Biomed Pharmacother"},{"key":"ref16","doi-asserted-by":"crossref","first-page":"759188","DOI":"10.3389\/fimmu.2022.759188","type":"journal-article","article-title":"An Artifact in Intracellular Cytokine Staining for Studying T Cell Responses and Its Alleviation","volume":"13","author":"Gong","year":"2022","unstructured":"Gong, Z., Li, Q., Shi, J., Ren, G. An Artifact in Intracellular Cytokine Staining for Studying T Cell Responses and Its Alleviation. Front Immunol. 13, 759188 (2022).","journal-title":"Front Immunol"},{"issue":"12","key":"ref17","doi-asserted-by":"crossref","first-page":"100350","DOI":"10.1016\/j.crmeth.2022.100350","type":"journal-article","article-title":"Ex vivo assays show human gamma-delta T cells specific for common allergens are Th1-polarized in allergic donors","volume":"2","author":"Yu","year":"2022","unstructured":"Yu, E. D. et al. Ex vivo assays show human gamma-delta T cells specific for common allergens are Th1-polarized in allergic donors. Cell Rep Methods. 2 (12), 100350 (2022).","journal-title":"Cell Rep Methods"},{"issue":"1","key":"ref18","doi-asserted-by":"crossref","first-page":"5427","DOI":"10.1038\/s41467-018-07815-5","type":"journal-article","article-title":"Broad CD8(+) T cell cross-recognition of distinct influenza A strains in humans","volume":"9","author":"Grant","year":"2018","unstructured":"Grant, E. J. et al. Broad CD8(+) T cell cross-recognition of distinct influenza A strains in humans. Nat Commun. 9 (1), 5427 (2018).","journal-title":"Nat Commun"},{"issue":"2","key":"ref19","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/S1046-2023(03)00124-5","type":"journal-article","article-title":"Quantification and characterization of specific T-cells by antigen-specific cytokine production using ELISPOT assay or intracellular cytokine staining","volume":"31","author":"Letsch","year":"2003","unstructured":"Letsch, A., Scheibenbogen, C. Quantification and characterization of specific T-cells by antigen-specific cytokine production using ELISPOT assay or intracellular cytokine staining. Methods. 31 (2), 143-149 (2003).","journal-title":"Methods"},{"issue":"5","key":"ref20","doi-asserted-by":"crossref","first-page":"979","DOI":"10.4049\/jimmunol.2101147","type":"journal-article","article-title":"Duck CD8(+) T Cell Response to H5N1 Highly Pathogenic Avian Influenza Virus Infection In Vivo and In Vitro","volume":"209","author":"Dai","year":"2022","unstructured":"Dai, M. et al. Duck CD8(+) T Cell Response to H5N1 Highly Pathogenic Avian Influenza Virus Infection In Vivo and In Vitro. J Immunol. 209 (5), 979-990 (2022).","journal-title":"J Immunol"},{"issue":"1","key":"ref21","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1189\/jlb.0206118","type":"journal-article","article-title":"The generation and modulation of antigen-specific memory CD8 T cell responses","volume":"80","author":"Jabbari","year":"2006","unstructured":"Jabbari, A., Harty, J. T. The generation and modulation of antigen-specific memory CD8 T cell responses. J Leukoc Biol. 80 (1), 16-23 (2006).","journal-title":"J Leukoc Biol"},{"issue":"2","key":"ref22","doi-asserted-by":"crossref","first-page":"e2103","DOI":"10.21769\/BioProtoc.2103","type":"journal-article","article-title":"Isolation of Peripheral Blood Mononuclear Cells Using Vacutainer Cellular Preparation Tubes (CPT(TM))","volume":"7","author":"Puleo","year":"2017","unstructured":"Puleo, A., Carroll, C., Maecker, H. T., Gupta, R. Isolation of Peripheral Blood Mononuclear Cells Using Vacutainer Cellular Preparation Tubes (CPT(TM)). Bio Protoc. 7 (2), e2103 (2017).","journal-title":"Bio Protoc"},{"issue":"4","key":"ref23","doi-asserted-by":"crossref","first-page":"e0231132","DOI":"10.1371\/journal.pone.0231132","type":"journal-article","article-title":"A protocol for rapid monocyte isolation and generation of singular human monocyte-derived dendritic cells","volume":"15","author":"Chometon","year":"2020","unstructured":"Chometon, T. Q. et al. A protocol for rapid monocyte isolation and generation of singular human monocyte-derived dendritic cells. PLoS One. 15 (4), e0231132 (2020).","journal-title":"PLoS One"},{"key":"ref24","doi-asserted-by":"crossref","first-page":"114557","DOI":"10.1016\/j.jpba.2021.114557","type":"journal-article","article-title":"Subcellular metabolomics: Isolation, measurement, and applications","volume":"210","author":"Qin","year":"2022","unstructured":"Qin, S., Zhang, Y., Tian, Y., Xu, F., Zhang, P. Subcellular metabolomics: Isolation, measurement, and applications. J Pharm Biomed Anal. 210, 114557 (2022).","journal-title":"J Pharm Biomed Anal"},{"issue":"3","key":"ref25","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/0022-1759(80)90209-4","type":"journal-article","article-title":"Separation of human monocytes on density gradients of Percoll","volume":"33","author":"Pertoft","year":"1980","unstructured":"Pertoft, H., Johnsson, A., W\u00e4rmeg\u00e5rd, B., Seljelid, R. Separation of human monocytes on density gradients of Percoll. J Immunol Methods. 33 (3), 221-229 (1980).","journal-title":"J Immunol Methods"},{"issue":"1","key":"ref26","doi-asserted-by":"crossref","first-page":"52","DOI":"10.4049\/jimmunol.1300802","type":"journal-article","article-title":"Mixed proteasomes function to increase viral peptide diversity and broaden antiviral CD8+ T cell responses","volume":"191","author":"Zanker","year":"2013","unstructured":"Zanker, D., Waithman, J., Yewdell, J. W., Chen, W. Mixed proteasomes function to increase viral peptide diversity and broaden antiviral CD8+ T cell responses. J Immunol. 191 (1), 52-59 (2013).","journal-title":"J Immunol"},{"issue":"3","key":"ref27","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1053\/j.gastro.2012.12.002","type":"journal-article","article-title":"A dominant CD4(+) T-cell response to Helicobacter pylori reduces risk for gastric disease in humans","volume":"144","author":"Chen","year":"2013","unstructured":"Chen, L. et al. A dominant CD4(+) T-cell response to Helicobacter pylori reduces risk for gastric disease in humans. Gastroenterology. 144 (3), 591-600 (2013).","journal-title":"Gastroenterology"}],"container-title":["Journal of Visualized Experiments"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/app.jove.com\/t\/68291\/in-vitro-culture-for-h5n1-specific-duck-t-cells-detection-immune","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T15:20:42Z","timestamp":1792596042000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.jove.com\/t\/68291\/in-vitro-culture-for-h5n1-specific-duck-t-cells-detection-immune"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,30]]},"references-count":27,"journal-issue":{"issue":"219","published-online":{"date-parts":[[2025]]}},"URL":"https:\/\/doi.org\/10.3791\/68291","relation":{},"ISSN":["1940-087X"],"issn-type":[{"value":"1940-087X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,30]]}}}