Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infection

Avian Leukosis Virus (ALV) can induce tumorigenesis and immune suppression by acting on lymphocytes in the bone marrow. In this study, single-cell RNA sequencing (scRNA-seq) was used to analyze chicken bone marrow lymphocytes under Avian Leukosis Virus subtype J (ALV-J) infection. Using subgroup-spe...

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Main Authors: Cheng Liu, Yu Zhang, Ruyu Liao, Lecheng Wang, Xinyi Zhou, Min Tan, Keyun Xu, Haiwei Wang, Qigui Wang, Yongju Zhao, Zhifu Cui, Xi Lan
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:Poultry Science
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Online Access:http://www.sciencedirect.com/science/article/pii/S0032579125002342
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author Cheng Liu
Yu Zhang
Ruyu Liao
Lecheng Wang
Xinyi Zhou
Min Tan
Keyun Xu
Haiwei Wang
Qigui Wang
Yongju Zhao
Zhifu Cui
Xi Lan
author_facet Cheng Liu
Yu Zhang
Ruyu Liao
Lecheng Wang
Xinyi Zhou
Min Tan
Keyun Xu
Haiwei Wang
Qigui Wang
Yongju Zhao
Zhifu Cui
Xi Lan
author_sort Cheng Liu
collection DOAJ
description Avian Leukosis Virus (ALV) can induce tumorigenesis and immune suppression by acting on lymphocytes in the bone marrow. In this study, single-cell RNA sequencing (scRNA-seq) was used to analyze chicken bone marrow lymphocytes under Avian Leukosis Virus subtype J (ALV-J) infection. Using subgroup-specific marker genes and cell state analysis, we identified 18 distinct cell clusters, including 8 T cell clusters, 2 B cell clusters, 5 tumor-like cell clusters, and 3 unidentified clusters. Gene expression analysis revealed that in the 10 T/B lymphocyte clusters, the differentially expressed genes in double-positive T cells, B1-like B cells, and cytotoxic T cells were highly enriched in pathways related to viral infection and immune response. These three cell populations exhibited high proportions and significant changes after infection, suggesting a strong immune response to ALV-J infection. Additionally, during ALV-J infection, the proportion of regulatory T cells and CTLA4 T cells increased, while immune suppressive factors TGFB1 and IL16 were highly expressed across the cell populations, indicating an immune-suppressive state in bone marrow lymphocytes. Moreover, ALV-J infected all cell populations; however, within the same cluster, only a fraction of the cells expressed ALV-J viral genes. Notably, in all cells expressing ALV-J viral genes, the ''Rho family GTPase signaling pathway'' associated with antiviral responses was activated. The Rho family, which is a key regulator of cytoskeletal reorganization and cell polarity, also plays a critical role in tumor cell proliferation and metastasis. Further analysis using Ingenuity Pathway Analysis (IPA) software predicted key upstream regulators of immune response, such as MYC and MCYN. In conclusion, this study identifies key genes and signaling pathways involved in immune responses of different lymphocyte subpopulations triggered by ALV-J infection in bone marrow. These findings contribute to a better understanding of the immune mechanisms in ALV-J-infected bone marrow lymphocytes and provide insights for discovering breeding loci for ALV-J resistance.
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publisher Elsevier
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series Poultry Science
spelling doaj-art-a3f7d676c8f14ddf9fca16e11ca035222025-08-20T02:30:59ZengElsevierPoultry Science0032-57912025-05-01104510499510.1016/j.psj.2025.104995Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infectionCheng Liu0Yu Zhang1Ruyu Liao2Lecheng Wang3Xinyi Zhou4Min Tan5Keyun Xu6Haiwei Wang7Qigui Wang8Yongju Zhao9Zhifu Cui10Xi Lan11College of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaChongqing Academy of Animal Science, Chongqing, ChinaChongqing Academy of Animal Science, Chongqing, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, ChinaCollege of Animal Science and Technology, Southwest University, Chongqing 400715, China; Corresponding authors.College of Animal Science and Technology, Southwest University, Chongqing 400715, China; Corresponding author at: No. 2 Tiansheng Road, College of Animal Science and Technology, Southwest University, Chongqing 400715, China.Avian Leukosis Virus (ALV) can induce tumorigenesis and immune suppression by acting on lymphocytes in the bone marrow. In this study, single-cell RNA sequencing (scRNA-seq) was used to analyze chicken bone marrow lymphocytes under Avian Leukosis Virus subtype J (ALV-J) infection. Using subgroup-specific marker genes and cell state analysis, we identified 18 distinct cell clusters, including 8 T cell clusters, 2 B cell clusters, 5 tumor-like cell clusters, and 3 unidentified clusters. Gene expression analysis revealed that in the 10 T/B lymphocyte clusters, the differentially expressed genes in double-positive T cells, B1-like B cells, and cytotoxic T cells were highly enriched in pathways related to viral infection and immune response. These three cell populations exhibited high proportions and significant changes after infection, suggesting a strong immune response to ALV-J infection. Additionally, during ALV-J infection, the proportion of regulatory T cells and CTLA4 T cells increased, while immune suppressive factors TGFB1 and IL16 were highly expressed across the cell populations, indicating an immune-suppressive state in bone marrow lymphocytes. Moreover, ALV-J infected all cell populations; however, within the same cluster, only a fraction of the cells expressed ALV-J viral genes. Notably, in all cells expressing ALV-J viral genes, the ''Rho family GTPase signaling pathway'' associated with antiviral responses was activated. The Rho family, which is a key regulator of cytoskeletal reorganization and cell polarity, also plays a critical role in tumor cell proliferation and metastasis. Further analysis using Ingenuity Pathway Analysis (IPA) software predicted key upstream regulators of immune response, such as MYC and MCYN. In conclusion, this study identifies key genes and signaling pathways involved in immune responses of different lymphocyte subpopulations triggered by ALV-J infection in bone marrow. These findings contribute to a better understanding of the immune mechanisms in ALV-J-infected bone marrow lymphocytes and provide insights for discovering breeding loci for ALV-J resistance.http://www.sciencedirect.com/science/article/pii/S0032579125002342ALV-JscRNA-seqBone marrowLymphocytesImmune suppression
spellingShingle Cheng Liu
Yu Zhang
Ruyu Liao
Lecheng Wang
Xinyi Zhou
Min Tan
Keyun Xu
Haiwei Wang
Qigui Wang
Yongju Zhao
Zhifu Cui
Xi Lan
Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infection
Poultry Science
ALV-J
scRNA-seq
Bone marrow
Lymphocytes
Immune suppression
title Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infection
title_full Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infection
title_fullStr Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infection
title_full_unstemmed Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infection
title_short Single-cell RNA sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup J infection
title_sort single cell rna sequencing of bone marrow reveals the immune response mechanisms of lymphocytes under avian leukosis virus subgroup j infection
topic ALV-J
scRNA-seq
Bone marrow
Lymphocytes
Immune suppression
url http://www.sciencedirect.com/science/article/pii/S0032579125002342
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