Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows

ABSTRACT: Nutritional and metabolic state in dairy cows are important determinants of the immune response. During the periparturient period, a state of negative energy balance in the cow increases plasma concentrations of fatty acids (FA), which are associated with inflammation. Among immune cells,...

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Main Authors: Bingbing Zhang, Jianan Wen, Ming Li, Jingjing Wang, Ziwei Ji, Xinquan Lv, Muhammad Usman, John Mauck, Juan J. Loor, Wei Yang, Guihua Wang, Jinzhu Ma, Chuang Xu
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:Journal of Dairy Science
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Online Access:http://www.sciencedirect.com/science/article/pii/S0022030224012438
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author Bingbing Zhang
Jianan Wen
Ming Li
Jingjing Wang
Ziwei Ji
Xinquan Lv
Muhammad Usman
John Mauck
Juan J. Loor
Wei Yang
Guihua Wang
Jinzhu Ma
Chuang Xu
author_facet Bingbing Zhang
Jianan Wen
Ming Li
Jingjing Wang
Ziwei Ji
Xinquan Lv
Muhammad Usman
John Mauck
Juan J. Loor
Wei Yang
Guihua Wang
Jinzhu Ma
Chuang Xu
author_sort Bingbing Zhang
collection DOAJ
description ABSTRACT: Nutritional and metabolic state in dairy cows are important determinants of the immune response. During the periparturient period, a state of negative energy balance in the cow increases plasma concentrations of fatty acids (FA), which are associated with inflammation. Among immune cells, CD4+ T are able to function under high-FA conditions, but the underlying mechanisms regulating these events remain unclear. The objective of this study was to clarify the functional mechanisms of CD4+ T cells under high-FA conditions. The effects of glycolysis and calcium release-activated calcium modulator 1 (ORAI1) on migration of CD4+ T cells exposed to high FA were investigated in vivo and in vitro. The CD4+ T cells were isolated from peripheral blood of healthy (n = 9) and high-FA (n = 9) Holstein cows (average 2.5 ± 0.2 lactations [SE], 12.3 ± 0.8 DIM). In the first experiment, real-time quantitative PCR was used to assess chemokine receptors in isolated CD4+ T cells and migration capacity. The relative mRNA measurements results revealed downregulation of CCR1 and CXCR2, and upregulation of CCR2, CCR4, CCR5, CCR7, CCR8, CCR10, CXCR1, CXCR3, CXCR4, and CX3CR1. Among them, the expression of CXCR4 was relatively high. Therefore, CXCL12, a ligand chemokine of CXCR4, was an inducer of CD4+ T cell migration. The CD4+ T cells were inoculated in the upper chamber and CXCL12 (100 ng/mL, Peprotech) in RPMI1640 was added to the lower chamber and transmigrated for 3 h at 37°C and 5% CO2. The cell migration assay revealed that the migration capacity of CD4+ T cells from high-FA cows was greater. Real-time-qPCR indicated greater abundance of the glycolysis-related targets HIF1A, HK2, PKM2, Glut1, GAPDH, and LDHA and Western blotting indicated greater abundance of the glycolysis-related targets HIF1A, HK2, PKM2, Glut1, GAPDH, and LDHA in CD4+ T cells of high-FA cows. To characterize specific mechanisms of CD4+ T cell migration in vitro, cells from the spleens of 3 newborn (1 d old, 40–50 kg) healthy female Holstein calves were isolated after euthanasia. Inhibition of glycolysis attenuated the migration ability of cells, but had no effect on the protein and mRNA abundance of store-operated Ca2+ entry (SOCE)-associated calcium release-activated calcium modulator 1 (ORAI1) and stromal interaction molecule 1 (STIM1). In contrast, ORAI1 was upregulated in CD4+ T cells of cows exposed to high FA. To explore the potential mechanisms whereby an active glycolytic metabolism affects CD4+ T cells under high-FA conditions, we knocked down ORAI1 using small interfering RNA (siORAI1). Isolated CD4+ T cells from high-FA cows with the siORAI1 had an attenuated glycolytic metabolism and migration capacity. Taken together, these data suggested that calcium ions in CD4+ T cells from cows with high FA regulate glycolytic metabolism and influence cell migration at least in part by modulating ORAI1. Thus, these studies identified a novel mechanism of Ca2+ regulation of CD4+ T cell glycolytic metabolism affecting their migration through the SOCE pathway.
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spelling doaj-art-4c524b0aed6249398bc2a8aa4cd6a4622025-08-20T02:39:08ZengElsevierJournal of Dairy Science0022-03022025-01-01108185686710.3168/jds.2024-24845Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cowsBingbing Zhang0Jianan Wen1Ming Li2Jingjing Wang3Ziwei Ji4Xinquan Lv5Muhammad Usman6John Mauck7Juan J. Loor8Wei Yang9Guihua Wang10Jinzhu Ma11Chuang Xu12College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, ChinaCollege of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China; College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 032699, ChinaCollege of Veterinary Medicine, China Agricultural University, Beijing 100000, ChinaCollege of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, ChinaCollege of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, ChinaCollege of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, ChinaMammalian Nutri Physio Genomics, Department of Animal Sciences and Division of Nutritional Sciences, University of Illinois, Urbana, IL 61801Mammalian Nutri Physio Genomics, Department of Animal Sciences and Division of Nutritional Sciences, University of Illinois, Urbana, IL 61801Mammalian Nutri Physio Genomics, Department of Animal Sciences and Division of Nutritional Sciences, University of Illinois, Urbana, IL 61801College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, ChinaCollege of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, ChinaCollege of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, ChinaCollege of Veterinary Medicine, China Agricultural University, Beijing 100000, China; Corresponding authorABSTRACT: Nutritional and metabolic state in dairy cows are important determinants of the immune response. During the periparturient period, a state of negative energy balance in the cow increases plasma concentrations of fatty acids (FA), which are associated with inflammation. Among immune cells, CD4+ T are able to function under high-FA conditions, but the underlying mechanisms regulating these events remain unclear. The objective of this study was to clarify the functional mechanisms of CD4+ T cells under high-FA conditions. The effects of glycolysis and calcium release-activated calcium modulator 1 (ORAI1) on migration of CD4+ T cells exposed to high FA were investigated in vivo and in vitro. The CD4+ T cells were isolated from peripheral blood of healthy (n = 9) and high-FA (n = 9) Holstein cows (average 2.5 ± 0.2 lactations [SE], 12.3 ± 0.8 DIM). In the first experiment, real-time quantitative PCR was used to assess chemokine receptors in isolated CD4+ T cells and migration capacity. The relative mRNA measurements results revealed downregulation of CCR1 and CXCR2, and upregulation of CCR2, CCR4, CCR5, CCR7, CCR8, CCR10, CXCR1, CXCR3, CXCR4, and CX3CR1. Among them, the expression of CXCR4 was relatively high. Therefore, CXCL12, a ligand chemokine of CXCR4, was an inducer of CD4+ T cell migration. The CD4+ T cells were inoculated in the upper chamber and CXCL12 (100 ng/mL, Peprotech) in RPMI1640 was added to the lower chamber and transmigrated for 3 h at 37°C and 5% CO2. The cell migration assay revealed that the migration capacity of CD4+ T cells from high-FA cows was greater. Real-time-qPCR indicated greater abundance of the glycolysis-related targets HIF1A, HK2, PKM2, Glut1, GAPDH, and LDHA and Western blotting indicated greater abundance of the glycolysis-related targets HIF1A, HK2, PKM2, Glut1, GAPDH, and LDHA in CD4+ T cells of high-FA cows. To characterize specific mechanisms of CD4+ T cell migration in vitro, cells from the spleens of 3 newborn (1 d old, 40–50 kg) healthy female Holstein calves were isolated after euthanasia. Inhibition of glycolysis attenuated the migration ability of cells, but had no effect on the protein and mRNA abundance of store-operated Ca2+ entry (SOCE)-associated calcium release-activated calcium modulator 1 (ORAI1) and stromal interaction molecule 1 (STIM1). In contrast, ORAI1 was upregulated in CD4+ T cells of cows exposed to high FA. To explore the potential mechanisms whereby an active glycolytic metabolism affects CD4+ T cells under high-FA conditions, we knocked down ORAI1 using small interfering RNA (siORAI1). Isolated CD4+ T cells from high-FA cows with the siORAI1 had an attenuated glycolytic metabolism and migration capacity. Taken together, these data suggested that calcium ions in CD4+ T cells from cows with high FA regulate glycolytic metabolism and influence cell migration at least in part by modulating ORAI1. Thus, these studies identified a novel mechanism of Ca2+ regulation of CD4+ T cell glycolytic metabolism affecting their migration through the SOCE pathway.http://www.sciencedirect.com/science/article/pii/S0022030224012438high FACD4+ T cellsglycolysisSOCEmigration
spellingShingle Bingbing Zhang
Jianan Wen
Ming Li
Jingjing Wang
Ziwei Ji
Xinquan Lv
Muhammad Usman
John Mauck
Juan J. Loor
Wei Yang
Guihua Wang
Jinzhu Ma
Chuang Xu
Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows
Journal of Dairy Science
high FA
CD4+ T cells
glycolysis
SOCE
migration
title Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows
title_full Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows
title_fullStr Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows
title_full_unstemmed Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows
title_short Fatty acids promote migration of CD4+ T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows
title_sort fatty acids promote migration of cd4 t cells through calcium release activated calcium modulator orai1 sensitive glycolysis in dairy cows
topic high FA
CD4+ T cells
glycolysis
SOCE
migration
url http://www.sciencedirect.com/science/article/pii/S0022030224012438
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