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In Vitro Supplementation of Copper Modulates the Functional Th1/Th2 Phenotype of Peripheral Blood Mononuclear Cells in Cattle

Bunting, M; Challice, B; van Winden, S

Authors

M Bunting

B Challice

S van Winden



Abstract

Simple Summary This study investigated the association of copper levels and the appearance of blood monocytes, a white blood cell. One type of monocytes (M1) supports cellular immunity and the M2 monocyte helps the immune system through antibodies. Five samples of cow monocytes (PBMCs) were incubated in different levels of copper; 0, 4, 8 and 16 mu M. After stimulation under three different conditions, we stained them for CD14 and CD16 to allow typing of the monocytes (M1 and M2). M1 function was also measured through nitric oxide (NO) production. The results showed a significant reduction in viability of the monocytes with increased copper (p < 0.001). Increasing copper resulted in more M1 type monocytes in cows older than 4 years (p = 0.001). CD14 expression affected both CD16 (M2) expression and NO production. For CD16 expression, there was a further significant negative effect of copper levels in cows older than 4 years, whereas NO was not affected by the varying copper levels. In our small sample, monocytes incubating in a higher copper environment showed a stronger M1 support for better cellular immunity containing intracellular infections more effectively. In the live animal low copper levels could possibly affect progression of a bacterial infection to clinical disease. This study investigated the association of copper levels and monocyte plasticity between M1 (CD14(+) CD16(-)) and M2 (CD14(-) CD16(++)) phenotypes. Five samples of female bovine PBMCs were incubated in 0, 4, 8 and 16 mu M copper and stimulated (PPD-A, TLR- 2 ligand (Pam(3)CSK(4)), or media alone) before they were washed and stained for cell surface expression analysis by flow cytometry. M1 function was measured through nitric oxide production using a Griess assay. Flow cytometry analysis showed a significant reduction in viability with increased copper (p < 0.001). Increasing copper had a significant impact on CD14 expression (p = 0.026) and in cows older than 4 years copper levels positively affected CD14 expression (p = 0.001), whereas in animals of four years or younger, Cu did not affect the CD14 expression (p = 0.701 and 0.939, respectively). CD14 expression affected both CD16 expression and NO production. For CD16 expression, there was a further significant negative effect of copper levels in cows older than 4 years, NO was not affected by varying copper levels. In our small sample, monocytes in the presence of a higher copper environment showed a stronger M1 support for better cellular immunity which might contain intracellular infections more effectively. To test this, a randomised clinical trial will be required to determine whether copper supplementation could prevent progression to Johne's disease in MAP infected cows.

Citation

Bunting, M., Challice, B., & van Winden, S. (2021). In Vitro Supplementation of Copper Modulates the Functional Th1/Th2 Phenotype of Peripheral Blood Mononuclear Cells in Cattle. Animals, 11(9), https://doi.org/10.3390/ani11092739

Journal Article Type Article
Acceptance Date Sep 16, 2021
Publication Date 2021
Deposit Date Nov 18, 2021
Publicly Available Date Mar 29, 2024
Print ISSN 2076-2615
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 11
Issue 9
DOI https://doi.org/10.3390/ani11092739
Keywords cattle; paratuberculosis; MAP; Johne's disease; copper; PBMC; AVIUM SUBSP-PARATUBERCULOSIS; MYCOBACTERIUM-PARATUBERCULOSIS; MACROPHAGE; SURVIVAL; GROWTH; ZINC
Public URL https://rvc-repository.worktribe.com/output/1553838