Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes
Abstract Background Dairy cows experiencing ketosis after calving suffer greater disease incidence and are at greater risk of leaving the herd. In vitro administration of beta-hydroxybutyric acid (BHBA; the primary blood ketone) has inhibitory effects on the function of bovine leukocytes. BHBA is a...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-03-01
|
| Series: | Journal of Animal Science and Biotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40104-024-00999-5 |
| _version_ | 1797266637606354944 |
|---|---|
| author | Laman K. Mamedova Kirby C. Krogstad Paiton O. McDonald Laxman Pokhrel Duy H. Hua Evan C. Titgemeyer Barry J. Bradford |
| author_facet | Laman K. Mamedova Kirby C. Krogstad Paiton O. McDonald Laxman Pokhrel Duy H. Hua Evan C. Titgemeyer Barry J. Bradford |
| author_sort | Laman K. Mamedova |
| collection | DOAJ |
| description | Abstract Background Dairy cows experiencing ketosis after calving suffer greater disease incidence and are at greater risk of leaving the herd. In vitro administration of beta-hydroxybutyric acid (BHBA; the primary blood ketone) has inhibitory effects on the function of bovine leukocytes. BHBA is a ligand of HCAR2 and the activation of these receptors promotes an anti-inflammatory response which may be related with immunosuppression observed in transition dairy cattle. The objective of this study was to identify and test antagonists for HCAR2 in bovine immune cells cultured with BHBA. Results We observed expression of HCAR2 at the protein level within lymphocytes, monocytes, and granulocytes. The proportion of cells expressing HCAR2 tended to be greater in mid-lactation compared to early lactation cows; the increase was a result of increased proportion of T and B cells expressing HCAR2. Stimulation of HCAR2 with niacin or BHBA promoted Ca2+ mobilization in neutrophils and mononuclear cells. Mononuclear cells treated with BHBA had diminished intracellular Ca2+ responses when HCAR2 was knocked down by siRNA silencing, indicating Ca2+ mobilization was mediated by HCAR2 signaling. Two candidate antagonists for HCAR2, synthesized from niacin (NA-1 and NA-5), were tested; monocytes and neutrophils pre-treated with NA-1 and NA-5 had reduced Ca2+ mobilization after incubation with BHBA. Furthermore, NA-5 but not NA-1 prevented BHBA-associated reductions in cyclic AMP. Conclusions We demonstrated that HCAR2 is present on bovine leukocytes and has greater expression later in lactation. We confirmed that BHBA and niacin derived HCAR2 antagonists alter bovine leukocyte activity. Our results demonstrate that both BHBA and niacin affect bovine leukocyte Ca2+ mobilization in a HCAR2-dependent manner. |
| first_indexed | 2024-04-25T01:03:52Z |
| format | Article |
| id | doaj.art-6dd0be8d368f41e5b9bb82ad18a24cd9 |
| institution | Directory Open Access Journal |
| issn | 2049-1891 |
| language | English |
| last_indexed | 2024-04-25T01:03:52Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Animal Science and Biotechnology |
| spelling | doaj.art-6dd0be8d368f41e5b9bb82ad18a24cd92024-03-10T12:18:51ZengBMCJournal of Animal Science and Biotechnology2049-18912024-03-0115111110.1186/s40104-024-00999-5Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytesLaman K. Mamedova0Kirby C. Krogstad1Paiton O. McDonald2Laxman Pokhrel3Duy H. Hua4Evan C. Titgemeyer5Barry J. Bradford6Department of Animal Science, Michigan State UniversityDepartment of Animal Science, Michigan State UniversityComparative Medicine and Integrative BiologyDepartment of Chemistry, Kansas State UniversityDepartment of Chemistry, Kansas State UniversityDepartment of Animal Sciences and Industry, Kansas State UniversityDepartment of Animal Science, Michigan State UniversityAbstract Background Dairy cows experiencing ketosis after calving suffer greater disease incidence and are at greater risk of leaving the herd. In vitro administration of beta-hydroxybutyric acid (BHBA; the primary blood ketone) has inhibitory effects on the function of bovine leukocytes. BHBA is a ligand of HCAR2 and the activation of these receptors promotes an anti-inflammatory response which may be related with immunosuppression observed in transition dairy cattle. The objective of this study was to identify and test antagonists for HCAR2 in bovine immune cells cultured with BHBA. Results We observed expression of HCAR2 at the protein level within lymphocytes, monocytes, and granulocytes. The proportion of cells expressing HCAR2 tended to be greater in mid-lactation compared to early lactation cows; the increase was a result of increased proportion of T and B cells expressing HCAR2. Stimulation of HCAR2 with niacin or BHBA promoted Ca2+ mobilization in neutrophils and mononuclear cells. Mononuclear cells treated with BHBA had diminished intracellular Ca2+ responses when HCAR2 was knocked down by siRNA silencing, indicating Ca2+ mobilization was mediated by HCAR2 signaling. Two candidate antagonists for HCAR2, synthesized from niacin (NA-1 and NA-5), were tested; monocytes and neutrophils pre-treated with NA-1 and NA-5 had reduced Ca2+ mobilization after incubation with BHBA. Furthermore, NA-5 but not NA-1 prevented BHBA-associated reductions in cyclic AMP. Conclusions We demonstrated that HCAR2 is present on bovine leukocytes and has greater expression later in lactation. We confirmed that BHBA and niacin derived HCAR2 antagonists alter bovine leukocyte activity. Our results demonstrate that both BHBA and niacin affect bovine leukocyte Ca2+ mobilization in a HCAR2-dependent manner.https://doi.org/10.1186/s40104-024-00999-5Flow cytometryGPR109AHM74AKetosis |
| spellingShingle | Laman K. Mamedova Kirby C. Krogstad Paiton O. McDonald Laxman Pokhrel Duy H. Hua Evan C. Titgemeyer Barry J. Bradford Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes Journal of Animal Science and Biotechnology Flow cytometry GPR109A HM74A Ketosis |
| title | Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes |
| title_full | Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes |
| title_fullStr | Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes |
| title_full_unstemmed | Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes |
| title_short | Investigation of HCAR2 antagonists as a potential strategy to modulate bovine leukocytes |
| title_sort | investigation of hcar2 antagonists as a potential strategy to modulate bovine leukocytes |
| topic | Flow cytometry GPR109A HM74A Ketosis |
| url | https://doi.org/10.1186/s40104-024-00999-5 |
| work_keys_str_mv | AT lamankmamedova investigationofhcar2antagonistsasapotentialstrategytomodulatebovineleukocytes AT kirbyckrogstad investigationofhcar2antagonistsasapotentialstrategytomodulatebovineleukocytes AT paitonomcdonald investigationofhcar2antagonistsasapotentialstrategytomodulatebovineleukocytes AT laxmanpokhrel investigationofhcar2antagonistsasapotentialstrategytomodulatebovineleukocytes AT duyhhua investigationofhcar2antagonistsasapotentialstrategytomodulatebovineleukocytes AT evanctitgemeyer investigationofhcar2antagonistsasapotentialstrategytomodulatebovineleukocytes AT barryjbradford investigationofhcar2antagonistsasapotentialstrategytomodulatebovineleukocytes |