Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming
(1) Background: The latest research illustrates that microglia phenotype is not the binary ‘resting’ and ‘activated’ profiles. Instead, there is wide diversity in microglia states. Similarly, when testing different stimulation protocols for BV2 microglia, we discovered differences in the response of...
| Main Authors: | , , , , |
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| Format: | Article |
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Multidisciplinary Digital Publishing Institute
2023
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| Online Access: | http://psasir.upm.edu.my/id/eprint/108722/ |
| _version_ | 1848865198155235328 |
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| author | Pan, Meng Liy Ahmad Puzi, Nur Nabilah Ooi, Yin Yin Ramasamy, Rajesh Vidyadaran, Sharmili |
| author_facet | Pan, Meng Liy Ahmad Puzi, Nur Nabilah Ooi, Yin Yin Ramasamy, Rajesh Vidyadaran, Sharmili |
| author_sort | Pan, Meng Liy |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | (1) Background: The latest research illustrates that microglia phenotype is not the binary ‘resting’ and ‘activated’ profiles. Instead, there is wide diversity in microglia states. Similarly, when testing different stimulation protocols for BV2 microglia, we discovered differences in the response of the cells in terms of the production of intracellular ROS (iROS), nitric oxide (NO), CD40 expression, and migratory capacity. (2) Methods: BV2 microglia were treated with single interferon gamma (IFN-γ) stimulation, LPS/IFN-γ co-stimulation, and priming with IFN-γ followed by stimulation with LPS for 24 h. The responses of BV2 microglia were then assessed using the H2DCFDA test for iROS, the Griess assay for NO, immunophenotyping for CD40/CD11b/MHC II, and migration using a transwell apparatus. (3) Results: Single stimulation with IFN-γ induced NO but not ROS in BV2 microglia. Co-stimulation with LPS200IFN-γ2.5 induced a higher iROS production (a 9.2-fold increase) and CD40 expression (28031 ± 8810.2 MFI), compared to priming with primedIFN-γ50LPS100 (a 4.0-fold increase in ROS and 16764 ± 1210.8 MFI of CD40). Co-stimulation also induced cell migration. On the other hand, priming BV2 microglia (primedIFN-γ50LPS100) resulted in a higher NO production (64 ± 1.4 µM) compared to LPS200IFN-γ2.5 co-stimulation (44 ± 1.7 µM). Unexpectedly, priming inhibited BV2 migration. (4) Conclusions: Taken together, the findings from this project reveal the ability of co-stimulation and priming in stimulating microglia into an inflammatory phenotype, and the heterogeneity of microglia responses towards different stimulating approaches. |
| first_indexed | 2025-11-15T14:00:53Z |
| format | Article |
| id | upm-108722 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T14:00:53Z |
| publishDate | 2023 |
| publisher | Multidisciplinary Digital Publishing Institute |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1087222024-09-26T08:36:49Z http://psasir.upm.edu.my/id/eprint/108722/ Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming Pan, Meng Liy Ahmad Puzi, Nur Nabilah Ooi, Yin Yin Ramasamy, Rajesh Vidyadaran, Sharmili (1) Background: The latest research illustrates that microglia phenotype is not the binary ‘resting’ and ‘activated’ profiles. Instead, there is wide diversity in microglia states. Similarly, when testing different stimulation protocols for BV2 microglia, we discovered differences in the response of the cells in terms of the production of intracellular ROS (iROS), nitric oxide (NO), CD40 expression, and migratory capacity. (2) Methods: BV2 microglia were treated with single interferon gamma (IFN-γ) stimulation, LPS/IFN-γ co-stimulation, and priming with IFN-γ followed by stimulation with LPS for 24 h. The responses of BV2 microglia were then assessed using the H2DCFDA test for iROS, the Griess assay for NO, immunophenotyping for CD40/CD11b/MHC II, and migration using a transwell apparatus. (3) Results: Single stimulation with IFN-γ induced NO but not ROS in BV2 microglia. Co-stimulation with LPS200IFN-γ2.5 induced a higher iROS production (a 9.2-fold increase) and CD40 expression (28031 ± 8810.2 MFI), compared to priming with primedIFN-γ50LPS100 (a 4.0-fold increase in ROS and 16764 ± 1210.8 MFI of CD40). Co-stimulation also induced cell migration. On the other hand, priming BV2 microglia (primedIFN-γ50LPS100) resulted in a higher NO production (64 ± 1.4 µM) compared to LPS200IFN-γ2.5 co-stimulation (44 ± 1.7 µM). Unexpectedly, priming inhibited BV2 migration. (4) Conclusions: Taken together, the findings from this project reveal the ability of co-stimulation and priming in stimulating microglia into an inflammatory phenotype, and the heterogeneity of microglia responses towards different stimulating approaches. Multidisciplinary Digital Publishing Institute 2023-09-27 Article PeerReviewed Pan, Meng Liy and Ahmad Puzi, Nur Nabilah and Ooi, Yin Yin and Ramasamy, Rajesh and Vidyadaran, Sharmili (2023) Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming. Biomedicines, 11 (10). art. no. 2648. pp. 1-14. ISSN 2227-9059 https://www.mdpi.com/2227-9059/11/10/2648 10.3390/biomedicines11102648 |
| spellingShingle | Pan, Meng Liy Ahmad Puzi, Nur Nabilah Ooi, Yin Yin Ramasamy, Rajesh Vidyadaran, Sharmili Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming |
| title | Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming |
| title_full | Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming |
| title_fullStr | Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming |
| title_full_unstemmed | Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming |
| title_short | Response profiles of BV2 microglia to IFN-γ and LPS co-stimulation and priming |
| title_sort | response profiles of bv2 microglia to ifn-γ and lps co-stimulation and priming |
| url | http://psasir.upm.edu.my/id/eprint/108722/ http://psasir.upm.edu.my/id/eprint/108722/ http://psasir.upm.edu.my/id/eprint/108722/ |