Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures
Human whole blood cultures are widely used for the investigation of physiological pathways and drug effects in vitro. Detailed information on the effect of “sample aging” (the time span between blood collection and experimental start) on the experimental outcome is not readily available in the publi...
Saved in:
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2018-01-01
|
| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2018/8901485 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832554787274489856 |
|---|---|
| author | H. W. Grievink M. Moerland |
| author_facet | H. W. Grievink M. Moerland |
| author_sort | H. W. Grievink |
| collection | DOAJ |
| description | Human whole blood cultures are widely used for the investigation of physiological pathways and drug effects in vitro. Detailed information on the effect of “sample aging” (the time span between blood collection and experimental start) on the experimental outcome is not readily available in the public domain. We studied the effect of sample aging on the ability of immune cells to respond to cell-specific immune triggers (LPS, PMA/ionomycin, and SEB). Sample aging at room temperature profoundly inhibited the LPS-induced monocytic cytokine release in minimally diluted whole blood cultures. The reduction ranged from 20–50% after 30 minutes to 80–100% after 10 hours and differed between cytokines (IL-1β, IL-2, IL-6, IFNγ, and TNFα). Sample storage at 4°C or 37°C even worsened this. PMA/ionomycin- and SEB-induced cytokine release, both mainly T-cell-driven, were also reduced by sample aging but to a lesser extent (20–50% after 24 hours). Intracellular cytokine staining revealed that the number of LPS-responding cells was not impacted by sample aging and reduced LPS responsivity could also not be explained by apoptosis or downregulated TLR4 expression. Thus, we speculate that sample aging induces an inhibitory pathway downstream from TLR4 in monocytes. These results underline the importance of quick sample handling when investigating innate immune responses in whole blood, especially for monocyte responses. |
| format | Article |
| id | doaj-art-6fe536c7297a4a72937b3d6246a5c382 |
| institution | Kabale University |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2018-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-6fe536c7297a4a72937b3d6246a5c3822025-02-03T05:50:33ZengWileyJournal of Immunology Research2314-88612314-71562018-01-01201810.1155/2018/89014858901485Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood CulturesH. W. Grievink0M. Moerland1Centre for Human Drug Research, Zernikedreef 8, 2333 CL Leiden, NetherlandsCentre for Human Drug Research, Zernikedreef 8, 2333 CL Leiden, NetherlandsHuman whole blood cultures are widely used for the investigation of physiological pathways and drug effects in vitro. Detailed information on the effect of “sample aging” (the time span between blood collection and experimental start) on the experimental outcome is not readily available in the public domain. We studied the effect of sample aging on the ability of immune cells to respond to cell-specific immune triggers (LPS, PMA/ionomycin, and SEB). Sample aging at room temperature profoundly inhibited the LPS-induced monocytic cytokine release in minimally diluted whole blood cultures. The reduction ranged from 20–50% after 30 minutes to 80–100% after 10 hours and differed between cytokines (IL-1β, IL-2, IL-6, IFNγ, and TNFα). Sample storage at 4°C or 37°C even worsened this. PMA/ionomycin- and SEB-induced cytokine release, both mainly T-cell-driven, were also reduced by sample aging but to a lesser extent (20–50% after 24 hours). Intracellular cytokine staining revealed that the number of LPS-responding cells was not impacted by sample aging and reduced LPS responsivity could also not be explained by apoptosis or downregulated TLR4 expression. Thus, we speculate that sample aging induces an inhibitory pathway downstream from TLR4 in monocytes. These results underline the importance of quick sample handling when investigating innate immune responses in whole blood, especially for monocyte responses.http://dx.doi.org/10.1155/2018/8901485 |
| spellingShingle | H. W. Grievink M. Moerland Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures Journal of Immunology Research |
| title | Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures |
| title_full | Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures |
| title_fullStr | Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures |
| title_full_unstemmed | Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures |
| title_short | Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures |
| title_sort | sample aging profoundly reduces monocyte responses in human whole blood cultures |
| url | http://dx.doi.org/10.1155/2018/8901485 |
| work_keys_str_mv | AT hwgrievink sampleagingprofoundlyreducesmonocyteresponsesinhumanwholebloodcultures AT mmoerland sampleagingprofoundlyreducesmonocyteresponsesinhumanwholebloodcultures |