Effect of storage conditions on subpopulations of peripheral blood T lymphocytes isolated from naïve cattle and cattle infected with foot-and-mouth disease virus
© 2016 American Society for Veterinary Clinical Pathology.
| Veröffentlicht in: | Veterinary clinical pathology. - 1975. - 45(2016), 1 vom: 14. März, Seite 110-5 |
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| 1. Verfasser: | |
| Weitere Verfasser: | , , , |
| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2016
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| Zugriff auf das übergeordnete Werk: | Veterinary clinical pathology |
| Schlagworte: | Journal Article Research Support, Non-U.S. Gov't Cryopreservation flow cytometry immunophenotyping peripheral blood mononuclear cells storage |
| Zusammenfassung: | © 2016 American Society for Veterinary Clinical Pathology. BACKGROUND: Immunophenotyping of blood lymphocytes by flow cytometry is important in infectious disease research. In animal experiments and other longitudinal studies, the processing, prompt staining, and analysis of fresh samples is a logistical challenge and daily assay variation can confound data interpretation OBJECTIVE: This study examined the feasibility of cryopreservation and deferred analysis of bovine peripheral blood T lymphocytes from normal or infected animals METHODS: Peripheral blood mononuclear cells were collected from 4 naïve Holstein steers and 4 steers infected with foot-and-mouth-disease virus serotype Asia1. Identical aliquots were labeled and analyzed immediately, labeled for deferred analysis, or stored at -70°C or over liquid nitrogen for up to 3 weeks before labeling RESULTS: Freezing of unlabeled cells induced statistically significant changes in phenotypic recognition. In infected animals, the γδ T-cell population increased by 28% and CD8(+) αβT cells by 32%, while total CD3(+) cells decreased by 16%, and CD4(+) αβT cells decreased by 12%. Subsequent storage of frozen cells for the duration of the study, however, had no significant effect. There was less than 20% relative change in subpopulation sizes, and storage at -70°C or over liquid nitrogen was equivalent CONCLUSIONS: Depending on the objectives and practical limitations of a study, deferred labeling of peripheral blood lymphocytes can be a viable option. Although frozen storage of lymphocytes can introduce some artifactual distortion of relative cell populations, frozen cells can be maintained in storage until all samples in a longitudinal study can be analyzed in batch under standardized conditions and without introducing further bias |
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| Beschreibung: | Date Completed 28.02.2018 Date Revised 28.02.2018 published: Print-Electronic Citation Status MEDLINE |
| ISSN: | 1939-165X |
| DOI: | 10.1111/vcp.12327 |