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通过流式细胞术对mHLA-DR和nCD64进行定量以研究宿主反应失调:QuantiBRITE™ PE微球的应用及其稳定性。

Quantitation of mHLA-DR and nCD64 by Flow Cytometry to Study Dysregulated Host Response: The Use of QuantiBRITE™ PE Beads and Its Stability.

作者信息

Sanju S, Jain Paresh, Priya Veeraraghavan Vishnu, Varma Praveen K, Mony Ullas

机构信息

Centre for Nanosciences and Molecular Medicine, Amrita VishwaVidyapeetham, Kochi, 682041, Kerala, India.

BD Biosciences, Singapore, Singapore.

出版信息

Appl Biochem Biotechnol. 2023 Sep;195(9):5747-5752. doi: 10.1007/s12010-022-03819-6. Epub 2022 Jan 11.

Abstract

Quantitation of mHLA-DR and nCD64 is useful in understanding the dysregulated host response. The down regulation of HLA-DR expression on the circulating monocytes (mHLA-DR) is associated with anti-inflammatory response, and an increased expression of CD64 on neutrophil surface (nCD64) is associated with pro-inflammatory response. Quantitation of these antigen expression using beads (QuantiBRITE™ PE) is a precision technique. These beads are reported to be stable for 24 h after reconstitution. We report the results of our investigation examining the stability of QuantiBRITE PE beads over a period of 4-week post-reconstitution. The data suggest that reconstituted QuantiBRITE PE beads, if stored in dark at 2-8 °C, can be effectively used for up to 2 weeks for determining nCD64 and mHLA-DR antibody bound per cell (ABC) values.

摘要

定量检测可溶性人类白细胞抗原DR(mHLA-DR)和中性粒细胞表面CD64(nCD64)有助于理解失调的宿主反应。循环单核细胞上HLA-DR表达下调(mHLA-DR)与抗炎反应相关,而中性粒细胞表面CD64表达增加(nCD64)与促炎反应相关。使用微珠(QuantiBRITE™ PE)定量这些抗原表达是一项精确技术。据报道,这些微珠复溶后24小时内稳定。我们报告了复溶后4周内对QuantiBRITE PE微珠稳定性进行研究的结果。数据表明,复溶后的QuantiBRITE PE微珠若在2-8°C避光保存,可有效用于测定每细胞结合的nCD64和mHLA-DR抗体(ABC)值,最长可达2周。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/888e/8748532/f4ea0397072f/12010_2022_3819_Fig1_HTML.jpg

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