Development of a High-Color Flow Cytometry Panel for Immunologic Analysis of Tissue Injury and Reconstruction in a Rat Model.

IF 2.9 4区 生物学 Q1 ANATOMY & MORPHOLOGY
Kenneth M Adusei, Tran B Ngo, Andrea Lucia Alfonso, Ravi Lokwani, Sabrina DeStefano, Maria Karkanitsa, Jacquelyn Spathies, Stephen M Goldman, Christopher L Dearth, Kaitlyn Noelle Sadtler
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引用次数: 3

Abstract

The rat model is an important resource in biomedical research due to its similarities to the human immune system and its use for functional studies. However, because of the preponderance of mouse models in foundational and mechanistic immunological studies, there is a relative lack of diverse, commercially available flow cytometry antibodies for immunological profiling in the rat model. Available antibodies are often conjugated to common fluorophores with similar peak emission wavelengths, making them hard to distinguish on conventional flow cytometers and restricting more comprehensive immune analysis. This can become a limitation when designing immunological studies in rat injury models to investigate the immune response to tissue injury. In addition, this lack of available antibodies limits the number of studies that can be done on the immune populations in lymphoid organs in other research areas. To address this critical unmet need, we designed a spectral flow cytometry panel for rat models. Spectral cytometry distinguishes between different fluorophores by capturing their full emission spectra instead of their peak emission wavelengths. This flow cytometry panel includes 24 distinct immune cell markers to analyze the innate and adaptive immune response. Importantly, this panel identifies different immune phenotypes, including tolerogenic, Type 1, and Type 2 immune responses. We show that this panel can identify unique immune populations and phenotypes in a rat muscle trauma model. We further validated that the panel can identify distinct adaptive and innate immune populations and their unique phenotypes in lymphoid organs. This panel expands the scope of previous rat panels providing a tool for scientists to examine the immune system in homeostasis and injury while pairing mechanistic immunological studies with functional studies.

用于大鼠组织损伤和重建模型免疫分析的高彩色流式细胞仪面板的研制。
大鼠模型是生物医学研究的重要资源,因为它与人类免疫系统相似,可用于功能研究。然而,由于小鼠模型在基础和机制免疫学研究中的优势,相对缺乏用于大鼠模型免疫学分析的多样化、市售的流式细胞术抗体。现有抗体通常与具有相似峰值发射波长的常见荧光团结合,这使得它们难以在常规流式细胞仪上区分,并限制了更全面的免疫分析。当设计大鼠损伤模型的免疫学研究以研究组织损伤的免疫反应时,这可能成为一个限制。此外,这种抗体的缺乏限制了其他研究领域对淋巴器官免疫群体的研究数量。为了解决这一关键的未满足需求,我们为大鼠模型设计了一个光谱流式细胞仪面板。光谱细胞术通过捕获它们的全发射光谱而不是它们的峰值发射波长来区分不同的荧光团。流式细胞术面板包括24种不同的免疫细胞标记物来分析先天和适应性免疫反应。重要的是,该小组确定了不同的免疫表型,包括耐受性,1型和2型免疫反应。我们表明,该面板可以在大鼠肌肉创伤模型中识别独特的免疫群体和表型。我们进一步证实,该小组可以识别出不同的适应性和先天免疫群体及其在淋巴器官中的独特表型。该小组扩大了以前的大鼠小组的范围,为科学家提供了一种工具来检查免疫系统的稳态和损伤,同时将机制免疫学研究与功能研究结合起来。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Cells Tissues Organs
Cells Tissues Organs 生物-发育生物学
CiteScore
4.90
自引率
3.70%
发文量
45
审稿时长
6-12 weeks
期刊介绍: ''Cells Tissues Organs'' aims at bridging the gap between cell biology and developmental biology and the emerging fields of regenerative medicine (stem cell biology, tissue engineering, artificial organs, in vitro systems and transplantation biology). CTO offers a rapid and fair peer-review and exquisite reproduction quality. Special topic issues, entire issues of the journal devoted to a single research topic within the range of interests of the journal, are published at irregular intervals.
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