Multicolor flow cytometry-based immunophenotyping for preclinical characterization of nanotechnology-based formulations: an insight into structure activity relationship and nanoparticle biocompatibility profiles

Frontiers in Allergy

Allergy

Jenny Zhang¹  Edward Cedrone²  Hannah S. Newton²  Jeffrey D. Clogston²  Alison Vermilya²  Jie Xu²  Marina A. Dobrovolskaia²  Ragi Unnithan³  Duncan Donohue³  Tyler Malys³ 

[1] Agilent Technologies, Santa Clara, CA, United States;Nanotechnology Characterization Laboratory, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research Sponsored by the National Cancer Institute, Frederick, MD, United States;Statistics Department, Frederick National Laboratory for Cancer Research Sponsored by the National Cancer Institute, Frederick, MD, United States;

关键词: nanomaterials;    immunophenotyping;    flow cytometry;    lymphocytes;    monocytes;    nanoparticles;   

DOI  :  10.3389/falgy.2023.1126012

 received in 2022-12-16, accepted in 2023-06-14,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionImmunophenotyping, which is the identification of immune cell subsets based on antigen expression, is an integral technique used to determine changes of cell composition and activation in various disease states or as a response to different stimuli. As nanoparticles are increasingly utilized for diagnostic and therapeutic applications, it is important to develop methodology that allows for the evaluation of their immunological impact. Therefore, the development of techniques such as immunophenotyping are desirable. Currently, the most common technique used to perform immunophenotyping is multicolor flow cytometry.MethodsWe developed two distinct multicolor flow cytometry immunophenotyping panels which allow for the evaluation of the effects of nanoparticles on the composition and activation status of treated human peripheral blood mononuclear cells. These two panels assess the presence of various lymphoid and myeloid-derived cell populations as well as aspects of their activation statuses—including proliferation, adhesion, co-stimulation/presentation, and early activation—after treatment with controls or nanoparticles. To conduct assay performance qualification and determine the applicability of this method to preclinical characterization of nanoparticles, we used clinical-grade nanoformulations (AmBisome, Doxil and Feraheme) and research-grade PAMAM dendrimers of different sizes (G3, G4 and G5) and surface functionalities (amine-, carboxy- and hydroxy-).Results and DiscussionWe found that formulations possessing intrinsic fluorescent properties (e.g., Doxil and AmBisome) interfere with accurate immunophenotyping; such interference may be partially overcome by dilution. In the absence of interference (e.g., in the case of dendrimers), nanoparticle size and surface functionalities determine their effects on the cells with large amine-terminated dendrimers being the most reactive.

【 授权许可】

Unknown   
© 2023 Newton, Zhang, Donohue, Unnithan, Cedrone, Xu, Vermilya, Malys, Clogston and Dobrovolskaia.