Immunophenotyping of PBMCs by flow cytometry

WM Weijie Ma
JZ Jie Zeng
SC Shuai Chen
YL Yue Lyu
KT Kyra A. Toomey
CP Chinh T. Phan
KY Ken Y. Yoneda
TL Tianhong Li
ask Ask a question
Favorite

Fresh or thawed PBMCs were immunophenotyped for T cell subsets and reported as percentages of total PBMCs and as percentages of total T cells in the case of T cell subtypes. Specifically, PBMCs were stained with well-characterized antibodies against markers of interest, including CD3, CD4 plus CD8 according to standard protocols. Results were analyzed using a BD Fortessa multi-color flow cytometer and FlowJo 7.6.1 program (Ashland, OR). Further, immunophenotypic changes of major innate and adaptive immune cells of ICI were evaluated using a minimum of a million PMBCs collected before and after cancer therapy for individual lung cancer patients by a 24-color antibody panel using the “Aurora” Spectral cytometer (Cytek Biosciences, CA). The data analysis and statistical evaluations of this complex data set were performed using the Cytobank (Cytobank Inc., CA) [27].

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A