Public Library of Science
ppat.1011811.g001.tif (1.28 MB)

Sorting and proportion of FMDV serotype A-specific B cells via different bait antigens by flow cytometry.

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posted on 2023-11-20, 18:46 authored by Kun Li, Yong He, Li Wang, Pinghua Li, Huifang Bao, Shulun Huang, Shasha Zhou, Guoqiang Zhu, Yali Song, Ying Li, Sheng Wang, Qianliang Zhang, Pu Sun, Xingwen Bai, Zhixun Zhao, Zhiyong Lou, Yimei Cao, Zengjun Lu, Zaixin Liu

Bovine PBMCs (A) were analyzed by 3-color flow cytometry and the gate P1 (B) was selected to exclude cells debris with lower values of SSC-A and FSC-A, and further analyzed singlets in gate P2 (C) based on diagonal streak of the FSC-A and FSC-H plot. The class-switched (IgM-) B cells in gate P3 were used to check the distribution of CD21 and FMDV-specific cells, in the presence of FMDV bait antigen A/GDMM/2013 (D) and A/AF72 (E), respectively. The demarcation line between FMDV-specific binding and nonbinding B cells was set according to the FMO control (F) in absence of bait antigen. The CD21+/- Antigen+ B cells in gate P4 were separately sorted for preparation of bovine mAbs. Appropriately one million bovine PBMCs were collected to analyze the distribution proportion of A/GDMM/2013-binding (G) and A/AF72-binding (H) B cells in circulating peripheral blood.