Computational protocol: Changes in Circulating B Cell Subsets Associated with Aging and Acute SIV Infection in Rhesus Macaques

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Protocol publication

[…] Multiparameter flow cytometry was performed using a BD FACSFortessa cell sorter operated by BD FACSDiva software (BD Biosciences, San Jose, CA). PBMC from rhesus macaques or human donors were stained with directly conjugated monoclonal antibodies against human (rhesus macaque cross-reactive) CD3 (clone SP34-2), CD20 (clone L27), CD21 (clone B-ly4), CD27 (clone M-T7271), IgM (clone G20-127), and IgG (clone G18-145) (all purchased from BD Biosciences) or directly conjugated polyclonal goat antibodies against human (rhesus macaque cross-reactive) IgD and IgA (both purchased from Southern Biotech, Birmingham, AL). Staining for intracellular immunoglobulins was performed with the Fixation/Permeabilization solution kit (BD Biosciences) according to the manufacturer’s instructions after surface staining with the appropriate antibodies using protocols previously described []. Data were analyzed and illustrated using FlowJo software (Tree Star Inc., Ashland, OR). The FACS data generated from post SIV infection samples can be found at FlowRepository (ID: FR-FCM-ZZVV, FR-FCM-ZYZZ, and FR-FCM-ZYZY). […]

Pipeline specifications

Software tools BD FACSDiva, FlowJo
Application Flow cytometry
Organisms Simian immunodeficiency virus, Macaca mulatta, Homo sapiens
Diseases Communicable Diseases, Infection, HIV Infections