Blood stage Plasmodium falciparum antigens induce immunoglobulin class switching in human enriched B cell culture.
This study aimed to demonstrate class switch recombination (CSR) in heavy chain expressing immunoglobulin G (IgG) and IgE in human B cells after exposure to Plasmodium falciparum schizont lysate. Human B cells (CD20+CD27-) were cultured with crude P. falciparum antigen (cPfAg) and anti-CD40. On Day 4 post-exposure, total RNA from B cells was prepared and the occurrence of CSR from IgM to IgG and/or IgE was investigated by reverse transcription-polymerase chain reaction. Molecular markers to detect active CSR included enzyme activation-induced cytidine deaminase mRNA, gamma and epsilon-germline transcripts (gamma, epsilon-GLT), circle transcript (CT) and mature transcript (gamma and epsilon-mRNA) expression. On Day 7 and Day 14 after exposure, levels of Igs in the culture supernatant were determined by enzyme-linked immunosorbent assay. Our findings showed that we could demonstrate cPfAg-stimulated B cells undergoing CSR by use of the expressed CSR markers and the increase in specific IgG and IgE indicating the potential of this approach in the study of CSR in P. falciparum-stimulated B cells.
Potup
Pachuen
P
Department of Tropical Radioisotopes, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Kumsiri
Ratchanok
R
Kano
Shigeyuki
S
Kalambaheti
Thareerat
T
Looareesuwan
Sornchai
S
Troye-Blomberg
Marita
M
Maneerat
Yaowapa
Y
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology / Q000469:parasitology*; D015415:Biomarkers; D002478:Cells, Cultured; D006801:Humans; D017578:Immunoglobulin Class Switching; D007073:Immunoglobulin E / Q000235:genetics / Q000276:immunology; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D010963:Plasmodium falciparum / Q000276:immunology*; D012333:RNA, Messenger / Q000032:analysis / Q000276:immunology
D000953:Antigens, Protozoan; D015415:Biomarkers; D007074:Immunoglobulin G; D012333:RNA, Messenger; D007073:Immunoglobulin E