Active EAE and adoptive transfer EAE
In the active EAE model, FACS-sorted MP CD4+ T cells (γδTCR–NK1.1–Vβ11–TCRβ+CD4+CD1d tetramer–Foxp3–CD62LlowCD44high, 5 × 105) from Foxp3-GFP mice were adoptively transferred into 5-week-old female C57BL/6 mice. After the transfer, the mice were immunized with 200 μg of MOG35-55 peptide in complete Freund’s adjuvant (Chondrex, Inc., Woodinville, WA, USA). At 0 and 48 h after immunization, the mice were intraperitoneally treated with 500 ng of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA). The animals were scored daily for clinical disease.
In another EAE model established by adoptive transfer, naive (CD4+Vβ11+CD25–CD62LhighCD44low) CD45.1− T cells (1–5 × 104) from 2D2 TCR-transgenic mice were transferred into Rag–/– mice with or without WT CCR6high, WT CCR6low, Bhlhe40–/– CCR6high, GM-CSF–/– CCR6high, GM-CSF–/– CCR6low, Il1r1–/– CCR6high or Il1r1–/– CCR6low MP CD4+ T cells (CD45.1+γδTCR–NK1.1–Vβ11–TCRβ+CD4+CD1d tetramer–CD25–CD62Llow CD44high, 1.0 × 105). Before the transfer, CCR6high or CCR6low MP CD4+ T cells were primed in vitro with IL-7 (10 ng/mL) and IL-1β (20 ng/mL) for 5–7 days. After the transfer, the mice were immunized with 100 μg of MOG35−55 peptide in complete Freund’s adjuvant. At 0 and 48 h after immunization, the mice were intraperitoneally injected with 200 ng of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA). …
Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)