Anti-Toxin A from Clostridium difficile (Goat)

EAE induction

For EAE induction, mice were injected subcutaneously at two sites on the back with 200 mg MOG35-55peptide (synthesised by Toray Research Center, Tokyo, Japan) and 1 mg heat-killed Mycobacterium tuberculosis H37RA emulsified in complete Freund's adjuvant (Difco, KS, USA). Four hundred nanograms of Pertussis toxin (List Biological Laboratories, CA, USA) were injected intraperitoneally on days 0 and 2 after immunisation. ...

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)

LFNEffector intoxication of T84 cells:

... For intoxication of cells, media was replaced over 105 cells previously seeded in a 12-well plate. 31.7 nM PA alone (List Labs, #171E) or in the presence of 13.6nM LFN, LFN-ACD, or LFN-RID was added to media and cells were incubated for 20 hr at 37C in the presence of 5% CO2. For co-intoxication of 13.6 nM LFNACD and LFN-RID, 3x excess PA was used to ensure equal translocation of both effectors.

• Product #171 - Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Protein expression and purification:

...Protein was detected by immunoblotting using commercial polyclonal goat anti-toxin A (List Biological Laboratories, Campbell, CA)...

Immunization Regimen:

...To perform a direct comparison to the formalin-inactivated toxoid vaccine currently in human clinical trials, we immunized one cohort of mice with the toxoid preparation by the i.p. route using alum as the adjuvant. Toxoid A and toxoid B from C. difficile...were purchased from List Biological Laboratories.