Citations

Plasma antibody measurements

Pertussis antigen-specific antibody responses were quantified in human plasma by performing an indirect serological assay with xMAP Microspheres (details described in xMAP Cookbook, Luminex 5th edition). Pertussis, Tetanus, and Diphtheria antigens (PT, PRN, Fim2/3, TT, and DT (all from List Biological Laboratories) …

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)

• Product #186 – Fimbriae 2/3 from B. pertussis

• Product #187 – Pertactin from B. pertussis (69 kDa Protein)

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

• Product #190 – Tetanus Toxin from Clostridium tetani

Induction and assessment of EAE

Mice were subcutaneously injected with 200 mg MOG (myelin oligodendrocyte glycoprotein) 35–55 (MEVGWYRSPFSPVVHLYRNGKCOOH, eurofins) peptide emulsified in complete Freund’s adjuvant containing 400 mg of Mycobacterium tuberculosis H37Ra (Chondrex), in addition to two intraperitoneal injections of 200 ng of pertussis toxin (List Laboratories) on days 0 and 3. …

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)

4.1. Cell Line and Reagents for Development of the AP-1 Reporter Assay
Murine hepatoma Hepa1c1c7 cells (CRL-2026™) purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) were cultured in an alpha minimum essential medium (MEM), supplemented with 10% FBS, 100 IU/mL penicillin, 100 µg/mL streptomycin, and 2 mM L-glutamine. Cultures were maintained at 37 °C in an incubator with 5% CO2. Lyophilized recombinant protective antigen (PA) and lethal factor (LF) were purchased from List Biological Laboratories, Inc. (Campbell, CA, USA), and reconstituted in sterile water to make stock solutions with a final concentration of 1 mg/mL for LF and 2 mg/mL for PA. The stock solutions were then diluted 10 times with 50% glycerol in sterile water to make stock solutions with a final concentration of 0.1 mg/mL for LF and 0.2 mg/mL for PA. …

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

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… Pertussis toxin (200 ng/mouse, List Biological Laboratories, Inc., Campbell, CA, USA) in sterile saline was administered via the tail vein on day 0 and day 2 after MOG35-55 immunization. …

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)

CT intoxication of human monolayers

… After 3 days, the monolayers were challenged with 2 μg mL−1 CT (List Labs) on the apical side. …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Limbus processing procedures

… 0.1 nM cholera toxin (List Biological Laboratories, Campbell, California, USA), …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

… Pertussis toxin (List Biologicals) …

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)

4.2. Induction and Evaluation of EAE

We induced EAE by subcutaneous (s.c.) sensitization with 100 nmol/mouse of modified PLP139–151 peptide (VSLGKWLGHPDKF, United BioSystems, Herndon, VA, USA) in SJL/J mice, MOG92–106 peptide (DEGGYTCFFRDHSYQ, United BioSystems) in SJL/J mice or A.SW mice, or MOG35–55 (MEVGWYRSPFSRVVHLYRNGK, United BioSystems) peptide in C57BL/6 mice, in which myelin peptides were emulsified in CFA composed of incomplete Freund’s adjuvant (BD, Franklin Lakes, NJ, USA] and Mycobacterium tuberculosis H37 Ra (BD)…. To induce chronic MOG35–55-induced EAE, we injected C57BL/6 mice twice with MOG35–55 s.c. on days 0 and 19 and 300 ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA) i.p. on days 0 and 2 [63]. …

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)

EAE induction

All experiments were conducted under the approval of the Harvard Medical School Animal Care committee and the Animal Care. SJL/J mice were immunized subcutaneously in two sites (left and right flank) with 150 μg of PLP139-151 (New England Peptide LLC., Gardner, MA) emulsified in complete Freund’s adjuvant (CFA, Sigma Aldrich, Saint Louis, MO) containing 200 μg Mycobacterium Tuberculosis (Difco Laboratories, Detroit, MI). Mice received 200 ng pertussis toxin (PT, List Biological Laboratories Inc., Campbell, CA) in 0.2 ml PBS (Lonza, Walkersville, MD) intraperitoneally (ip) at the time of immunization and 48 h later. Control mice were immunized with CFA followed by PT (12). The mice were sacrificed at different time points. For MOG 35–55 (M-E-V-G-W-Y-R-S-P-F-S-R-O-V-H-L-Y-R-N-G-K) corresponding to mouse sequence is synthesized by QCB Inc. Division of BioSource International (Hopkinton, MA), and purified to > 99% by HPLC. Mice are immunized subcutaneously in the flanks with 150–200 μg of MOG peptide in 0.1 ml PBS and 0.1 ml CFA containing 0.4 mg Mycobacterium Tuberculosis (H37Ra, Difco, Detroit, Michigan) and injected i.p. with 200 ng Pertussis toxin (List Laboratories, Campbell, California) on the day of immunization and 2 days later. …

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)

Lipopolysaccharide, Bleomycin, House Dust Mite Administration

Lipopolysaccharide (LPS, Escherichia coli O111: B4, List Biological Laboratories, Campbell, CA, USA) was administered by oropharyngeal aspiration under isoflurane anesthesia at a dose of 2.0 mg/kg of mouse body weight in 50μL of PBS. …

• Product #201 – LPS from Escherichia coli O111:B4