Citations

Diet and treatment

Breeding cages were randomly assigned to one of the experimental isocaloric diets (AIN93G, BioServ): standard (Stand, BioServ #S7628), n-3 long chain (LC)-PUFA enriched (n-3, BioServ #S7630; soybean oil replaced with Menhaden Fish oil) or n-6 LC-PUFA supplemented (n-6, BioServ #S7629; based on corn oil). The detailed composition of the diets is presented in our previous publication (Chumak et al., 2022). The day of birth was defined as postnatal day (P) 0. At P9 pups received a single intraperitoneal injection of lipopolysaccharide (LPS from E. coli O55:B5, 1mg/kg, Cat# 423, List labs) or equivalent volume of saline. The dams remained on the experimental diet until offspring sacrifice (P10-P12). Efforts were taken to include equal number of pups of both sexes in each experimental group.

• Product #423 – ULTRA PURE LPS from Escherichia coli O55:B5

Diphtheria toxin (DT) injection

Mice received 500 ng diphtheria toxin (DT, Cat# 150, List Biological Laboratory) in 100 µL distilled, 486 deionized water (ddH2O) through intraperitoneal injection (i.p.) (Binnewies et al., 2019) on 31, 32 487 and 34 dpi (days post-injury), …

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

… 2) Diphtheria Toxin: DTx (List Labs, cat# 150) was suspended in filter sterilized phosphate buffered saline (PBS) to create stock aliquots of 2 mg/mL solution, stored at -80 °C. …

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

… The following antigens were used: … tetanus toxin (TTx) (List Biological Laboratories, Campbell, USA), …

• Product #190 – Tetanus Toxin from Clostridium tetani

Induction and evaluation of active EAE

For the construction of chimeric mice, Rag2-/- mice were adoptively transferred with 107 wild-type (Wt) T cells, Mid1-/- T cells, or a mixture of CD45.1-expressing Wt and CD45.2-expressing Mid1-/- T cells at a 1:1 ratio. Wt, Mid1-/-, or chimeric mice were then used for the induction of active EAE as previously reported 25. Briefly, mice were subcutaneously injected with a 200 μL emulsion that contains 1 mg/ml MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK), 2.5 mg/ml Mycobacteria tuberculosis H37Ra (B.D.), and 100 μL Freund’s adjuvant (Sigma). On the day of immunization and two days after immunization, mice were intraperitoneally injected with 200 ng of pertussis toxin (List Biological Laboratories Inc.).

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)

Processing of material for light microscopy.

… Subsequently, for fluorescence microscopy, slices were stained for GAD67 with mouse
anti-GAD67 (MAB5406; 1:2,000, Millipore), for VGLuT2 with guinea-pig anti-VGLuT2 (AB2251-I; 1:2,000, Millipore), for
CTB with goat anti-CTB (703; 1:2,000, List Labs), …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

Sample collection

Subjects underwent a first bronchoscopy (28 ± 2 days after placebo treatment) to collect pre-challenge baseline BAL from a segment of the left lower lobe using 100 mL of pre-warmed saline. Two baseline mucosal biopsies from the anterior segment of the left lower lobe and the right lower lobe were sampled. Segmental challenge with LPS (40 endotoxin units per kg body weight diluted in 10 mL of saline; endotoxin from E. coli Type O113; List Biological Laboratories Inc., Campbell, California, USA) was performed in the medial segment of the middle lobe and 10 mL saline (0.9%) was applied in the medial segment of the lingula as control. …

• Product #433 – LIST™ HPT™ from Escherichia coli O113

Induction of EAE

Experimental animals were subcutaneously injected with 200 μL of myelin oligodendrocyte glycoprotein35-55 (MOG) peptide at 1 mg/mL (Koma Biotech, Seoul, Korea) emulsified in complete Freund’s adjuvant (CFA; Sigma-Aldrich, St. Louis, MO, USA) supplemented with Mycobacterium tuberculosis H37Ra (5 mg/mL; Difco Laboratories, Franklin Lakes, NJ, USA). All mice were injected intraperitoneally with 500 ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA) on the day of induction and again 2 days later. After immunization, all mice were weighed and observed daily for clinical signs of EAE, …

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)

… 0.2 µg/ml pertussis toxin (List Biological …

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)

Adjuvant-immunized EAE and ESS mouse model

The induction of EAE was described in our previous studies.77,78 Briefly, female C57/BL6 mice at 8 weeks of age were induced by
subcutaneous immunization in the back region with 200 mg of the MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK, GL Biochem)
in complete Freund’s adjuvant (CFA, Chondrex) containing 20 mg/mL Mycobacterium tuberculosis H37Ra (BD difco). At 2 h and 48 h
postimmunization, 200 mg doses of pertussis toxin (List Biological Laboratories) were diluted in PBS and intravenously injected into EAE model mice.

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)