Citations

… Goat anti-CTb (1:5000) List Biological Laboratories Inc., CA, USA. Cat#703; …

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

C. difficile cytotoxin assay:

Cytotoxicity assay was performed as previously described with the following modifications 189. Briefly, ATCC CCL-81 Vero cells were grown to confluence in Dulbecco modified Eagle medium (Gibco 11965) supplemented with 10% fetal bovine serum (16140) and 1% Penicillin streptomycin (Gibco 15070). Cells were plated to a density of 105 cells/well. Mouse cecal content was diluted 1:10 in sterile PBS, passed 46 through a 0.22m filter, and serially diluted to 10-6 . Filtered samples were tested in duplicate with a corresponding control which both antitoxin (Techlab T5000) and sample were added. A positive control of C. difficile TcdA (List Biologicals 152C) was used. Samples were incubated overnight at 37C and the cytotoxic titer was determined as the reciprocal of the highest dilution that produced 80% cell rounding.

• Product #152C – Toxin A from Clostridium difficile

Cell Culture:

… cholera toxin (100 ng/ml; List Biological Laboratories …

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

Test substances:

… Clostridium difficile Toxin A (ToxA) was purchased from List Laboratories, Inc. (Campbell, CA). …

Evaluating endpoint intestinal monolayer barrier integrity and IEC viability:

… Cytotoxic responses were also examined for BSA, ToxA, and WGA along with positive and negative controls. As expected, TX-100 caused complete loss of T84 monolayer viability even upon a single exposure. EDTA and BSA did not elicit any cytotoxicity, nor did a single exposure to WGA or ToxA (Fig. 5H). Repeated exposures to WGA caused a significant degree of loss in MTT conversion indicating loss of cell viability. Repeated exposures to ToxA caused complete loss of T84 monolayer viability, a strikingly different response than observed with a single exposure to ToxA, which elicited no change in T84 monolayer viability (Fig. 5H). …

• Product #152C – Toxin A from Clostridium difficile

… Along with immunization and two days post immunization, 100 ng pertussis toxin (List-Biological-Laboratories) in Dulbecco’s phosphate buffered saline (DPBS; Sigma Aldrich) was administered by intraperitoneal injection …

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)

 EAM Induction

To induce EAM, we injected mice with 125 g myosin heavy chain peptide MyHC_614-629 (Ac-SLKLMATLFSTYASAD; Genscript) (Pummerer et al., 1996) emulsified in CFA (Sigma-Aldrich) supplemented with 5mg/mL heat-killed Mycobacterium tuberculosis strain H37Ra (Difco) on days 0 and 7. On the first day of immunization, mice also received a dose of 500 ng pertussis toxins intraperitoneally (List Biological Laboratories) (Cihkov et al., 2004).

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)

… Cholera toxin (Lot#10067A1) was obtained from List Biological Laboratories, Inc. (Campbell, USA. …

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

… On the day of immunization and 2 days post-immunization, each mouse received 200 ng of Pertussis toxin (List Biological Laboratories, 180) as an adjuvant by intraperitoneal injection. …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

Animal Procedures:

A schematic representation of the experimental design is shown in Figure 1. On Experimental Days 0, 7, 14, 21 and 28, mice (n = 8/group) were orally sensitized to 20 mg of the hens egg protein OVA (grade V; Sigma-Aldrich, Zwijndrecht, The Netherlands) dissolved in 0.5 mL phosphate-buffered saline (PBS) containing 10 µg cholera toxin (CT; List Biological Laboratories, Campbell, CA, USA) as an adjuvant. The PBS-sensitized control mice (n = 6) received CT alone (10 µg/0.5 mL PBS). …

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

Reagents:

… Cholera toxin was from list biological laboratories; …

Results:

Cyclic AMP signaling reprograms cDC2 cells from a pro-Th2 to a pro-Th17 phenotype We previously reported that cAMP signaling in CD11c+ bone marrow derived DCs (BMDCs), i.e., BM-derived antigen presenting cells(BM-APCs), affectsthe differentiation of CD4+ T cells(Datta et al., 2010; Lee et al., 2015). Low cAMP levels, as occurs in GnasCD11c mice (generated by breeding of floxed Gnas with CD11c-Cre deleter strains), provoke a Th2 polarization that leads to an allergic phenotype (Lee et al., 2015), while cholera toxin (CT) and other treatmentsthat increase cAMP levels in CD11c+ cells, induce differentiation to Th17 cells (Datta et al., 2010). Given these observations, it was important to investigate how this phenotypic reprogramming occurs in bona fide DCs.

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