Bacterial Toxin Research Citations

Reagents and antibodies (Abs)

LPS from Escherichia coli O55:B5 and recombinant mouse CX3CL1 were purchased from LIST Biological Laboratories (Campbell, CA) and R&D Systems (Minneapolis, MN), respectively. …

Animal model:

LPS-induced PTL was conducted similarly as described previously [16]. In brief, two female mice were co-housed with a male one of the same genotype and checked for vaginal plugs as the evidence of mating at the next morning. The day of vaginal plug detection was designated as gd 0.5 of pregnancy, and the pregnant mice were removed from the male. All pregnant mice delivered their pups on gd 19 or 20. At gd 15.5, the pregnant mice were intraperitoneally administered with LPS (25 g in 200 l of PBS) or PBS 200 l, followed by the observation until delivery. The presence of intact or partial fetal tissue in the cage was noted as the evidence for delivery, and the delivery before gd 18.5 was judged as pre-term labor, and pups born before gd 18.5 could not survive >24 h after birth [30]. In our dose, 25 g was the highest dose of LPS that reproducibly induced PTL without significant morbidity or mortality in gravid C57BL/6 females, based on the established method [31, 32]. In some experiments, pregnant WT mice were i.p. injected with anti-CX3CL1 Ab (200 g/mouse) or control IgG at 3 h after LPS injection, and were observed until delivery. The percentage of preterm labor was calculated as follows: percentage (%) = 100 x (number of LPS-treated mice that delivered until gd 18.5/number of LPS-treated pregnant mice). All animal experiments were approved by the Committee on Animal Care and Use at Wakayama Medical University.

Blood and tissue sampling:

At gd 5.5, 10.5, 15.5, 18.5, or 6 h after LPS or PBS injection, mice were anesthetized by the intraperitoneal injection of Avertin (tribromoethanol, 20 g/g), and whole blood samples were taken, followed by centrifugation (3,000 x g for 15 min) to collect serum. In each pregnant mouse, uterus was removed, and viable fetuses were dissected from the amniotic sac and umbilical cord, to weigh the obtained fetuses and placentas. Gestational tissues including uterus (from between implantation sites), placenta and amnion were stored at -80C until processing for the subsequent analyses.

• Product #203A – LPS from Escherichia coli O55:B5

… Migration was calculated by dividing the number of migrated cells by the number of input cells. In some studies, cells were preincubated for 1 hour at 37°C with PTX (100 ng/ml; 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)

… The following reagents were purchased from commercial vendors: … Ctx (List Biological Laboratories, #101C, Campbell, CA), … 

cAMP analysis:

Cellular concentrations of cAMP were examined using the Direct Immunoassay Detection Kit (Abcam, ab138880, Cambridge, MA). Briefly, cells were treated with Ctx (50 µg/mL, 4 h), lysed, and the protein amounts were measured. The cell lysates were loaded into anti-cAMP antibody-coated plates. The HRP-linked cAMP was added to compete with the cellular cAMP. The activity of HRP-cAMP conjugate was measured using a microplate reader. A calibration curve of free cAMP was analyzed in parallel. The cAMP concentrations in different cells were normalized by the total protein amount in cell lysates.

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

Experimental procedures:

Male TLR4^+/+ C57BL/6 mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA). TLR4-null (TLR4^-/-) mice were bred at our facility on a C57BL/6 background [53]. Mice were allowed access to rodent chow and water ad libitum and used at the age of 8 and 12 weeks. Since numerous commercial LPS preparations contain measurable contaminating proteins, for this study we utilized ultra-purified LPS (from Escherichia coli O111:B4) purchased from List Biological Laboratories, Inc. (Campbell, CA). …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

… At the same time mice were also administered 250 ng of pertussis toxin (List Biological Laboratories, Campbell, California) i.p. and again on d2. …

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)

Experimental autoimmune encephalomyelitis:

EAE was induced by immunization with 100µg per mouse of MOG₃₅₅₅peptide emulsified in CFA (Difco Laboratories) and i.p. injection of pertussis toxin (150ng per mouse; List Biological Laboratories, Inc.) on days 0 and 2. …

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)

… Experimental autoimmune encephalitis (EAE) was induced in control and Gch1;Lck mice by immunization with an emulsion of 100µg myelin oligodendrocyte glycoprotein (MOG)3555 in complete Freunds adjuvant … We injected 100 µl MOG/CFA subcutaneously above the inguinal lymph node on both sides of the mouse. We then injected 200µl pertussis toxin/PBS … List Biological Labs) intraperitoneally per mouse on days zero and one. Scoring for EAE was performed as described over the course of 45 days. …

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)

… Single retrograde tracings were carried out with cholera toxin B subunit (CTB; List Biological Laboratories: 104) labeling, while doubleretrograde tracings used CTB and Fluorogold (FG; Fluorochrome). …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt

… Pertussis toxin List Biological Cat# 181 …

In vivo suppression assay:

EAE was induced by subcutaneous immunization with 100 ug myelin oligodendrocytepeptide (MOG_35-55; Anaspec) in complete Freunds adjuvant, with 200 ng intraperitoneal pertussis toxin (List Biological) given on days 0 and 2 after immunization. …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

Active induction of EAE:

Mice were anesthetized with 200 mg/kg tribromomethanol (1.5% Avertin) injected i.p. Active EAE was induced by s.c. injections into the flanks with 200µg of mouse myelin oligodendrocyte glycoprotein₃₅₅₅ (MOG_p3555) (MEVGWYRSPFSRVVHLYRNGK; CS Bio Menlo Park, CA, USA) emulsified in complete Freund’s adjuvant (CFA) (DIFCO Laboratories, Detroit, MI, USA) containing 400 g of heat inactivated Mycobacterium tuberculosis (Difco, Detroit, MI, USA). Mice also received i.p. injections of 200 ng pertussis toxin on days 0 and 2 (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)