Bacterial Toxin Research Citations

Nasal vaccination:

Mice were anesthetized with isourane before vaccination. Vaccines were administered intranasally in 10 ml (5ml per nostril). Nonchimeric mice were vaccinated on days 0, 7, and 21. Chimeric mice were vaccinated on days 0, 7, 21, and 42. For each chimeric study replicate, mice were divided into groups of 4 to 8. Vaccine groups included PBS alone, recombinant LF (rLF) alone (#172; List Biological Laboratories Cambell, CA; 4 endotoxin units [EU]/mg), rLF + 4 mg IL-1a (400-ML; 0.00144 EU/mg; R&D, Minneapolis, MN), and rLF + 1 mg cholera toxin (CT) (EU/mg unknown; List #100B). For three of the four Il1r1 chimeric mouse experiments, mice were vaccinated with 25 mg LF for the day 0, 7, and 21 immunizations and 50 mg LF on day 42. For the remaining experiment, mice were immunized with 50 mg LF each time. The calculated endotoxin delivery per mouse was 0.20 (LF alone) to 0.22 EU (LF + IL-1a). Data from each group for all repeat experiments were combined and analyzed together. Mice vaccinated with LF + CT served as a positive control, and LF + CT-vaccinated mice responded similarly in all four chimeric groups. …

• Product #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Cells and Reagents:

… Stimulations were performed using 100 ng/mL of Salmonella minnesota ultra pure lipopolysaccaride (LPS) (List Biological Laboratories), …

• Product #434 – ULTRA PURE LPS from Salmonella minnesota R595 (Re)

… PAMP exposures were performed for 6 h, except in the time-course experiments. PAMPs used for BMDM and cell line exposures were from InvivoGen, except for E. coli O111:B4 LPS, which was from List Biological Laboratories. …

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

Animal maintenance and neuron ablation

… All except Tph2^lox/lox mice were on the C57Bl/6 background (> 9 generations backcrossed); Tph2^lox/lox mice were on a mixed 129/Sv x C57Bl/6 background. Agrp^DTR/DTR male mice were bred with Slc17a6^lox/lox female mice to generate Agrp^DTR/+; Slc17a6^lox/+ mice, which were further bred to each other to create Agrp^DTR/+; Slc17a6^lox/lox mice and Agrp^DTR/+; Slc17a6^lox/+ control mice. Similar breeding strategy was adopted when generating Agrp^DTR/+; Tph2^lox/lox mice and Agrp^DTR/+; Grin1^lox/lox mice. Mice were group housed with standard chow diet (LabDiet 5053) and water provided ad libitum until the beginning of the experiments. All experiments were performed with ~8-wk-old male mice. To ablate AgRP neurons, mice carrying the Agrp^DTR allele, were injected twice with DT (50 g/kg; intramuscular; 2 d apart; List Biological Laboratories, Campbell, CA)⁴. The extent of ablation (>95%) was determined by immunohistochemistry⁴⁷.

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

Adoptive transfer, EAE challenge, and TGF- neutralization:

To isolate T_reg cells, CD4⁺CD25 and CD4⁺CD25⁺ T cells were obtained from spleens and head and neck lymph nodes (HNLNs) from H696- or H647-vaccinated mice. …

To induce EAE, SJL females were given s.c 200 µg of the encephalitogenic PLP peptide (PLP₁₃₉₁₅₁; HSLGKWLGHPDKF, HPLC-purified to >90%; Biosynthesis, Inc., Lewisville, TX) in 200 µl of complete Freunds adjuvant. On days 0 and 2, mice received i.p. 200 ng of Bordetella pertussis toxin (List Biological Laboratories; Campbell, CA), as previously described …

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)

Sample Preparation:

To provide a source of data that was rich in content for functional annotation, we prepared a complex sample of soluble proteins from neutrophils activated by lipopolysaccharide (LPS). The human promyelocytic HL-60 cell line (ATCC) was differentiated in culture using 1 M alltrans retinoic acid (Sigma-Aldrich), 6 pM 1,25-Dihydroxyvitamin D3 (Sigma-Aldrich), and 30 ng/mL granulocyte-colony stimulatory factor (Invitrogen) in Iscove’s Modified Dulbecco’s medium (Mediatech, Inc.) supplemented with 20% FBS and 4 mM L-glutamine.³¹ Cells were activated via treatment with 100 ng/mL of lipopolysaccharide (LPS) from E. coli O111:B4 (List Biological Laboratories) for 30 minutes. …

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

EAE induction, disability evaluation and tissue harvesting:

… For the mouse model, 8-10 weeks-old female C57BL/6 mice (Taconic, Hudson, NY) were immunized subcutaneously in the flank with 200 µg myelin oligodendrocyte glycoprotein peptide 3555 (MEVGWYRSPFSRVVHLYRNGK; Stanford PAN Facility, Stanford, CA) emulsified in CFA that had been supplemented with M. tuberculosis strain H37RA (MT; Difco Laboratories, Detroit, MI) at 7 mg/ml. All mice also received an intraperitoneal injection of 350 ng pertussis toxin (List Biological, Campbell, CA) at 0 and 48 h post-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)

… Cholera toxin B (CTB) subunit-biotin conjugates was from List Biological Laboratories, Inc. (Campbell, CA). …

• Product #112 – Cholera Toxin B Subunit Biotin Conjugate>

Flow cytometry:

… Medium alone and 10 g/ml tetanus toxoid protein (List Biological Laboratories, Inc.) were used as negative controls, …

• Product #191 – Tetanus Toxoid from Clostridium tetani

Experimental protocols:

After completion of the surgical procedures, anesthetized rats were allowed to stabilize for 60 min before initiation of experimental protocols. As described by Cui et al. (4, 6), rats received LeTx [LF (100 g/kg) + PA (200 g/kg) infused at 0.5 ml/h] or vehicle (PA + PBS with 1% BSA or PBS with 1% BSA infused at 0.5 ml/h). LF and PA are recombinant proteins prepared from B. anthracis and purchased from List Biological Laboratories. Experiments were completed in intact and SAD rats. LeTx or vehicle was infused intravenously via a femoral venous catheter, and the dose of LeTx was chosen on the basis of the time course of physiological responses to intravenous LeTx infusions as reported by Cui and colleagues (6). MAP, HR, and SND were recorded continuously during intravenous LeTx or vehicle infusions. The maximum infusion duration was 6 h; however, because of LeTx-induced reductions in arterial blood pressure, the majority of experiments were terminated before 6 h. Additional experiments involved the intravenous infusion of sodium nitroprusside (SNP) at a dose (20 g/kg) that reduced basal levels of MAP to 5060 mmHg. At the end of experiments, rats were euthanized by an overdose of methohexital sodium (150 mg/kg iv).

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

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