Bacterial Toxin Research Citations

Cell lines, bacterial strains, toxin, and antibodies:

…Purified C. difficile toxin B was purchased from List Biological Laboratories (Campbell, CA). …

RTCA version 2 assay:

…After the CI reached a plateau, the CI for each sample well was normalized to one at the last measurement time point prior to toxin treatment and inoculated with eluted samples or purified CDT to be tested. The nCI was then automatically monitored at 5-min intervals for 36 h… Each run included one positive control with 100 ng/ml purified CDT in duplicate and one negative control in quadruplicate.

The time point at which the nCI dropped by 30% was considered a positive step time (PST). PST was used to calculate the toxin concentrations in samples by comparing standard curves which were developed by monitoring the nCI at serial 3-fold dilutions of purified CDT ranging from 0.01 to 787.32 ng/ml and plotting the PST versus the purified CDT concentration. The limit of detection of the RTCA version 2 assay was determined by testing 11 3-fold serial dilutions of purified CDT (concentrations ranging from 0.01 to 787.32 ng/ml). All testing was done in triplicate.

• Product #155 – Toxin B from Clostridium difficile

Intravitreal Injections and Factors:

Balb/c mice were anesthetized as described, and the vitreous of the right eye (treated) was injected with Rho GTPases activator or inactivators and/or growth factors (GFs), …Other factors injected included the general inhibitor for Rho GTPase, Toxin A, from C. difficile (10 ng/eye, No. 152C; List Biological Laboratories, Inc., Campbell, CA, USA), …

• Product #152C – Toxin A from Clostridium difficile

Iso- lated epithelial cells were suspended in a keratinocyte culture medium (KCM) composed of a basal mixture of 3 parts Dulbeccos modified Eagles medium (DMEM) and …1 nM cholera toxin (List Biological Laboratories, Campbell, CA), …

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

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

Reagents:

…LPS from E. coli serotype 0111:B4 was purchased from List Biological Laboratories (Campbell, CA). …

Animal protocols:

C57BL/6 and TLR-4^/ mice (78 weeks age) …Using methods that were previously described, the intratracheal (IT) instillation of live E. coli, E. coli 0111:B4 LPS (1 g/g), or PBS was performed in mice anesthetized with 34% isoflurane …

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

EAE:

…Mycobacterium tuberculosis H37Ra (Difco). Each mouse was subsequently administered 500 ng pertussis toxin (List Biological Laboratories) ip on days 0 and 2 after MOG 3555 peptide immunization. The EAE clinical manifestations…

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:

… Pertussis toxin (400 ng) (List Biological Laboratories, Campbell, CA, USA) in PBS and 50 mM NaCl was administered ip on the day of immunization and again after 24 h. For the treatment of EAE, EB (25 and 50 mg/kg) or PBS (vehicle control) was administered po daily started …

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)

Induction and evaluation of EAE:

Mice were immunized for the induction of EAE using either a 1 or 2 immunization protocol. …On the day of immunization, each mouse received 200 ng of PTX (List Biological Laboratories, Campbell, CA, USA) by intervenous 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)

In vivo cellular depletion:

For cDC depletion, 25 ng/g of diphtheria toxin (DT; List Biological Laboratories Inc.) was administered intraperitoneally (IP) into the chimeric CD11c-DTR mice. Mice were sacrificed 12 hours later and bone marrow, blood, liver, spleen and lung were harvested for further analysis. For pDC depletion, mice were injected IP with 500 g 120G8 every other day for 14 days. 120G8 and control antibody were purchased from Imgenex. Mice were sacrificed 24 hours after the last injection and livers were harvested. For Foxp3+CD4+T cell depletion, DT was injected IP at a dose of 50ng/g for 2 consecutive days into Foxp3-DTR mice. Mice were sacrificed 48h after the last dose.

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

DT/Diphtheria Toxin Mutant Administration:

CD11b-DTR mice were treated with DT, 15 ng/g body weight (List Biological Laboratories, Campbell CA), by intraperitoneal injection. Control CD11b-DTR mice received the same amount of mutated DT (DTm) (List Biological Laboratories), which does not bind to the DTR. For the current study, we determined the optimal dose schedule of DT administration that could be safely used to evaluate skeletal muscle regeneration allowing long-term survival of the mice. Mice were divided into groups and received one dose of DT from 15 to 35 ng/g body weight in 5 ng/g steps in dosages between groups. The higher DT dose group (range, 20 to 35 ng/g body weight) resulted in 75% mortality at days 7 to 12 after DT injection. By contrast, mice in the low-dose DT group (15 ng/g body weight) had a mortality rate of 17% and could therefore be used in experiments requiring a 21-day time point. To determine whether multiple doses could be used, a cohort of mice was given two to three doses of DT (10 to 15 ng/g body weight) at least 1 week apart. Most of these mice died at days 7 to 12. Therefore, we chose a single dose of 15 ng/g body weight DT to ablate CD11b⁺ cells. These conditions allowed for the survival of the animals through the course of the experiment (21 days) while temporarily ablating the monocyte/macrophage population within a specific time frame. Previous work and our results in the kinetics of DT ablation of monocyte/macrophage populations in multiple tissues including regenerating skeletal muscle and blood indicate that ablation occurs within 12 hours, lasts 24 hours, with recovery generally occurring 48 hours after DT administration.^{7 ;  15} Single injections of DT were administered at various times (0.5, 0, 1, 2, and 4 days) relative to the injection of CTX to transiently ablate monocyte/macrophages at different time points during skeletal muscle regeneration.

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #149 – Diphtheria Toxin CRM₁₉₇, Mutant