Bacterial Toxin Research Citations

Liver Metastasis Model:

Tumor cells (5 10⁵/100 L phosphate-buffered saline [PBS]) were injected into the spleens of C57BL/6, CCR2 knockout (KO), severe combined immunodeficiency (SCID), or CD11b-diphtheria toxin receptor (DTR) mice. …CD11b-DTR or C57BL/6 mice received diphtheria toxin (7.5 ng DT/g body weight; List Biological Laboratories)…

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

Dieser Effekt konnte unabhngig vom Dt-Hersteller (Merck, List Biological Laboratories, Sigma-Aldrich) und in unterschiedlich hohen Dosen (1 g, 0,5 g sowie 0,1 g pro Maus) demonstriert werden. …

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

Liver Fibrosis Models:

Adult male mice at least 6 wk of age were used. Hepatic fibrosis was induced by two times per week i.p. CCl₄ (0.4 L/g; Sigma) diluted 1:3 in olive oil (Sigma) for 4 wk (nine injections). Animals were culled at stated time points after the final CCl₄ injection. For depletion studies, DT (10 ng/g in PBS; List Biological Laboratories) or PBS control was administered to fibrotic CD11B-DTR or WT mice i.v. through the tail vein at the stated time points. …

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

Administration of diphtheria toxin in vivo:

Diphtheria toxin was purchased from one of two sources: … or List Biological Laboratories. …

Adult mice (69weeks of age) received two intramuscular injections of DT at 50ng/g, spaced 2 days apart.  Each injection contained 1 g of DT dissolved in 50 l of saline.  Mice received 0.4 ml of lactated Ringers solution by subcutaneous injection once or twice daily on days 36 after the first DT injection.  Between days 1 and 6 after the first DT injection, food was supplemented with high-calorie gel (Tomlyn/Vetoquinol from Nutri-Cal).

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

In vivo cell depletion:

Treg cells were depleted by administration of diphtheria toxin (DT) to Foxp3^DTR mice (List Biological Laboratories). Fifty micrograms per kilogram of DT was administered intraperitoneally (i.p.) in a 200 L volume every other day for 12 days unless otherwise noted. Control animals received 200 L sterile PBS ip. DT administration results in greater than 95% reduction specifically of Treg cells in the spleen, LN’s, peripheral blood, and bone marrow (data not shown). DT treatment of control (C57BL/6) mice failed to induce any signs of B-cell activation, terminal differentiation, T_FH development, or autoantibody production (Supporting Information Fig. 1). CD4 depletion was achieved by administration of 250 g GK1.5 ip every 3rd day (Bio X Cell).

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

Mouse Treatments and Immunizations:

For immunization with OVA in complete Freunds adjuvant (OVA/CFA), OVA was emulsified in CFA at final concentration of 1 mg/ml OVA, and mice received hind footpad injection of 20 l of OVA/CFA as described (Webster et al., 2006).

Foxp3-DTR mice express simian diphtheria toxin (DT) receptor that allows for depletion of Foxp3⁺ regulatory T cells upon DT administration (Kim et al., 2007; Chinen et al., 2010). For regulatory T cell depletion, wild-type control or Foxp3-DTR mice were injected intraperitoneally with 1.5 g of DT … List Biological Laboratory, Campbell, CA, USA) on days 0, 1, 4, and 7 and examined on day 8. CD4⁺, CD25⁺, Foxp3⁺ regulatory T cells in lymph nodes were depleted by at least 12-fold by day 8.

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

Mice and Immunizations:

… In CD11c-DTR mice, DCs were depleted by i.p. injection of 8 ng diphtheria toxin (DT) (List Biological Labs) per gram body weight, in combination with rear footpad injection of 10 ng of DT each, all 12 hours before VRP injection.

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

Experimental Procedures 

Mice, Virus, and DT Administration:

… Diphtheria toxin (100 ng) (List Biological Laboratories, Campbell, CA) was administered intraperitoneally on days 5 and 7 after LCMV-Cl 13 infection.

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

ELISA:

… An in-house ELISA was used for measuring anti-diphtheria toxoid (DT). In brief, plates (Medisorp, Thermo Scientific Nunc) were coated with 50l/well DT (1g/ml in PBS) (List Biological Laboratories, Campbell, CA, USA) and blocked with 3% BSA in T-PBS. The sera samples were diluted 1:100 in 3% BSA in T-PBS. Anti-human IgG antibody conjugated with HRP and a substrate was used in the manner described above.

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

In vivo depletion experiments:

To deplete CD207+ DCs or LCs, 1 g of DT (List Biological Laboratories) was injected i.p. in Lang-DTR mice at day 3 or 15, respectively. DEREG mice were depleted of CD4+Foxp3+ Tregs (8) by i.p. injection of 1 g DT at days 2 or 1 with respect to day 0 of LC transfer.

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