Venous blood samples were drawn from one local control, the brother, the parents, and the patient. Samples were collected in heparin-containing tubes and processed as previously described5 . Briefly, blood samples were diluted 1:2 in RPMI-1640 medium (Gibco) and supplemented with penicillin (100 U/mL) and streptomycin (Gibco, 100 μg/mL). The samples were then dispensed into a 48-well plate (1 mL/well), either left non-stimulated or stimulated with live Mycobacterium bovis BCG Pasteur substrain at a multiplicity of infection (MOI) of 20 BCG cells per leukocyte alone, or with BCG plus recombinant human IL-12 (R&D Systems, Cat: 219-IL, 20 ng/mL). As a positive control, cells in separate wells were stimulated with phorbol 12-myristate 13-acetate (PMA, Sigma, Cat: P8139, 40 ng/mL) and ionomycin (Sigma, Cat: I9657, 10 μM). After 48 hours of stimulation, secreted cytokine levels in the supernatants were determined with the human IFN-γ ELISA Ready-SET-Go kit (Invitrogen, Cat: 50–173-27) and the human IL-12p40 ELISA kit (Invitrogen, Cat: KAC1561).
Bcg plus recombinant human il 12
Manufactured by R&D Systems
BCG plus recombinant human IL-12 is a laboratory product formulated with Bacillus Calmette-Guérin (BCG) and recombinant human interleukin-12 (IL-12). The core function of this product is to provide a combination of these two components for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Ionomycin, by Merck Group (2 mentions)
Human il 12p40 elisa kit, by Thermo Fisher Scientific (2 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Human ifn γ elisa kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using bcg plus recombinant human il 12
1
Cytokine Response to Mycobacterial Stimulation
2
Cytokine Response to BCG Stimulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Venous blood samples were drawn from one local control, one travel control, the two Iranian patients, and their father. Samples were collected in heparin-containing tubes and processed as previously described (Boisson-Dupuis et al., 2018 (link)). Briefly, blood samples were diluted 1:2 in RPMI-1640 medium (Gibco) and supplemented with penicillin (100 U/ml) and streptomycin (Gibco, 100 μg/ml). The samples were then dispensed into a 48-well plate (1 ml/well) and were either left non-stimulated or were stimulated with live M. bovis BCG Pasteur substrain at a multiplicity of infection of 20 BCG cells per leukocyte or with BCG plus recombinant human IL-12 (Cat: 219-IL; R&D Systems, 20 ng/ml). As a positive control, cells in separate wells were stimulated with phorbol 12-myristate 13-acetate (PMA, Cat: P8139; Sigma-Aldrich, 40 ng/ml) and ionomycin (Cat: I9657; Sigma-Aldrich, 10 μM). After 48 h of stimulation, the levels of secreted cytokines in the supernatant were determined with the human IFN-γ ELISA kit (Cat: 50-173-27; Invitrogen) and the human IL-12p40 ELISA kit (Cat: KAC1561; Invitrogen), according to the manufacturer’s instructions.
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