D-Galacto-D-mannan Modulates IFNγ Secretion

D-galacto-D-mannan-mediated interferon γ (IFNγ) secretion with or without antigen (O PA2 or A YC) was assayed using commercial enzyme-linked immune absorbent spot (ELISpot) assay kits (R&D Systems, MN, USA) as per the manufacturer’s recommendations. Murine PECs or porcine PBMCs (5 × 10⁵ cells/well) were cultured in 96-well polyvinylidene fluoride-backed microplates containing a monoclonal capture antibody specific for murine or porcine IFNγ and stimulated with 2 μg/mL (final concentration) of antigen mixed with 0.625, 1.25, 2.5, and 5 μg/mL of D-galacto-D-mannan sequentially, at 37°C and 5% CO₂ for 18 h. Antigen and PBS were used as positive (PC) and negative controls (NC), respectively. Data were obtained using the ImmunoSpot ELISpot reader (AID iSpot Reader System; Autoimmune Diagnostika GmbH, Strassberg, Germany). The results were presented as spot-forming cells (SFC) per number of cells added to the well (7 (link), 29 (link)).

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Kim H.W., Ko M.K., Park S.H., Shin S., Kim G.S., Kwak D.Y., Park J.H., Kim S.M., Lee J.S, & Lee M.J. (2024). D-galacto-D-mannan-mediated Dectin-2 activation orchestrates potent cellular and humoral immunity as a viral vaccine adjuvant. Frontiers in Immunology, 15, 1330677.

Publication 2024

ELISpot) assay kits

Corresponding Organization :

Other organizations : Animal and Plant Quarantine Agency, Chungnam National University

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Variable analysis

independent variables

Concentration of D-galacto-D-mannan (0.625, 1.25, 2.5, and 5 μg/mL)

dependent variables

Interferon γ (IFNγ) secretion

control variables

Cell type (murine PECs or porcine PBMCs)
Cell density (5 × 10^5 cells/well)
Incubation time (18 h)
Incubation temperature (37°C)
Incubation atmosphere (5% CO2)

controls

Positive control: Antigen (O PA2 or A YC)
Negative control: Phosphate-buffered saline (PBS)

Annotations

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