Inflammatory Marker Quantification in Tissue

We selected 16 inflammatory markers based on previous reports.21 (link)23 (link) The protein concentrations of tissue extracts were determined using the Pierce 660 nm Protein Assay Kit (Thermo Fisher Scientific ., Waltham, MA, USA). Enzyme-linked immunosorbent assays for IL-22 and human neutrophil elastase (HNE) were performed with commercially available kits (all from R&D Systems, Minneapolis, MN, USA). Multiplex cytokine analysis kits (IL-1β, IL-5, IL-6, IL-8, IL-17A, IFN-γ, eotaxin-3, myeloperoxidase [MPO], matrix metalloproteinase [MMP]-9, TGF-β1, and periostin) were purchased from R&D Systems, and data were collected using a Luminex 100 reader (Luminex, Austin, TX, USA). Data analysis was conducted using MasterPlex QT version 2.0 (MiraiBio, Alameda, CA, USA). The levels of total IgE, specific IgE to staphylococcal enterotoxins (SE-IgE [SEA, SEB, SEC, and TSST-1]), and eosinophil cationic protein (ECP) in nasal tissue homogenates were measured using the ImmunoCAP assay (Thermo Fisher Scientific). A SE-IgE level > 0.35 IU/mL for at least one SE-specific IgE (SEA, SEB, SEC, and TSST-1) was defined as SE-specific IgE positive. All assay procedures mentioned were run in duplicate according to the manufacturer’s protocol. All protein levels in the tissue homogenates were normalized to the concentration of total protein. Further details are provided in the Supplementary Data S1.

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Min J.Y., Kim J.Y., Sung C.M., Kim S.T., Cho H.J., Mun S.J., Cho S.W., Hong S.D., Ryu G., Cho K.R., Kim Y.H., Park S.K., Kim D.K., Lee D.H., Heo S.J., Lee K.I., Kim S.J., Lee S., Mo J.H., Shin S.H, & Kim D.W. (2023). Inflammatory Endotypes of Chronic Rhinosinusitis in the Korean Population: Distinct Expression of Type 3 Inflammation. Allergy, Asthma & Immunology Research, 15(4), 437-450.

Publication 2023

Pierce 660 nm Protein Assay Kit Multiplex cytokine analysis kits Luminex 100 reader ImmunoCAP assay

Assay Austin Cytokine Enzyme linked immunosorbent assays Eosinophil cationic protein Eotaxin 3 Human neutrophil elastase Ifn γ Il 17a Il 1β Il 22 Inflammatory Matrix metalloproteinase 9 Myeloperoxidase Nasal Periostin Protein Staphylococcal enterotoxins Tgf β1 Tissue Tissue extracts Tsst 1

Corresponding Organization :

Other organizations : Kyung Hee University, Dongguk University Ilsan Hospital, Chonnam National University Hospital, Gachon University, Gyeongsang National University Hospital, Pusan National University Yangsan Hospital, Seoul National University Bundang Hospital, Samsung Medical Center, Sungkyunkwan University, Inje University Sanggye Paik Hospital, Chungnam National University, Chuncheon Sacred Heart Hospital, Hallym University, Chonnam National University Hwasun Hospital, Kyungpook National University, Konyang University, National Medical Center, Seoul National University, Dankook University, Catholic University of Daegu, Seoul Metropolitan Government, Boramae Medical Center

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Variable analysis

independent variables

Inflammatory markers (16 markers)

dependent variables

Protein concentrations of tissue extracts
Levels of IL-22 and human neutrophil elastase (HNE)
Levels of IL-1β, IL-5, IL-6, IL-8, IL-17A, IFN-γ, eotaxin-3, myeloperoxidase (MPO), matrix metalloproteinase (MMP)-9, TGF-β1, and periostin
Levels of total IgE, specific IgE to staphylococcal enterotoxins (SE-IgE [SEA, SEB, SEC, and TSST-1]), and eosinophil cationic protein (ECP) in nasal tissue homogenates

control variables

Total protein concentration in tissue homogenates (used for normalization of protein levels)

positive controls

Commercially available kits for IL-22 and human neutrophil elastase (HNE) assays
Multiplex cytokine analysis kits for other inflammatory markers

negative controls

None explicitly mentioned

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