{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T23:47:55Z","timestamp":1788220075219,"version":"build-2803163510"},"reference-count":51,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2023,11,28]],"date-time":"2023-11-28T00:00:00Z","timestamp":1701129600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Glanbia Nutritionals"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Whey protein isolate (WPI) consists of an array of proteins and peptides obtained as a byproduct of the cheesemaking process. Research suggests that WPI, along with its peptides such as glycomacropeptide (GMP), possesses immunomodulatory properties. These properties hold potential for alleviating the adverse effects of inflammatory conditions such as inflammatory bowel disease. Although promising, the immunoregulatory properties of the digested forms of WPI and GMP\u2014those most likely to interact with the gut immune system\u2014remain under-investigated. To address this knowledge gap, the current study examined the effects of in vitro-digested WPI and GMP, in vivo-digested WPI, and undigested WPI and GMP on the secretion of pro-inflammatory cytokines (TNF-\u03b1 and IL-1\u03b2) in lipopolysaccharide-stimulated macrophage-like cells. Our results indicate that digested WPI and GMP reduced the expression of TNF-\u03b1 and IL-1\u03b2, two pro-inflammatory cytokines. Whole WPI had no effect on TNF-\u03b1 but reduced IL-1\u03b2 levels. In contrast, in vivo-digested WPI reduced TNF-\u03b1 but increased IL-1\u03b2. Undigested GMP, on the other hand, increased the secretion of both cytokines. These results demonstrate that digestion greatly modifies the effects of WPI and GMP on macrophages and suggest that digested WPI and GMP could help mitigate gastrointestinal inflammation. Further clinical studies are necessary to determine the biological relevance of WPI and GMP digestion products within the gut and their capacity to influence gut inflammation.<\/jats:p>","DOI":"10.3390\/nu15234942","type":"journal-article","created":{"date-parts":[[2023,11,28]],"date-time":"2023-11-28T07:40:01Z","timestamp":1701157201000},"page":"4942","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Macrophage-Immunomodulatory Actions of Bovine Whey Protein Isolate, Glycomacropeptide, and Their In Vitro and In Vivo Digests"],"prefix":"10.3390","volume":"15","author":[{"given":"Wyatt","family":"Olsen","sequence":"first","affiliation":[{"name":"Department of Food Science & Technology, Oregon State University, Corvallis, OR 97331, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ningjian","family":"Liang","sequence":"additional","affiliation":[{"name":"Nutrition Program, College of Health, Oregon State University, Corvallis, OR 97331, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9696-0967","authenticated-orcid":false,"given":"David C.","family":"Dallas","sequence":"additional","affiliation":[{"name":"Department of Food Science & Technology, Oregon State University, Corvallis, OR 97331, USA"},{"name":"Nutrition Program, College of Health, Oregon State University, Corvallis, OR 97331, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1016\/j.foodres.2007.07.005","article-title":"Bovine Whey Proteins\u2014Overview on Their Main Biological Properties","volume":"40","author":"Madureira","year":"2007","journal-title":"Food Res. Int."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"10863","DOI":"10.1021\/acs.jafc.3c02085","article-title":"Comparison of Whey Proteome and Glycoproteome in Bovine Colostrum and Mature Milk","volume":"71","author":"Zhang","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"389S","DOI":"10.1080\/07315724.2001.10719173","article-title":"Anti-inflammatory Activities of Lactoferrin","volume":"20","author":"Conneely","year":"2001","journal-title":"J. Am. Coll. Nutr."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1314","DOI":"10.3945\/ajcn.112.055202","article-title":"Sequential Release of Milk Protein\u2013Derived Bioactive Peptides in the Jejunum in Healthy Humans","volume":"97","author":"Boutrou","year":"2013","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1016\/j.foodchem.2017.04.056","article-title":"Milk Bioactive Peptide Database: A Comprehensive Database of Milk Protein-Derived Bioactive Peptides and Novel Visualization","volume":"232","author":"Nielsen","year":"2017","journal-title":"Food Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"6902","DOI":"10.3168\/jds.2016-11186","article-title":"Isolation and Characterization of Anti-inflammatory Peptides Derived from Whey Protein","volume":"99","author":"Ma","year":"2016","journal-title":"J. Dairy Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.nutres.2017.01.005","article-title":"Whey Protein Hydrolysate and Branched-Chain Amino Acids Downregulate Inflammation-Related Genes in Vascular Endothelial Cells","volume":"38","author":"Bigo","year":"2017","journal-title":"Nutr. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.idairyj.2005.05.003","article-title":"Precipitation Behaviour of Caseinomacropeptides and their Simultaneous Determination with Whey Proteins by RP-HPLC","volume":"16","author":"Krause","year":"2006","journal-title":"Int. Dairy J."},{"key":"ref_9","unstructured":"Batt, C.A., and Tortorello, M.L. (2014). Encyclopedia of Food Microbiology, Academic Press. [2nd ed.]."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1093\/jn\/138.2.316","article-title":"Dietary Glycomacropeptide Supports Growth and Reduces the Concentrations of Phenylalanine in Plasma and Brain in a Murine Model of Phenylketonuria","volume":"138","author":"Ney","year":"2008","journal-title":"J. Nutr."},{"key":"ref_11","first-page":"2025","article-title":"Properties of Glycomacropeptide and Para-K-casein Derived from Human K-Casein and Comparison of Human and Bovine K-Caseins as to Susceptibility to Chymosin and Pepsin","volume":"48","author":"Azuma","year":"1984","journal-title":"Agric. Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Qu, Y., Kim, B.-J., Koh, J., and Dallas, D.C. (2021). Analysis of Bovine Kappa-Casein Glycomacropeptide by Liquid Chromatography\u2013Tandem Mass Spectrometry. Foods, 10.","DOI":"10.3390\/foods10092028"},{"key":"ref_13","first-page":"1818","article-title":"The Effect of Glycosylation on the Interfacial Properties of Bovine Caseinomacropeptide","volume":"23","author":"Strixner","year":"2009","journal-title":"Food Hyrdocoll."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"C\u00f3rdova-D\u00e1valos, L.E., Jim\u00e9nez, M., and Salinas, E. (2019). Glycomacropeptide Bioactivity and Health: A Review Highlighting Action Mechanisms and Signaling Pathways. Nutrients, 11.","DOI":"10.3390\/nu11030598"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1017\/S0022029900031034","article-title":"Inhibition of Proliferative Responses of Mouse Spleen Lymphocytes and Rabbit Peyer\u2019s Patch Cells by Bovine Milk Caseins and their Digests","volume":"62","author":"Otani","year":"1995","journal-title":"J. Dairy Res."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Qu, Y., Park, S.H., and Dallas, D.C. (2023). The Role of Bovine Kappa-Casein Glycomacropeptide in Modulating the Microbiome and Inflammatory Responses of Irritable Bowel Syndrome. Nutrients, 15.","DOI":"10.3390\/nu15183991"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1232","DOI":"10.1111\/j.1476-5381.2009.00195.x","article-title":"Bovine Glycomacropeptide Induces Cytokine Production in Human Monocytes Through the Stimulation of the MAPK and the NF-\u03baB Signal Transduction Pathways","volume":"157","author":"Requena","year":"2009","journal-title":"Br. J. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5803639","DOI":"10.1155\/2020\/5803639","article-title":"Activation of the NF-\u03baB and MAPK Signaling Pathways Contributes to the Inflammatory Responses, but Not Cell Injury, in IPEC-1 Cells Challenged with Hydrogen Peroxide","volume":"2020","author":"Xiao","year":"2020","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_19","first-page":"89","article-title":"Milk-derived Proteins and Peptides of Potential therapeutic and Nutritive Value","volume":"6","author":"Zimecki","year":"2007","journal-title":"J. Exp. Ther. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Feeney, S., Ryan, J.T., Kilcoyne, M., Joshi, L., and Hickey, R. (2017). Glycomacropeptide Reduces Intestinal Epithelial Cell Barrier Dysfunction and Adhesion of Entero-hemorrhagic and Entero-pathogenic Escherichia coli In Vitro. Foods, 6.","DOI":"10.3390\/foods6110093"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1017\/S0007114520003736","article-title":"Casein Glycomacropeptide is Well Tolerated in Healthy Adults and Changes Neither High-sensitive C-reactive Protein, Gut Microbiota Nor Faecal Butyrate: A Restricted Randomised Trial","volume":"125","author":"Wernlund","year":"2021","journal-title":"Br. J. Nutr."},{"key":"ref_22","unstructured":"Li, E.W.Y. (2003). Recovery of Bovine Glycomacropeptide as Nutraceutical and its Enhancement in Human Immunity. [Ph.D. Thesis, University of Guelph]."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1016\/j.sjbs.2015.06.005","article-title":"Milk Derived Bioactive Peptides and their Impact on Human Health\u2014A Review","volume":"23","author":"Mohanty","year":"2016","journal-title":"Saudi J. Biol. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.idairyj.2007.07.008","article-title":"Immunomodulating Properties of a Whey Protein Isolate, its Enzymatic Digest and Peptide Fractions","volume":"18","author":"Gauthier","year":"2008","journal-title":"Int. Dairy J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.idairyj.2003.08.003","article-title":"Immunomodulating Effects of Whey Proteins and their Enzymatic Digests","volume":"14","author":"Mercier","year":"2004","journal-title":"Int. Dairy J."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1093\/jn\/nxab400","article-title":"Top-Down Glycopeptidomics Reveals Intact Glycomacropeptide Is Digested to a Wide Array of Peptides in Human Jejunum","volume":"152","author":"Koh","year":"2022","journal-title":"J. Nutr."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1038\/ncpgasthep0528","article-title":"Mechanisms of Disease: Pathogenesis of Crohn\u2019s Disease and Ulcerative Colitis","volume":"3","author":"Sartor","year":"2006","journal-title":"Nat. Clin. Pract. Gastroenterol. Hepatol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1756","DOI":"10.1053\/j.gastro.2011.02.016","article-title":"Proinflammatory Cytokines in the Pathogenesis of Inflammatory Bowel Diseases","volume":"140","author":"Strober","year":"2011","journal-title":"Gastroenterology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1016\/j.crohns.2011.10.004","article-title":"Anti-TNF-\u03b1 Antibodies Improve Intestinal Barrier Function in Crohn\u2019s Disease","volume":"6","author":"Noth","year":"2012","journal-title":"J. Crohn\u2019s Colitis"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2311","DOI":"10.3748\/wjg.v10.i16.2311","article-title":"Drug Therapy for Ulcerative Colitis","volume":"10","author":"Xu","year":"2004","journal-title":"World J. Gastroenterol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1007\/BF00310046","article-title":"The Anatomical Basis for the Immune Function of the Gut","volume":"176","author":"Pabst","year":"1987","journal-title":"Anat. Embryol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1530","DOI":"10.1016\/j.immuni.2022.08.005","article-title":"Macrophages in the Gut: Masters in Multitasking","volume":"55","author":"Delfini","year":"2022","journal-title":"Immunity"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"9653","DOI":"10.3748\/wjg.v20.i29.9653","article-title":"Intestinal Antigen-presenting Cells in Mucosal Immune Homeostasis: Crosstalk Between Dendritic Cells, Macrophages and B-cells","volume":"20","author":"Mann","year":"2014","journal-title":"World J. Gastroenterol."},{"key":"ref_34","first-page":"e8668","article-title":"The Identification of Markers of Macrophage Differentiation in PMA-Stimulated THP-1 Cells and Monocyte-Derived Macrophages","volume":"5","author":"Daigneault","year":"2010","journal-title":"Public Libr. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.cellimm.2018.07.008","article-title":"Similarities and Differences in Surface Receptor Expression by THP-1 Monocytes and Differentiated Macrophages Polarized Using Seven Different Conditioning Regimens","volume":"332","author":"Forrester","year":"2018","journal-title":"Cell. Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"173090","DOI":"10.1016\/j.ejphar.2020.173090","article-title":"Macrophage M1\/M2 Polarization","volume":"877","author":"Yunna","year":"2020","journal-title":"Eur. J. Pharmacol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2733","DOI":"10.3389\/fimmu.2018.02733","article-title":"Origin, Differentiation, and Function of Intestinal Macrophages","volume":"9","author":"Bain","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1039\/C3FO60702J","article-title":"A Standardised Static In Vitro Digestion Method Suitable for Food\u2014An International Consensus","volume":"5","author":"Minekus","year":"2014","journal-title":"Food Funct."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Qu, Y., Kim, B.-J., Koh, J., and Dallas, D.C. (2023). Comparison of Solid-Phase Extraction Sorbents for Monitoring the In Vivo Intestinal Survival and Digestion of Kappa-Casein-Derived Caseinomacropeptide. Foods, 12.","DOI":"10.3390\/foods12020299"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1093\/jn\/nxab350","article-title":"Peptides Derived from In Vitro and In Vivo Digestion of Human Milk Are Immunomodulatory in THP-1 Human Macrophages","volume":"152","author":"Liang","year":"2022","journal-title":"J. Nutr."},{"key":"ref_41","unstructured":"Riss, T.L., Moravec, R.A., Niles, A., Duellman, S., Benink, H.A., Worzella, T.J., and Minor, L. (2013). Assay Guidance Manual, Eli Lilly & Company and the National Center for Advancing Translational Sciences."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1016\/j.idairyj.2009.02.010","article-title":"Effect of Feeding Whey Peptide Fractions on the Immune Response in Healthy and Escherichia coli Infected Mice","volume":"19","author":"Gauthier","year":"2009","journal-title":"Int. Dairy J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"228","DOI":"10.4028\/www.scientific.net\/AMR.345.228","article-title":"Immunomodulating Effects of the Peptides Derived from Whey Protein","volume":"345","author":"Fan","year":"2012","journal-title":"Adv. Mater. Res."},{"key":"ref_44","unstructured":"Ali, A., Ain, Q., Saeed, A., Khalid, W., Ahmed, M., and Bostani, A. (2021). New Advances in the Dairy Industry, IntechOpen."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"351","DOI":"10.3390\/foods3020351","article-title":"The Effect of Oral Intake of Low-Temperature-Processed Whey Protein Concentrate on Colitis and Gene Expression Profiles in Mice","volume":"3","author":"Jayatilake","year":"2014","journal-title":"Foods"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"G590","DOI":"10.1152\/ajpgi.00211.2015","article-title":"Glycomacropeptide is a Prebiotic that Reduces Desulfovibrio Bacteria, Increases Cecal Short-Chain Fatty Acids, and is Anti-inflammatory in Mice","volume":"309","author":"Sawin","year":"2015","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.1093\/jn\/135.5.1164","article-title":"Bovine Glycomacropeptide is Anti-inflammatory in Rats with Hapten-Induced Colitis","volume":"135","author":"Daddaoua","year":"2005","journal-title":"J. Nutr."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1202","DOI":"10.1017\/S0007114513003590","article-title":"Validation of Bovine Glycomacropeptide as an Intestinal Anti-inflammatory Nutraceutical in the Lymphocyte-Transfer Model of Colitis","volume":"111","author":"Requena","year":"2014","journal-title":"Br. J. Nutr."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2704","DOI":"10.1021\/jf0355102","article-title":"Immunoenhancing Effects of Bovine Glycomacropeptide and Its Derivatives on the Proliferative Response and Phagocytic Activities of Human Macrophagelike Cells, U937","volume":"52","author":"Li","year":"2004","journal-title":"J. Agric. Food Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1017\/S0007114522001787","article-title":"Effects of Whey and Soy Protein Supplementation on Inflammatory Cytokines in Older Adults: A Systematic Review and Meta-Analysis","volume":"129","author":"Prokopidis","year":"2023","journal-title":"Br. J. Nutr."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Qu, Y., Park, S.H., and Dallas, D.C. (2023). Evaluating the Potential of Casein Glycomacropeptide in Adult Irritable Bowel Syndrome Management: A Pilot Study. Nutrients, 15.","DOI":"10.3390\/nu15194174"}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/15\/23\/4942\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:32:33Z","timestamp":1760131953000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/15\/23\/4942"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,28]]},"references-count":51,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["nu15234942"],"URL":"https:\/\/doi.org\/10.3390\/nu15234942","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,28]]}}}