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Flad, L.M.* ; Weisser, T.* ; Liedl, J.* ; Gard, F.* ; Petrera, A. ; von Toerne, C. ; Hauck, S.M. ; Deeg, C.A.* ; Kleinwort, K.J.H.*

Inhibitory impact of A1 and A2 cow's milk on human peripheral blood mononuclear cells identifies TOM1 as a candidate anti-inflammatory mediator.

Front. Immunol. 17:1728935 (2026)

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Introduction: Cows milk has long been discussed as a modulator of immune response, with a particular focus on different health effects of its beta-casein variants A1 and A2. To date, the factors in milk which may be responsible for effects on the human immune system remain unknown, revealing a lack of information that warrants further investigations. We previously found that both A1 and A2 milk inhibit Concanavalin A-induced human peripheral blood mononuclear cells (PBMC) proliferation. Accordingly, this study aimed to investigate the underlying mechanisms of the inhibitory effect. Methods: In a first step, we narrowed down the size-range of possible causative factors in A1 and A2 milk samples by separating each milk type in a high (> 100 kDa; HWF) and a low molecular weight fraction (< 100 kDa, LWF) and used them in co-incubation with Concanavalin A stimulated human PBMC. In in vitro proliferation assays, we identified HWFs as the inhibitory fractions and analyzed the PBMCs supernatant proteomes using high-resolution LC-MS/MS to assess changes in the cellular response to HWFs. Olink proteomics was used to asses the influence of the candidate protein on Immune-Response-Marker release of PBMC. Results: HWFs of milk exert an inhibitory effect on human PBMC. Among several candidates, Target of Myb protein 1 (TOM1) emerged as the only protein markedly increased in the supernatant proteomes after co-incubation with the HWFs of A1 and A2 milk. Further in vitro proliferation assays revealed an inhibitory impact of TOM1 on stimulated humane PBMC. Olink analysis of PBMC Immune-Response-Marker secretion reinforced a possible role for TOM1 as anti-inflammatory mediator in milk involved in the regulation of immune response via Interleukin-1 beta-associated signaling. Discussion: Our findings provide new insights into milk-induced immune regulation and identify TOM1 as a promising anti-inflammatory factor.

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Publication type Article: Journal article

Document type Scientific Article

Keywords A1/A2 milk fractions; ConA; immunonutrition; Olink; PBMC; proliferation; supernatant proteome; TOM1; Beta-casein Variants; Bovine-milk; In-vitro; Stimulation; Beta-casomorphin-7; Suppression; Activation; Proteins; Release

ISSN (print) / ISBN 1664-3224

e-ISSN 1664-3224

Journal Frontiers in Immunology

Quellenangaben Volume: 17, Article Number: 1728935

Publisher Frontiers

Publishing Place Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland

Reviewing status Peer reviewed

Institute(s) CF Metabolomics & Proteomics (CF-MPC)

Grants Bayerischer Milchfrderungsfond