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Milk-derived supplement inhibits in vitro lymphocyte proliferation and IL-2 production

Published online by Cambridge University Press:  12 May 2008

S. Marín-Gallén
Affiliation:
Department of Physiology, Faculty of Pharmacy, University of Barcelona, Barcelona, Spain
F. J. Pérez-Cano
Affiliation:
Department of Physiology, Faculty of Pharmacy, University of Barcelona, Barcelona, Spain
M. Castell
Affiliation:
Department of Physiology, Faculty of Pharmacy, University of Barcelona, Barcelona, Spain
C. Castellote
Affiliation:
Department of Physiology, Faculty of Pharmacy, University of Barcelona, Barcelona, Spain
A. Franch
Affiliation:
Department of Physiology, Faculty of Pharmacy, University of Barcelona, Barcelona, Spain
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Abstract

Type
1st International Immunonutrition Workshop, Valencia, 3–5 October 2007, Valencia, Spain
Copyright
Copyright © The Authors 2008

Recently, significant progress has been made in the characterisation of milk components affecting the function of the immune system. The aim of the present report was to study the effect of a premium ultra-grade freeze-dried bovine whey-protein concentrate (WPC) on the functional capacity of lymphocytes. Thus, the immunomodulating properties of this dairy extract were evaluated in relation to the proliferative response of spleen T and B lymphocytes. Splenocytes from adult Wistar rats were incubated in a medium supplemented with WPC (60–480 μg protein/ml), which contains (g/kg)<0.1 fat, 750–900 proteins and 35–50 carbohydrates, and also lactoferrin 9.2; IgG 300–600, IgA 50–70 and IgM 70–90 and high proportions of active compounds, e.g. natural growth factors and hormones, vitamins and amino acids. Standard commercial infant formula (SIF; 40–415 μg protein/ml) and BSA (250–500 μg protein/ml) were also added as milk-derived and inert control proteins respectively. Concanavalin A (ConA; specific stimulus for T-cells) or pokeweed (Phytolacca americana) mitogen (specific for B-cells) were added to the cell culture for 72 h. Proliferating cells were quantified by means of the BioTrak™ cell proliferation system (BioTrak, Carlsbad, CA, USA) based on 5-bromo-2′-deoxyuridine incorporation. IL-2 levels were quantified by ELISA in 24 h-culture supernatant fractions obtained from ConA-stimulated lymphocytes. Statistical analysis was performed by conventional ANOVA and when an experimental group variable had a significant effect on the dependent variable post hoc comparisons (LSD test) were performed. Significant differences were accepted at P<0.05.

Results showed a dose–dependent inhibitory effect of WPC on both spleen B- and T-lymphocyte proliferative response induced by mitogen stimulation (see Figure). The inhibitory effect on T- and B-cell proliferation was approximately 55% and approximately 40% respectively. In parallel, a WPC dose-dependent inhibition of IL-2 production was also found (approximately 80%). Cell viability was not modified by WPC addition. SIF produced similar inhibitory effects. However, non-milk-derived proteins such as BSA did not modify these lymphocyte responses. WPC and SIF, both milk-derived components, inhibited lymphocyte proliferation and IL-2 production in vitro. This immunomodulatory effect may prevent the newborn from over-reacting immunologically to the environmental antigens during early life.