British Journal of Nutrition

Nutritional Immunology

Immunostimulatory effect of faecal Bifidobacterium species of breast-fed and formula-fed infants in a peripheral blood mononuclear cell/Caco-2 co-culture system

T. Pozo-Rubioa1, J. R. Mujicoa1, A. Marcosa1, E. Puertollanoa2, I. Nadala3, Y. Sanza3 and E. Novaa1 c1

a1 Immunonutrition Group, Metabolism and Nutrition Department, Instituto de Ciencia y Tenología de Alimentos y Nutrición (ICTAN-CSIC), C/Jose Antonio Novais, 10, 28040 Madrid, Spain

a2 Division of Microbiology, Department of Health Sciences, Faculty of Experimental Sciences, University of Jaén, Jaén, Spain

a3 Microbial Ecophysiology and Nutrition Group, Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Valencia, Spain


Bifidobacterium spp. typical of the human intestinal microbiota are believed to influence the balance of immune responses in the intestinal mucosa. The aim of the present study was to investigate the effect of different bifidobacterial species and their mixtures in in vitro experiments with peripheral blood mononuclear cells (PBMC) and Caco-2 cells. Bifidobacterium adolescentis, B. angulatum, B. breve, B. catenulatum, B. infantis, B. longum and two combinations of these bifidobacteria simulating the species composition found in faecal samples from breast-fed (BF) and formula-fed (FF) infants were used. The levels of several cytokines were measured by direct stimulation of PBMC and by stimulation of a Caco-2/PBMC co-culture with bifidobacteria. B. catenulatum and B. breve were the strongest enhancers of interferon-γ (IFN-γ) production by direct stimulation of PBMC. B. longum was the highest inducer of IL-10 and the lowest TNF-α stimulus. In the Caco-2/PBMC system, B. breve was the highest inducer of IL-8 production by Caco-2 cells, significantly different from B. infantis, B. adolescentis and the FF mixture (P < 0·05). IFN-γ produced by PBMC stimulated with the BF mixture (containing 22 % B. breve, compared with 7 % in the FF mixture) was significantly higher compared with B. adolescentis, B. infantis and B. longum. B. adolescentis also inhibited IFN-γ production compared with the FF mixture and B. longum. The proportion of different Bifidobacterium strains seems to be an important determinant of the cytokine balance in the simulated intestinal environment studied. B. breve and the combination of the Bifidobacterium species typically found in the microbiota of BF infants have shown the most significant effects.

(Received September 24 2010)

(Revised January 19 2011)

(Accepted February 28 2011)

(Online publication May 31 2011)

Key Words:

  • Bifidobacterium spp.;
  • Caco-2 cells;
  • Peripheral blood mononuclear cells;
  • Cytokines;
  • Breast-feeding;
  • Formula feeding;
  • Infant's microbiota


c1 Corresponding author: E. Nova, fax +34 915493627, email


Abbreviations: BF, breast-fed; FF, formula-fed; IEC, intestinal epithelial cells; IFN-γ, interferon-γ; PBMC, peripheral blood mononuclear cells