Postpartum changes in the concentrations of IL-1beta, IL-6, tumor necrosis factor-alpha (TNF-alpha), transforming growth factor-beta1 (TGF-beta1), TGF-beta2, and prostaglandin E2 in 257 human milk samples collected longitudinally from 49 healthy mothers during the first 12 wk of lactation were determined by ELISA or RIA. The proinflammatory cytokines IL-1beta, IL-6, and TNF-alpha were present in only a proportion of samples, and there was a wide range of concentrations detected at each time in the present study (IL-1beta, <15-400 pg/mL; IL-6, <15-1032 pg/mL; TNF-alpha, <15-2933 pg/mL). Concentrations of prostaglandin E2 increased after the first week and remained elevated for the remainder of the study (range, < 10-9966 pg/mL). The antiinflammatory cytokines TGF-beta1 (range, 43-7108 pg/mL) and TGF-beta2 (range, 208-57935 pg/mL) were present in substantial quantities in all samples, and there was little change in the mean concentration during 12 wk of lactation. The present study shows that immunomodulating agents are normally present in human milk in physiologically relevant quantities for at least the first 3 mo of the breast-fed infant's life.

Breast milk contains many immunologically active components that influence the development of the immune system of the breast-fed infant. The purpose of this study was to investigate the difference in specific lymphocyte subsets between breast-fed and formula-fed 6-mo-old infants. Peripheral blood samples were collected from 79 breast-fed (< 120 mL formula/wk) and 69 formula-fed (breast-fed < 4 wk) infants at 6 mo. All infants had been born at term and had no known illness at the time of blood collection. Packed cells from whole blood were incubated with fluorochrome-labeled monoclonal antibodies, followed by erythrocyte lysis. Washed lymphocytes were analyzed by two-color direct immunofluorescence on a flow cytometer. The percentage of T and B lymphocytes in the peripheral blood of 6-mo-old infants was the same, regardless of feeding regimen. However, the relative frequency of natural killer (NK) cells was greater in breast-fed infants than in formula-fed infants (9.7% vs 7.1%; p < 0.001). The percentage of cells expressing CD4 was lower in breast-fed infants than in formula-fed infants (47.3% vs 50.9%; p < 0.005), and that of cells expressing CD8 was greater (18.0% vs 16.4%; p < 0.05). As a result, the CD4:CD8 ratio in breast-fed infants was lower than that in formula-fed infants (2.8 vs 3.3; p < 0.005). The absolute size of the lymphocyte subpopulations T, B, and CD8+ was the same for each of the two populations of infants. However, breast-fed infants had fewer CD4+ T cells (p < 0.05) and a greater number of NK cells (p < 0.01) than the age-matched formula-fed infants. The immunophenotypic differences between breast-fed and formula-fed infants are consistent with reported age-related changes, suggesting greater maturity in the development of the immune system of breast-fed infants.

n-3 Fatty acid-enriched eggs may provide a means of increasing dietary DHA during the second 6 mo of life. Egg yolks may also be a useful source of iron during the weaning period and can be safely included in the weaning diet with no perturbations in plasma cholesterol.

Objective: To examine the effect of nucleotide (NT)-supplemented cow's milk-based formula on growth and biochemical indices of immune function in healthy infants. Design: Randomized controlled trial (RCT) of formula-fed term infants allocated to control formula with an innate level of NT at 10 mg/l (n ¼ 102), or formula fortified with NT at 33.5 mg/l (n ¼ 98). A parallel group of 125 breastfed infants followed the same protocol as a reference. Outcome measures: Growth was assessed at enrolment, 7 weeks, 4 months and 7 months of age. Natural killer cell activity, cytokine production and lymphocyte subpopulations were assessed at 7 weeks of age. Antibody responses to diphtheria toxoid, tetanus toxoid and Haemophilus influenzae type b (Hib) immunizations were measured at 7 months of age. Results: NT supplementation did not influence the growth of formula fed infants or any markers of immunity measured at 7 weeks of age. Antibody responses to tetanus toxoid were higher in the NT-supplemented group (n ¼ 68) compared with the control group (n ¼ 70) at 7 months of age (median (5th, 95% percentile): 1.57(0.42, 3.43) vs 1.01(0.41, 4.66) IU/ml, Po0.03). A difference between treatments was seen in response to diphtheria toxoid but this effect disappeared when adjusted for hepatitis B immunization at birth. There was no effect of treatment on antibody responses to Hib immunization. Conclusions: Supplementation of formulas with NT at 33.5 mg/l resulted in a modest improvement in antibody response consistent with RCTs that used higher levels of NT supplementation. Whether this translates to clinical benefits in well-nourished infants requires further study.