Activation of human NK cells by staphylococci and lactobacilli requires cell contact-dependent costimulation by autologous monocytes.

NK cells are instrumental in innate immune responses, in particular for the early production of gamma interferon (IFN-gamma) and other cytokines necessary to control certain bacterial, parasitic, and viral infections. NK cell-mediated effector functions are controlled by a fine balance between distinct receptors mediating activating and inhibitory signals; however, little is known about activating receptors on NK cells and their corresponding ligands. Several studies have shown that commensal lactobacilli isolated from the human gastrointestinal tract activate human mononuclear cells and are potent inducers of IFN-gamma and monocyte-derived interleukin 12 (IL-12). NK cell activation was shown for Lactobacillus johnsonii La1. In this study the cellular mechanisms of in vitro NK cell activation by gram-positive bacteria were analyzed. Staphylococcus aureus- and L. johnsonii La1-mediated activation of CD3(-) CD16(+) CD56(+) human peripheral blood NK cells, including expression of the activation antigen CD69 and secretion of IFN-gamma, required cell contact-dependent costimulation by autologous monocytes. S. aureus- and L. johnsonii-preactivated monocytes retained their capacity to induce NK cell activation. In contrast, cytokine-primed monocytes completely failed to induce NK cell activation unless bacteria were present. This suggests that phagocytosis of bacteria provided additional coactivation signals on accessory cells that may differ from those induced by tumor necrosis factor and IFN-gamma. Blocking of costimulatory molecules by B7.1, B7.2, and IL-12 but not CD14 monoclonal antibodies inhibited S. aureus- and L. johnsonii-induced effector function of NK cells. Our data suggest an important role for accessory cell-derived signals in the process of NK cell activation by gram-positive bacteria.

Bioactive molecules in milk and their role in health and disease: the role of transforming growth factor-beta.

Human breast milk is rich in nutrients, hormones, growth factors and immunoactive molecules, which influence the growth, development and immune status of the newborn infant. Although several of these factors are also present in bovine milk, the greater susceptibility of the formula-fed infant to infection and disease and the development of allergy is often attributed to the reduced level of protective factors in milk formulas. Nevertheless, modifying manufacturing processes may preserve the biological activity of some bioactive molecules in end products. Transforming growth factor (TGF)-beta is one such molecule. TGF-beta is a polypeptide, which has been described in both human and bovine milk. It is implicated in many processes, including epithelial cell growth and differentiation, development, carcinogenesis and immune regulation. The present article discusses the biological activity of TGF-beta2 that has been preserved and activated in a cow's milk-based product. More specifically, it addresses possible mechanisms of action in the intestinal lumen and speculates on how milk products containing naturally occurring TGF-beta2 could be exploited in functional foods for the infant or as therapies for specific intestinal diseases.

Modulation of nonspecific mechanisms of defense by lactic acid bacteria: effective dose.

The purpose of this study was to determine the effect of a fermented milk product containing Lactobacillus johnsonii La1 (formerly known as Lactobacillus acidophilus La1) on the phagocytic activity of peripheral blood leukocytes in healthy adult volunteers. Furthermore, we sought to define the effective doses of the bacteria, examine the effect on respiratory burst activity, and, finally, examine the contribution made by the starter culture to the biological effects. Volunteers were randomly distributed among three groups; each subject received one pot (150 ml) of fermented milk each day for 3 wk. The first two groups received a freshly prepared product fermented by Streptococcus thermophilus (group A) alone or S. thermophilus and 10(7) cfu/ml L. johnsonii La1 (group B). Group C received a product stored for a period of 21 to 28 d and that contained S. thermophilus and 10(6) cfu/ml of L. johnsonii La1. Ingestion of L. johnsonii La1 did not significantly increase fecal lactobacilli counts. However, L. johnsonii La1 was able to survive intestinal transit and was only recovered from the feces of the volunteers of groups B and C. The fermented base alone showed a weak effect on respiratory burst but not on phagocytic activity. However, the product containing 10(7) cfu/ml L. johnsonii La1 significantly enhanced both functions. The product containing 10(6) cfu/ml of L. johnsonii La1 had no significant effect on either function. These results suggest that fecal persistence may not necessarily reflect in vivo colonization and may not be a prerequisite for all forms of immune reactivity.

A simple rapid method to slice biological specimens: an application for non-embedded and embedded Golgi-stained tissue.

A rotary wire saw is faster and simplifies markedly the slicing of soft and hard tissues without apparent loss of quality. Two applications for the Golgi-Cox method (using the rotary wire saw) are described: one eliminates specimen freezing and embedding while the other uses LR-White instead of celloidin, reducing preparation time. Sections of 20 microns to several mm thick can be obtained.