Immunomodulatory Effect of Bovine Lactoferrin Pepsin Hydrolysate on Murine Splenocytes and Peyer's Patch Cells

Abstract

The effects were examined of a pepsin hydrolysate of bovine lactoferrin on the proliferation of murine splenocytes. The hydrolysate enhanced [3H]thymidine uptake by splenocytes, but undigested bovine lactoferrin exerted an inhibitory effect. The hydrolysate had the ability to inhibit the blastogenesis that was induced by mitogens such as concanavalin A, phytohemagglutinin, and lipopolysaccharide; inhibition was similar to that with undigested lactoferrin. These results suggested that the hydrolysate contained both immunostimulatory and immunoinhibitory peptides. The stimulatory effect of the hydrolysate in the absence of mitogens was then explored in more detail using nonadherent splenocytes. The proliferative response of splenocytes to the hydrolysate was much greater in the fraction that was enriched with B cells than in the fraction that was enriched with T cells. The hydrolysate did not affect thymocyte proliferation. These data indicated that the adherent cells resembling macrophages and found among the splenocytes were not the target cells of the hydrolysate. The stimulatory effect of the hydrolysate was due to the activation of B cells by the hydrolysate and enhanced immunoglobulin production by splenocytes. Because the hydrolysate also enhanced the proliferation and immunoglobulin A production of Peyer's Patch cells, the immunostimulatory effect of the hydrolysate in vivo was examined using mice that had been orally immunized with cholera toxin. The concentrations of immunoglobulin A conjugated against cholera toxin in bile and in the intestinal contents of mice fed liquid diets containing 1% (wt/vol) lactoferrin hydrolysate were greater than those of mice fed control diets. This result suggested that the use of the lactoferrin hydrolysate is beneficial to enhance mucosal immunity.

Immunomodulatory peptides obtained by the enzymatic hydrolysis of whey proteins

“…WPCs and PP fractions prepared from human and bovine milk have no effect on IgA production in murine Peyer's patch cell cultures (Shida et al, 1992), whereas GMP suppresses in vitro antibody production (plaque-forming cell, PFC) to sheep RBC in murine spleen cell cultures . A pepsic H of bLF containing LFc also significantly enhances immunoglobulin (IgM, IgG and IgA) production in cultured murine splenocytes and IgA production in Peyer's patch cells (Miyauchi et al, 1997).…”

“…This study highlighted the importance of measuring LPS contamination in commercial whey protein products before studying their immunomodulatory properties. Miyauchi, Kaino, Shinoda, Fukuwatari, and Hayasawa (1997) showed that bovine lactoferrin (bLF) inhibits the proliferation of murine splenocytes while a peptic H of bLF promotes cell proliferation, especially in a B-cellenriched fraction as well as in Peyer's patch cells. They also showed that the peptic H inhibits the blastogenesis induced by mitogens (concanavalin A (ConA), phytohemagglutinin (PHA), LPS), as observed for intact bLF.…”

A rapid method for purifying osteopontin from bovine milk and interaction between osteopontin and other milk proteins

“…Notable among them are LF and LPO, which are basic proteins with respective pI values of 8-9 and 9.62 (Hahn, Schulz, Schaupp, & Jungbauer, 1998) and are bound to OPN in physiologically conditioned PBS with dissociation constants of 1.3 Â 10 À6 and 2.5 Â 10 À6 , respectively. LF functions as an antimicrobial agent for host defense and as a physiological regulator with respect to inflammatory and immune responses (Brock, 2002;Tomita, Wakabayashi, Yamauchi, Teraguchi, & Hayasawa, 2002;Miyauchi, Kaino, Shinoda, Fukuwatari, & Hayasawa, 1997), inhibiting complement activity (Kijlstra & Jeurissen, 1982), and activating NK cells, lymphokineactivated killer cells (Shau, Kim, & Golub, 1992), polymorphonuclear leukocytes and macrophages (Gahr, Speer, Damerau, & Sawatzki, 1991). LPO, a hemecontaining oxidation-reduction enzyme, is part of an antimicrobial system that converts thiocyanate to hydrothiocyanate which can kill both Gram-positive (Steele & Morrisons, 1969) and Gram-negative (Bjo¨rck, Rosen, Marshall, & Reiter, 1975) bacteria.…”

Potential use of bovine lactoferrin as adjuvant for the modulation of the specific immune response in rainbow trout (Oncorhynchus mykiss W.)

“…Zimecki et al (1991) observed that the humoral immune response against sheep red blood cells increases up to five times in mice treated with a single injected dose of human LF, stimulating the immune response leading to T cell differentiation. Miyauchi et al (1997) demonstrated that a pepsin LF hydrolysate activates murine B cells, enhancing Ig secretion by splenocytes and IgA production in Peyer's Patch cells.…”