Whey proteins and peptides: beneficial effects on immune health

Citation metadata

Date: Jan. 2006
From: Therapy(Vol. 3, Issue 1)
Publisher: Future Medicine Ltd.
Document Type: Article
Length: 7,355 words

Document controls

Main content

Full Text: 

Author(s): Josée Beaulieu 1 , Claude Dupont 2 , Pierre Lemieux 3 [[dagger]]

Keywords:

immune; inflammation; peptide; protein; whey

Until recently, whey proteins have been mainly studied for their positive effects on endurance by increasing muscle protein balance during sport training [1] . Many studies have also demonstrated that whey proteins possess an excellent lowering effect on serum lipid levels and blood pressure [2] . The discovery of the antioxidant potential of whey protein products [3] was one of the principal studies demonstrating the potential health benefit of these products for cancer and the immune system. Subsequently, many studies have reported a protective effect against different types of cancer by the whey proteins including an important review by Bounous [4] .

Currently, most research focuses on the effects whey proteins and peptides have on immune health and their protective roles in immune disorders. Whey is composed mainly of proteins as well as many peptides derived from these proteins. The proteins found in whey are principally [beta]-lactoglobulin, [alpha]-lactalbumin, lactoferrin, immunoglobulin (Ig) and bovine serum albumin (BSA). Their biochemical properties, as well as their concentrations in milk and whey, are summarized in Table 1. All of these proteins and peptides possess specific effects on immunity. These effects will be discussed with relation to specific associated proteins or peptides, and are summarized in Table 2.

[beta]-lactoglobulin

[beta]-lactoglobulin is the most abundant protein found in whey, comprising between 55 and 65% of the total whey protein content. This protein possesses numerous physicochemical attributes such as acting as an emulsifier, as well as many reported health effects.

In 1985 Bounous and Kongshavn were among the first researchers to demonstrate an immunoenhancing effect of [beta]-lactoglobulin [5] . They showed that the whey proteins (principally [beta]-lactoglobulin and BSA) increase the number of antibody-forming cells in the spleen of mice immunized with trinitrophenylated ficoll. Subsequently, the team of Wong and colleagues demonstrated that the stimulatory effects of [beta]-lactoglobulin on normal murine spleen cells are more intense than those observed with BSA [6] . The immune effects on murine spleen cells are postulated to be due to an increase in glutathione (GSH) production by splenocytes. This has been confirmed by the addition of S -(n-butyl)homocysteine sulfoxamine, an inhibitor of glutamylcysteine synthetase, which in this case, blocked the stimulatory effect [6] .

Controversially, another study demonstrated that the observed effects of [beta]-lactoglobulin on spleen cells to produce T-helper (Th)-cell 1 cytokines are due to endotoxin contamination [7] . This study was conducted with a commercial [beta]-lactoglobulin product and utilization of a specific isolation technique led to the presence of an endotoxin in the preparation. However, the team of Wong and colleagues have proven that the effect observed with [beta]-lactoglobulin is not due to endotoxin contamination but the utilization of polymixin B, a lipopolysaccharide (LPS) inhibitor [6] . The utilization of this LPS inhibitor markedly reduced the effect of LPS; however, it had no effect on [beta]-lactoglobulin. This study confirms that the [beta]-lactoglobulin really does possess an immunomodulatory effect. It is important to note that preparation is essential in the quality and reliability of results, as demonstrated by Brix and colleagues [7] . An additional control should be included in the study to confirm its validity.

Peptides from [beta]-lactoglobulin

Many studies have been conducted with specific peptides from [beta]-lactoglobulin; however, due to the complex nature of the protein, some of the peptides derived from it are still unknown. Of the [beta]-lactoglobulin peptides, four exhibit antimicrobial activities [8] . [beta]-lactoglobulin is also a carrier of many small hydrophobic molecules, such as retinoic acid, a modulator of lymphocyte responses [9] .

A peptide from [beta]-lactoglobulin, known as [beta]-lactotensin, has demonstrated an effect on the contraction of the ileum longitudinal muscle [10] . A study completed with [beta]-lactotensin showed similar results - that this peptide possesses a stimulating effect on smooth muscle in vitro [11] . Smooth muscle is responsible for the contractility of hollow organs, such as blood vessels, the gastrointestinal tract, the bladder and the uterus. Many diseases, including allergy, asthma and atherosclerosis, are related to dysfunctions of smooth muscle. Although the role of smooth muscle cells is contraction, they also exhibit extensive phenotypic diversity during normal development, repair of vascular injury and in disease states [12] . The stimulating effect of [beta]-lactotensin on smooth muscle provides information that this peptide can enhance general health.

[beta]-lactoglobulin is the protein responsible for milk allergies in children (2-3% of children suffer from milk allergies); however, in 80% of cases allergy symptoms disappear before the age of 3 years [13] . It has been demonstrated that the peptides from [beta]-lactoglobulin induce oral tolerance and consequently, diminish IgE production specific to [beta]-lactoglobulin [14] . This observation suggests that the consumption of peptides from [beta]-lactoglobulin possesses an important protective role against milk allergies.

[alpha]-lactalbumin

[alpha]-lactalbumin is the second major protein present in whey, accounting for 15 to 25% of the whey protein content. This protein is rich in essential amino acids and has a low immunogenenicity, indicating that [alpha]-lactalbumin is a good nutrient for children [15] . It has also been shown that a diet supplemented with [alpha]-lactalbumin increases resistance against acute infection caused by Escherichia coli [16] .

In the 1980s, many studies demonstrated an immunoenhancing effect of [alpha]-lactalbumin in mice [17,18] . Despite these studies, it is surprising to observe an absence of research on the immunomodulatory effects of [alpha]-lactalbumin. It has been clear since 1997 that [alpha]-lactalbumin possesses stimulatory effects. The production of IL-1[beta] by sheep macrophages issued from bronchoalveolar lavage is increased by the presence of this protein [19] .

The team of Svanborg and colleagues has identified a variant of [alpha]-lactalbumin similar to the modified [alpha]-lactalbumin found in the stomach of nursing children, which possess interesting immune effects [20,21] . This protein can protect against cancer by the induction of apoptosis in tumor and immature cells, but has no effect on healthy cells [21] .

Peptides from [alpha]-lactalbumin

Hydrolyzed [alpha]-lactalbumin, which contains unidentified peptides, has demonstrated a modulation of B- as well as T-lymphocyte activities [5] . After this observation, research into the identification of peptides from [alpha]-lactalbumin as well as its associated immunomodulatory effects, have been undertaken.

An immunomodulatory tripeptide derived from [alpha]-lactalbumin has been discovered [22] . This tripeptide, glycyl-leucyl-phenylalanine (GLF), stimulates the adherence and phagocytosis of human monocytes and macrophages, apparently by a mechanism of binding to the specific receptors for GLF on these cells [22,23] . The same research team evaluated the properties of this tripeptide on human and rat polymorphonuclear (PMN) cells. GLF increases the production of the superoxide anion by human PMNs, increasing the oxidative burst response in the presence of GLF. Phosphoinositide metabolism is also increased in the presence of GLF, which can be seen by the liberation of a second messenger, IP3, leading to an enhancement in membrane fluidity [24] . Moreover, a study carried out with another peptide (Tyr-Gly-Gly) from [alpha]-lactalbumin revealed a stimulation of human peripheral blood lymphocytes in the presence of this peptide [25] . In relation to antimicrobial activities, Pelligrini and colleagues have identified that these three peptides from [alpha]-lactalbumin possess interesting antimicrobial activities [26] .

Lactoferrin

Lactoferrin, an antioxidant glycoprotein with iron-binding properties, is the most studied of the whey proteins and also appears to be the best immunomodulator. The antioxidant capacity of lactoferrin is due to its iron-binding site that participates in the generation of hydroxyl radicals [27] . This iron-binding capacity is also partly responsible for its antimicrobial potential [28] . Lactoferrin is found in colostrums [29] , mucosal secretions [30] and neutrophil granules [31] . The localization of lactoferrin in important sites implicated in acquiring immunity, suggests an important contribution of lactoferrin in immune health. In whey, the proportion of lactoferrin is 1 to 2%. Numerous studies on the roles of lactoferrin in immunity have been conducted. This paper will review the most significant observations reported to date.

Lactoferrin exhibits a known immunomodulatory potential by its capacity to act as both an immunosuppressive as well as an immunostimulatory agent. Certain studies revealed the anti-inflammatory potential of lactoferrin in some circumstances, while in others the immune status demonstrated its stimulatory potential.

Lactoferrin possesses an anti-inflammatory potential in part by inhibiting cytokines such as tumor necrosis factor (TNF)-[alpha] and interleukin (IL)-1[beta], which are key inflammatory cytokines [32] . Many animal models demonstrate that lactoferrin can protect against inflammatory conditions. A study by Dial and colleagues shows that lactoferrin protects against gastritis induced by Helicobacter felis bacterium in mice [33] . Moreover, local administration of lactoferrin in inflamed joints of two different arthritis models in mice shows a strong local anti-inflammatory effect not related to the reduction of the proinflammatory cytokine IL-6 [34] . The effect of this protein on arthritis has also been reported in another study in which lactoferrin protected against arthritis in rats induced by adjuvant administration in the right hind paw [35] . In this model, the protective effect against arthritis is due to a downregulation of the proinflammatory cytokine TNF-[alpha] and an upregulation of the anti-inflammatory cytokine IL-10 [35] .

Lactoferrin also possesses the capacity to inhibit atopic contact dermatitis induced by oxazolone. In this study, lactoferrin applied topically prior to oxazolone sensitization prevents, in a dose-dependent manner, the migration of Langerhan's cells into the lymph nodes and subsequently activates cytotoxic T-cells and the accumulation of dendritic cells in inflamed sites. The mechanism of action seems to be the inhibition of TNF-[alpha] production by keratinocytes - the cytokine responsible for delivering the activation signal to Langerhan's cells [36] . Lactoferrin has the ability to bind to the CpG motifs via charge-charge interactions of the N-terminal sequence of lactoferrin. This binding of CpG on lactoferrin inhibits the stimulatory effects of these motifs on immune cells [37] . The binding of CpG and LPS suggests another mechanism for the anti-inflammatory effect of lactoferrin.

The immunosuppressive potential of lactoferrin has also been observed in lymphocyte cells as it demonstrates an inhibitory effect on lymphocyte proliferation upon stimulation by mitogens, as well as on IFN-γ production by these cells [19] . Other studies have reported the regulatory potential of lactoferrin on myelopoiesis by its suppressive activities on many cells including lymphocytes, macrophages and monocytes [38-40] . These processes are regulated by cytokines, indicating that lactoferrin could act as a regulatory nutrient by controlling cytokine production. The potential of lactoferrin to modulate immunity via regulation of cytokines has previously been demonstrated [41] .

Lactoferrin also exhibits an immunostimulatory potential. It has been demonstrated that this protein promotes the differentiation of T- and B-lymphocytes [42,43] . The incubation of immature T-lymphocytes in the presence of lactoferrin allows for the differentiation of these cells in CD4+ helper T-cells and the immune response of sheep red blood cells increases [42] . Natural killer (NK) cells as well as CD8+ T-cells have also increased in circulating cells after consumption of lactoferrin. This observation reveals a protective role in the control of tumor metastases since NK and CD8+ T-cells have important role in tumor inhibition [44,45] . Debbabi and colleagues have investigated the immune responses induced by repeated oral administration of lactoferrin in mice. IgA and IgG secretions are enhanced in the Peyer's patches and spleen from lactoferrin-fed mice but not in serum [46] . Wang and colleagues have also confirmed these observations in a study in which the CD4+ , CD8+ , asialoGM1+ (marker of NK cells), IgA+ and IgM+ B-cells have increased in the small intestine of mice treated with lactoferrin [47] . In this study, lactoferrin enhanced production of IL-18, IFN-γ and caspase-1, leading to an important stimulation of intestinal immunity. These results suggested that lactoferrin could act as an immunostimulatory factor on the mucosal immune system [46] . Immune effects occur as a result of the action through gut-associated lymphoid tissue (GALT), where lactoferrin may bind to epithelial cells or interact with M-cells in the Peyer's patches. These interactions of GALT may lead to the production of cytokines being released in the circulation and act systemically on circulating leukocytes.

Controversially, lactoferrin shows an absence of stimulation in B- or T-cells or in cytokine production (IL-6, TNF-[alpha], INF-γ) in Peyer's patches of newborn mice [48] . The consumption of nutrients by newborns in these studies probably leads to oral tolerance due to the supposed absence of early consumption of whey proteins prior to the experiment. The immune system of newborns was not completely developed and they do not have effective cells for the development of the same immunomodulation as an adult mouse [49] .

The phagocytic activity of human neutrophils is increased by the presence of lactoferrin. This activation appears to be a result of the specific binding of lactoferrin to neutrophils [50] . Following binding, lactoferrin is thought to be transported into the nucleus where it activates gene expressions responsible for the activation of the phagocytosis mechanism. It is also demonstrated that lactoferrin stimulates the production of IL-8 (a chemokine implicated in the activation of neutrophil activities) from human neutrophils [51] . In contrast, consumption of lactoferrin by newborn calves does not seem to change the production of the superoxide by PMN cells [52] . However, this study was completed with newborn calves that received lactoferrin only over a 9-day period; it is possible that the immunity of these young animals was not stimulated to activate their PMN cells over such a brief time period.

Peptides from lactoferrin

The main lactoferrin-derived peptide studied is lactoferricin, which corresponds to 25 amino acids from its N-terminus. This peptide appears to be responsible for the majority of immune benefits reported for lactoferrin. The activation of phagocytosis, as well as the production of IL-8 from neutrophils when treated with lactoferrin, is also observed when treated with the lactoferricin fraction [50,51] . It is mentioned above that the production of IL-18, IFN-γ and caspase-1 is increased by lactoferrin; a response also observed with lactoferricin [47] . Lactoferricin is capable of binding CpG motifs and preventing their immunostimulatory effects on B-cells [37] , suggesting a potential anti-inflammatory role of lactoferricin similar to lactoferrin. Moreover, the lactoferricin peptide has been found to suppress the IL-6 response in a monocytic cell line upon stimulation by lipopolysaccharide [53] .

Lactoferricin demonstrates an induction of apoptotic in human monocytic leukemic cells in a dose- and time-dependent manner. The apoptosis effect of lactoferricin remains present even if various cytokines and mitogens are added. This effect is correlated with high levels of intracellular reactive oxygen species (ROS), suggesting that the apoptosis-inducing activity is related to the production of intracellular ROS by phagocytic cells [54] .

Immunoglobulins

Igs are present in whey at a rate of approximately 10%. IgG1 is the major Ig found in whey ([proportional to]75%), followed by IgM, IgA and IgG2. In general, the Igs possess several immune benefits. The principal role of Igs is to defend organisms against pathogens and viruses. They are responsible for the activation of complement, increasing phagocytosis by leukocytes, preventing adhesion of microbes and neutralizing viruses and toxins [55] . All of these activities can likely be attributed to whey Igs. Effectively, Roos and colleagues have demonstrated that ingested Igs retain these immunological activities in the human ileum [56] . It has also been demonstrated that an Ig-like receptor isolated from milk inhibits HIV integrase as well as HIV protease [57] .

The immunoenhancing effects often demonstrated in animals fed with whey proteins can be attributed in part to an increase in intracellular gluthatione [58] . A study of animals fed with defined whey products has demonstrated that the product containing a higher proportion of Igs (and BSA) exhibited a better GSH-enhancing effect [3] . GSH is an intracellular antioxidant, which requires cysteine, glycine and glutamine for its synthesis. It is important to note that the Igs possess high levels of cysteine and glutamine.

Bovine serum albumine

BSA is a good source of essential amino acids and is found in approximately 5 to 10% of the whey protein content. Only a few studies have demonstrated the immunologic effects of BSA. GSH production increases when animals are fed with a whey diet containing a higher proportion of BSA. This explains the positive effect of BSA since GSH is responsible for some of the immune benefits observed with whey products [3] . As with the Igs, serum albumin fractions contain glutamine and cysteine amino acids that provide an enhancing effect on GSH production.

Moreover, as with [beta]-lactoglobulin, BSA increases the amount of antibody-forming cells in the spleen of mice immunized with trinitrophenylated ficoll [5] .

Glycomacropeptide

Glycomacropeptide (GMP) is released during the digestion of casein with the chymosin enzyme has been found in whey at a rate of approximately 10%. This is the only casein-derived peptide to be found in whey; all of the other peptides derived from casein remain in the cheese fraction. The GMP is present in whey only when chymosin is used during the cheese fabrication process. GMP is known to inhibit proliferation of mouse splenocytes as well as in cells from Peyer's patches isolated from a rabbit upon stimulation with LPS and phytohemmaglutinin (PHA) [59-61] . These results indicate that this peptide downregulates the immune system by suppressing T-lymphocytes (stimulated by PHA) as well as B-lymphocytes (stimulated by LPS). Li and Mine demonstrated that GMP is a potent immunoenhancer of macrophage proliferation as well as phagocytic activity in vitro [62] . The second part of this study demonstrated that the immunomodulatory effects of GMP are essentially due to sialic acid. Effectively, when enzymatic treatment removes this fragment, the effects are abolished.

Other effects of whey proteins

The majority of the studies concerning the immunomodulatory potential of whey proteins used whey directly and not one of individual whey proteins; the effects are therefore associated with whey proteins in general and not a specific protein.

The immunoenhancing effect of whey proteins on the formation of specific antibodies is well documented [5,63-66] . For example, one study demonstrated that mice fed with whey protein concentrate (WPC) express an elevated level of antibodies after the administration of different vaccines via different routes [66] . The same immunoenhancing effect was observed by Wong and Watson, when they reported higher concentrations of anti-ovalbumin antibodies in the presence of whey proteins [65] . The same study also demonstrated an increase in cell-mediated immunity after a period of 5 weeks in contrast with another study [5] . This difference; however may be as a result of different whey sources as well as the dose and duration of feeding. Variable effects of whey can be attributed to the whey source [67] . It is also demonstrated that whey enhances the proliferation of non-stimulating (mitogens) splenocytes [68] .

A hypothesis suggests that whey proteins may act via the GALT system for the production of antibodies. This hypothesis is supported by results demonstrating that WPC increases the antibody response in the intestinal tract of mice upon immunization with ovalbumin [69] .

Roth and colleagues demonstrated that ultrafiltered bovine whey increases the in vitro neutrophil functions of cattle cells as well as cytochromeC reduction in cells of dexamethasone-treated cattle [70] . This finding indicates that whey proteins increase the oxidative metabolism of cattle treated with dexamethasone. However, in this study, the whey came from hyperimmunized cows and it is possible that these effects are due to the cytokines present and not by the whey proteins.

Until now, the only immunostimulating properties of whey proteins have been discussed; however, some studies indicate that whey can be an immunosuppressive agent in some circumstances. In vitro studies show an immunosuppresion of T- and B-lymphocyte proliferation upon stimulation with mitogens [19,71,72] . Whey proteins also show an immunoregulatory effect in spleen cells, since the activation of these cells is downregulated by oral administration of whey [73] . The increase in transforming growth factor (TGF)-[beta] production in cells after whey treatment may explain this regulation. TGF-[beta] is a cytokine secreted during the tolerance induction and is responsible for the downregulation against food antigens [49] . These results show that whey proteins might be a good nutritive supplement due to their capabilities to modulate immunity, which may help in many different immune diseases.

Other effects of whey peptides

Some effects of whey peptides have been reported; however, the characterization of these peptides have not yet been investigated, thus the protein associated with the effective peptides is unknown. A hydrolyzed whey protein isolate composed of [beta]-lactoglobulin, [alpha]-lactalbumin and GMP has been shown to possess a stimulatory effect on lymphocyte proliferation in vitro [68] . Another peptide, a glycophosphopeptide isolated from cheese whey, has demonstrated a mitogenic activity on splenocytes [74] . It has also been demonstrated that the peptides issued from whey and milk possess a good immunostimulatory effect on keratinocyte growth in vitro [75] .

Some whey peptides also possess an inhibitory effect on the angiotensin-converting enzyme (ACE) [76] . ACE is responsible for inactivation of bradykynin. Bradykinin exhibits an essential role in inflammatory defence by the stimulation of macrophage and cytokine production [55] . Consequently, the peptides that act as ACE inhibitors stimulate the immune system via macrophage activation.

As mentioned previously, [beta]-lactoglobulin is principally responsible for milk allergies in children. It has been postulated that peptides from [beta]-lactoglobulin diminish the allergic reaction to this protein [14] . Many studies have also demonstrated a preventive role against milk allergies from hydrolyzed whey proteins [77-79] .

Moreover, many other constituents in milk and whey are known to possess immunomodulatory effects, such as cytokines, growth factors, enzymes and hormones [80-82] . A growth factor derived from whey has been shown to have a protective effect in a colitis animal model [83,84] . Francis and colleagues have demonstrated that a growth factor isolated from whey improved the growth rate of cells such as epithelials and fibroblasts [85,86] . These results led them to postulate that whey could be an interesting product against many diseases, not only due to the protein or peptide content.

Expert commentary & outlook

It has been known for many years that the consumption of nutraceuticals is important in maintaining general health. Statistics confirming the important rise in the consumption of nutraceutical products by the general population show that this is indeed the case [87,88] . For example, some reviews have already demonstrated the immunologic effects of milk-derived products such as yogurt [89] .

This article, which emphasizes the reported immune effects of whey proteins and derived peptides, demonstrates an important therapeutic potential of whey products, not only for maintaining good general health but also in guarding against many immune diseases, such as inflammatory disorders and autoimmune diseases. All the proteins and peptides present in whey possess an immunomodulatory potential, in both innate and acquired immunity, which provide an excellent mechanism of defence against all infections. The potential of enhancing GSH, NK cells, cytotoxic T-cells and the phagocytic process, leads to an enhancement in its capacity to defend against cancer.

Moreover, it is now reported that lactoferrin (and lactoferricin) provides whey with an important effect against bacterial and autoimmune inflammatory diseases. Many studies have proven that whey (and especially lactoferrin) plays a protective role against gastritis, asthma, colitis, arthritis and atopic contact dermatitis. The antiallergy properties of whey with regard to milk reactions were also reported in a study involving peptides derived from [beta]-lactoglobulin. These peptides appear to protect against allergies by the stimulation of oral tolerance. This observation is very important, as it presents an encouraging perspective that children, having consumed whey early on in life, can consume milk (an important nutrient) without the onset of allergies.

Whey products appear to have essential properties and could be used in the treatment of many diseases. Their effects could be comparable with the effects observed with many medicines and drugs, whilst still acting as a unique natural product. Further studies are needed to compare whey products with medicines in the treatment and prevention of disease.

These products or derived products from part of a future solution in obtaining and maintaining a healthy general immune system and in providing a natural, nutritive and non-chemical supplement against numerous diseases and immune disorders.

Table 1. Specific properties of whey proteins.

Protein MW (g/mol) IP Concentration Other structural properties

Milk (g/l)

Whey(g/l)

Whey (%)

[beta]-lactoglobulin

18,400

5.35-5.49

2.0-4.0

3.3

55-65

Presence of a hydrophobic region capable of binding to vitamin A and helping its absorption.

[alpha]-lactalbumin

14,200

4.2-4.5

1.0-1.5

1.2

15-25

Presence of a hydrophobic region that binds galactosyltransferase and aids lactose biosynthesis.

Lactoferrin

80,000

8.4-9.0

0.2

0.2

1-2

Presence of ferric ion binding sites, which permits Lf binding and the transport of iron - an important role in iron assimilation.

Immunoglobulin

80-900,000

5.5-8.3

0.4-1.0

0.5

10-15

Five categories of IGs: IgG1 (0.3-0.6 g/l), IgG2 (0.05-0.1 g/l), IgA (0.05-0.15 g/l), IgM (0.05-0.1 g/l) and IgE.

BSA

69,000

4.7-4.9

0.1-0.4

0.3

5-10

Important capabilities to bind fatty acids.

BSA: Bovine serum albumin; Ig: Immunoglobulin; IP: Isoelectric point; Lf: Lactoferrin; MW: Molecular weight. Adapted from [90-92] .

Table 2. Immunologic effects of whey proteins and peptides.

Whey fraction Reported effect Ref.

[beta]-lactoglobulin (proteins)

Stimulatory effect on splenocytes

[5,6]

Increase GSH

[3]

[beta]-lactoglobulin (peptides)

Carrier of retinoic acid

[9]

Contraction of ileum muscle

[10,11]

Increase oral tolerance to whey

[14]

[alpha]-lactalbumin (proteins)

Increase IL-1[alpha] production by macrophages

[19]

Induce apoptosis in tumor and immature cells

[20,21]

[alpha]-lactalbumin (peptides)

Modulation of B- and T-lymphocyte activities

[5,25]

Stimulation of adherence and phagocytosis of macrophages

[22,23]

Stimulation of oxidative burst response

[22]

Lactoferrin (proteins)

Inhibition of cytokines TNF-[alpha], IL-1[alpha] and IFN-γ

[19,32,35,36]

Anti-inflammatory effect on animal model

[33-36]

Upregulation of cytokine IL-10

[35]

Stimulatory effect on lymphocyte proliferation

[19]

Regulatory effect on myelopoiesis

[38-40]

Promote the differentiation of T- and B-lymphocytes

[42,43]

Increase NK cells, CD8+ cells, CD4+ cells

[44,45,47]

Bind to CpG and prevent stimulatory effect on B-cells

[37]

Stimulation of mucosal immunity through Peyer's patches

[46,47]

Increase phagocytosic activity of neutrophils as well as IL-8

[50,51]

Lactoferrin (peptides)

Increase phagocytosic activity of neutrophils as well as IL-8

[50,51]

Bind to CpG and prevent stimulatory effect on B-cells

[37]

Inhibition of IL-6 production by LPS

[53]

Increase of apoptosis in leukemic cells lines via production of ROS by phagocytic cells

[54]

Immunoglobulins

Increase of GSH

[58]

Activation of complement, increase of phagocytosis, prevent adhesion of microbes, neutralize viruses and toxins

[55]

BSAs

Increase of GSH

[3]

Stimulatory effect on splenocytes

[5]

GMPs

Suppress proliferation of cells upon mitogens stimulation

[59-61]

Stimulation of macrophages proliferation and phagocytosis

[62]

BSA: Bovine serum albumin; GSH: Glutathione; GMP: Glycomacropeptide; Ig: Immunoglobulin; IL: Interleukin; IFN: Interferon; LPS: Lipopolysaccharide; NK: Natural killer cell; ROS: Reactive oxygen species; TNF: Tumor necrosis factor.

Highlights

* The most important immune effect reported for the majority of whey proteins and peptides is the stimulation of innate immunity via an increase in macrophage activity and interleukin-8 production.

* Lactoferrin is the most studied of the whey proteins and its immune effects are diverse and dependent on the conditions for which it is used. For example, lactoferrin stimulates innate immunity in some conditions and could also could exert anti-inflammatory effects in others.

* Lactoferrin also exhibits effect on mucosal immunity via stimulation of Peyer's patches. This mucosal immunomodulation could be responsible for multiple reported systemic effects.

* The ability of whey proteins and peptides to enhance glutathione, natural killer cells, cytotoxic T-cells and the phagocytic process, fortifies the immune system against the development of cancers.

* Many studies demonstrated that whey plays a protective role against diseases such as gastritis, asthma, colitis, arthritis and atopic contact dermatitis.

Bibliography

Papers of special note have been highlighted as of interest (*) or of considerable interest (**) to readers.

1 Tipton KD, Elliott TA, Cree MG, Wolf SE, Sanford AP, Wolfe RR. Ingestion of casein and whey proteins results in muscle anabolism after resistance exercise. Med. Sci Sports Exerc. 36(12), 2073-2081 (2004).

2 Kawase M, Hashimoto H, Hosoda M, Morita H, Hosono A. Effect of administration of fermented milk containing whey protein concentrate to rats and healthy men on serum lipids and blood pressure. J. Dairy Sci. 83(2), 255-263 (2000).

3 Bounous G, Gold P. The biological activity of undenatured dietary whey proteins: role of glutathione. Clin. Invest. Med. 14(4), 296-309 (1991).

* Good introduction to GSH properties and its role in immunity as well as the effect of whey on GSH production.

4 Bounous G, Molson JH. The antioxidant system. AntiCancer Res. 23(2B), 1411-1415 (2003).

5 Bounous G, Kongshavn PA. Differential effect of dietary protein type on the B-cell and T-cell immune responses in mice. J. Nutr. 115(11), 1403-1408 (1985).

6 Wong KF, Middleton N, Montgomery M, Dey M, Carr RI. Immunostimulation of murine spleen cells by materials associated with bovine milk protein fractions. J. Dairy Sci. 81(7), 1825-1832 (1998).

7 Brix S, Bovetto L, Fritsche R, Barkholt V, Frokiaer H. Immunostimulatory potential of [beta]-lactoglobulin preparations: effects caused by endotoxin contamination. J. Aller. Clin. Imm. 112(6), 1216-1222 (2003).

8 Pellegrini A, Dettling C, Thomas U, Hunziker P. Isolation and characterization of four bactericidal domains in the bovine [beta]-lactoglobulin. Biochim. Biophys. Acta. 1526(2), 131-140 (2001).

9 Guimont C, Marchall E, Girardet JM, Linden G. Biologically active factors in bovine milk and dairy by-products: influence on cell culture . Crit. Rev. Food Sci. Nutr. 37(4), 393-410 (1997).

10 Yamauchi K. Biologically functional proteins of milk and peptides derived from milk proteins. International Dairy Federation Bulletin 272, 51-58 (1992).

11 Pihlanto-Leppala A, Paakkari I, Rinta-Koski M, Antila P. Bioactive peptide derived from in vitro proteolysis of bovine [beta]-lactoglobulin and its effect on smooth muscle . J. Dairy Res. 64(1), 149-155 (1997).

12 Yoshida T, Owens GK. Molecular determinants of vascular smooth muscle cell diversity. Circ Res. 96(3), 280-291 (2005).

13 Host A. Frequency of cow's milk allergy in childhood. Ann. Allergy Asthma Immunol. 89(6 Suppl. 1), 33-37 (2002).

14 Pecquet S, Bovetto L, Maynard F, Fritsche R. Peptides obtained by tryptic hydrolysis of bovine [beta]-lactoglobulin induce specific oral tolerance in mice. J. Allergy Clin. Immunol. 105(3), 514-521 (2000).

* Shows that peptides from [beta]-lactoglobulin can protect against allergies to the protein.

15 Matsumoto H, Shimokawa Y, Ushida Y, Toida T, Hayasawa H. New biological function of bovine [alpha]-lactalbumin: protective effect against ethanol- and stress-induced gastric mucosal injury in rats. Biosci. Biotechnol. Biochem. 65(5), 1104-1111 (2001).

16 Bruck WM, Kelleher SL, Gibson GR, Nielsen KE, Chatterton DEW, Lonnerdal B. rRNA probes used to quantify the effects of glycomacropeptide and [alpha]-lactalbumin supplementation on the predominant groups of intestinal bacteria of infant rhesus monkeys challenged with enteropathogenic Escherichia coli. J. Ped. Gastr. Nut. 37(3), 273-280 (2003).

17 Bounous G, Stevenson MM, Kongshavn PA. Influence of dietary lactalbumin hydrolysate on the immune system of mice and resistance to salmonellosis . J. Infect. Dis. 144(3), 281 (1981).

18 Bounous G, Letourneau L, Kongshavn PA. Influence of dietary protein type on the immune system of mice. J. Nutr. 113(7), 1415-1421 (1983).

19 Wong CW, Seow HF, Husband AJ, Regester GO, Watson DL. Effects of purified bovine whey factors on cellular immune functions in ruminants. Vet. Immunol. Immunopathol. 56(1-2), 85-96 (1997).

20 Hakansson A, Andreasson J, Zhivotovsky B, Karpman D, Orrenius S, Svanborg C. Multimeric [alpha]-lactalbumin from human milk induces apoptosis through a direct effect on cell nuclei. Exp. Cell Res. 246(2), 451-60 (1999).

21 Svensson M, Hakansson A, Mossberg AK, Linse S, Svanborg C. Conversion of [alpha]-lactalbumin to a protein inducing apoptosis. Proc. Natl. Acad. Sci. USA. 97(8), 4221-6 (2000).

22 Gattegno L, Migliore-Samour D, Saffar L, Jolles P. Enhancement of phagocytic activity of human monocytic macrophagic cells by immunostimulating peptides from human casein. Immunol. Lett. 18(1), 27-31 (1988).

23 Jaziri M, Migliore-Samour D, Casabianca-Pignede MR, Keddad K, Morgat JL, Jolles P. Specific binding sites on human phagocytic blood cells for Gly-Leu-Phe and Val-Glu-Pro-Ile-Pro-Tyr, immunostimulating peptides from human milk proteins. Biochim. Biophys. Acta. 1160(3), 251-261 (1992).

* Molecular mechanism by which peptides can exert their effects on immune cells.

24 Migliore-Samour D, Roch-Arveiller M, Tissot M et al. Effects of tripeptides derived from milk proteins on polymorphonuclear oxidative and phosphoinositide metabolisms. Biochem. Pharmacol. 44(4), 673-680 (1992).

25 Kayser H, Meisel H. Stimulation of human peripheral blood lymphocytes by bioactive peptides derived from bovine milk proteins. FEBS Lett. 383(1-2), 18-20 (1996).

26 Pellegrini A, Thomas U, Bramaz N, Hunziker P, von Fellenberg R. Isolation and identification of three bactericidal domains in the bovine [alpha]-lactalbumin molecule. Biochim. Biophys. Acta. 1426(3), 439-448 (1999).

* Shows antimicrobial activities of peptides isolated from [alpha]-lactalbumin.

27 Baldwin DA, Jenny ER, Aisen P. The effect of human serum transferrin and milk lactoferrin on hydroxyl radical formation from superoxide and hydrogen peroxide. J. Biol. Chem. 259(21), 13391-13394 (1984).

28 Bezwoda WR, Mansoor N. Lactoferrin from human breast milk and from neutrophil granulocytes. Comparative studies of isolation, quantitation, characterization and iron binding properties. Biomed. Chromatogr. 3(3), 121-126 (1989).

29 Masson PL, Heremans JF. Lactoferrin in milk from different species . Comp. Biochem. Physiol. B. 39(1), 119-129 (1971).

30 Masson PL, Heremans JF, Ferin J. Presence of an iron-binding protein (lactoferrin) in the genital tract of the human female I. Its immunohistochemical localization in the endometrium. Fertil. Steril. 19(5), 679-689 (1968).

31 Masson PL, Heremans JF, Schonne E. Lactoferrin, an iron-binding protein in neutrophilic leukocytes. J. Exp. Med. 130(3), 643-658 (1969).

32 Machnicki M, Zimecki M, Zagulski T. Lactoferrin regulates the release of tumour necrosis factor-[alpha] and interleukin-6 in vivo. Int. J. Exp. Pathol. 74(5), 433-439 (1993).

33 Dial EJ, Romero JJ, Headon DR, Lichtenberger LM. Recombinant human lactoferrin is effective in the treatment of Helicobacter felis -infected mice. J. Pharm. Pharmacol. 52(12), 1541-1546 (2000).

34 Guillen C, McInnes IB, Vaughan D, Speekenbrink AB, Brock JH. The effects of local administration of lactoferrin on inflammation in murine autoimmune and infectious arthritis. Arthritis Rheum. 43(9), 2073-2080 (2000).

35 Hayashida K, Kaneko T, Takeuchi T, Shimizu H, Ando K, Harada E. Oral administration of lactoferrin inhibits inflammation and nociception in rat adjuvant-induced arthritis. J. Vet. Med. Sci. 66(2), 149-154 (2004).

** Describes mechanism of lactoferrin exerting its anti-inflammatory activity against arthritis.

36 Kimber I, Cumberbatch M, Dearman RJ, Headon DR, Bhushan M, Griffiths CE. Lactoferrin: influences on Langerhans cells, epidermal cytokines, and cutaneous inflammation. Biochem. Cell Biol. 80(1), 103-107 (2002).

** Utilization of lactoferrin by topical contact instead of orally.

37 Britigan BE, Lewis TS, Waldschmidt M, McCormick ML, Krieg AM. Lactoferrin binds CpG-containing oligonucleotides and inhibits their immunostimulatory effects on human B-cells. J. Immunol. 167(5), 2921-2928 (2001).

38 Broxmeyer HE, DeSousa M, Smithyman A et al. Specificity and modulation of the action of lactoferrin, a negative feedback regulator of myelopoiesis. Blood 55(2), 324-333 (1980).

39 Bagby GC, Jr., Rigas VD, Bennett RM, Vandenbark AA, Garewal HS. Interaction of lactoferrin, monocytes, and T-lymphocyte subsets in the regulation of steady-state granulopoiesis in vitro. J. Clin. Invest. 68(1), 56-63 (1981).

40 Bagby GC, Jr. Regulation of granulopoiesis: the lactoferrin controversy . Blood Cells 15(2), 386-399 (1989).

41 Crouch SP, Slater KJ, Fletcher J. Regulation of cytokine release from mononuclear cells by the iron-binding protein lactoferrin. Blood 80(1), 235-240 (1992).

42 Zimecki M, Mazurier J, Machnicki M, Wieczorek Z, Montreuil J, Spik G. Immunostimulatory activity of lactotransferrin and maturation of CD4- CD8- murine thymocytes. Immunol. Lett. 30(1), 119-123 (1991).

43 Zimecki M, Mazurier J, Spik G, Kapp JA. Human lactoferrin induces phenotypic and functional changes in murine splenic B-cells. Immunology 86(1), 122-127 (1995).

44 Sekine K, Ushida Y, Kuhara T et al. Inhibition of initiation and early stage development of aberrant crypt foci and enhanced natural killer activity in male rats administered bovine lactoferrin concomitantly with azoxymethane. Cancer Letter 121(2), 211-216 (1997).

45 Iigo M, Kuhara T, Ushida Y, Sekine K, Moore MA, Tsuda H. Inhibitory effects of bovine lactoferrin on colon carcinoma 26 lung metastasis in mice . Clin. Exp. Metastasis 17(1), 35-40 (1999).

46 Debbabi H, Dubarry M, Rautureau M, Tome D. Bovine lactoferrin induces both mucosal and systemic immune response in mice. J. Dairy Res. 65(2), 283-293 (1998).

* Shows the effect on mucosal immunity from lactoferrin consumption.

47 Wang WP, Iigo M, Sato J, Sekine K, Adachi I, Tsuda H. Activation of intestinal mucosal immunity in tumor-bearing mice by lactoferrin . Jpn. J. Cancer Res. 91(10), 1022-1027 (2000).

48 Griffiths EA, Duffy LC, Schanbacher FL et al. In vivo effects of bifidobacteria and lactoferrin on gut endotoxin concentration and mucosal immunity in balb/c mice. Dig. Dis. Sci. 49(4), 579-589 (2004).

49 Weiner HL. Oral tolerance: immune mechanisms and treatment of autoimmune diseases. Immunol. Today 18(7), 335-343 (1997).

50 Miyauchi H, Hashimoto S, Nakajima M, Shinoda I, Fukuwatari Y, Hayasawa H. Bovine lactoferrin stimulates the phagocytic activity of human neutrophils: identification of its active domain. Cell. Immunol. 187(1), 34-37 (1998).

51 Shinoda I, Takase M, Fukuwatari Y, Shimamura S, Koller M, Konig W. Effects of lactoferrin and lactoferricin on the release of interleukin-8 from human polymorphonuclear leukocytes. Biosci. Biotechnol. Biochem. 60(3), 521-523 (1996).

52 Dawes ME, Lakritz J, Tyler JW et al. Effects of supplemental lactoferrin on serum lactoferrin and IgG concentrations and neutrophil oxidative metabolism in Holstein calves . J. Vet. Int. Med. 18(1), 104-108 (2004).

53 Mattsby-Baltzer I, Roseanu A, Motas C, Elverfors J, Engberg I, Hanson LA. Lactoferrin or a fragment thereof inhibits the endotoxin-induced interleukin-6 response in human monocytic cells. Pediatr. Res. 40(2), 257-262 (1996).

54 Yoo YC, Watanabe R, Koike Y et al. Apoptosis in human leukemic cells induced by lactoferricin, a bovine milk protein-derived peptide: involvement of reactive oxygen species . Biochem. Biophys. Res. Commun. 237(3), 624-628 (1997).

55 Janeway CA, Travers P. Immunobiologie (2nd Edition). De Boeck et Larcier, Paris, France, (1996).

56 Roos N, Mahe S, Benamouzig R, Sick H, Rautureau J, Tome D. 15 N -labeled immunoglobulins from bovine colostrum are partially resistant to digestion in human intestine. J. Nutr. 125(5), 1238-1244 (1995).

57 Ng TB, Ye XY. A polymeric immunoglobulin receptor-like milk protein with inhibitory activity on human immunodeficiency virus type 1 reverse transcriptase. Int. J. Bioch. Cell. Biol. 36(11), 2242-2249 (2004).

58 Bounous G, Batist G, Gold P. Immunoenhancing property of dietary whey protein in mice: role of glutathione. Clin. Invest. Med. 12(3), 154-161 (1989).

59 Otani H, Monnai M, Hosono A. Bovine kappa-casein as inhibitor of the proliferation of mouse splenocytes induced by lipopolysaccharide stimulation. Milchwissenschaft 47, 512-515 (1992).

60 Otani H, Hata I. Inhibition of proliferative responses of mouse spleen lymphocytes and rabbit Peyer's patch cells by bovine milk caseins and their digests. J. Dairy Res. 62(2), 339-348 (1995).

61 Otani H, Monnai M, Kawasaki Y, Kawakami H, Tanimoto M. Inhibition of mitogen-induced proliferative responses of lymphocytes by bovine κ-caseinoglycopeptides having different carbohydrate chains . J. Dairy Res. 62(2), 349-357 (1995).

62 Li EW, Mine Y. Immunoenhancing effects of bovine glycomacropeptide and its derivatives on the proliferative response and phagocytic activities of human macrophagelike cells, U937. J. Agric. Food. Chem. . 52(9), 2704-2708 (2004).

63 Bounous G, Shenouda N, Kongshavn PA, Osmond DG. Mechanism of altered B-cell response induced by changes in dietary protein type in mice . J. Nutr. 115(11), 1409-1417 (1985).

64 Bounous G, Kongshavn PA, Gold P. The immunoenhancing property of dietary whey protein concentrate. Clin. Invest. Med. 11(4), 271-278 (1988).

65 Wong CW, Watson DL. Immunomodulatory effects of dietary whey proteins in mice. J. Dairy Res. 62(2), 359-368 (1995).

66 Low PPL, Rutherfurd KJ, Gill HS, Cross ML. Effect of dietary whey protein concentrate on primary and secondary antibody responses in immunized BALB/c mice. Int. Immunopharmacol. 3(3), 393-401 (2003).

* Describes the potential of whey to act as an oral adjuvant in vaccination.

67 Middleton N, Reid JR, Coolbear T, Jelen P. Proliferation and intracellular glutathione in Jurkat T-cells with concentrated whey protein products . Int. Dairy J. 13(7), 565-573 (2003).

68 Mercier A, Gauthier SF, Fliss L. Immunomodulating effects of whey proteins and their enzymatic digests. Int. Dairy J. 14(3), 175-183 (2004).

69 Low PPL, Rutherfurd KJ, Cross ML, Gill HS. Enhancement of mucosal antibody responses by dietary whey protein concentrate. Food Agri. Imm. 13, 255-264 (2001).

70 Roth JA, Frank DE, Weighner P, Weighner M. Enhancement of neutrophil function by ultrafiltered bovine whey. J. Dairy Sci. 84(4), 824-829 (2001).

71 Otani H, Odashima M. Inhibition of proliferative responses of mouse spleen lymphocytes by lacto- and ovotransferrins. Food Agri. Imm. 9, 193-201 (1997).

72 Cross ML, Gill HS. Modulation of immune function by a modified bovine whey protein concentrate. Immunol. Cell. Biol. 77(4), 345-350 (1999).

73 Penttila IA, Zhang MF, Bates E, Regester G, Read LC, Zola H. Immune modulation in suckling rat pups by a growth factor extract derived from milk whey. J. Dairy Res. 68(4), 587-599 (2001).

74 Yun SS, Sugita-Konishi Y, Kumagai S, Yamauchi K. Isolation of mitogenic glycophosphopeptides from cheese whey protein concentrate. Biosci. Biotechnol. Biochem. 60(3), 429-433 (1996).

75 Amiot J, Germain L, Turgeon S, Lemay M, OrySalam C, Auger FA. Peptides from milk protein hydrolysates to improve the growth of human keratinocytes in culture. Int. Dairy J. 14(7), 619-626 (2004).

76 Murakami M, Tonouchi H, Takahashi R et al. Structural analysis of a new antihypertensive peptide ([beta]-lactosin B) isolated from a commercial whey product. J. Dairy Sci. 87(7), 1967-1974 (2004).

77 Han YS, Park HY, Ahn KM, Lee JS, Choi HM, Lee SI. Short-term effect of partially hydrolyzed formula on the prevention of development of atopic dermatitis in infants at high risk. J. Korean Med. Sci. 18(4), 547-551 (2003).

* Clincal study that proves that hydolyzed whey formula can protect against allergies.

78 Tanabe S, Tesaki S, Watanabe J, Fukushi E, Sonoyama K, Kawabata J. Isolation and structural elucidation of a peptide derived from Edam cheese that inhibits [beta]-lactoglobulin transport. J. Dairy Sci. 86(2), 464-468 (2003).

79 Peng HJ, Su SN, Tsai JJ, Tsai LC, Kuo HL, Kuo SW. Effect of ingestion of cow's milk hydrolysed formulae on whey protein-specific Th2 immune responses in naive and sensitized mice. Clin. Exp. Allergy 34(4), 663-670 (2004).

80 Grosvenor CE, Picciano MF, Baumrucker CR. Hormones and growth factors in milk. Endocr. Rev. 14(6), 710-728 (1993).

81 Garofalo RP, Goldman AS. Cytokines, chemokines, and colony-stimulating factors in human milk: the 1997 update. Biol. Neonate 74(2), 134-142 (1998).

82 Rowlands JC, He L, Hakkak R, Ronis MJ, Badger TM. Soy and whey proteins downregulate DMBA-induced liver and mammary gland CYP1 expression in female rats. J. Nutr. 131(12), 3281-3287 (2001).

83 Procaccino F, Reinshagen M, Hoffmann P et al. Protective effect of epidermal growth factor in an experimental model of colitis in rats. Gastroenterology 107(1), 12-17 (1994).

84 Porter SN, Howarth GS, Butler RN. An orally administered growth factor extract derived from bovine whey suppresses breath ethane in colitic rats . Scand. J. Gastroenterol. 33(9), 967-974 (1998).

85 Francis GL, Regester GO, Webb HA, Ballard FJ. Extraction from cheese whey by cation-exchange chromatography of factors that stimulate the growth of mammalian cells. J. Dairy Sci. 78(6), 1209-1218 (1995).

86 Rogers ML, Belford DA, Francis GL, Ballard FJ. Identification of fibroblast growth factors in bovine cheese whey. J. Dairy Res. 62(3), 501-507 (1995).

87 Hilliam M. The market for functional foods. Int. Dairy J. 8, 349-353 (1998).

88 Milner JA. Functional foods: the US perspective. Am. J. Clin. Nutr. 71(6 Suppl), 1654S-1659S (2000).

89 Meydani SN, Ha WK. Immunologic effects of yogurt. Am. J. Clin. Nutr. 71(4), 861-872 (2000).

90 de Wit JN. Marschall Rhone-Poulenc Award Lecture. Nutritional and functional characteristics of whey proteins in food products. J. Dairy Sci. 81(3), 597-608 (1998).

** Good overview of the characteristics of the different whey proteins.

91 Marshall KR, Harper WJ. Trends in utilization of whey and whey derivatives. Bulletin of the International Dairy Federation 233, 21-32 (1988).

92 Cayot P, Lorient D. Structures et technofonctions des protéines du lait . Arilait Recherches, Paris, France, 37-182 (1998).

Author Affiliation(s):

1 1 Institut National de la Recherche Scientifique, Institut Armand-Frappier, 531 Blvd. Des Prairies, Laval, Quebec, H7V 1B7, Canada

2 2 Technologie Biolactis, 531 Blvd. Des Prairies, Building 18, Laval, Quebec, H7V 1B7, Canada. plemieux@biolactis.com

Author Note(s):

[dagger] Author for correspondence

Source Citation

Source Citation   

Gale Document Number: GALE|A225319428