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Immune Components of Colostrum and Milk—

A Historical Perspective

Article in Journal of Mammary Gland Biology and Neoplasia · January 2008

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J Mammary Gland Biol Neoplasia
DOI 10.1007/s10911-007-9051-7

Immune Components of Colostrum

and Milk—A Historical Perspective
Thomas T. Wheeler & Alison J. Hodgkinson &
Colin G. Prosser & Stephen R. Davis

Received: 2 October 2007 / Accepted: 12 October 2007

# Springer Science + Business Media, LLC 2007

Abstract Key developments in the understanding of the Introduction

immune functions of milk and colostrum are reviewed,
focusing on their proteinaceous components. The topics The concept that milk, mammary secretions and the
covered include the immunoglobulins, immune cells, mammary gland have major roles in immune defense is
immunomodulatory substances, and antimicrobial proteins. an old one. The bactericidal property of milk was recorded
The contributions of new technologies and the introduction in the scientific literature in the late nineteenth century [1,
of fresh approaches from other fields are highlighted, as are 2]. Moreover, observations at this time on the ability of
the contributions that mammary biology research has made milk to provide immunity to the newborn [3] played a key
to the development of other fields. Finally, a summary of role in the development of modern immunology. The aim of
some current outstanding questions and likely future this review is to provide a modern audience with a timeline
directions of the field are given. for the key discoveries in milk immunology, illustrating
how the current understanding of the immune function of
Keywords Lactation . Mammary . Antimicrobial milk evolved, and to offer some pointers for the future
direction of the field.
Abbreviations This review is divided into three sections: the immuno-
BPI bactericidal/permeability increasing protein globulins and immune cells, immunomodulatory compo-
DHA docosahexaenoic acids nents, and antimicrobial components, covering elements of
EPA eicosapentaenoic acid both innate and adaptive immunity, immune defenses in the
IgA immunoglobulin A mammary gland and the participation of the mammary
IgG immunoglobulin G gland in the mucosal defense system. In this we focus
LBP lipopolysaccharide binding protein largely on the proteinaceous components of milk, some of
LC-PUFA long chain-polyunsaturated fatty acids which are depicted in Fig. 1. Reviews describing physical
barriers, the role of probiotics, and the carbohydrate and
lipid components of milk that have host defense functions
T. T. Wheeler (*) : A. J. Hodgkinson
Dairy Science and Technologies Section,
have recently been presented elsewhere [4, 5].
AgResearch, Ruakura Research Centre,
Private Bag 3123,
Hamilton, New Zealand Immunoglobulins in Milk
e-mail: [email protected]

C. G. Prosser Today it is largely forgotten that the immune properties of

Dairy Goat Co-operative (NZ) Ltd., milk helped lay the foundation of modern immunology. In
Hamilton, New Zealand 1892, Paul Ehrlich demonstrated that mice immunized
S. R. Davis
against plant toxins passed immunity to the fetus in utero as
Vialactia Biosciences Ltd., well as via the milk. These observations were subsequently
P. O. Box 109-185, Newmarket, Auckland, New Zealand shown to be due to an ammonium sulfate precipitable
 J Mammary Gland Biol Neoplasia

blood IgG (substances causing agglutination of cells) in the blood,

IgA
basolateral derived from the mother’s milk [11].
membrane It was appreciated early on that there were significant
Immune cells
differences between mammals in the immunological func-
tion of the mammary gland. For example, it was noted that
colostrum was important for immune transfer to the newborn
only in mammals with multi-layered placentae (for example
mammary
epithelial cell the ungulates) [12–14]. In these species, the antibodies were
shown to be absorbed into the bloodstream via the neonatal
IgG sIgA proteases intestinal mucosa in the first 24–48 h of life [15]. Thus, by
complement lactoferrin 1930, the role of the mammary gland in conferring
cytokines antimicrobial acute phase protection to the neonate through the accumulation and
peptides proteins
secretion of agglutinating antibodies directed against
lysozyme, oxidases,
alveolar lumen peroxidases pathogens was firmly established.
Fig. 1 Schematic showing some of the principal known proteinaceous The bivalent structure of antibodies and the nature of
components of the host defense system in milk and colostrum. their interaction with antigens were gradually elucidated
through the 1940s. The highly specific nature of the
substance, termed an antibody [6]. Ehrlich extended this antibodies in mammary secretions was demonstrated
work to develop the concept of passive and active through analysis of the antibodies in colostrum fed to
immunisation via antibodies, for which he was awarded calves that had died from infections with different strains of
the Nobel Prize in 1908. Many other researchers made E. coli [16]. While the presence of antibodies in milk and
important contributions through this early period (see colostrum was indisputable, at this time there was still
Campbell and Petersen [7] for an excellent review of the debate over their source and the nature of the therapeutic
history of immune milk). For example, Famulener [8] benefit that could be derived from them [7, 17].
showed that goats immunized prior to parturition could The development of polyacrylamide gel electrophoresis
transmit this immunity to the sera of their offspring via and size-exclusion chromatography by the 1960s greatly
colostrum and that this transfer could also take place if goat accelerated the characterisation of the immune components
serum was fed. Further experiments around this time of milk. These techniques led to the definition of individual
established the concept of passive immunisation through classes of immunoglobulins (i.e. the agglutinating globulins)
the transfer of milk-derived antibodies from the intestinal as well as comparisons between species. Thus, in sheep the
lumen into the bloodstream. Colostrum was shown to predominant antibody in colostrum was shown to be IgG1
contain a greater concentration of antibodies than mature [18], while in humans it was IgA [19]. Moreover,
milk, and the origin of these substances from serum was mammary secretions and serum were shown to have
confirmed [9]. The importance of colostrum in providing distinct immunoglobulin compositions (see Table 1). De-
protection from bacterial infection was shown by Smith and spite much additional data being accumulated, there were
Little [10] who reported that a calf deprived of colostrum still questions about the fundamental reason for the
lacks “something” which allowed intestinal bacteria to difference in milk immunoglobulins between species.
invade the body and multiply in various organs. Allied In the 1960s, Campbell and Petersen [7] were enthusiastic
work showed that this “something” was specific agglutinins advocates for the therapeutic benefits of colostrum and milk

Table 1 Comparison of levels of immunoglobulin in human and cow colostrum, milk and serum.

Species Ig Concentration (g/l) % of total Ig

Colostrum Milk Serum Colostrum Milk Serum

Humana IgG 0.43 0.04 12.10 2.0 3.0 78.0

IgA 17.35 1.00 2.50 90.0 87.0 16.0
IgM 1.59 0.10 0.93 8.0 10.0 6.0
Cowb IgG1 46.40 0.58 11.20 75.5 71.6 47.0
IgG2 2.87 0.06 9.20 4.7 7.4 38.6
IgA 5.36 0.08 0.37 8.8 9.9 1.6
IgM 6.77 0.09 3.05 11.0 11.1 12.8

Ig immunoglobulin, a [20], b [21].

J Mammary Gland Biol Neoplasia

from immunized cows. In particular, and most controver- presence of the prolactin release inhibitor, bromoergocryp-
sially, systemic responses to oral consumption of immune tine, colostrum-like secretion continued post-partum for
milk were claimed, with beneficial effects on symptoms of several days and mammary IgG1 receptor activity was
arthritis and hay fever [7]. This work was commercialized maintained [26]. Addition of prolactin to mammary cells in
by Ralph Stolle, whose company (http://www.smbimilk. culture resulted in down-regulation of the receptor [29].
com) still markets ‘hyperimmune milk’ to the present Thus, it seems that prolactin plays a role in regulating the
day. IgG1 receptor during Stage 2 lactogenesis. However,
The origin of the immunoglobulins in milk was the regulation of the induction of IgG receptor during Stage 1
source of some controversy. Campbell and Petersen lactogenesis remains poorly understood.
believed that the majority of antibodies in bovine mammary Advances in the knowledge of cellular immunity in
secretions were synthesized within the udder. They based mucosal tissues led to greater understanding of the origins
this premise, in part, on their observation that ‘adequate’ of plasma cells in the mammary gland. These cells were
numbers of plasma cells are present in the udder immedi- shown to migrate into the mammary gland from the gut-
ately before parturition [20]. However, later workers found associated lymphoid tissues (GALT) [30]. Adoptive transfer
only low levels of plasma cells [21]. These differences may studies revealed that lymphocytes from the GALT populate
have been due to differences in the health status of the many mucosal effector sites including the mammary gland.
gland between the experiments. Others favoured a humoral The precursors of plasma cells destined to produce IgA
source of the immunoglobulins in colostrum and milk [17], were shown to originate from GALT and traffic into the
based on the observed decrease in serum immunoglobulin mammary gland near the time of parturition as well as in
levels before parturition and a corresponding increase in the middle and late lactation [31]. In the late 1970s the concept
udder [22]. While IgG was shown to be transported to the of the ‘common mucosal immune system’ was proposed
mammary gland from the serum, IgA was found to be [32] in which the antigenic experience at one mucosal
synthesized within the mammary gland by plasma cells surface was deemed to lead to effector responses at a distant
which had migrated into the gland from the gastrointestinal mucosal tissue, a concept that was supported by the
tract (see below). It was found that in species where IgG is discovery of cell surface receptors and cytokines. Thus,
transferred to the foetus prior to birth (e.g. humans), IgA is the idea emerged that cellular immune defense in the
the predominant immunoglobulin in colostrum, whereas in mammary gland is a local feature of an organism-wide
those born with no circulating IgG (e.g. cattle and sheep), integrated system.
IgG is the predominant immunoglobulin in colostrum. A In the mouse, T-cell migration to the mammary gland
10-fold increase in the IgG1:IgG2 ration in bovine colos- was shown to be mediated by mucosal addressin cell-
trum over that in serum suggested a specific IgG1 transport adhesion molecule-1 (MAdCAM-1) expressed on the
mechanism [23]. mammary vascular endothelium [33], while IgA plasma
By the 1970s, the mechanism for the transport of IgG cell recruitment was found to be facilitated by vascular cell-
into the mammary gland had been elucidated. The IgG adhesion molecule (VCAM-1) [34]. These adhesion mole-
receptor, FcRn, was identified and shown to be present on cules were found to be present in different proportions in
the apical surface of the gut of the suckling rat [24] as well the ruminant mammary gland [35], possibly accounting for
as on the basolateral surface of the secretory epithelial cell some of the differences between species of the immuno-
during colostrogenesis [25]. Immunohistochemical analysis globulin isotypes in colostrum or milk. Recent studies have
showed that FcRn expression coincided with Stage 1 established a role for chemokines in directing immune cells
lactogenesis (the onset of colostrogenesis) and was de- to the mammary gland. Mouse mammary tissue has been
creased during stage 2 lactogenesis (the onset of copious shown to express the CCL28 (MEC) chemokine receptor,
secretion; [26]). which binds to the CCR10 ligand on IgA lymphocytes [36].
The hormonal regulation of immunoglobulin transport Despite these advances, the nature of the stimulus for the
into colostrum and milk has been investigated but remains trafficking of IgA plasma cells from the intestine to the
incompletely described. Smith and co-workers [27] sug- mammary gland, particularly in late-pregnancy, is as yet
gested that changing serum concentrations of estrogen and unknown.
progesterone in late pregnancy exerted a controlling Transport of IgA from immune cells in the mammary
influence on the selective transport of IgG1 to bovine gland into colostrum and milk has been elucidated. Key to
lacteal fluid and thus colostrum formation. However, others this was the identification of the polymeric immunoglobulin
suggested that the rate of IgG1 transfer was a consequence receptor (pIgR), a transmembrane glycoprotein selectively
of mammary gland development [28], which is itself under expressed on mucosal and secretory epithelial basolateral
the control of estrogen and progesterone. Investigation into cell surfaces. Earlier studies had identified a ‘secretory
the role of prolactin in IgG transport showed that in the component’ that was linked to dimeric IgA following its
 J Mammary Gland Biol Neoplasia

secretion into mucosal fluids [37]. This secretory compo- immunoglobulins and complement present in milk. By the
nent was later identified as the extracellular domain of 1970s, most components of the complement system were
pIgR. The transport of IgA involved binding to pIgR at the described in human [50] and bovine milk and colostrum
basolateral membrane, passage to the apical membrane then [51].
release into the alveolar lumen by cleavage of the complex, The immune cells in milk may also modulate the
with IgA remaining linked to secretory component [38]. neonatal immune system. This idea was first raised as a
The pIgR protein was found to function as a sacrificial result of observing an increased level of neutrophils in the
receptor, with one molecule of pIgR synthesised for every colostrum of non-nursing mothers in the absence of
molecule of immunoglobulin secreted. The production of infection [52]. Evidence of transfer of tuberculin sensitivity
pIgR appears to limit the amount of IgA that is transported. to suckling infants [53] was used to support this hypothesis,
Transgenic mice overexpressing pIgR in their mammary as was transfer of partial tolerance and graft-versus-host
glands had double the IgA concentration in their milk reactions in rodents [54]. Subsequent studies have con-
compared with non-transgenic mice [39]. The expression of firmed that milk cells can traverse the neonatal intestinal
pIgR was shown to be under endocrine control in some epithelium in a range of species [55, 56]. However, other
tissues [40]. Prolactin and glucocorticoids were shown to studies found no evidence for transfer of cells across the gut
enhance mammary pIgR protein and mRNA levels during [57] or tuberculin-sensitivity via milk [58].
lactation in the sheep [41]. A more recent explanation for modulation of the neonatal
Secretory component was found to be important for the immune system via the cells themselves, is the production of
function of IgA. When bound to IgA, it conferred soluble immune mediators. These “lymphokines” were first
protection from proteolytic degradation in the intestine described in the 1980s and shown to stimulate the immune
[42, 43], and its presence facilitated localization of IgA to cells in the suckling animal [58, 59]. Many studies have
mucus [44]. Free secretory component has antimicrobial since revealed the large range of these proteins (now termed
properties in its own right. It was first isolated from bovine “cytokines”) present in human colostrum or milk. These
milk [45] and subsequent studies have demonstrated that it include IL-1β, IL-6, IL-8, IL-10, IL-12, IL-18, IFN-γ,
binds to fimbrial colonization factors on bacterial surfaces, TNF-α, TGF-β, G-CSF, M-CSF, GM-CSF. These cyto-
thereby reducing their pathogenicity [46]. Free secretory kines are thought to regulate the neonatal immune system
component also binds the cytokine IL-8 thereby modifying in a variety of ways. For example, TGF-β has been
the pro-inflammatory effects of this cytokine [47]. proposed to reduce inflammatory reactions in the gut [60],
to reduce allergy [61] and to stimulate intestinal IgA
production [62]. Cyokines are also present in cow’s milk,
Immunomodulatory Properties of Milk with increased levels during infection (reviewed in [63]).
The advent of recombinant technology enabled the
The immune cells in milk are an important aspect of the production of cytokines such as Il-1, IL-2, IFN-γ and
immunomodulatory activity of milk. The presence of cells GM-CSF which induce higher numbers of neutrophils and
in milk and their association with mammary infection was macrophages when infused into the bovine udder. This
described in the early twentieth century [48, 49]. However, finding has led to the idea that cytokines may be used as
the types of cells, their origin and their role in prevention of immunotherapy for prevention of mastitis in cattle, but to
infection within the gland were debated for many years, date none have been commercialized for routine use.
particularly in non-bovine species. Significant advances in In vitro studies carried out over the last 20 years suggest
immunological analytical techniques eventually led to that the repertoire of immune factors in milk also includes
characterisation of the cell types in milk of humans as well immunomodulatory peptides derived from caseins or whey
as several other species. It is now clear that the cells in proteins. These peptides are receiving much attention as
mammary secretions of all species studied consist of possible sources of ‘natural’ bioactivity with health benefits
neutrophils, macrophages, lymphocytes and a smaller for the consumer (see [64]). Casein peptides have also been
percentage of epithelial cells. In general, macrophages are used to stimulate the innate immune system within the
the major cell population in milk from the healthy lactating mammary gland and prevent infections within the udder of
mammary gland, whereas neutrophils predominate during cows at drying off [65]. However, peptides from the A1
early inflammation. These cells play an important role in variant of β-casein have been proposed to be involved in
signalling the presence of pathogens to the systemic the development of type 1 diabetes, an auto-immune
immune system and thereby mounting a local immune related disease, largely on the basis of epidemiological
response against pathogens. evidence [66].
Neutrophils and macrophages also phagocytose and kill In the 1980s, nucleotides were found in milk and
bacteria directly, and this activity is enhanced by opsonic proposed as potential immune regulators. By the mid
J Mammary Gland Biol Neoplasia

1980s, dietary nucleotide/nucleosides were shown to the reduction in exposure of the infant’s immune system to
modulate cell-mediated immunity and prevent infection in foreign antigens at critical time points may be as important
animals [67]. Since then several infant feeding studies using as the supply of immune factors from milk in the
formula supplemented with nucleotides suggest they subsequent development of allergy in the infant. Much
can influence immune cell development and reduce the further work in these areas will be required to unravel the
incidence of diarrhea. role of immune factors in colostrum and milk and
Long chain poly-unsaturated fatty acids (LC-PUFA) development of mucosal immunity in infants.
such as DHA (docosahexaenoic acids) and EPA (eicosa-
pentaenoic acid) were also identified in the 1980s as
possible candidates for immune modulation. This notion Antimicrobial Proteins in Milk
was based on the observation that the diets of Eskimo
children have a high proportion of fish containing these Initial characterisation of the bactericidal components in
fatty acids, as well as a low incidence of asthma [68]. milk was carried out in the 1920s. In 1922, Alexander
Subsequent studies have shown that infants fed formula Fleming [77] described a bacteriolytic activity which was
supplemented with DHA also have decreased incidence of present in a number of biological fluids, which he named
bronchitis than infants fed un-supplemented formula [69]. lysozyme. Not long after, this activity was also described in
Supplementation of a mother’s diet with LC-PUFA to milk [78]. About this time, what appeared to be a distinct
enhance the immunomodulatory profile of breast milk is antimicrobial action in milk was also described. This
also receiving interest as a means to modify the infant’s activity against streptococci was termed “lactenin” [79].
immune system [70]. Improved techniques for protein analysis in the 1940s
A common feature of the studies over the years is that and 1950s led to the characterisation of these activities. The
human milk is a rich source of many of the above- bactericidal activity of lysozyme was shown to be largely
mentioned immunomodulatory components. The elucida- due to its ability to digest the complex polysaccharide
tion of many of these compounds was driven by the idea component of the bacterial cell wall (reviewed in [80]). The
that breast-feeding prevents gastrointestinal and respiratory isolation of lysozyme from human and cow’s milk was
infectious disease in infants, as initially documented in reported in the 1960s [81, 82], with human milk having by
1930s [71]. This concept has now been broadened to far the greater abundance. Milk was shown to contain at
include potential role of immunomodulatory elements in least two distinct lactenins [83]. One of these was an
development of atopic disease (allergy) in children. Con- oxidase, termed lactoperoxidase [84], which had earlier
centrations of LCPUFA, cytokines, nucleotides and poly- been purified from cow’s milk [85]. Lactoperoxidase was
amines in breast milk have all been associated with subsequently shown to have antimicrobial activity against
development of atopy in infants. In addition, the proportion streptococci in the presence of peroxide and thiocyanate
of neutrophils is significantly higher and macrophages is [86], thereby contributing to the antimicrobial activity of
lower in milk from mothers with an infant suffering from milk.
atopic dermatitis [72]. As human milk cells are a major Other milk antimicrobial activities were also discovered. A
source of some of the cytokines, it is possible that this fraction of cow’s milk with a distinctive red colour was
finding could explain the link between cytokines and described in 1939 [87] and iron-chelating activity in milk was
allergy. In some studies, an infant’s risk of developing noted in 1951 [88]. Later workers characterised the protein
allergy has been linked to the mother, but not the father, responsible and termed it red milk protein, lactotransferrin,
having an allergy [73]. Thus, the association of a specific lactosiderophilin or lactoferrin. Lactoferrin was shown to
cytokine or fatty acid profile in breast milk with subsequent have a bacteriostatic effect against E. coli [89] as well as the
development of atopy in infants may simply relate to the fungus, Candida albicans [90]. The antimicrobial properties
mother’s history of atopy. of lactoferrin were attributed to its ability to sequester iron
Interestingly, one study has suggested a reduction in from the surrounding solution, thereby depriving bacteria of
atopic dermatitis in at risk infants (when at least one parent a mineral necessary for its growth. Xanthine oxidoreductase
has an allergy) after 6 months feeding of a partially activity was first reported in milk, in 1902 [91]. Its
hydrolysed infant formula as opposed to breast milk [74]. antimicrobial activity, when supplied with exogenous sub-
In addition, it has been suggested that prolonged strate, was demonstrated in 1943, and was attributed to the
(>7.5 months) exclusive breast-feeding of high risk infants formation of peroxide [92, 93].
has been associated with increased risk of IgE mediated As a result of these studies, by the mid 1960s, there was
food allergy, asthma and atopy, and atopic dermatitis [75], an appreciation that the defense property of milk was more
although shorter periods of breast-feeding are likely to be complicated than simply the presence of immunoglobulins
protective [76]. These observations support the notion that [94, 95]. Nevertheless, immunoglobulin transfer to the
 J Mammary Gland Biol Neoplasia

neonate still dominated most studies on this topic at that action of a range of antimicrobial proteins in milk. Yet, the
time. understanding of host defense in milk was essentially the
Further work in the 1970s provided more detail as to the same as in the 1960s. However, around this time significant
function and biological role of antimicrobial milk proteins. new insights were gained into the nature of innate
Lactoferrin was found to be present in a number of immunity in vertebrates. These included the extensive
additional secretions and fluids that are subject to patho- cross-talk between the innate and acquired immune system,
genic challenge [96], as well as in neutrophils [97], which the first elucidation of the molecular mechanisms for
play an important role in host defense. The level of pathogen recognition and identification of components of
lactoferrin in cow’s milk was shown to rise significantly the signal transduction cascade leading to specific effector
during mastitis [98]. The iron binding properties, predom- responses (reviewed in [117]). Identification of some of the
inance of the apo form of the protein in milk and the iron components of host defense mechanisms in plants, insects
sequestering mechanism for its bacteriostatic effect were all and amphibians led to the discovery of equivalent systems
corroborated [99, 100]. Lactoferrin was also shown to have in mammals, including the β-defensin and cathelicidin
bactericidal as well as bacteriostatic activity, and that it families of cationic antimicrobial peptides (reviewed in
could act on a wider range of microbes than just E. coli [118, 119]). This rapid series of discoveries energized the
[101]. Its antimicrobial activity was found to be due in part field of mammalian innate immunity, and provided a fresh
to alternative mechanisms such as membrane disruption, approach to investigating the host defense properties of
which was attributed to proteolytic cleavage products of milk.
lactoferrin, termed lactoferricins [102]. Lactoferrin has also As a result, additional immune components of milk were
been shown to have activity against a range of viruses, an identified. Members of the β-defensin family were found to
activity that appears largely due to its binding to viral be expressed by mammary epithelial cells with expression
particles, thereby blocking virus entry to the cell (reviewed of their genes induced during mastitis [120, 121]. Members
in [103]. of the β-defensin and cathelicidin families were also found
The bactericidal actions of the enzymes in milk have been in milk and some at elevated levels in colostrum [122, 123].
characterised in greater detail. The substrates for lactoperox- A variant of an acute phase protein, serum amyloid A3, was
idase, hydrogen peroxide and thiocyanate, were shown to be found to be expressed in mammary cells in response to
produced by streptococci and liver detoxification pathways pathogens or pathogen-derived lipoteichoic acid and to be
acting on dietary glucosinolates, respectively [104]. Lacto- present in milk during mastitis [124], suggesting a role for
peroxidase was shown to produce a range of highly reactive this milk protein in host defense.
groups which were thought to react with and disrupt the The repertoire of putative immune factors in milk
bacterial cell membranes (reviewed in [105, 106]). In continues to grow. A member of the RNAse family of
addition to streptococci, this antimicrobial system was also proteins, angiogenin, has been known to be present in milk
found to act against a range of coliforms and Pseudomonas since the 1980s [125]. More recently, a host defense role
species, providing there was a source of peroxide [107, 108]. was claimed for this protein, based on the discovery that
Xanthine oxidoreductase was shown to contribute to the mouse and human angiogenins have antimicrobial activity
lactoperoxidase antimicrobial system by supplying it with [126]. This role in milk is still to be verified, but is
hydrogen peroxide [109]. Further evidence for the host supported by the observation that other members of the
defense role of xanthine oxidoreductase included: its RNase family are found in neutrophils and have antimicro-
production of a range of potentially bactericidal reactive bial as well as antiviral activities [127, 128].
oxygen species [110]; its increased abundance in neutrophils
during infection [111]; and its inhibition being associated
with an increase in microbial activity [112]. Lysozyme was Future Directions
shown to attack the cell walls of Gram positive bacteria by
cleaving the glycosidic bond of N-acetylmuramic acid within It is almost axiomatic that much remains to be discovered
the peptidoglycan molecule [113]. Lysozyme was identified about the innate host defense system in the mammary gland
in the secretory granules of neutrophils [114], associated and its secretions. The potential of the recently developed
with lactoferrin [115]. Proteolytic fragments of the caseins in technologies of genomics, transcriptomics and proteomics
milk were also shown to have antimicrobial activity to shed light on the biology of milk and the mammary
(reviewed in [116]), suggesting that the proteases in milk gland have yet to be fully realised. Gene expression studies
as well as the major milk proteins themselves may also play using microarrays and proteomics published to date have
a role in host defense. revealed a hitherto hidden complexity of the host defense-
By the late 1990s, a considerable body of knowledge associated proteins in milk and the immune response of the
had been accumulated as to the composition and mode of mammary gland [129–134]. These techniques are likely to
J Mammary Gland Biol Neoplasia

result in the identification of yet more components marketed as a product to prevent travellers’ diarrhea. IgA
contributing to the host defense property of milk and has perhaps even more potential in this area, as it has better
colostrum. stability in the GI tract [137]. In the future, milk proteins
A few fundamental questions remain to be answered. with intrinsic bactericidal activity may be alternatives or
The regulation of the milk host defense system is not well supplements to traditional antibiotics, for which there is an
understood. For example, the mechanisms responsible for increasing need [138]. It has also been suggested that the
differences in the composition of colostrum and milk LPS-binding activity of lactoferrin could be used therapeu-
between species and between stages of lactation have yet tically [139]. Other potential uses are in dairying. For
to be described. It is conceivable that these might involve example, antimicrobial proteins have been expressed in
epigenetic mechanisms. Also, the mechanisms by which cow’s milk, either through production of transgenic cattle or
pathogens are recognised by the host defense system is still by transfection of mammary cells in vivo, thereby enhanc-
a relatively unexplored area. Proteins such as Lipopolysac- ing the animal’s resistance to mastitis [140, 141]. It is
charide Binding Protein (LBP) and Bactericidal/Permeability- conceivable that in the future, the level of the endogenous
Increasing Protein (BPI) have a key role in the systemic host defense components in milk could be enhanced by
response to pathogens by binding to pathogen-derived inducing their over-expression through animal management
compounds and presenting them to receptors on immune approaches. Finally, detection of components of the host
cells. It seems likely that an analogous system for pathogen defense in milk could conceivably be used to detect mastitis
recognition exists in milk, but to date this has not been in dairy cows. Similarly, their genes could be used as the
described. basis for genetic selection for resistance to mastitis. Each of
The biological roles of many of the known milk host these applications has particular functional requirements
defense components have still to be fully elucidated. The that will need to be met. Future work will determine to
multifunctional nature of some of these has recently been what extent these potential uses are realised.
suggested, for example the immunomodulatory properties
of lactoferrin and some antimicrobial peptides known to be
present in milk [135, 136]. To date most work on the Acknowledgement The authors wish to acknowledge the efforts of
Claire Miller and the staff at the AgResearch, Ruakura Information
proteins in mammary secretions has concentrated on the Services section in procuring the sometimes obscure original
suppression of pathogens, while other possible functions literature.
have been under-represented in the literature. The immune
system is known to play a role in mammary development
and involution, as well as responding to neoplasia. It will be References
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the extent of co-operativity and complementarity between Hyg Infektionskrankh 1889;5:491–6.
the various components, as well as the nature of the 2. Fokker AP. Ueber bacterienvernichtende Eigenschaften der
Milch. Z Hyg Infektionskrankh 1890;9:41–55.
mechanisms by which collectively they suppress the 3. Ehrlich P. Ueber Immunität durch Vererbung und Säugung. Z
viability, growth, or virulence of pathogens of the mamma- Hyg Infektionskr 1892;12:183–203.
ry gland and neonatal GI tract. The possible synergies 4. Isaacs CE. The antimicrobial function of milk lipids. Adv Nutr
between the different proteins have not been thoroughly Res 2001;10:271–85.
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