Antibodies and Bioactive Peptides
Immune System
• Composed of a complex constellation of cells, organs and tissues
• Arranged in an elaborate and dynamic communication network • Equipped to optimize the response against invasion by pathogenic organisms and foreign materials
Immune systems
– Antigen specific: recognize and act against particular antigen
– Systemic: not confined to the initial infection site, but work throughout the body
– Memory: recognize antigens and mount an even stronger attack to the same antigen the next time. Memory B and T cells
Dual Natures of Immune System
•
•
•
•
•
•
Self/non‐self recognition: the most important
General/Specific
Natural/adaptive = innate/acquired
Cell‐mediated/humoral
Active/passive
Primary/secondary
Self/non‐self Recognition
• Self recognition is achieved by a marker based on major histocompatibility complex (MHC)
• Any cell not displaying this marker is treated as non‐self and attacked
• Undigested proteins are treated as antigens
• Autoimmune diseases: attack of self cells (multiple sclerosis, arthritis, diabetes etc.)
• Allergy: An inappropriate immune response to harmless substances
– Allergen: Substances that elicits allergy
Transport Systems for the Agents of the Immune System
• Lymphocytes circulate in the blood and lymph systems • Lymph systems:
– Primary organs: bone marrow and thymus gland – Secondary organs: At possible portal of entry for pathogens
• Adenoids, tonsil, spleen, lymph nodes, Peyer’s patches (within the intestines), the appendix
Blood System
7% of body weight
52‐62% liquid plasma and 38‐48% cells
Manufactured by stem cells in bone marrow
Erythrocytes (RBC)
Leucocytes (WBC)
Granulocytes
Neutrophils (55‐70%), ‐polymorphonuclear leucocytes
Eosinophils (1‐3%), Basophils (0.5‐1.0%) Agranulocytes
Lymphocytes (20‐40%); B‐cells, T‐cells Monocytes (1‐6%) mature into macrophage (big eater)
Thrombocytes (platelets)
macrophage
From:http://uhaweb.hartford.edu/BUGL/immune.htm
Basic Strategies of Immune System
• Creating and maintaining a barrier that prevents bacteria and viruses from entering the body. • If a pathogen gets into the body, the innate immune system detect or eliminate the invader before it is able to reproduce and cause potentially serious injury to the host. • If a pathogen is able to successfully evade the innate immune cells, the immune system activates a second, adaptive immune response against the pathogen. Innate Immunity
• The basic resistance to disease that a species possesses • The first line of defense against infection. • The characteristics of the innate immune response: – Responses are Broad‐Spectrum (non‐specific) – No memory or lasting protective immunity – All antigens are attacked equally – Bind to antigen using pattern‐recognition receptors
– Encoded in the gene and passed from generation to generation Functions of the innate immune system
• Recruiting immune cells to sites of infection, through the production of chemical factors ‐ cytokines. • Activation of the complement cascade to identify bacteria, activate cells, and to promote clearance of dead cells or antibody complexes. • Identification and removal of foreign substances by white blood cells. • Activation of the adaptive immune system
Adaptive immune response
– Composed of highly specialized systemic cells and processes – Eliminate or prevent pathogenic challenges – Activated by the “non‐specific” innate immune system
– Provides the immune system with the ability to recognize and remember specific pathogens (to generate immunity)
– Mount an even stronger attack each time the pathogen is encountered
– The system is highly adaptable
– Allows a small number of genes to generate a vast number of different antigen receptors, which are then uniquely expressed on each individual lymphocyte
Antibodies are produced by B lymphocytes
Clonal Selection
EM of resting and activated B cells
• Millions of B cell clones w/ specific cell‐surface receptors Activated: Extensive rough ER for antibody production/secretion
• Activation of B cell clones by specific target antigen
• Activated B cells secrete specific antibodies
Adaptive or Acquired Immune Systems
Two systems: cell‐mediated and humoral
1. Cell‐mediated immunity: • by T‐cell (80% of lymphocytes) – Non‐antibody producing lymphocytes: activated by antigen presenting macrophages and recognition by T cell – Produced in the stem cells of the bone marrow but sensitized in the thymus – T‐cells constitute the basis of cell‐mediated immunity
– Produce cytokine cytotoxin Humoral immunity
2. By B‐cells (15% of lymphocytes)
– Produced in the stem cells of the bone marrow. – Produce antibody and oversee humoral immunity
– The antibodies inactivate antigens by
• Complement fixation (proteins attached to antigen surface and cause cell lysis)
• Neutralization (binding to specific sites to prevent attachment)
• Agglutination (clumping)
• Precipitation (forcing insolubility)
From
bone marrow stem cells
From
bone marrow stem cells
Thymus processing
Bursa processing
T-lymphocyte
B-lymphocyte
Carried to secondary
lymphoid organs
Carried to secondary
lymphoid organs
Lymphoblasts
Plasma cells
Cell-mediated reaction
Humoral antibody synthesis
Antigen stimulation
Antigen stimulation
Clones of T cells
Clones of B cells
Cytokine
cytotoxin
Antibody
From:http://uhaweb.hartford.edu/BUGL/immune.htm
Antibodies
• Molecules with high specificity for binding and inactivating foreign substances, such as toxic or antigenic molecules that invade the body
• Used for – Diagnostic purposes
– Cancer diagnosis and therapy
– Passive immunotherapy against infectious diseases
– Inactivation of toxic substances
Antibodies
• In active immunisation (vaccination), the body is primed to make its own antibodies and confers life‐
long immunity • In passive immunity, antibodies are passed and provide immediate, but short‐lived protection against specific diseases • Most neonates receive passive immunity from their mother either as colostrum (IgA) in mammals or as egg yolks (IgY) in reptiles and birds General Roles of Antibodies
ƒ Neutralization of toxin and virus
ƒ Opsonization: marking of a pathogen for ingestion and destruction by a phargocytes
ƒ Agglutination (clumping)
ƒ Precipitation (immune complex)
Antibodies in the Immune System
Types of Antibodies
IgM
• A basic antibody that is present on B cells
• The primary antibody against A and B antigen on red blood cells
• Produced after an animal has been exposed to an antigen for an extended time or when an animal is exposed to an antigen for the second time
• Forms polymers where multiple immunoglobulins are covalently linked together with disulfide bonds, mostly as a pentamer or a hexamer
• Due to its polymeric nature, IgM possesses high avidity, and is particularly effective at complement activation
ABO blood group antigens present on red blood cells
IgG
• Involved in the secondary immune response
• The presence of specific IgG corresponds to maturation of the antibody response
• The only isotype that can pass through the human placenta ‐
providing protection to the fetus in utero
• Colostrum contains a high percentage of IgG, especially in bovine colostrum
• Plays an important role in Antibody‐dependent cell‐mediated cytotoxicity (ADCC).
Mammal Immunoglobulins (IgG)
Monomer structure
- 2 light chains + 2 heavy chains
ƒ Variable domain
a
g
nt i
an t
en
ig e
n
- antigen-binding sites
- neturalize, ppt or agglutinate
ƒ Can bind to many kinds of pathogens
Fab
(viruses, bacteria, and fungi)
ƒprotects the body against them by
ƒ agglutination and immobilization,
ƒ complement activation (classical
pathway),
ƒ opsonization for phagocytosis
ƒ neutralization of their toxins.
Fc
Fc
Production of Antibody
• To raise antibodies in an animal, the host animal should be challenged with an antigen. • A repeated exposure of antigen (immunization) elicits antibodies in the host animal. • If the injecting substance is identical or similar to the host, it will be a poor immunogen, resulting in low or no antibodies. • Many highly conserved proteins such as insulin and cytochrome c are poor antigens in hosts such as rabbits or goats. Production of Antibody from Chicken
• In some cases it may help to change the host animal as birds are more distant in evolution than some mammals. • The fact that many mammalian proteins are less conserved in birds; therefore, chickens may be more desirable as an antibody host. • Every foreign protein or microorganism that the chicken has been exposed to, whether by vaccination or by natural exposure, is processed and antibodies raised against it • These antibodies are then placed in the egg yolk at differing titers depending upon the degree of exposure
Polyclonal antibody production
• Uses
– Laboratory animals: rabbits, rats, mice and guinea pigs
– larger mammals: horses, sheep’s and goats
• Involves – i) immunization – ii) bleeding
• Cause suffering for the animals
Antibody Production 1. Inject antigen (i.e. purified protein) into animal (i.e. mouse, rabbit, chicken)
2. Animal produces antibodies that recognize antigen
3. Antigen injected more than once: response heightened in subsequent injections
Producing antibodies to a specific antigen
Polyclonal antibodies: Derived from multiple B‐cell clones,
recognize multiple epitopes on antigens
Inject with antigen
collect blood serum
purify antibodies w/ affinity chromatography
using antigen attached to beads
Linear epitope
Conformational
epitope
“epitope” = unique part of antigen recognized by antibody
Producing antibodies to a specific antigen
Monoclonal antibodies:
• Derived from B‐cell clone “Hybridoma”
• Recognize single epitope on antigen
Uses of antibodies in molecular biology
Applications:
Western blotting (Immunoblotting) ‐ Identification of protein antigen following SDS‐PAGE
Immunoprecipitation ‐ Isolation of specific proteins + binding partners
Immunofluorescence microscopy
‐ Localization of specific proteins in cells
ELISA (Enzyme‐Linked Immunosorbent Assay)
‐ Detection of proteins in a sample
Detection of specific proteins:
SDS‐PAGE and Western blot
1.
2.
3.
4.
5.
6.
Separate proteins by SDS PAGE
Transfer proteins to membranes (i.e. Nitrocellulose)
Block non‐specific sites on membrane
Incubate with primary antibody, wash
Incubate with secondary antibody, wash
Detect secondary antibody
Western blotting
From Lodish et al. Molecular Cell Biology 4th edition.
Indirect immunodetection
Immunopreciptation: Identification of protein‐protein interactions
Steps:
1. Attach antibody to beads via protein A
2. Lyse cells to release antigen and its binding partners
3. Mix cell lysate + antibody‐coated beads (antibody binds antigen)
4. Purify antigen and its binding partners by centrifugation
bead
protein A
primary
antibody
Immunofluorescence Microscopy
ELISA
(Enzyme‐Linked Immunosorbent Assay)
Detection of proteins (i.e. cytokines, HIV antigens) in samples
Chicken Antibodies
• The chicken is an optimum animal to produce antibodies. • While the cow or other mammals produce colostrum at parturition, the amount of antibody in milk at other times is minimal. • Yolk immunoglobulins (IgY) are an important group of antibodies which are found in the egg yolk after specific antigens are injected into the hen. • IgY can successfully compete with antibodies produced in the blood of mammals (IgG). Use of chickens for antibody production
• A reduction and a refinement in animal use
– A reduction in animal use: hens produce larger amounts of antibodies then laboratory rodents.
– Collection of blood can be replaced by egg yolk: minimizing the distress. •
•
•
•
High immunoglobulin content (200 mg/egg yolk)
Isolation method is easy and fast
Excellent alternative to mammalian antibodies Good for animals welfare
Comparision of Mammal IgG and IgY
ƒDifferent M.W (150 vs 180 kDa), PI, and binding behavior with complement
ƒLimited flexibility of IgY due to lack of hinge region ƒLess binding activity of IgY with Fc receptor on the cell surface Immunology today, 16 (8) :393
IgY
• The molecular mass of IgY is ~ 180 kD (light chain ~25 kilo Dalton [kD] each: heavy chain ~ 65‐68 [kD] each). • Yolk antibodies are highly resistant to acid and heat
– IgY lost little of antibody activity by incubation for 10 min at pH 7.2 at 60 °C or for 10 min at 40 °C above pH 4.0. – Antibody activity were significantly diminished by incubation above 65 °C or below pH 4.0. – 30‐50% (w/v) sucrose reduced the heat denaturation of IgY at 75‐
80 °C – The addition of high levels of sucrose, maltose, glycerol or glycine displayed effective additional protection against thermal denaturation of IgY.
IgY
• Antibodies are naturally protected by the yolk granules. • If encapsulated, they are particularly resistant to pH and digestive enzymes. • Obtained GRAS status from both the USDA and the FDA
• Approval of individual products by the FDA for the use of egg antibodies in human patients is relatively easy.
IgY
• The major serum antibody in chicken is IgG, but IgG is also transported to egg in a manner similar to the placental transfer of IgG in mammals. • The protection of relatively immuno‐incompetent newly hatched chicks from pathogens is through the transmission of antibodies from the mother via the egg. • In the egg, chicken IgG is found mainly in egg yolk, and the concentration in egg white is very low. How is immunoglobulin Y generated?
Oviduct
Albumin
Yolk IgG
Yolk
IgA IgM
Ovum
Chicken Serum IgG, IgA, IgM Egg
Use of IgY
• Oral administration of antibodies has been successfully used to prevent specific diseases in infants, fish and small animals
– E. coli‐induced diarrhoea – Rotavirus diarrhoea – Travelers’ diarrhea – Helicobacter
– anti‐Streptococcus mutans IgY for oral care
• Encapsulation of IgY with egg lecithin/cholesterol liposomes reduced the activity loss of IgY under gastric conditions Use of IgY
• Cancer therapy and as a biochemical tool for protein characterization. • Specific IgYs have been used for characterizing the high molecular weight mucin‐like glycoprotein‐A of human milk.
• Egg anti‐inflammatory compounds can be used to treat and prevent inflammation, particularly arthritis and autoimmune diseases • The effect of passive immunization with IgY against the virus responsible for Marek’s disease delayed the development of Marek’s disease lesions.
• Antibodies against Pseudomonas aeruginosa ‐ prevents the colonization of these bacteria in the lungs of patients with cystic fibrosis. Use of IgY
• The IgY obtained from hens immunised with S. mutans inhibited the adherence of dental plaque in humans and in animals • Antibody was raised against bacteria that cause mastitis in dairy cows. • The whole egg containing the antibodies against E. coli 0157:H7 can be utilized in foods.
• Several commercial antibody companies in the US and European Union (EU) market place produce polyclonal IgY
Advantage of egg yolk antibodies over mammalian sources 1
• Better immune responsiveness to mammalian antigens
– A phylogenic distance between chickens and mammals – The production of IgY antibodies against conserved mammalian antigens in general are more successful in chickens than in other mammals – The antibodies are directed against more antigenic epitopes and recognize the same proteins in several species, making them more widely useful – The evolutionary spread also means that there is no immunological cross reactivity between chicken IgY and mammalian IgG – Possible to obtain a strong immune response with small amounts of antigen in hens thus reducing the amount of antigen required for producing antibodies
Advantage of Chicken Antibodies 2
• IgY reduces interference problems in immunological assay
– Chicken antibodies do not react with either rheumatoid factor (RF) or human anti‐mouse IgG antibody (HAMA) – Thus, eliminate interference due to these factors – Chicken antibodies neither activate nor recognize the mammalian (human) complements – Thus, prevent false positive responses
Advantage of Chicken Antibodies 3
• No binding to bacterial or mammalian Fc receptors
– immunological difference in the Fc region from that of IgG
• A simple and economical isolation process
• Resistant to extreme pH and temperature
– IgY is stable at pH 4–9 and up to 65 C in aqueous condition • Storability
– Stable in 0.9% NaCl, 0.02% NaN3 at 4°C for 20 years without any significant loss of antibody titer – Affinity‐purified and biotinylated antibodies retained high activity after 5 years of storage at 4°C
Disadvantages of Chicken Antibodies
• IgY does not bind to protein A and G: precludes the use of these proteins in IgY isolation. • Less able to precipitate antigen compared to mammalian IgG • Better precipitation can sometimes be achieved by increasing the salt concentration in the buffer solution
Modification of IgY
• IgY from egg yolks of chickens immunized with formalin‐treated E. coli was able to aggregate bacterial cells and inhibit their growth in vitro. • Conjugation of antibodies and drugs may be an important agent for cancer treatment. • An appreciable IgY activity is retained after trypsin and chymotrypsin digestion of IgY. • Possible to use IgY for fortification of food products, especially infant formula
IgY Theraphy
• Enteric Infections
– Bovine and human rotaviruses
– Diarrhoea due to enterotoxigenic • Escherichia coli • Salmonella ssp.
• An alternative to antibiotics in a group of patient with cystic fibrosis (CF)
• Respiratory infections
• Helicobacter
• Dental cary
Bioactive Proteins and Peptides in Milk
Functions of Proteins
• Bioactivity
– Protective functions
– Regulation of biological functions
• Nutritional value
– Source of nitrogen for protein synthesis
– Source of energy
• Technological value
– Functional (physicochemical) properties
– Sensory properties
Bioactive Proteins ‐ Definition
• Food components that can affect biological
processes or substrates
• Have an impact on body function or condition and
ultimately health
– Should impact a measurable biological effect at a
physiologically realistic level
– The health effects should be beneficial
– Thus, potentially damaging effects (such as toxicity,
allergenicity and mutagenicity) should be excluded
Bioactive Proteins in Milk
•
Caseins
–
–
–
–
•
Whey proteins
–
–
–
–
–
•
αs1‐casein
αs2‐casein
β‐casein
κ‐casein
β‐lactoglobulin
α‐lactalbumin
BSA
Immunoglobulins
Lactoferrin
Minor proteins
–
–
–
–
Growth factors and growth hormones (IGF‐1, TGF−β2, GH)
Milk basic protein and cytokines
Milk Fat Globule membrane proteins
Enzymes (Lactoperoxidase glutathionperoxidase)
Protein Composition of Milk
COMPONENT COW MILK
content (g/l)
COLOSTRUM
content (g/l)
CASEINS αs1‐CN αs2‐CN β‐CN κ‐CN 26 10 2.6 9.3 3.3 27
10.3
2.7
9.5
3.4
WHEY PROTEINS β‐lactoglobulin α‐lactalbumin Bovine serum albumin Immunoglobulins Lactoferrin 6.3 3.2 1.2 0.4 0.6‐1.0 0.1 34
8.0
3.0
1.3
2.0‐15.0
1.5
MFGM PROTEINS 0.4 0.4
Lactalbumin
• Rich in double‐bonded cystine which promotes the production of glutathione, the major intracellular endogenous antioxidant and detoxicant. • Help raise serotonin in deficiency states and lower cortisol when in excess.
Beta‐Lactoglobulin
• Bioactive functions
Transport of retinol (vitamin A)
Antioxidant (potential)
Antimicrobial and Antiviral effect
Pathogenic adhesion effect
Controlling enzymes in mammary secretory vesicles
Mammalian cell growth factor activity, mitogenic activity
– Anticarcinogenic activity
–
–
–
–
–
–
Bovine Colostrum:
Natural Healing Miracle
• Nature’s first and complete food for newborn mammals • Considered essential for the life of newborn
• Contains essential nutrients, immune factors and growth factors
Colostrum’s Immune Factors
Immunoglobulins
Lactalbumin
Lactoferrin
Lysozymes
Glycoproteins
Proline‐rich Polypeptides (PRP)
Many other peptides, glycoconjugates, phospholipids, cytokines, complement system, etc with biological activities
• Transfer factors (TF)
•
•
•
•
•
•
•
Immunoglobulins in Colostrum
• Predominantly IgA with trace amounts of IgD, IgE, IgG, and IgM
• Convey passive immunity, especially enterically
• Colostrum contains specific antibodies to more than 19 specific disease‐causing pathogen
• lgA protects mucosal surface against viruses (e.g. polioviruses,
influenza A viruses and herpes simplex virus) and bacteria (E coli, salmonella, streptococcus) • IgD stimulates B‐cells to produce antibodies • IgE (associated with allergic reaction) and IgA are highly antiviral • IgG enhances phagocytosis, neutralizes toxins and microbes in the lymph and circulatory system (80‐85%)
• IgM Enhances phagocytosis, especially against microorganisms (5‐
10%)
Immunoglobulin Concentration
Ig
Colostrum
mg/ml
Milk
mg/ml
IgG1
52.0-87.0
0.31-0.40
IgG2
1.6-2.1
0.03-0.08
IgM
3.7-6.1
0.03-0.06
IgA
3.2-6.2
0.04-0.06
Lactoferrin
• Bioactive functions
– Regulation of iron transport
– Antimicrobial activity
– Antiinflammatory and Immunomodulatory activity
– Antioxidative effect
– Anti‐inflammatory and Anticarcinogenic activity
– Stimulation of cell proliferation
Lactoferrin in biological fluid
Biological Fluid
Human Colostrum
Tear Fluid
Bovine Colostral Whey
Human Milk
Seminal Plasma
Bovine Milk
Saliva
From Steijns and van Hooijdonk 2000
Amounts Reported
>7 mg/ml
>2.2 mg/ml
>1.5 mg/ml
>1‐2 mg/ml
>0.4‐1.9 mg/ml
>20‐200 μg/ml
>7‐10 μg/ml
Proline‐rich Polypeptides (PRP)
•
Small, very low weight molecules (6,000 Da) •
Helps to regulate thymus gland (body’s central command for the immune system)
•
Can both stimulate immune system and/or down‐regulate an overactive immune system, as in the case of many autoimmune diseases
•
Described as “biological response modulators, “info‐peptides”, “transfer factors” and “colostrinin”. •
Induces maturation and differentiation of murine thymocytes and affects humoral and cellular immune reactions
•
Colostrinin and colostrinin‐derived nonapeptide facilitate learning and memory in rats (Popik, 2001) Lysozymes and Glycoproteins
• Lysozymes contain enzymes that can attach to and digest bacteria cell walls, thus destroying them.
• Glycoproteins are sugar‐amino complexes – act as protease and trypsin inhibitors
– protecting the immune and growth factors in colostrum which are otherwise vulnerable to degradation via enzymatic action.
Cytokines
• Integral to intercellular communications that regulate immune activity and related down stream inflammatory responses. • Include the interleukins, the lymphokines, and
interferon. • Colostrum contains many cytokines (IL)1β, IL‐6, IL‐10, tumor necrosis factor α, granulocyte, macrophage, granulocyte‐macrophage colony stimulating factors
GSH Antioxidant System
• The foremost among cellular protective system and biosynthesis is depending up the availability of cysteine.
• Remove reactive oxygen species during increased cellular activity • Could occur in lymphocytes during the development of the immune response
• Abundant in whey protein and easily readily enters into cells. • Anticancer properties
Colostrum’s Growth Factors
• Insulin‐like Growth Factor I and I (IGF‐I / IGF‐II) – Help regulate lipid, protein and carbohydrate anabolism. – IGF‐I promotes the growth and repair of DNA and RNA.
• Epithelial Growth Factor: – Enhances dermal anabolism: used as a cosmeceutical ingredient.
• Transforming Growth Factors A & B – Promote mesenchymal cellular proliferation. – Has potential for assisting bone and cartilage repair, deep wound healing, and restoring intestinal integrity in “leaky gut” syndromes.
• Platelet‐Derived Growth Factor
– Promotes growth not only in connective tissue such as fibroblasts and smooth muscle, – Has some promise in sparing and regenerating nerve tissue as well.
“Leaky Gut” or “leaky gut syndrome” (LGS) • Definition – An increase in permeability of the intestinal mucosa to luminal macromolecules, antigens and toxins associated with inflammatory degenerative and/or atrophic mucosal damage. – Represents a hyperpermeable intestinal lining.
– Large spaces develop between the cells of the gut wall, and bacteria, toxins and food leak in. “Leaky Gut”
• Additional space allows toxic substance such as bacteria, viruses, parasites and other harmful factors to enter the bloodstream and reach every part of the body. In healthy gut, these toxins are eliminated.
• Undigested foods and proteins can escape into the intestinal wall, resulting in allergic responses
• Dyspepsia (indigestion) is a major health care problem – 40% of general population
– Gastro‐esophageal reflux, peptic ulceration, non‐ulcer dyspepsia, constipation, irritable bowel syndrome and inflammatory bowel disease
How does the “gut” become leaky?
• Once the gut lining becomes inflamed or damaged, this disrupts the functioning of the system. • The spaces open up and allow large food antigens, for example, to be absorbed into the body. What leads to “leaky gut”?
• Antibiotics because they lead to the overgrowth of abnormal microflora in the gastrointestinal tract (bacteria, parasites, candida, fungi)
• Alcohol and caffeine (strong gut irritants) • Foods and beverages contaminated by parasites
• Foods and beverages contaminated by bacteria like helicobacter pylori, klebsiella, citrobacter, pseudomonas and others • Chemicals in fermented and processed food (dyes, preservatives, peroxidized fats) • Enzyme deficiencies (e.g. celiac disease, lactase deficiency causing lactose intolerance) • NSAIDs (non‐steroidal anti‐inflammatory drugs) like ASA, ibuprofen, indomethacin, etc. • Prescription corticosteroids
• High refined carbohydrate diet (e.g. candy bars, cookies, cake, soft drinks, white bread) – the cause of Syndrome X
• Prescription hormones like the birth control pill
• Mold and fungal mycotoxins in stored grains, fruit and refined carbohydrates • Indigeston and gastrointestinal Disease
• Chemo‐therapy and radiation therapy causing immune overload NSAIDs
• Non‐steroidal anti‐inflammatory drugs (NSAIDs) such as aspirin and ibuprofen (e.g. Motrin and Advil) are some of the most commonly prescribed medicines
• Although NSAIDs are effective analgesics and that some even reduce heart attack risk, chronic administration of NSAIDs results in both gastric and intestinal damage including peptic ulceration and injury to the small and large intestine causing increased permeability with blood and protein loss and structure formation
Growth Factors
• Colostrum and its growth factors stimulate – muscle protein synthesis, – inhibit protein breakdown, – benefit GI tract structure and function, – act on the immune system to inhibit tissue inflammation and allergic response.
• Growth factors stimulate – normal growth,
– help regenerate and accelerate the repair of aged and injured muscle, skin collagen, bone, cartilage and nerve tissues
• Growth factors help body to – burn fat for fuel – build lean muscle
Epithelial Growth Factor ‐ EGF
• 53 amino acids
• Present in human colostrum (200 ng/ml) and milk (30‐50 ng/ml)
• Stimulate normal skin growth and repair cellular tissue including GI track
• Play role in gut development
• Other EGF Family: amphiregulin, epiregulin, heparin binding EFG‐like growth factor (HB‐EGF), NTAK (neutral‐ and thymus‐
derived activator for erbB kinase), neuregulin gene product, and TGF‐α
Growth Factor – IGF, TGF
• Insulin‐like Growth Factor (IGF‐I, IGF‐II)
– Most abundant growth factor in bovine colostrum (560‐
890 ng/ml, 6h; 490 ng/ml, 11‐20 hr) and milk.
– Both about 7500kDa, binding protein (about 45kDa), – IGF‐I (pI=8.2), IGF‐II (pI=6.4)
• Transforming growth factor (TGF‐α, and TGF‐β )
– 25 kDa + binding protein (80 kDa)
– TGF‐β2 (85‐95%), – Bovine colostrum (4.3 ng/ml) and milk (3.7 ng/ml)
Conclusions
• Bovine colostral whey is a rich source of nutrients, growth factors and immune factors
• Defatted bovine colostral whey may reduce the rise in the gut permeability (LGS)
• Defatted bovine colostral whey and its fractions could be used nutraceutical and functional food ingredients for the immune compromised person to enhance immunity
Bioactive Peptides
• Biological active peptides or functional peptides are food derived peptides that in addition to their nutritional value exert a physiological effect in the body
– Defined sequences of amino acids which are inactive within the original protein, but which display specific properties once they are released by enzymatic hydrolysis (Vermeirssen et al 2004)
Production of Bioactive Peptides
• In the cow
– Naturally occurring peptides
– Proteolysis in the udder
• Dairy product processing
– Enzymatic hydrolysis
– Proteolytic action of microorganisms
• In human after ingestion
– Digestion in the intestinal tract
Controlling Hypertension
• Diet/Exercise
• Ca++ Channel Blockers
• Beta Blockers
• Angiotensin Converting Enzyme (ACE) Inhibitors
• Angiotensin Receptor Blockers
ACE Inhibitors and Angiotensin Receptor Blockers
ACE-Inhibiting Activity of Enzyme-Treated Egg Albumen ACE-Inhibition
Percentage (%)
Enzyme treatment
Egg Albumen
Peptide Fractions
Pepsin to Egg Albumen
85.1a
61.8a
Trypsin to Egg Albumen
23.3d
36.8ab
Chymotrypsin to Egg Albumen
65.0b
44.6ab
Themolysin to Egg Albumen
85.8a
72.4a
Fermented Soy powder to egg
albumen
25.71c
18.45b
Fermented Soy Product
64.9b
50.7ab