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Presented By:
Supta Sarkar
HHM-2013-10
M.Sc FN- 2nd Yr.
 Introduction
 Definition
 Types of prebiotics
 Mechanism for the effect of prebiotics on
immunity
 Structure & role of different prebiotics
 Conclusion
 Reference
Introduction
• The concept of prebiotic was introduced by Gibson & Roberfroid,
1995.
• Prebiotics is a general term to refer to chemicals that induce the
growth and/or activity of commensal microorganisms (e.g., bacteria
and fungi) that contribute to the well-being of their host.
• The criteria which must be met in order to be classified a prebiotic,
as defined by Gibson & Roberfroid,1995.
-Resistance to hydrolysis or absorption in the upper gastrointestinal
tract.
-Fermented by the intestinal microbiota.
-Selectively stimulate the growth and/or activity of beneficial intestinal
bacteria, such as Lactobacillius species and Bifidobacterium species.
Definition
• Gibson & Roberfroid, 1995:
‘Prebiotics are non-digestible food ingredients that
beneficially affect the host by selectively stimulating the
growth, and/or activity, of one or a limited number of
beneficial bacteria in the colon and thus improve host
health’.
• Roberfroid, 2007:
‘A prebiotic is a selectively fermented ingredient that
allows specific changes, both in the composition and/or
activity in the gastrointestinal microflora that confers
benefits upon host well-being and health’.
TYPES OF PREBIOTICS
Prebiotic sources:
• Only two particular prebiotics then fully met the refined
definition of prebiotics: Inulin(IN) and Trans-
galactooligosaccharide (Roberfroid, 2007).
• Other authorities also classify resistant starch,
fructooligosaccharide (FOS) and lactulose as prebiotics.
Mannan Oligosaccharides (MOS) have been termed as
prebiotics but would more correctly be termed
immunosaccharides (Roberfroid, 2007).
• Carbohydrates including gluco-oligosaccharides,
isomalto-oligosaccharides, nigero-oligosaccharides, oat
b-glucans, raffinose, soyabean oligosaccharides (SOS),
and xylo-oligosaccharides are considered as candidate
prebiotics.
Mechanism for the effect of prebiotics on immunity
• The underlying mechanisms of how prebiotics modulate the
immune system are not known at present.
• However, experimental data suggests that these compounds
exert effects in the GALT and also point to a few different
mechanisms that might explain these effects:
1. Selective changes in bacterial composition and bacterial
products which modulate cytokine and antibody production;
2. Production of SCFAs and their interactions with leukocytes
3. Modulated mucin production
4. Interaction with carbohydrate receptors of pathogens inhibiting
their enhancement to epithelial cells as well as receptors on
immune cells.
A. SELECTIVE CHANGES IN BACTERIAL COMPOSITION
AND BACTERIAL PRODUCTS:
 It is well known that prebiotics increase the no. of
beneficial bacterial (i.e bifidobacteria and lactobacilli).
 Probiotics when administered orally are known to
increase the secretion of IgA in the small intestine and
the feces and to stimulate PP B lymphocyte IgA
production.
 They are also known to exert effects on systemic immune
functions and various immune parameters in the lungs,
spleen and peritoneal cells.
 Intestinal epithelial cells are involved in both innate
and adaptive immune responses and act by
transducing signals from luminal pathogens to
adjacent immune cells of the intestinal immune
system, via specific germinal-encoded pattern-
recognition receptors, such as Toll-like receptors
(TLRs) and cytoplasmic receptors.
 TLRs are able to discriminate between the normal
commensal biota and pathogens and induce the
transcriptional activation of a no. of genes mediating
immune and inflammatory responses.
 Pathogen-associated molecular pattern (PAMPs) [eg.,
endotoxin(lipopolysaccharide), lipoproteins, lipopeptides
and imidazoquinolines] present on diverse microbes are
initially recognized by TLRs and their interaction result in
the activation of intracellular signaling pathways, nuclear
translocation of transcription factor NF-κB and the
transcription of pro-inflammatory cytokines.
 The changes that occur in the composition of the
intestinal microbiota due to prebiotic fermentation could
potentially reduce the presence of PAMPs and thereby
exert a positive effects on the immune system.
 Prebiotics also promote an increase in the bacterial
cell-wall components that are recognized by TLRs
and in DNA derived from luminal bacteria that, in
turn, stimulate the intestinal immune system.
 Cytoplasmic components and cell-free extracts of
probiotics have also been demonstrated to produce
some of the same immune effects (eg, IgA
production by PP and macrophage stimulation) as
live bacteria.
B. PRODUCTION OF SCFAs:
• The major end products of carbohydrate fermentation
are SCFAs, of which acetate, propionate and butyrate
are quantitively the most important in the human
colon.
• The production of SCFAs in the colon averages 400
mmol/day, with a range of 150-600 mmol/day.
• All SCFAs are rapidly absorbed from the large
intestine and stimulate salt and water absorption:
principally, the gut epithelium, liver and muscle
metabolize them, with virtually none appearing in the
urine and only small amounts appearing in the feces.
• The three major SCFAs are trophic when infused into the
colon and these trophic properties have important
physiological implications in addition to maintaining the
mucosal defense barrier against invading organisms.
• Butyrate is known to suppress lymphocyte proliferation, inhibit
cytokine production of Th1-lymphocytes and upregulates IL-
10 production.
• It also suppresses expression of the transcription factor NF-
κB and upregulates TLR expression.
• Butyrate is also believed to protect against colon cancer as it
inhibits DNA synthesis and induces cell differentiation.
• It has been demonstrated in a rat model that supplementing total
parenteral nutrition with a SCFA mixture results in increased NK cell
activity(Pratt et al.,1996).
• Pharmacological doses of acetate administered intravenously to
both healthy individuals and cancer patients also increased NK cell
activity and peripheral blood antibody production.
• In addition, it has been shown that serum glutamine levels are
raised following lactulose administration and suggested that
increased SCFA levels were responsible for this (glutamine is a
preferred substrate for lymphatic tissue).
• Therefore, SCFA production in the large intestine could potentially
reduce the requirement of epithelial cells for glutamine, making it
available to the cells of the immune system.
C. MUCIN PRODUCTION:
• The first line of defense of the mucosa against luminal
contents is the mucous layer, which is mainly composed of
high-molecular-weight glycoprotein (mucins) that are secreted
by goblet cells.
• The effect of prebiotics on mucin production has been
reported in a study, where it was shown that inulin
administration resulted in increased mucin production in rats.
• Greater mucin production was found to be associated with a
lower incidence of bacterial translocation across the mucosa
following dietary fibre supplementation.
• It has been shown in a perfused rat colon model that the
production of acetate and butyrate from the fermentation
of dietary fibre stimulates mucin secretion, but fibers do
not have the same effect on their own.
• Pathogens and beneficial commensal bacteria are able
to modulate mucin synthesis by regulating some of the
mucin genes.
• Currently there are 16 identified mucin genes, but further
work is needed to fully explain the function of each of
them and to identify new genes.
D. CARBOHYDRATE
RECEPTORS:
 Studies suggest that some prebiotics are directly involved
in protecting the gut from infection and inflammation by
inhibiting the attachment of pathogenic bacteria or their
toxins to the colonic epithelium.
 This attachment is necessary before pathogens can
colonize and cause disease and it is mediated by
glycoconjugates on glycoproteins and lipids present on
the microvillus membrane.
 Certain prebiotic oligosachharides contain structures, similar
to those found on the microvillus membrane, that interfere with
the bacterial receptors by binding to them and thus preventing
bacterial attachment to the same sugar on microvillus
glycoconjugates.
 For example, α-linked TOS, present in human milk, are known
to have anti-adhesive properties and be capable of toxin
neutralization.
 Recently, a novel TOS mixture, which contains an
oligosachharide alpha anomeric configuration, was shown to
significatntly decrease the attachment of enteropathogenic
Escherichia coli (EPEC) and salmonella enterica serovar
Thyphimurium in vitro.
 In addition, immune cells also express specific carbohydrate
receptors which mediate various cellular reactions when
activated. For example, C-type receptors expressed on
phagocytic cells.
 It is hypothesised that carbohydrate moieties on the prebiotic
may interact with receptors on immune cells.
 Although a specific fructose receptor has not yet been
identified, receptors for b-glucan and mannose have been
identified on immune cells, and in vitro, fructose has been
shown to alter non-opsonic phagocytosis, suggesting that a
receptor for fructose on immune cells may exist.
 In addition, some oligosaccharides, for example OF, can bind to
receptors on pathogenic bacteria and prevent them from
attaching to this same sugar on the epithelial membrane, thus
preventing adherence.
Structure & role of different prebiotics
• β2-1 Fructans, which include inulin (IN) and fructo-
oligosaccharides (FOS), fulfil the criteria for prebiotics.
(Gibson, et al.,2004).
• According to Roberfroid, 2007, only two particular
prebiotics then fully met the refined definition of
prebiotics: Inulin(IN) and Trans-galactooligosaccharide
(TOS).
INULIN:
• Inulin is the most common type of FOS.
• IN is a linear carbohydrate molecule which contains β-(2→1)
fructosyl–fructose linkages with a terminal glucose.
• IN may contain between two and sixty fructose residues with an
average of twelve. (fig.1)
• Partial enzymatic hydrolysis of IN yields a FOS known as
oligofructose(OF), which can have a terminal glucose or fructose
residue (fig.1)
• In OF there can be two to eight (average five) fructose residues with a
terminal glucose residue or a chain of three to eight (average five)
fructose residues.
FIG.1:
Dietary Source of Inulin (IN):
• IN is found naturally in a variety of plant foods such as
bananas, barley, chicory, garlic, artichoke, leeks, onions
and wheat.
• Oligosaccharides, including some believed to be prebiotics,
are present in human breast milk.
• The presence of oligosaccharides in large amounts in
breast milk suggests that these compounds may play an
important role in early infant development, perhaps of the
gut, its microbiotia and the immune system.
Role of Inulin on immunity:
• The effect of β2-1 fructans upon macrophage
number and function has been studied, with the
results suggesting that macrophage functions
are enhanced by the addition of β2-1 fructans to
the diet.
• Peritoneal macrophage phagocytic activity was
increased in rodents given IN or OF for varying
periods of time (Kelly-Quaglian et al.,2003).
• In rats, OF-enriched IN (100g/kg) increased caecal
secretory IgA concentrations(Roller et al., 2004).
• The number of T cells in the MLN of rats was increased
upon 10% OF or IN supplementation and also the blood
IL-2 and IL-4 concentrations were increased upon the
supplementation for 4 months (Trushina et al., 2005).
• A study shows the percentage of blood B cells (defined as
CD19+) was increased in young male adults after
consumption of a bread containing IN (Seidel et al.,
2007).
REFERENCE Prebiotics
dose used
Animal
studied
Immune effect
Stillie et
al, 2005
Inulin
(4·8 %
w/w
diet)
Rats (21 d
old, male
& female);
diabetes
resistant or
diabetes
prone
In diabetes-resistant rats:
↑Small intestine length
↓ Number of splenocytes
↑CD8+ Lymphocytes in PP
↑ Proliferation of splenocytes and MLN
cells to mitogens
↓Production of IL-4 and
↑ production of IL-10 by stimulated
splenocytes
In diabetes-prone rats:
↓Number of splenocytes
↑IgA+ cells in jejunal lamina propria
↑B lymphocytes in PP
↓Production of IL-4 and
↑production of IL-10 by stimulated
splenocyte
FRUCTOOLIGOSACCHARIDES (FOS):
STRUCTURE:
• FOS consists of several β(1-2) or β(1-6) linked fructose
units which may be linked to glucose residues.
DIETARY SOURCE:
• They can be found naturally in some cereal crops as well
as fruits and vegetables such as bananas, onions,
chicory root, garlic, asparagus, onion and leeks.
Role of FOS on immunity:
• They stimulate the growth of bifido bacteria and to inhibit
growth and multiplication of potentially pathogenic
bacteria such as enterobacteria, clostridia and salmonella.
• Fermentation of FOS leads to the production of short
chain fatty acids, which is substrate for energy
metabolism in the colonic mucosa stimulating epithelial
cell growth.
• Short chain FOS can enhance IgA content and thus play a
major role in immunity.
• A study conducted by Bourgot et al.,2014 says that maternal scFOS
supplementation modified the intestinal immune functions in piglets in
association with increased colostral immunity.
• Thirty-four sows received a standard or a scFOS supplemented diet (10
g scFOS/d) for the last 4 weeks of gestation and the 4 weeks of
lactation.
• Colostrum and milk immunoglobulins (Ig) and TGFβ1 concentrations
were evaluated on the day of delivery and at d 6 and d 21 postpartum.
• Colostral IgA (P<0.05) significantly increased because of scFOS and
TGFβ1 concentrations tended to improve (P<0.1).
• Such results underline the key role of maternal nutrition in supporting the
postnatal development of mucosal immunity.
Another study shows dietary supplement of FOS to
the Caspian roach (Rutilus rutilus) fry increase the
serum Ig levels, lysozyme activity and alternative
complement activity (ACH50) and thus improving
the innate immune response (Soleimani et
al.,2012).
TRANS-GALACTOOLIGOSACCHARIDES (TOS):
STRUCTURE:
• Galacto-oligosaccharides (GOS), also known as
oligogalactosyllactose, oligogalactose, oligolactose or
transgalactooligosaccharides (TOS), belong to the group of
prebiotics.
• GOS/TOS generally comprise a chain of galactose units that
arise through consecutive transgalactosylation reactions, with
a terminal glucose unit. However, where a terminal galactose
unit is indicated, hydrolysis of GOS formed at an earlier stage
in the process has occurred.
• The degree of polymerization of GOS/TOS can vary quite
markedly, ranging from 2 to 8 monomeric units.
Role of TOS/GOS on immunity:
• Galactooligosaccharides, GOS can positively influence
the immune system—indirectly through the production of
antimicrobial substances as the result of
galactooligosaccharide fermentation, that can reduce the
proliferation of pathogenic bacteria, and directly by
interaction with immune cells.
• In infants the usage of GOS has been shown to have a
potential role in allergy prevention and reduction of
infectious diseases.
• Several studies have shown that GOS and FOS
fermentation can lead to the production of butyrate,
the principal fuel for colonic epithelial cells. This SCFA
also stimulates apoptosis (Rowland 1998) and may be
a protective factor in carcinogenesis (Scheppach and
Weiler 2004).
• Propionate is also produced from GOS, and has been
shown to be anti-inflammatory with respect to colon
cancer cells (Nurmi et al. 2005).
Role of TOS/GOS on Mucosal
Immune System:
• The intestinal mucosa contains large amounts of sIgA which has a
protective role against adherence and invasion by harmful bacteria
and viruses.
• Levels of sIgA in faeces have been found to correlate with an
increased ability to neutralize and clear viruses.
• Infants have an immature immune system, and because of the lack of
transfer of immunoglobulins from breast milk, formula-fed babies have
low levels of sIgA, which can lead to increased risk of gastrointestinal
infection.
• Thus, the addition of prebiotics such as GOS alone or in combination
with probiotic bifidobacteria or lactobacilli with immunomodulatory
abilities that are able to utilize the substrate, has potential to increase
production of sIgA in the body.
ROLE OF OTHER PREBIOTICS ON IMMUNITY:
OLIGOFRUCTOSE:
• The studies conducted with recognized prebiotic
fibres (oligofructose) have shown increased
lymphocyte and/or leucocyte numbers in GALT
(Gaskins et al., 1996; Pierre et al., 1997; Field et
al.,1999) and peripheral blood (Kaufhold et al.,
2000).
PECTIN:
• More CD4+ T cells have been found in mesenteric lymph
nodes in rats fed a diet containing 5% pectin, compared
to less fermentable cellulose (Lim et al., 1997).
LACTULOSE:
• Studies have documented that feeding lactulose is
associated with increases in IgA secretion or IgA+
cells in GALT (Kudohet al.1998; Kudohet al.1999), a
decrease in the CD4+/CD8+ ratio in the spleen
(Kudoh et al. 1998), and an increase in the
phagocytic function of intraperitoneal macrophages
(Nagendra & VenkatRao, 1994).
• Immunological effects have been observed after
adding prebiotics such as FOS and lactulose to the
diet, including increases in mucosal immunoglobulin
production, mesenteric lymph nodes, Peyer’s
patches, and altered cytokine formation and
lymphocyte numbers in the spleen and intestinal
mucosa (Schley and Field, 2002).
REFERENCE SUBJECTS EXPERIMENTAL
FIBRE DOSE
CONTROL DIET/
FIBRE DOSE
IMMUNE EFFECT
Field et al.,
1999
Adult
mongrel
dogs
Fermentable
fibre mixture
(beet pulp,
Oligofructose,
gum arabic),
8·7 g/kg
Cellulose, 8·3
g/kg
• ↑ CD8+ cells in Intra
Epithelial Lymphocytes(IEL),
PP and LP
• ↑ CD4+ cells in MLN and
peripheral blood.
• ↑ T cell mitogen responses
in MLN and IEL
Lim et
al.1997
Sprague-
Dawley rats
Pectin, konjak
mannin, or
chitosan, 5 %
w/w
cellulose, 5 %
w/w
• ↓serum and MLN IgE (all
fibres)
• ↑serum IgA and IgG
(pectin)
• ↑MLN IgA and IgG (pectin,
chitosan)
• ↑CD4+ T cells in MLN
(pectin)
• ↑INF-γ in MLN (pectin)
REFERENCE SUBJECTS EXPERIMENTAL
FIBRE DOSE
CONTROL DIET/
FIBRE DOSE
IMMUNE EFFECT
Pierre et
al.1997
Min mice Oligofructose
(from sucrose),
wheat bran, or
resistant
starch, 5·8 %
w/w
Cellulose, 2 %
w/w
• ↑ number of PP in small
intestine (short-chain FOS)
Yamada et
al., 1999
Sprague-
Dawley
rats
PHGG, guar
gum, HM
pectin, or
glucomannan,
5 % w/w
cellulose, 5 %
w/w
• ↑IgA in spleen and MLN
(all fibres).
• ↑ IgG in spleen
(glucomannan, pectin) and
MLN (all fibres).
• ↑ serum IgA (guar gum,
glucomannan, pectin) and
IgM (glucomannan).
Yun et al.,
1997
C57BL/6
mice
(immunos
uppresse
d
Oat b-glucan, 3
mg every 48 hr
Diet not
specified
• ↑non-specific and
antigen-specific IgG in
serum.
• ↑ IFN-γ-and IL-4-secreting
cells in spleen and MLN.
***
Conclusion
 Chemicals or food ingredients, selectively fermentable.
 supports the beneficial bacteria.
 Plays major role in immunity
 Mainly 2 types of prebiotic fulfill all the criteria.
 Prebiotics can be combined with probiotics to form
Synbiotics which can be given as nutritional
supplements to get synergic effect.
Reference:
TEXT BOOK:
• Gary B. Huffnagle, Mairi Noverr, 2008, GI Microbiota and Regulation of the
Immune System, springer.
JOURNALS:
• Bourgot,CL, Stéphanie Ferret-Bernard, Laurence Le Normand, Gérard
Savary, Enrique Menendez-Aparicio, Sophie Blat, Emmanuelle Appert-
Bossard, Frédérique Respondek, Isabelle Le Huërou-Luron, 2014, Maternal
short-chain fructooligosaccharide supplementation influences intestinal
immune system maturation in piglets. PLoS ONE. 9(9): pp e107508.
• Field CJ, McBurney MI, Massimino S, Hayek MG & Sunvold GD (1999) The
fermentable fiber content of the diet alters the function and composition of
canine gut associated lymphoid tissue.
• Veterinary Immunology and Immunopathology72, 325– 341
• Gibson GR & Roberfroid MB (1995) Dietary modulation of the human
colonic microbiota: introducing the concept of prebiotics.J Nutr, 125: 1401 –
1412
• Gibson GR, Probert HM, Van Loo J, Rastall RA & Roberfroid MB (2004)
Dietary modulation of the human colonic microbiota: updating the concept
of prebiotics.Nutr Res Rev17, 259 – 275.
• Inan, M.S., Rasoulpour, R.J., Yin, L., Hubbard, A.K., Rosenberg, D.W. and
Giardina, C. (2000) The luminal short-chain fatty acid butyrate modulates
NF-κB activity in a human colonic epithelial cell line. Gastroenterology 118,
724–734.
• Kelly-Quagliana K, Nelson P & Buddington R (2003) Dietary oligofructose and
inulin modulate immune functions in mice. Nutr Res23, 257 – 267
• Lim, B.O., Yamada, K., Nonaka, M., Kuramoto, Y., Hung, P. and Sugano, M.
(1997) Dietary fibers modulate indices of intestinal immune function in rats. J
Nutr 127, 663–667.
• Nurmi, J., Puolakkainen, P. and Rautonen, N. (2005) Bifidobacterium lactis
sp. 420 up-regulates cylooxygenase (Cox) 1 and down-regulates COX-2 gene
expression in a Caco-2 cell culture model. Nutr Can 51, 83–92.
• Pierre F, Perrin P, Champ M, Bornet F, Meflah K & Menanteau J (1997) Short-
chain fructo-oligosaccharides reduce the occurrence of colon tumors and
develop gut-associated lymphoid tissue in min mice.Cancer Research57,
225– 228.
• Roberfroid,M.B., 2007. "Prebiotics: The Concept Revisited". J Nutr. 137 (3
Suppl 2): 830S–7S
• Roller M, Rechkemmer G & Watzl B (2004) Prebiotic inulin enriched with
oligofructose in combination with the probiotics Lactobacillus rhamnosus and
Bifidobacterium lactismodulates intestinal immune functions in rats. J Nutr134,
153 – 156.
• Rowland, I.R. (1998) Role of the Gut Flora in Toxicity and Cancer. London:
Academic Press.
• Schley, P.D. and Field, C.J. (2002) The immune-enhancing effects of dietary fibres
and prebiotics. Br J Nutr 87, S221–S230.
• Seidel C, Boehm V, Vogelsang H,et al.(2007) Influence of prebiotics and
antioxidants in bread on the immune system, antioxidative status and
antioxidative capacity in male smokersand non-smokers.Br J Nutr97, 349 –356.
• Shadid R, Haarman M, Knol Jet al.(2007) Effects of galactooligosaccharide and
long-chain fructooligosaccharide supplementation during pregnancy on
maternal and neonatal microbiota and immunity – a randomized, double-blind,
placebo-controlled study.Am J Clin Nutr86, 1426– 1437.
• Stillie RM, Bell RC & Field CJ (2005) Diabetes-prone BioBreeding rats do not have
a normal immune response when weaned to a diet containing fermentable
fibre.Br J Nutr93, 645 – 653
• Soleimani,N., Seyed Hossein Hoseinifar, , , Daniel L. Merrifield,
Mohsen Barati, Zohreh Hassan Abadi, 2012. Dietary
supplementation of fructooligosaccharide (FOS) improves the
innate immune response, stress resistance, digestive enzyme
activities and growth performance of Caspian roach (Rutilus
rutilus) fry. Fish & Shellfish Immunology.32(2):316–321.
• Trushina EN, Martynova EA, Nikitiuk DB, Mustafina OK &
Baigarin EK (2005) The influence of dietary inulin and
oligofructose on the cell-mediated and humoral immunity in
rats. Vopr Pitan74, 22– 27
• Yamada K, Tokunaga Y, Ikeda A, Ohkura K, Mamiya S, Kaku
S,Sugano M & Tachibana H (1999) Dietary effect of guar gum
and its partially hydrolyzed product on the lipid metabolism
and immune function of Sprague-Dawley rats. Bioscience,
Biotechnology and Biochemistry63, 2163– 2167.
• Yun C-H, Estrada A, Van Kessel A, Gajadhar AA, Redmond MJ &
Laarveld B (1997) B-(1!3, 1!4) oat glucan enhances resistance
to Eimeria vermiformisinfection in immunosuppressed mice.
International Journal for Parasitology 27, 329– 337.
Prebiotics & Immunity
Prebiotics & Immunity

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Prebiotics & Immunity

  • 2.  Introduction  Definition  Types of prebiotics  Mechanism for the effect of prebiotics on immunity  Structure & role of different prebiotics  Conclusion  Reference
  • 3. Introduction • The concept of prebiotic was introduced by Gibson & Roberfroid, 1995. • Prebiotics is a general term to refer to chemicals that induce the growth and/or activity of commensal microorganisms (e.g., bacteria and fungi) that contribute to the well-being of their host. • The criteria which must be met in order to be classified a prebiotic, as defined by Gibson & Roberfroid,1995. -Resistance to hydrolysis or absorption in the upper gastrointestinal tract. -Fermented by the intestinal microbiota. -Selectively stimulate the growth and/or activity of beneficial intestinal bacteria, such as Lactobacillius species and Bifidobacterium species.
  • 4. Definition • Gibson & Roberfroid, 1995: ‘Prebiotics are non-digestible food ingredients that beneficially affect the host by selectively stimulating the growth, and/or activity, of one or a limited number of beneficial bacteria in the colon and thus improve host health’. • Roberfroid, 2007: ‘A prebiotic is a selectively fermented ingredient that allows specific changes, both in the composition and/or activity in the gastrointestinal microflora that confers benefits upon host well-being and health’.
  • 6. • Only two particular prebiotics then fully met the refined definition of prebiotics: Inulin(IN) and Trans- galactooligosaccharide (Roberfroid, 2007). • Other authorities also classify resistant starch, fructooligosaccharide (FOS) and lactulose as prebiotics. Mannan Oligosaccharides (MOS) have been termed as prebiotics but would more correctly be termed immunosaccharides (Roberfroid, 2007). • Carbohydrates including gluco-oligosaccharides, isomalto-oligosaccharides, nigero-oligosaccharides, oat b-glucans, raffinose, soyabean oligosaccharides (SOS), and xylo-oligosaccharides are considered as candidate prebiotics.
  • 7. Mechanism for the effect of prebiotics on immunity • The underlying mechanisms of how prebiotics modulate the immune system are not known at present. • However, experimental data suggests that these compounds exert effects in the GALT and also point to a few different mechanisms that might explain these effects: 1. Selective changes in bacterial composition and bacterial products which modulate cytokine and antibody production; 2. Production of SCFAs and their interactions with leukocytes 3. Modulated mucin production 4. Interaction with carbohydrate receptors of pathogens inhibiting their enhancement to epithelial cells as well as receptors on immune cells.
  • 8. A. SELECTIVE CHANGES IN BACTERIAL COMPOSITION AND BACTERIAL PRODUCTS:  It is well known that prebiotics increase the no. of beneficial bacterial (i.e bifidobacteria and lactobacilli).  Probiotics when administered orally are known to increase the secretion of IgA in the small intestine and the feces and to stimulate PP B lymphocyte IgA production.  They are also known to exert effects on systemic immune functions and various immune parameters in the lungs, spleen and peritoneal cells.
  • 9.  Intestinal epithelial cells are involved in both innate and adaptive immune responses and act by transducing signals from luminal pathogens to adjacent immune cells of the intestinal immune system, via specific germinal-encoded pattern- recognition receptors, such as Toll-like receptors (TLRs) and cytoplasmic receptors.  TLRs are able to discriminate between the normal commensal biota and pathogens and induce the transcriptional activation of a no. of genes mediating immune and inflammatory responses.
  • 10.  Pathogen-associated molecular pattern (PAMPs) [eg., endotoxin(lipopolysaccharide), lipoproteins, lipopeptides and imidazoquinolines] present on diverse microbes are initially recognized by TLRs and their interaction result in the activation of intracellular signaling pathways, nuclear translocation of transcription factor NF-κB and the transcription of pro-inflammatory cytokines.  The changes that occur in the composition of the intestinal microbiota due to prebiotic fermentation could potentially reduce the presence of PAMPs and thereby exert a positive effects on the immune system.
  • 11.  Prebiotics also promote an increase in the bacterial cell-wall components that are recognized by TLRs and in DNA derived from luminal bacteria that, in turn, stimulate the intestinal immune system.  Cytoplasmic components and cell-free extracts of probiotics have also been demonstrated to produce some of the same immune effects (eg, IgA production by PP and macrophage stimulation) as live bacteria.
  • 12. B. PRODUCTION OF SCFAs: • The major end products of carbohydrate fermentation are SCFAs, of which acetate, propionate and butyrate are quantitively the most important in the human colon. • The production of SCFAs in the colon averages 400 mmol/day, with a range of 150-600 mmol/day. • All SCFAs are rapidly absorbed from the large intestine and stimulate salt and water absorption: principally, the gut epithelium, liver and muscle metabolize them, with virtually none appearing in the urine and only small amounts appearing in the feces.
  • 13. • The three major SCFAs are trophic when infused into the colon and these trophic properties have important physiological implications in addition to maintaining the mucosal defense barrier against invading organisms. • Butyrate is known to suppress lymphocyte proliferation, inhibit cytokine production of Th1-lymphocytes and upregulates IL- 10 production. • It also suppresses expression of the transcription factor NF- κB and upregulates TLR expression. • Butyrate is also believed to protect against colon cancer as it inhibits DNA synthesis and induces cell differentiation.
  • 14. • It has been demonstrated in a rat model that supplementing total parenteral nutrition with a SCFA mixture results in increased NK cell activity(Pratt et al.,1996). • Pharmacological doses of acetate administered intravenously to both healthy individuals and cancer patients also increased NK cell activity and peripheral blood antibody production. • In addition, it has been shown that serum glutamine levels are raised following lactulose administration and suggested that increased SCFA levels were responsible for this (glutamine is a preferred substrate for lymphatic tissue). • Therefore, SCFA production in the large intestine could potentially reduce the requirement of epithelial cells for glutamine, making it available to the cells of the immune system.
  • 15. C. MUCIN PRODUCTION: • The first line of defense of the mucosa against luminal contents is the mucous layer, which is mainly composed of high-molecular-weight glycoprotein (mucins) that are secreted by goblet cells. • The effect of prebiotics on mucin production has been reported in a study, where it was shown that inulin administration resulted in increased mucin production in rats. • Greater mucin production was found to be associated with a lower incidence of bacterial translocation across the mucosa following dietary fibre supplementation.
  • 16. • It has been shown in a perfused rat colon model that the production of acetate and butyrate from the fermentation of dietary fibre stimulates mucin secretion, but fibers do not have the same effect on their own. • Pathogens and beneficial commensal bacteria are able to modulate mucin synthesis by regulating some of the mucin genes. • Currently there are 16 identified mucin genes, but further work is needed to fully explain the function of each of them and to identify new genes.
  • 17. D. CARBOHYDRATE RECEPTORS:  Studies suggest that some prebiotics are directly involved in protecting the gut from infection and inflammation by inhibiting the attachment of pathogenic bacteria or their toxins to the colonic epithelium.  This attachment is necessary before pathogens can colonize and cause disease and it is mediated by glycoconjugates on glycoproteins and lipids present on the microvillus membrane.
  • 18.  Certain prebiotic oligosachharides contain structures, similar to those found on the microvillus membrane, that interfere with the bacterial receptors by binding to them and thus preventing bacterial attachment to the same sugar on microvillus glycoconjugates.  For example, α-linked TOS, present in human milk, are known to have anti-adhesive properties and be capable of toxin neutralization.  Recently, a novel TOS mixture, which contains an oligosachharide alpha anomeric configuration, was shown to significatntly decrease the attachment of enteropathogenic Escherichia coli (EPEC) and salmonella enterica serovar Thyphimurium in vitro.
  • 19.  In addition, immune cells also express specific carbohydrate receptors which mediate various cellular reactions when activated. For example, C-type receptors expressed on phagocytic cells.  It is hypothesised that carbohydrate moieties on the prebiotic may interact with receptors on immune cells.  Although a specific fructose receptor has not yet been identified, receptors for b-glucan and mannose have been identified on immune cells, and in vitro, fructose has been shown to alter non-opsonic phagocytosis, suggesting that a receptor for fructose on immune cells may exist.  In addition, some oligosaccharides, for example OF, can bind to receptors on pathogenic bacteria and prevent them from attaching to this same sugar on the epithelial membrane, thus preventing adherence.
  • 20. Structure & role of different prebiotics • β2-1 Fructans, which include inulin (IN) and fructo- oligosaccharides (FOS), fulfil the criteria for prebiotics. (Gibson, et al.,2004). • According to Roberfroid, 2007, only two particular prebiotics then fully met the refined definition of prebiotics: Inulin(IN) and Trans-galactooligosaccharide (TOS).
  • 21. INULIN: • Inulin is the most common type of FOS. • IN is a linear carbohydrate molecule which contains β-(2→1) fructosyl–fructose linkages with a terminal glucose. • IN may contain between two and sixty fructose residues with an average of twelve. (fig.1) • Partial enzymatic hydrolysis of IN yields a FOS known as oligofructose(OF), which can have a terminal glucose or fructose residue (fig.1) • In OF there can be two to eight (average five) fructose residues with a terminal glucose residue or a chain of three to eight (average five) fructose residues.
  • 23. Dietary Source of Inulin (IN): • IN is found naturally in a variety of plant foods such as bananas, barley, chicory, garlic, artichoke, leeks, onions and wheat. • Oligosaccharides, including some believed to be prebiotics, are present in human breast milk. • The presence of oligosaccharides in large amounts in breast milk suggests that these compounds may play an important role in early infant development, perhaps of the gut, its microbiotia and the immune system.
  • 24. Role of Inulin on immunity: • The effect of β2-1 fructans upon macrophage number and function has been studied, with the results suggesting that macrophage functions are enhanced by the addition of β2-1 fructans to the diet. • Peritoneal macrophage phagocytic activity was increased in rodents given IN or OF for varying periods of time (Kelly-Quaglian et al.,2003).
  • 25. • In rats, OF-enriched IN (100g/kg) increased caecal secretory IgA concentrations(Roller et al., 2004). • The number of T cells in the MLN of rats was increased upon 10% OF or IN supplementation and also the blood IL-2 and IL-4 concentrations were increased upon the supplementation for 4 months (Trushina et al., 2005). • A study shows the percentage of blood B cells (defined as CD19+) was increased in young male adults after consumption of a bread containing IN (Seidel et al., 2007).
  • 26. REFERENCE Prebiotics dose used Animal studied Immune effect Stillie et al, 2005 Inulin (4·8 % w/w diet) Rats (21 d old, male & female); diabetes resistant or diabetes prone In diabetes-resistant rats: ↑Small intestine length ↓ Number of splenocytes ↑CD8+ Lymphocytes in PP ↑ Proliferation of splenocytes and MLN cells to mitogens ↓Production of IL-4 and ↑ production of IL-10 by stimulated splenocytes In diabetes-prone rats: ↓Number of splenocytes ↑IgA+ cells in jejunal lamina propria ↑B lymphocytes in PP ↓Production of IL-4 and ↑production of IL-10 by stimulated splenocyte
  • 27. FRUCTOOLIGOSACCHARIDES (FOS): STRUCTURE: • FOS consists of several β(1-2) or β(1-6) linked fructose units which may be linked to glucose residues. DIETARY SOURCE: • They can be found naturally in some cereal crops as well as fruits and vegetables such as bananas, onions, chicory root, garlic, asparagus, onion and leeks.
  • 28. Role of FOS on immunity: • They stimulate the growth of bifido bacteria and to inhibit growth and multiplication of potentially pathogenic bacteria such as enterobacteria, clostridia and salmonella. • Fermentation of FOS leads to the production of short chain fatty acids, which is substrate for energy metabolism in the colonic mucosa stimulating epithelial cell growth. • Short chain FOS can enhance IgA content and thus play a major role in immunity.
  • 29. • A study conducted by Bourgot et al.,2014 says that maternal scFOS supplementation modified the intestinal immune functions in piglets in association with increased colostral immunity. • Thirty-four sows received a standard or a scFOS supplemented diet (10 g scFOS/d) for the last 4 weeks of gestation and the 4 weeks of lactation. • Colostrum and milk immunoglobulins (Ig) and TGFβ1 concentrations were evaluated on the day of delivery and at d 6 and d 21 postpartum. • Colostral IgA (P<0.05) significantly increased because of scFOS and TGFβ1 concentrations tended to improve (P<0.1). • Such results underline the key role of maternal nutrition in supporting the postnatal development of mucosal immunity.
  • 30. Another study shows dietary supplement of FOS to the Caspian roach (Rutilus rutilus) fry increase the serum Ig levels, lysozyme activity and alternative complement activity (ACH50) and thus improving the innate immune response (Soleimani et al.,2012).
  • 31. TRANS-GALACTOOLIGOSACCHARIDES (TOS): STRUCTURE: • Galacto-oligosaccharides (GOS), also known as oligogalactosyllactose, oligogalactose, oligolactose or transgalactooligosaccharides (TOS), belong to the group of prebiotics. • GOS/TOS generally comprise a chain of galactose units that arise through consecutive transgalactosylation reactions, with a terminal glucose unit. However, where a terminal galactose unit is indicated, hydrolysis of GOS formed at an earlier stage in the process has occurred. • The degree of polymerization of GOS/TOS can vary quite markedly, ranging from 2 to 8 monomeric units.
  • 32. Role of TOS/GOS on immunity: • Galactooligosaccharides, GOS can positively influence the immune system—indirectly through the production of antimicrobial substances as the result of galactooligosaccharide fermentation, that can reduce the proliferation of pathogenic bacteria, and directly by interaction with immune cells. • In infants the usage of GOS has been shown to have a potential role in allergy prevention and reduction of infectious diseases.
  • 33. • Several studies have shown that GOS and FOS fermentation can lead to the production of butyrate, the principal fuel for colonic epithelial cells. This SCFA also stimulates apoptosis (Rowland 1998) and may be a protective factor in carcinogenesis (Scheppach and Weiler 2004). • Propionate is also produced from GOS, and has been shown to be anti-inflammatory with respect to colon cancer cells (Nurmi et al. 2005).
  • 34. Role of TOS/GOS on Mucosal Immune System: • The intestinal mucosa contains large amounts of sIgA which has a protective role against adherence and invasion by harmful bacteria and viruses. • Levels of sIgA in faeces have been found to correlate with an increased ability to neutralize and clear viruses. • Infants have an immature immune system, and because of the lack of transfer of immunoglobulins from breast milk, formula-fed babies have low levels of sIgA, which can lead to increased risk of gastrointestinal infection. • Thus, the addition of prebiotics such as GOS alone or in combination with probiotic bifidobacteria or lactobacilli with immunomodulatory abilities that are able to utilize the substrate, has potential to increase production of sIgA in the body.
  • 35. ROLE OF OTHER PREBIOTICS ON IMMUNITY: OLIGOFRUCTOSE: • The studies conducted with recognized prebiotic fibres (oligofructose) have shown increased lymphocyte and/or leucocyte numbers in GALT (Gaskins et al., 1996; Pierre et al., 1997; Field et al.,1999) and peripheral blood (Kaufhold et al., 2000). PECTIN: • More CD4+ T cells have been found in mesenteric lymph nodes in rats fed a diet containing 5% pectin, compared to less fermentable cellulose (Lim et al., 1997).
  • 36. LACTULOSE: • Studies have documented that feeding lactulose is associated with increases in IgA secretion or IgA+ cells in GALT (Kudohet al.1998; Kudohet al.1999), a decrease in the CD4+/CD8+ ratio in the spleen (Kudoh et al. 1998), and an increase in the phagocytic function of intraperitoneal macrophages (Nagendra & VenkatRao, 1994). • Immunological effects have been observed after adding prebiotics such as FOS and lactulose to the diet, including increases in mucosal immunoglobulin production, mesenteric lymph nodes, Peyer’s patches, and altered cytokine formation and lymphocyte numbers in the spleen and intestinal mucosa (Schley and Field, 2002).
  • 37. REFERENCE SUBJECTS EXPERIMENTAL FIBRE DOSE CONTROL DIET/ FIBRE DOSE IMMUNE EFFECT Field et al., 1999 Adult mongrel dogs Fermentable fibre mixture (beet pulp, Oligofructose, gum arabic), 8·7 g/kg Cellulose, 8·3 g/kg • ↑ CD8+ cells in Intra Epithelial Lymphocytes(IEL), PP and LP • ↑ CD4+ cells in MLN and peripheral blood. • ↑ T cell mitogen responses in MLN and IEL Lim et al.1997 Sprague- Dawley rats Pectin, konjak mannin, or chitosan, 5 % w/w cellulose, 5 % w/w • ↓serum and MLN IgE (all fibres) • ↑serum IgA and IgG (pectin) • ↑MLN IgA and IgG (pectin, chitosan) • ↑CD4+ T cells in MLN (pectin) • ↑INF-γ in MLN (pectin)
  • 38. REFERENCE SUBJECTS EXPERIMENTAL FIBRE DOSE CONTROL DIET/ FIBRE DOSE IMMUNE EFFECT Pierre et al.1997 Min mice Oligofructose (from sucrose), wheat bran, or resistant starch, 5·8 % w/w Cellulose, 2 % w/w • ↑ number of PP in small intestine (short-chain FOS) Yamada et al., 1999 Sprague- Dawley rats PHGG, guar gum, HM pectin, or glucomannan, 5 % w/w cellulose, 5 % w/w • ↑IgA in spleen and MLN (all fibres). • ↑ IgG in spleen (glucomannan, pectin) and MLN (all fibres). • ↑ serum IgA (guar gum, glucomannan, pectin) and IgM (glucomannan). Yun et al., 1997 C57BL/6 mice (immunos uppresse d Oat b-glucan, 3 mg every 48 hr Diet not specified • ↑non-specific and antigen-specific IgG in serum. • ↑ IFN-γ-and IL-4-secreting cells in spleen and MLN. ***
  • 39. Conclusion  Chemicals or food ingredients, selectively fermentable.  supports the beneficial bacteria.  Plays major role in immunity  Mainly 2 types of prebiotic fulfill all the criteria.  Prebiotics can be combined with probiotics to form Synbiotics which can be given as nutritional supplements to get synergic effect.
  • 40. Reference: TEXT BOOK: • Gary B. Huffnagle, Mairi Noverr, 2008, GI Microbiota and Regulation of the Immune System, springer. JOURNALS: • Bourgot,CL, Stéphanie Ferret-Bernard, Laurence Le Normand, Gérard Savary, Enrique Menendez-Aparicio, Sophie Blat, Emmanuelle Appert- Bossard, Frédérique Respondek, Isabelle Le Huërou-Luron, 2014, Maternal short-chain fructooligosaccharide supplementation influences intestinal immune system maturation in piglets. PLoS ONE. 9(9): pp e107508. • Field CJ, McBurney MI, Massimino S, Hayek MG & Sunvold GD (1999) The fermentable fiber content of the diet alters the function and composition of canine gut associated lymphoid tissue. • Veterinary Immunology and Immunopathology72, 325– 341 • Gibson GR & Roberfroid MB (1995) Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics.J Nutr, 125: 1401 – 1412 • Gibson GR, Probert HM, Van Loo J, Rastall RA & Roberfroid MB (2004) Dietary modulation of the human colonic microbiota: updating the concept of prebiotics.Nutr Res Rev17, 259 – 275.
  • 41. • Inan, M.S., Rasoulpour, R.J., Yin, L., Hubbard, A.K., Rosenberg, D.W. and Giardina, C. (2000) The luminal short-chain fatty acid butyrate modulates NF-κB activity in a human colonic epithelial cell line. Gastroenterology 118, 724–734. • Kelly-Quagliana K, Nelson P & Buddington R (2003) Dietary oligofructose and inulin modulate immune functions in mice. Nutr Res23, 257 – 267 • Lim, B.O., Yamada, K., Nonaka, M., Kuramoto, Y., Hung, P. and Sugano, M. (1997) Dietary fibers modulate indices of intestinal immune function in rats. J Nutr 127, 663–667. • Nurmi, J., Puolakkainen, P. and Rautonen, N. (2005) Bifidobacterium lactis sp. 420 up-regulates cylooxygenase (Cox) 1 and down-regulates COX-2 gene expression in a Caco-2 cell culture model. Nutr Can 51, 83–92. • Pierre F, Perrin P, Champ M, Bornet F, Meflah K & Menanteau J (1997) Short- chain fructo-oligosaccharides reduce the occurrence of colon tumors and develop gut-associated lymphoid tissue in min mice.Cancer Research57, 225– 228. • Roberfroid,M.B., 2007. "Prebiotics: The Concept Revisited". J Nutr. 137 (3 Suppl 2): 830S–7S
  • 42. • Roller M, Rechkemmer G & Watzl B (2004) Prebiotic inulin enriched with oligofructose in combination with the probiotics Lactobacillus rhamnosus and Bifidobacterium lactismodulates intestinal immune functions in rats. J Nutr134, 153 – 156. • Rowland, I.R. (1998) Role of the Gut Flora in Toxicity and Cancer. London: Academic Press. • Schley, P.D. and Field, C.J. (2002) The immune-enhancing effects of dietary fibres and prebiotics. Br J Nutr 87, S221–S230. • Seidel C, Boehm V, Vogelsang H,et al.(2007) Influence of prebiotics and antioxidants in bread on the immune system, antioxidative status and antioxidative capacity in male smokersand non-smokers.Br J Nutr97, 349 –356. • Shadid R, Haarman M, Knol Jet al.(2007) Effects of galactooligosaccharide and long-chain fructooligosaccharide supplementation during pregnancy on maternal and neonatal microbiota and immunity – a randomized, double-blind, placebo-controlled study.Am J Clin Nutr86, 1426– 1437. • Stillie RM, Bell RC & Field CJ (2005) Diabetes-prone BioBreeding rats do not have a normal immune response when weaned to a diet containing fermentable fibre.Br J Nutr93, 645 – 653
  • 43. • Soleimani,N., Seyed Hossein Hoseinifar, , , Daniel L. Merrifield, Mohsen Barati, Zohreh Hassan Abadi, 2012. Dietary supplementation of fructooligosaccharide (FOS) improves the innate immune response, stress resistance, digestive enzyme activities and growth performance of Caspian roach (Rutilus rutilus) fry. Fish & Shellfish Immunology.32(2):316–321. • Trushina EN, Martynova EA, Nikitiuk DB, Mustafina OK & Baigarin EK (2005) The influence of dietary inulin and oligofructose on the cell-mediated and humoral immunity in rats. Vopr Pitan74, 22– 27 • Yamada K, Tokunaga Y, Ikeda A, Ohkura K, Mamiya S, Kaku S,Sugano M & Tachibana H (1999) Dietary effect of guar gum and its partially hydrolyzed product on the lipid metabolism and immune function of Sprague-Dawley rats. Bioscience, Biotechnology and Biochemistry63, 2163– 2167. • Yun C-H, Estrada A, Van Kessel A, Gajadhar AA, Redmond MJ & Laarveld B (1997) B-(1!3, 1!4) oat glucan enhances resistance to Eimeria vermiformisinfection in immunosuppressed mice. International Journal for Parasitology 27, 329– 337.

Editor's Notes

  1. Modulate :To adjust or adapt to a certain proportion; regulate or temper. Cytokine:Any of several regulatory proteins, such as the interleukins and lymphokines, that are released by cells of the immune system and act as intercellular mediators in the generation of an immune response.
  2. igA:Immunoglobulin A (IgA, also referred to as sIgA) is an antibody that plays a critical role in mucosal immunity. Pp-peyer’s patch-organized lymphoid nodules found in the lowest portion of the small intestine, the ileum. located in the lamina propria layer of the mucosa and extending into the submucosa of the ileum. In adults, B lymphocytes are seen to predominate in the follicles' germinal centers. Systemic Inflammatory Response Syndrome (SIRS) is an inflammatory state affecting the whole body, frequently a response of the immune system to infection
  3. Toll-like receptors (TLRs) are a class of proteins that play a key role in the innate immune system. They are single, membrane-spanning, non-catalytic receptors usually expressed in sentinel cells such as macrophages and dendritic cells, that recognize structurally conserved molecules derived from microbes. Transcription is the first step of gene expression, in which a particular segment of DNA is copied into RNA
  4. Pathogen-associated molecular patterns, or PAMPs, are molecules associated with groups of pathogens, that are recognized by cells of the innate immune system. They are recognized by Toll-like receptors (TLRs). NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) is a protein complex that controls transcription of DNA.
  5. Interleukin-10 (IL-10), also known as human cytokine synthesis inhibitory factor (CSIF), is an anti-inflammatory cytokine.
  6. Luminal: The inner open space or cavity of a tubular organ, as of a blood vessel or an intestine TRANSLOCATION. : transfer of part of a chromosome to a different position especially on a nonhomologous chromosome; especially : the exchange of parts between nonhomologous chromosomes.
  7. Non-opsonic phagocytosis is typically mediated by cell surface receptors on leukocytes that recognize repeating carbohydrate subunits (comprising “molecular patterns”) onmicrobes. Opsonic phagocytosis is typically mediated by deposition of proteins (e.g., antibodies) on microbes that target them for recognition by specific phagocytic receptors onleukocytes.
  8. Caecal: a blind pouch connected to the large intestine between ileum and colon.
  9. A splenocyte can be any one of the different white blood cell types as long as it is situated in the spleen or purified from splenic tissue. CD(cluster of differentiation/classification determinant) is a protocol used for the identification and investigation of cell surface molecules providing targets for immunophenotyping of cells. MLN:Mesenteric lymph nodes, A mitogen is a chemical substance that encourages a cell to commence cell division, triggering mitosis. FUNCTION OF IL4(Interleukin)-including the stimulation of activated B-cell and T-cell proliferation, and the differentiation of B cells into plasma cells. It is a key regulator in humoral and adaptive immunity. Interleukin-10 (IL-10), also known as human cytokine synthesis inhibitory factor (CSIF), is an anti-inflammatory cytokine.
  10. TGF-B1- Transforming growth factor beta 1 or TGF-β1 is a polypeptide member of the transforming growth factor beta superfamily of cytokines.
  11. intraepithelial lymphocytes (IEL) , lamina propria (LP) Interferon Gamma (IFN-g)
  12. The function of Peyer's patches is to analyze and respond to pathogenic microbes in the ileum
  13. Synergy is the creation of a whole that is greater than the simple sum of its parts. meaning "working together"