GIN 6 – Gut Immunology Proscribe: Sanjita Ravishankar

September 13, 2007 Reviewer: Marisol Albuerne

Dr. Klimpel Chief Reviewer: Kimberlyn Fitchett
Gut Mucosa and Immunobiology
(A bullet change on page 3 in red has been made)
I. Some Trivia
• Surface area of the gut equals almost 3 tennis courts
• Gut is largest immunological organ in the body, containing 1012 lymphocytes
• Gut is home for 1014 bacteria (hence the need for all of the lymphocytes) – 1 kg, 400
different species
• Produces more antibody than the rest of the body
II. Functions of Gut Immunity
• Protect against enteric pathogens
• Maintain tolerance for other antigens such as self antigens and food
• Promote systemic immunity by sampling and presenting antigens
• The GI immune system has a spectrum of responses to different types of stimuli:
o Self/food antigens  clonal elimination, anergy, immune suppression (aka tolerance)
o Normal flora  Cell activation, cell recruitment, cytokines (perhaps a continuous
regulated immunological reaction?)
o Pathogens  Antibodies, cytokines, cell mediated lysis, inflammation (taking the bad
guys down)
III. Review of Immunology (memory lane of first half of PHD)
• Antibodies on the surface of B cells bind antigen  cell presents the antigen via MHC II
to a T cell  T cell activates the B cell  B cell becomes a plasma cell  antibodies
produced by plasma cell
• Cell types can be recognized by their surface markers
o All T cells express CD2 and CD3
o Helper T cells also express CD 4
o Cytotoxic T cells express CD 8
o Suppressor T cells express CD 8
• T cells can only recognize antigen if it is presented in the form of a small peptide on an
MHC complex by an antigen presenting cell (APC)
o CD 8 T cells recognize MHC Class I (8 x 1=8)
o CD 4 T cells recognize MHC Class II (4 x 2=8)
• CD 4 precursor cells can differentiate into different types of effector cells, each of which
has its own cytokine profile
o TH0 (naïve T cells) – IL 1, IL 4, IL 5, IFN γ, IL 10
o TH1 – IL 2, IFN γ, TNF β
o TH2 – IL 4, IL 5, IL 6, IL 10
o Memory cells – IL 2
o TH 1 responses are stimulated by intracellular pathogens: brucella, mycobacteria,
Borrelia, viruses, leishmania (cutaneous); allografts; encapsulated infectious agents
 Cytokines IL 12 and IFN γ promote TH1 and inhibit TH2
 Low dose antigen favors TH1 response
 Cutaneous immunization favors TH1
o TH 2 responses are stimulated by allergens and parasites/schistosomes, leishmania

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GIN 6 – Gut Immunology Proscribe: Sanjita Ravishankar
September 13, 2007 Reviewer: Marisol Albuerne
Dr. Klimpel Chief Reviewer: Kimberlyn Fitchett
 Cytokines IL 10 and IL 4 promote TH2 and inhibit TH1
 High dose antigen favors TH2
 Intestinal immunization favors TH2
o Remember that the responses aren’t exclusively TH 1 or TH 2, they are merely
skewing the overall immune response
• Immune effector mechanisms
o Neutralization (antibodies can neutralize toxins)
o Cytotoxicity (complement, cytotoxic T cells)
o Inflammation (lymphocytes, polys, macrophages)
o Cytostimulation (epithelial cells can be activated
to secrete protective substances)

IV. Gut Immunology

• General pathway:
• M cell  Peyer’s Patch  Memory T cells or naïve
B cells  Mesenteric lymph nodes  Thoracic duct
 Peripheral circulation  Homing to mucosa in
lamina propria (including back to the original section
of gut that picked up the antigen)
• M cells
o Doorway to the immune system of the gut,
responsible for antigen sampling
o Normal flora do not attach or enter M cell
o Have large involutions that surround other
immune cells, including macrophages and
lymphocytes
o Very different from normal
gut epithelium – do not
have microvilli/brush
border, reduced expression
of digestive enzymes
o Can express MHC I and II
o Transports antigens and passes them to macrophages to be
presented to T cells (may be able to present antigen themselves
as well)

• Peyer’s Patches (Gut associated lymph tissue/GALT)

o Help to initiate mucosal immune responses, the afferent response
o Contains macrophages, dendritic cells, immature T cells, B cells
o Site for lymphocyte activation, which then migrate via efferent
lymphatics to mesenteric lymph nodes and take residence in
lamina propria
• Lamina Propria

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GIN 6 – Gut Immunology Proscribe: Sanjita Ravishankar
September 13, 2007 Reviewer: Marisol Albuerne
Dr. Klimpel Chief Reviewer: Kimberlyn Fitchett
o More of the efferent limb of the immune response – contains already activated T cells
(larger cells than the GALT)
o Carry out specialized effector functions
 Helper cells activate B cells for antibody synthesis
 Cytokine production
 Cytolytic function
• Lymphocyte trafficking is directed by adhesion receptors
o Most important one we need to know is the interaction between α4,β7 on
lymphocytes that are homing to the GI tract with MAdCAM-1 on endothelial cells in
the gut
• Intraepthelial Lymphocytes (IEL) – “911 cells”
o Large granular lymphocyte morphology
o CD3+, CD8+
o HML-1 integrin expression
o γ,δ T cell receptors
o Alternative (non MHC) activation pathway
o Can recognize bacterial phospholipids
o Produces IL 2, IFN γ quickly
o Functions are cytotoxic and immunoregulatory
o Recognizes MIC-A and MIC-B that are produced by stressed epithelial cells (like
when they are infected), and induces apoptosis in those cells; may also help make
new cells to replace the apoptotic ones
• Intestinal Epithelial Cells
o Can be activated by cytokines
o Express MHC I and II, can express their own cytokines
to influence other cells
o IFN γ and TNF α increase paracellular permeability of
the tight cell junctions
 Also upregulate polymeric IgA receptor
o Other effectors increase chloride and mucus secretion
o Can act as antigen presenting cells via HLA-DR (small
bowel>colon, increases with inflammation); can
activate intraepithelial lymphocytes (CD8 T-cell)
• Antibodies
o The gut is the major source of IgA, and IgA is the
antibody produced most by the gut
o The majority of immunoglobulin secreting cells are
associated with the GI tract (70-80%)
o IgA B cell differentiation and maturation occur in two
sites – the Peyer’s patch and lamina propria
 IgM B cells become committed to producing IgA in
the Peyer’s patch, pass through the systemic
circulation, then become activated, become plasma
cells and start producing IgA in the lamina propria

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GIN 6 – Gut Immunology Proscribe: Sanjita Ravishankar
September 13, 2007 Reviewer: Marisol Albuerne
Dr. Klimpel Chief Reviewer: Kimberlyn Fitchett
o Polymeric IgA production/secretion (2 IgAs connected
by a J chain)
 Rough ER in epithelial cell produce secretory component that recognize the IgA
dimer and form disulfide bond with it
 The complex (secretory IgA) is endocytosed and transported in the epithelial cell
 The complex then goes through proteolytic cleavage and the dimer (polymeric
IgA) is released into the gut lumen
 Monomeric IgA is the main form found in serum
 Subclasses of IgA – products of two
different genes
o IgA1 – main form in serum
o IgA2 – main form in secretions
 Properties of IgA
o Relatively resistant to proteolysis
(IgA2>IgA1)  lives in the gut, so this
makes sense
o Poor activator of complement
o Inhibits bacterial adhesion,
macromolecule absorption,
inflammatory effects of other Igs
o Neutralizes viruses, toxins
o Enhances nonspecific defense
mechanisms
o Mediates antibody dependent
cytotoxicity
V. Oral Tolerance
• Necessary to prevent excessive response to normal flora and food antigens
• May be cause of poor or absent immune response to most antigens administered by oral
route
• May not be present for T independent antigens (bacterial polysaccharides)
• Several forms:
o Tolerance for all Ig classes
o Systemic tolerance, IgA immunity
• May be due to alterations of T and B cell function
o T cells – clonal deletion, clonal anergy, suppressor
cells
o B cells – clonal deletion, clonal anergy, anti-idiotype
(antibodies against antibodies)
• May depend on whether the antigen presenting cell has
been activated by a costimulatory molecule
• TH3 cells (suppressor cells) use TGF-β1 to suppress the
TH1 and TH2 immune responses to an antigen

VI. Inflammatory Bowel Disease (IBD)

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GIN 6 – Gut Immunology Proscribe: Sanjita Ravishankar
September 13, 2007 Reviewer: Marisol Albuerne
Dr. Klimpel Chief Reviewer: Kimberlyn Fitchett
• Major working hypothesis for IBD is that the disease is due to an abnormal and
uncontrolled immune response to one or more normally occurring gut constituents
• There are many models for experimental IBD
• The enteric bacterial environment has a major impact on disease expression:
o If you knockout a gene responsible for the immune reaction, but place the mouse in a
germ-free environment, it will not get IBD
o You can also reduce the disease by restricting bacterial colonization or giving
antibiotics to the knockout mouse.
o Note that different people have different normal flora.

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