Natural Killer (NK)

Cells and Mucosal

Immunity
MICP511M - Tuesday, 26 September 2023 (11:00-11:50 AM)
Yuko Ota, PhD
yota@som.umaryland.edu
 Learning Objectives
NK cells:
• Know the two main functions of the NK cell in host defense
• Understand how NK cells kill target cells
• Understand the similarity and differences between the activities of NK
cells and Cytotoxic T lymphocytes (CTL)

Mucosal immunity:
• Understand how the immune system maintains homeostasis in the
intestine in the presence of microbes
• Be able to identify key immune mediators protecting the oral cavity
NK cells are lymphocytes
B and T
Adaptive cells

NK

Innate cells
 NK cells

Important in antiviral and anti-tumor defense

 Natural Killer (NK) cells
• Kill virally infected cells
• Complement cytotoxic T cells
• Kill tumor cells
• Secrete cytokines (IFN gamma)
• NK cells are activated by IFNs (type I IFN alpha, beta), TNF, IL12
• 20-100 fold more deadly

Also activated by IFN gamma by TH1 cells

Mediate Antibody-dependent cellular cytotoxicity (ADCC) via Fc receptor (CD16)
Downregulation of MHC class I during viral infection
 Compensatory mechanism of cell mediated immunity

MHC class I

CTL killing

Or
Down-
regulation of
NK killing
MHC class I

NK cells kill virus-infected or some tumor cells by 2 mechanisms:
FAS-L-Fas and perforin-granzyme

Smyth et al. 2004. Mol. Immunology 42:501

 NK receptors
• are not Pathogen recognition receptors (PRR)
• are not generated by gene rearrangement (no GOD/ TCR, BCR)
• recognize self, not foreign
• 2 classes of receptors
• Activating receptors (ITAM) Kill
• Inhibitory receptors (ITIM) No Kill

• Also have Fc receptor (CD16: FcγRIII)  used for ADCC

 Antibody-Dependent Cell Cytotoxicity
(ADCC)

IgG binding CD16: FcgRIII perforin and Cell lysis

To the cell granzymes
surface
 Two types of NK receptors

Immunoreceptor tyrosine based activating

Two types of motif (ITAMs): Sequence motifs that are
Receptors phosphorylated and promote signaling
1) NKG2
2) KIR
(Killer cell Ig‐like Receptors) Important to note that these ITAMs are the same as
You learned for signaling molecules of TCRs and BCRs

Immunoreceptor tyrosine based inhibition

motif (ITIMs): Sequence motifs that are
phosphorylated and prevent signaling
NK cells are primed to kill, and kept in check
by inhibitory receptors

Inhibitory>activating

No inhibitory
NK Cell cytotoxic function is regulated by the BALANCE of activating
and inhibitory signals

Missing Self

More
More activating than
inhibitory
inhibitory signals
than
activating
“SOS” molecules signals
NK cells kill and they make cytokines
 TCR vs NK receptors

TCR MHC NK receptor

Gene Rearrangement Germ-line encoded, Germ-line encoded, alternatively

Polymorphic ligand spliced and/or polyhgenic

clonally expressed Codominant expression KIR variegated expression

MHC:peptide Binds peptids,TCR, NKR MHC-I:peptide or other molecules

TCR + co-stimulation Antigen presentation Balance of inhibitory and activating

activate T cells signals activate or inhibit NK cells
 Learning Objectives (check
points)
NK cells:
• Know the two main functions of the NK cell in host defense
• Understand how NK cells kill target cells
• Understand the similarity and differences between the activities of NK
cells and Cytotoxic T lymphocytes (CTL)
 What is the role of the systemic
immune compartment?
• Eliminate pathogens in the circulation or tissues that
breach the skin (lymph nodes) or enter the blood
(spleen)
Systemic Lymphoid Tissue vs. Mucosal Lymphoid Tissues

•Spleen •Tonsils, adenoids

•Peripheral LNs •Mesenteric LNs (MLN), Peyer’s patches
(PP), Appendix
What is meant by mucosal?
• Epithelial cell-lined tracts and ducts
• Oral
• Gastrointestinal
• Respiratory
• Urogenital
• Mucous blanket
•60-70% of total lymphocytes
• Largest immune compartment - 400 meter2
Major route of pathogen entry! Tissue is doing
something important physiologically and is exposed
to the environment; got to protect it.
What is the function of the mucosal
immune compartment?

• Preserve the intestinal barrier and prevent movement of

commensals into the circulation.
• Tolerate beneficial Ags from food and commensals.
• Protect mucosal tissues from invading pathogens.

Accomplishing all of this is complicated. Lots of potential for

pathology if something goes wrong: Coeliac, IBD, Allergy,
etc.
Benefits of our microbiota
• Mutualism: Symbiotic relationship beneficial to both
host and parasite
• 1014 organisms
• Commensal microbiota in body combined to weigh 1 kg
• Help prevent colonization of pathogens
• Provide nutrients to the host
• Vitamin K
• Microbiota can be a threat when barrier containment is
lost
GI tract
Mucosal Epithelial Cells:
(Single Layer of cells)
1. IEC  nutrient uptake & barrier
2. Goblet cells  mucins
3. Stem cells  regeneration
4. Paneth Cells  anti-microbial
peptides and cytokines
Tight junctions maintain
barrier function
Innate defenses of the intestinal
mucosa: Paneth Cells

a-defensins
Cathelicidins
Lysozyme

Paneth Cells: Specialized epithelial cells at base of the crypt that secrete antimicrobial
proteins into the lumen
Dimeric secretory IgA is a main effector
molecule of the mucosal immune system

• Plasma cells present in the lamina

propria
• 75,000 in normal intestine
• Secrete 3-4g IgA / day in absence of
infection
Mucosal transport of immunoglobulins- major
isotypes: dimeric IgA

NOTE: IgA and J chain are produced by the B-cell

The Poly-Ig receptor (PIgR) is produced by the epithelium
 Transport of IgA across the
epithelial barrier by transcytosis

Transport is
dependent
on B cell
produced
J-chain
 Functions of secretory
IgA

IgA is poor at
complement
activation and
opsonization
 No inflammation

Neutralization in Neutralization in Neutralization in

lumen endosomes lamina propria
 Summary: Local immune responses to
microbiota at homeostasis

Secreted IgA and antimicrobial peptides keeps microbiota from becoming too invasive
MALT: Mucosal-associated lymphoid tissue

GALT: Gut associated lymphoid tissue

BALT: bronchus-associated lymphoid tissue
NALT: nasopharinx-associated lymphoid tissue

“Common” mucosal immune system is MALT

Activated lymphocytes that originate in the GALT, NALT and BALT

inductive sites migrate via peripheral blood to proximal and distal
mucosal sites, ensuring that effector lymphocytes and secretory
antibodies are present across a range of mucosal membranes
Organization of the mucosal
immune system
• Primary lymphoid tissue
• None
• Secondary lymphoid tissues
• Mesenteric lymph nodes
• Tonsils
• Gut-associated lymphoid tissue (GALT): Anatomically
Compartmentalized structure typical of secondary
lymphoid tissue. Sites where mucosal immune responses
are initiated. Comprised of…
• Peyer’s patches (100-200)
• Isolated lymphoid follicles (thousands)
• Appendix
GI
Microbial sensing in the GALT

CD103+
CD11c+

CX3CR1+CD11b+CD11c+
Circulation of naïve lymphocytes in
the mucosal immune system
• Naïve B and T cells
• Trafficking to mucosal tissue not predetermined after exit from Bone
Marrow and Thymus
• Entry into Peyer’s patches through High Endothelial Venules
• CCL21/ CCR7 mediated chemotaxis
• L- selectin
• No antigen exposure
• Exit through the lymph back to blood stream
• Antigen exposure
• Lose CCR7 and L-selectin expression
• Not yet effector cells
• Exit through the lymph back to the blood stream
• Imprinted by DCs to return back to mucosal tissue
Effector cells are imprinted in PP
to return to mucosal tissue

Before Ag After Ag
CCR7 CCR9
L- Selectin a4b7 integrin
Mucosal Immune system is a
functional network
• Common mucosal immune system
• Endothelial cells lining mucosal blood vessels contain MAdCAM-1
• Lymphocytes primed in the gut can traffic to other mucosal sites
• Respiratory track
• Urinary tract
• Relevance to Vaccination
Organization of GALT in the small intestine

For oral: tonsils

Immunity in the oral cavity
• Diseases and disorders
caused by oral
microorganisms:
• Dental Caries
• Periodontitis
• Halitosis
• Tonsilitis

Moutsopoulos et al. Trends Imm. 2017

 Important components of oral
immunity
• Desquamation
• Oral microbiota
• Secreted IgA
• Saliva
• Antimicrobial peptides in Saliva
• Defensins
• Calprotectin
• Lactoferrin
• Enzymes
• Lysozyme
• Peroxidases
• Mucus Layer
• < 0.1 mM
• Acquired Pellicle
• 30-100 nM
Lymphoid tissues in oral cavity
 Learning Objectives

Mucosal immunity:
• Understand how the immune system maintains homeostasis in the
intestine in the presence of microbes
• Be able to identify key immune mediators protecting the oral cavity
Thank you for listening!