Pattern recognition receptors

What is Pattern?

Pattern: a regular and intelligible form or sequence discernible in the way in which
something happens or is done

Fibonacci number: a series or pattern of integers in Mathematics

Fn= Fn-1 +Fn-2

0,1,1,2,3,5,8,13,21, 34, 55, 89,144,…….

21 blue and 13 aqua spirals

 Pattern recognition and what to do with them

Machine learning: pattern recognition focuses on the recognition of patterns and

regularities in data

Immunology : Molecular patterns that are signature of microorganisms

Importantly, not present in host cells

Pathogen/Microbe Associated Molecular Patterns (P/MAMPS)

Pattern recognition receptors: present on Macrophages, DCs, Neutrophils etc.

 Pattern Recognition Receptors (PRRs)

Drosophila Toll : originally identified as a controller of

dorso-ventral patterning of the embryo
In adult fly it initiated the induction of host-defense
mechanisms including anti-microbial peptides and is
critical in defense against Gram-positive bacteria and
fungal pathogens

Jules Hoffman, 1996: discovered the pattern

recognition ability of the Drosophila Toll protein
Nobel Prize for Physiology and Medicine, 2011

First proposed by Charles Janeway, Jr. based on the requirement of adjuvants in driving
immune response to purified antigens

⮚ pathogen recognition by the innate immune system is specific

⮚ relies on germline encoded pattern-recognition receptors (PRRs)
⮚ PRRs detect components of foreign pathogens referred to as
PAMPs
 Pattern Recognition Receptors (PRRs): The Toll-like Receptors

Toll-like: homologues of Drosophilla Toll found in mammals

resistance to viral, bacterial and fungal infection

Plants: proteins with similar ligand binding domains to TLRs involved in production of
antimicrobial peptides

10 TLR genes in humans and 13 in mice

Each recognize a distinct set of molecular patterns on microbes

Initially called pathogen-associated molecular patterns (PAMPs)
microbes-associated molecular patterns (MAMPs)

⮚ Limited specificity
⮚ Found on a variety of cells including Macrophage, DC, B cells, stromal cells
and epithelial cells
⮚ initiation of anti-microbial responses in many tissues
Pattern Recognition Receptors (PRRs): The Toll-like Receptors
Division of labor:

Lipoteichoic acid of gram +ve

bacteria
LPS of gram –ve bacteria
Flagellin of Bacterial flagella
Unmethylated CpG motif of
bacterial DNA
DS RNA in viruses
 The Toll-like Receptors: recognition of the patterns

Indirect recognition: eg. Drosophila Toll

⮚ does not bind to the microbial product directly

⮚ binds to the cleaved self-protein Spatzle which binds to Toll and
activates the signaling cascade

TLRs: directly recognize the microbial pattern

TLR1 and TLR2 dimer with bound synthetic ligand

 The Toll-like Receptors: localization
Cell surface TLRs: TLR1,2,4,5,6,11
Intracellular: TLR3,7,8,9
❖ Transmembrane proteins
❖ 18-25 leucine rich domains
❖ cell-surface: detection of
extracellular pathogens
❖ intracellular location:
endosomal membranes
lysosomes
endolysosomes
detect pathogens that have been taken
up by phagocytosis, macropinocytosis,
receptor mediated endocytosis
❖ Homodimers or heterodimers
❖ signaling is induced when
ligands induce dimerization or
alteration in dimer conformation
 The Toll-like Receptors: ligand binding and signaling

TLR1, TLR2, TLR6:

⮚ lipoteichoic acid and diacyl and triacyl lipoproteins
⮚ ligand binding induces heterodimerization; TLR2:TLR1 and TLR2:TLR6
⮚ TLR2:TLR1 recognize triacylated lipopeptides from Gram-ve bacteria & mycoplasma
⮚ TLR2:TLR6 recognize diacylated lipopeptides from Gram+ve bacteria & mycoplasma
⮚
TLR1 and TLR6: differential ligand recognition

•2 out of 3 lipid side chains of the synthetic

lipid PAm3CSK4 (a triacylated lipopeptide)
interact with TLR2
•3rd chain binds the hydrophobic channel of
TLR1
•hydrophobic channel is absent from TLR6,
so the TLR2-TLR6 heterodimer does not
recognize triacylated lipopeptides

•different lipid-binding pockets of TLR1 and

TLR6 are responsible for the discrimination
between lipoproteins
 Toll-like Receptor 4: Heterodimers
C3H/Hej mice • LPS resistant (resistant to endotoxin)
• Very susceptible to Gram-ve bacteria infection
• Mutations in the lps allele

missense mutation in the third exon of the TLR4 gene

TLR4 : expressed on DC and Macrophages

Nobel Prize in Physiology and Medicine, 2011

 The Toll-like Receptors 5 and TLR11

TLR5: recognizes flagellin , a protein component of bacterial flagella

In the Lamina Propria of the small intestine, a DC subset CD11c+CD11b+ have high
expression of TLR5
These LP DCs are specially adapted in promoting Th1 and Th17 differentiation
Also causes diffrentiation of naïve B cells into IgA producing plasma cells in response to
flagellin

TLR 11:
• Only in mouse
• Kidney and Bladder have high expression of TLR11, related to TLR5
•Recognize uropathogenic bacterial components
• Recognize specific motif of protozoan profilin eg. From Toxoplasma gondii
 Nucleic acid sensing TLRs: TLR3
Expression: macrophage, cDCs, intestinal epithelial cells

Recognizes: double stranded RNA (replicative intermediate of many viruses)

synthetic analog of dsRNA, polyinosinic:polycytidylic acid (poly I:C)

Generation of ligand: endocytosis of viruses with ds-RNA genomes (Rotavirus)

phagocytosis of dying cells in which viruses are replicating
Encounters the TLR when the endocytic vesicle or phagosome fuses
with the TLR containing endosome
Function: Antiviral immune response through production of type 1 interferon and
inflammatory cytokines
 Nucleic acid sensing TLRs: TLR7
Expression: cDCs and plasmacytoid DCs
Recognizes:
• single stranded RNA derived from RNA viruses like vesicular stomatitis virus, influenza A
virus and HIV
• synthetic poly(U) RNA and certain siRNAs
• TLR7 on cDCs recognize RNA from Group B Streptococcus and induces type 1 interferon

Generation of ligand:
• Viruses are internalized and recruited to the endolysosomes where TLR7 mediated
recognition of ssRNA and the initiation of antiviral responses are triggered (therefore
recognition of the RNA viruses is replication-independent)
• via Autophagy in the double-membrane bound autophagosomes: (lysosomal degradation
of cellular proteins in double membrane autophagosome vesicles)

Function:
TLR7 serves as the sensor of infection with ssRNA viruses
 Nucleic acid sensing TLRs: TLR8 and TLR9
TLR8: recognized viral ssRNA
Highest expression on Monocytes
Mice lacking TLR8 respond normally to the agonist
Function not clearly established

TLR9: recognized unmethylated CpG dinucleotides (mammalian genome: CpG

dinucleotides are heavily methylated oon the cytosine by DNA methyltransferases;
CpG in bacteral and viral genomes remain unmethylated and therefore presents a
molecular pattern

CpG oligos directly activate DCs, Macrophage and B cells leading to a strong Th1
response
 TLRs: Downstream signaling

Type 1 interferon responses (anti-viral)

TLR signaling
Inflammatory cytokines (cell surface TLRs)

All mammalian TLRs have the TIR (Toll-IL-1 receptor domain) domain in their cytoplasmic tail
TLR TIR domains interact with other proteins containing TIR domains to transduce signal

TIR domain containing adaptor molecules: MyD88, TIRAP (MAL), TRIF, TRAM
Engagement of each of these adaptors leads to distinct signaling pathway

MyD88: universally used by all TLRs except TLR3

activates transcription factor NF-kB and MAPkinases to induce inflammatory cytokines

TRIF: used by TLR3 and TLR4 (MyD88-independent pathway)

activates transcription factor IRF3 and NF-kB leads to type1 interferon
and inflammatory cytokines
TRAM and TIRAP: sorting adaptors that recruit TRIF to TLR4 and MyD88 to TLR2
and TLR4, respectively