The document discusses molecular markers of innate immunity and inflammation and their role in therapeutic intervention. It covers topics such as pattern recognition receptors (PRRs) that detect pathogens and damage signals; classes of PRRs including Toll-like receptors; the signaling pathways of TLRs and their role in cancer and neurodegeneration. It also discusses the transcription factor NF-kB and proinflammatory cytokines such as TNF, IL-1, IL-6 that are involved in the inflammatory response. Specific conditions like meningococcal meningitis and gout are mentioned where blocking these cytokines has shown therapeutic potential.

1. Molecular markers of innate immunity and
inflammation in therapeutic intervention
by : Mahdi zarei
M.Sc. Student ,clinical
biochemistry
Ferdowsi university of mashhad

2. innate immune system& Inflammation response
 The innate immune system utilizes a variety of
transmembrane or secreted pattern-recognition receptors
(PRRs), which are vital for activation of complement and
coagulation
cascades,opsonization, phagocytosis, apoptosis and
induction of proinflammatory mediators. (Sylvia
Adams,2009)
 A typical inflammatory response consists of four
components: inflammatory inducers, the sensors that
detect them, the inflammatory mediators induced by the
sensors, and the target tissues that are affected by the
inflammatory mediators.( ruslan,2010)

3. ruslan,2010

4. PAMPs , DAMPs(inducers) and PRRs(sensors)
 Germline-encoded pattern recognition receptors (PRRs) are responsible
for sensing the presence of PAMPs & DAMPs.
 PAMPs : structures conserved among microbial species, which are called
pathogen-associated molecular patterns.
 DAMPs : endogenous molecules released from damaged cells, termed
damage-associated molecular patterns.(osamu ,2010)

5. Classes of PRRs
 four different classes of PRR families have
been identified :
 Toll-like receptors(TLRs)
 C-type lectin receptors (CLRs)
 RIG-I-like receptors (RLRs)
 NOD-like receptors (NLRs).

6. Toll like receptor
 In the late 90’s a protein was discovered in Drosophila named as
Toll. Toll is a transmembrane receptor that is required for the
establishment of proper dorso-ventral polarity during embryo
formation in Drosophila.(Hashimoto et al., 1988)
 Toll is responsible for the production of Drosomycin, antifungal
peptide .(Lemaitre et al., 1996).
 Toll homologues in mammals as well were revealed known as Toll
like receptors (TLRs).
 TLRs recognise PAMPs of diverse origin from bacteria, virus, fungi,
protozoa and others. TLRs can also sense the molecules that are
generated within the host cells alarming a sort of danger signal like
heat shock proteins (Hsp60, Hsp70, Hsp90), fibrinogen, surfactant
protein A, heparin sulphate and others.

7. TLR signaling pathway
1) MYD88-dependent pathway:MyD88 dependent
signalling pathway is found to be central to all TLRs except
TLR3.
2) TRIF-dependent pathway(MYD88-
independent):TLR3 and TLR4 activate TRIF-dependent
signaling, which activates NF-kB and IRF3 resulting in the
induction of proinflammatory cytokine genes and type I IFNs.

8. Osamu&shizuo ,
2010

9. Sylvia ,2009

10. TLRs in cancer and anti-cancer immunotherapy
 The TLR3 has been shown to be receptor for viral dsRNA, and also
seems to be potentially promising in anti-cancer therapy. Reports
have shown that cancer cells themselves express TLR3 in vivo and
agonist ploy (I:C) is activating the signalling pathway leading to the
anticancer effects (O'Neill et al., 2011)
 TLR agonists can induce differentiation, proliferation or activation of
Treg cells. Several TLR agonists such as Streptococcal agent OK-
432, double stranded RNA and CpG DNA have anti-tumour activity
(Chen & Oppenheim, 2009).
 Treg cells express TLR4, 5, 7 and 8 in mice. It has been reported that
transfer of Treg cells enhanced tumour growth in mice but it was
reversed upon stimulation of Treg cells with a TLR8 ligand.
Administration of LPS also abrogates Treg activity reveals latent anti-

11. Sylvia,2009

12. TLRs in neurodegeneration
 TLR signaling pathways are a potential therapeutic target in
AD(Alzheimer’s disease) ; however more work remains to
delineate the complex interaction of TLRs in Aβ deposition and
clearance and its precise role in AD development. (Okun et
al., 2009)
 Although TLRs often recognize pathogen associated molecular
patterns and protect the body from invasion of microbial
pathogens, the expression of TLRs within multiple sclerosis
suggests novel roles for these receptors in mediating neurological
disease and hence can be used as a biomarker of the
neurodegenerative disorders (Okun et al., 2009).
 Moreover, it is important to determine the precise role of distinct
TLRs in Aβ recognition and clearance, and the activation of glial

13. nuclear factor kB (NF-kB)
 NF-ĸB is a transcription factor that controls the expression of genes
involved in inflammation, immunity and apoptosis.
 About 100 genes are under the transcriptional control of NF-ĸB.
 NF-ĸB is an evolutionary conserved protein having five members in
mammals- p50,p52, RelA/p65, RelB and RelC.
 All these function either as homodimer or heterodimer for e.g. p50
homodimer and p50/p65 heterodimer.

14. Canonical and alternative pathways of NF-kB activation
Xavier, 2005

15. NF-κB and cancer therapy
 NF-kB regulates the expression of genes involved in many
processes that play a key role in the development and progression of
cancer such as proliferation, migration and apoptosis.
 constitutive NF-kB activation has been detected in many human
malignancies.
 some anti-inflammatory drugs may inhibit NF-kB by interfering with
IKK activity.
 Other substances such as curcumin, trans-resveratrol or
parthenolide are natural compounds that have been demonstrated to
inhibit IKK activity
 Another way to approach NF-kB inhibition is to target the process of
proteasome degradation. Proteasome inhibitors prevent NF-kB

16. Drug Molecular targets
(NF-kB-linked)
Clinical activity
Aspirin(NSAIDs)
Sulfasalazin
IKKβ inhibition colon, lung, prostate
cancer, Hodgkin’s
disease,
Glucocorticoids IⱪBα transcription Lymphoid leukemias,
Hodgkin’s disease,
multiple myeloma
Thalidomide/lenalido
mide
Macrolides
Arsenic trioxide
IKKβ inhibition multiple
myeloma,chronic
lymphocytic
leukaemia, various
solid tumor
Gene
therapy(siRNA)
Decreased p65
expression
not available
Sabine,2006

17. Proinflammatory cytokines
 Tumor necrosis factor(TNF)
 Interleukin-1
 Interleukin-6

20. Tumor necrosis factor(TNF) as therapeutic target
 In patients with meningococcal meningitis serum levels of TNF-α have
been shown to correlate with mortality(Arditi,1990) , but other authors have
failed to show significant increases in TNF-α in septic
patients(Casey,1993).
 In a rodent model, mortality after an injection of endotoxin was signifantly
decreased by pre-treatment with TNF-a antibodies.(Beutler,1987)
 the antibody approach has produced little evidence of benefit in man
although improvements in the haemodynamics of patients have been
observed.(Exley,1990)
 The use of recombinant soluble TNF-a receptors has been shown to
improve survival in mice, (Bertini,1993) but at present there have been no
similar results in man.
 So far the use of TNF-a or its soluble receptors has not proved to be useful
as a diagnostic marker.(Giannoudis,2004)

21. NLRP3
Inflammasome
Activation
Kate Schroder,2010

22. Interleukin-1 as terapiotic target
 Elevated local or systemic IL-1β has been linked to a number of human
hereditary or acquired diseases, and antagonists of IL-1β or its receptor are
proving successful treatments for a number of these diseases.(Kate,2010)
 Clinical trials are currently assessing efficacy of Anakinra and next-generation
IL-1β inhibitors for a wider range of diseases, including cryopyrin-associated
periodic syndromes (CAPS), gout, and type II diabetes (T2D).
(Hoffman,2009,…)
 Gout is an autoinflammatory disease characterized by severe joint
inflammation, resulting in arthropathy and considerable pain. Gout is strongly
associated with metabolic disturbances leading to elevated blood uric acid
levels (hyperuricemia) and the deposition of MSU crystals in joints. (Talbott,
1940)
 MSU is a potent activator of the NLRP3 inflammasome in vitro, and MSU-
dependent neutrophil recruitment is dependent on the ASC adaptor, caspase-1,
and IL-1R in vivo. (Martinon et al., 2006)

23. refrences
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25. Thank you