Memorias Conferencia Científica Anual sobre Síndrome Metabólico 2015 - Programa Científico - Dra. Julia Kzhyshkowska - Jefa del Departamento de Inmunidad Innata y Tolerancia, Facultad de Medicina, Universidad de Heidelberg, Alemania. Miembro del Editorial Board, Revista Immunobiology

1. PROGRAMMING OF INNATE IMMUNE CELLS IN
DIABETIC CONDITIONS:
EMERGING EPIGENETIC MECHANISMS
Prof. Dr. Julia Kzhyshkowska
Institute of Transfusion Medicine and Immunology,
Medical Faculty Mannheim, University of Heidelberg
Mexico 14.08.2015

2. Innate and adaptive immune system cooperate
Innate system:
Non-specific
“non-self” and
“unwanted-self” sensing,
local inflammation
Resolution of inflammation
Wound healing
Adaptive system:
Antigen specific
amplification of inflammation
Immunological memory
Antigen recognition
Activation, Recruitment,
Inhibition
Antigen presentation
Activation, Recruitment,
Inhibition
•Macrophages
•Mast cells
•Neutrophils
•Basophils & Eosinophils
•NK cells
Bone Osteoclasts
Brain Microglia
Liver Kupffer cells
Lung Alveolar macrophages
Skin Langerhans cells

3. Differentation of macrophages
© Prof. Dr. J. Kzhyshkowska
•PPSC – pluripotent stem cell
•GM-CFU – granulocyte-monocyte colony
forming unit
•M-CFU – monocyte colony forming unit
•GM-CSF - granulocyte-monocyte
stimulating factor
•CSF-1 – colony stimulating factor
•Monocytes and macrophages are
believed to be non-dividing cells
GM-SCF

4. Physiological functions of macrophages in the tissues
•Cytokine balance
•Regulation of tissue turn-over
•Inhibition of unwanted
inflammation
•Elimination of transformed and
dying cells
•Elimination of microorganisms
•Initiation und regulation of
inflammation
•Recruitmenat and activation of
adaptive immune cells, in particular T-
cells
•Phagozytosis of apoptotic and necrotic cells
•Stimulation of cell migration and
proliferation
•Induction and support of angiogenesis
•Inhibition of immune cell activities
Homeostasis Inflammation
Healing
Exogene effectors
Exogene effectors
Endogene effectors
Endogene effectors
Trauma
Pathogen
© Prof. Dr. J. Kzhyshkowska

5. Monocytes and macrophages in inflammation
© Prof. Dr. J. Kzhyshkowska
Bone marrow
Circulation Tissue
Resident
monocyte
Inflammatory
monocyte
Bone
marrow
Spleen?
Bone marrow
Spleen?
Micro- and
macrovascular
complications

6. Pathological functions of macrophages
Chronic Inflammation
Cancer
•Chemotaxis of monocytes, neutrophils , T-cells
and other immune cells
•Activation of innate and adaptive immune cells
•Inflammatory signals to neurons
•Stimulation of tumour cell proliferation
•Stimulation of tumour angiogenesis
•Inhibition of adaptive immune responses
•Support of intravasation
•Support of extravasation in metastatic
sites
•Production of pro-inflammatory
cytokines that modify neuron
functions
•Reduced scavenging function
leading to Abeta plaque formation
© Prof. Dr. J. Kzhyshkowska
Neurodegeneration
Cardiovascular disorders
Diabetic complications
•Endothelial inflammation
•Pathologic lipoprotein processing.
•Plaque instability

7. Factors that activate monocytes in the circulation
© Prof. Dr. J. Kzhyshkowska
Resident monocyte Inflammatory monocyte
Traumatic injury
Virus attack
Bacterial attack
Heat shock proteins
Cytokines/Chemokines
High glucose
Modified lipoproteins
Ischemia/tissue hypoxia
Inflammaging: recently discovered age-dependent process
Memory? Epigenetic fixation of activation status?

8. MACROPHAGES AS DIAGNOSTIC
BIOMARKERS AND CELL THERAPY TOOLS
Analysis of macrophage polarisation
Re-programming of macrophage
Immune cell therapy
Status of innate immune system activation in circulation and tissues
DiagnosticsDiagnostics
TherapyTherapy
Monocyte Macrophages

9. Phagocyte
“Whenever the organism enjoys immunity, the
introduction of infectious microbes is followed by the
accumulation of mobile cells, of white corpuscles of the
blood in particular which absorb the microbes and
destroy them. The white corpuscles and the other cells
capable of doing this have been designated
"phagocytes", i.e. devouring cells, and the whole
function that ensures immunity has been given the
name of "phagocytosis".
“It has been established as a general rule that in all
cases of immunity, natural or acquired, either by
preventive vaccination or following an attack of
infectious illness, phagocytosis takes place to a
marked degree, whereas in fatal or very dangerous
diseases, this phenomenon does not exist at all or is
attenuated.”
Nobel Lecture, December 11, 1908
Ilya Metchnikov

10. Activated macrophage
The mouse was found to be natively susceptible to Listeria
monocytogenes. Its susceptibility was attributed to the capacity of the
organism to survive and multiplying in host macrophages. During the first
3 days of a primary infection the bacterial populations of spleen and liver
were found to increase at a constant rate. On the 4th day of infection the
host became hypersensitive to Listeria antigens and at the same time
bacterial growth ceased. A rapid inactivation of the organism ensued.
Convalescent mice were resistant to challenge, but no protective factor
could be found in their serum. Histological evidence suggested that
acquired resistance was the result of a change occurring in the host's
mononuclear phagocytes. When challenged in vitro, the macrophages of
convalescent mice were found to resist infection with Listeria
monocytogenes. Listeria-resistant cells appeared during the course of
infection at a time which corresponded with the development of the
antibacterial mechanism in the spleen. They persisted for as long as the
antibacterial mechanism remained intact in this organ.
This period of absolute resistance to Listeria lasted about 3 weeks.
Thereafter, the host remained hypersensitive but unable to inactivate a
challenge inoculum of Listeria. However, it remained capable of
producing an accelerated response to reinfection. This was thought to
depend upon an ability to generate a new population of resistant cells
from a residuum of specifically sensitized macrophages or macrophage
precursors still surviving in the tissues as a result of the immunological
activation which occurred during the primary infection.
JEM 1962
George Mackaness

11. IFNgamma is the key factor activating macrophages
“IFN gamma activates human macrophage oxidative
metabolism and antimicrobial activity, and appeared to
be the only factor consistently capable of doing so in
the diverse lymphokine preparations tested”
JEM 1983
Carl Nathan

12. Alternative macrophage activation
Siamon Gordon
JEM 1992

13. Macrophage activation dichotomy (1992-1999)
Th1
IFNγ
LPS
GC
Th2
IL-4,
IL-10,
IL-13,
TGFβ
mannose receptor
scavenger receptor A
TNFα, IL-6, IL-1
FcγR I, II, III
NO, H2O2
Classically activated macrophage Mφ1
Alternatively activated macrophage Mφ2
AMAC-1
Stabilin-1

14. Design of the human monocyte-based ex vivo model system
IFNγ IL-4

15. Macrophage phenotypic diversity
IL-4DexTGFβ IFNγ LPS
CD163
Stab1
CD14
TLR4
TLR2
SI-CLP
MMR
FN1
βIGH3
MMP2
MMP12
TTG
AMAC-1
IL-1ra
FoxQ1
Stab1
NO
H2O2
FcγR
STAT1
MCP2
YKL40
TNFα
IL-1β
IL-12
IL-10
ID3
OLR1
HAMP
LTA4H
ALOX5AP
YKL39
FoxQ1
Phagocytosis
endocytosis
ECM
remodelling
Bactericidal
activity
Inflammatory
cytokine
production
Type 2 macrophages (M2) Type 1 macrophages (M1)
Each stimulation induces
individual set of macrophages
markers
Each stimulation induces
specific functions Interaction
with EC?
Lipoprotein
processing?
Technologies used:
2000: subtractive hybridization
2001-2003: Insyte microarrays
Since 2003: Affymetrix mircoarrays
and yeast two hybrid screening
Since 2010: antibody arrays
Scand J Immunol 2001, 2005
Journal of Leukocyte biology 2004
Immunobiology 2006,2012
Journal of Immunology 2006, 2008
Blood 2006, FASEB 2009
J of Immunology 2015

16. Macrophage phenotypic plasticity
IFNγIL-4
IFNγ
IL-4
antinflammatory
cytokines
Bactericidal
activity
TNFα
IL-1β
IL-10
TNFα
IL-1β
IL-12
IL-10
LPS LPS
Gratchev A, Kzhyshkowska J et al, Immunobiology. 2006;211(6-8):473-86
Over 600 papers at this topic
Gaps of Knowledge:
what are the frames
and limits of plasticity?
M2 M1

17. Signal transduction and vesicular transport define macrophage
function: example TGFbetaRII and stabilin-1
Inflammation Tolerance
Exogene effectors
Endogene effectors
Exogene effectors
Endogene effectors
Original papers:
Kzhyshkowska et al, J Leuk Biol, 2004
Kzhyshkowska et al J Immun. 2006
Kzhyshkowska et al J Immun. 2008
Gratchev; Kzhyshkowska et al, J Immun. 2008
Mosig et al, FASEB 2009
Zhang et al, Kzhyshkowska, MCB 2009
Schledzewski et al, Kzhyshkowska J Clin Invest 2011
Popova A et al, Immunobiology 2012
Ovsiy et al, manuscript in preparation
Nurgazieva te al, J Immun, 2015
Reviews:
Kzhyshkowska et al, J Cell Mol Med 2006
Kzhyshkowska et al, Immunobiology 2007;
Kzhyshkowska and Krussel, Immunobiology 2009
Kzhyshkowska, ScientificWorldJournal 2010
Kzhyshkowska et al, Immunobiology 2012
Gratchev et al, Kzhyshkowska, Immunobiology 2012
Gratchev et al, Kzhyshkowska, Curr Pharm Des. 2013

18. Orchestration of metabolism by macrophages
Biswas and Mantovani, Cell Metabolism, 2012

19. DIABETIC COMPLICATIONS
AND INFLAMMATION
microvascular complications
 diabetic retinopathy
 diabetic nephropathy
 diabetic neuropathy
macrovascular complications
 atherosclerosis
 cardio-vascular diseases
other chronic complications
 depression, dementia, sexual disfunction
 doubled over all risk of dying
 inflammatory processes involved
 not everybody suffer from complications
http://www.idf.org/sites/default/files/pictures/Complications-graphic.jpg

20. ATHEROSCLEROSIS
 Major Risk factors: Hypercholesterolemia/ dyslipedimia, hypertension, tabacco
consumption, diabetes mellitus
 Inflamed atherosclerotic plaques had a higher amount of macrophages and are thought to be at
higher risk to rupture [Puig 2011]
 LDL indices stabilin-1 and CD36 expression on monocytes promoting their adhesion to endothelium
[Mosig S 2009]
 TLR2 and 4 activation by modified LDL, Tenascin C or HMGB promotes foam cell formation [Howell
2011; Liu 2012; Tsung 2014]

21. Open question:
what are the mechanisms of long-term
pathological programming of innate
immune cells
by metabolic factors?

22. Epigenetic phenomena:
heritable alternative states of gene activity
that do not result from altered nucleotide
sequence

23. DNA methylation
Chromatin structure
Micro RNA
EPIGENETICS

24. DNA METHYLATION
In mammals only in CpG context
Blocking agent
for DNA methyl-transferase
For investogation or therapy

25. CpG ISLANDS
 CpG dinucleotides occur in clusters known as ‘CpG islands’.
 CpG islands are often associated with sites where the transcription of DNA
into RNA begins - the promoter regions.
 CpG islands are normally unmethylated, consistent with the ability of genes
to be transcribed in the presence of necessary transcription factors.
 Major function: regulation of gene expression and genome organization

26. DNA METHYLATION IN CANCER
 DNA hypomethylation contributes to cell transformation by 3 mechanisms:
 generation of chromosomal instability,
 reactivation of transposable elements,
 loss of imprinting.
I: Hypomethylation
II: Hypermethylation
 Hypermethylation of the CpG islands in the promoter regions of tumor-
suppressor genes (DNA-repair genes hMLH1, BRCA1, MGMT, p15, p16,
APC etc) is a major event in the origin of many cancers.

27. DNA METHYLATION IN CANCER

28. REGULATION OF DNA METHYLATION
 There are 2 major types of DNA methylation activity – maintenance
methylation and de novo methylation
 A critical step in DNA methylation involves DNMT (DNA methyl-
transferase), the enzymes that catalyze the methylation process (DNMT1,
DNMT3a, and DNMT3b).
 Loss of methylation requires active cell division

29. from Horn and Peterson Science, 2002
CHROMATIN AND HISTONES

30. Euchromatin
Transcriptionally active, less compacted
Heterochromatin
Less transcriptionally active, very compacted
a) constitutive heterochromatin
centromeres, telomeres
b) facultative heterochromatin
transposons, inactive X chromosome

31. Reddy and Natarajan,
Cardiovascular research, 2010

32. Latham and Dent
Nat Struct Mol Biol 2007
HISTONE MODIFICATIONS

33. “Histone Code” hypothesis
Modifications of the Histone tails act as marks that can
be read by other proteins to control the expression or
replication of chromosomal regions. The coding in the
histones may be heritable.
Generally, histone acetylation is associated with
transcriptionally active genes
Deacetylation is associated with inactive genes
(= gene silencing)
Role of methylation is context-specific

34. POI – protein of interest, may be histone or modified histone
CHIP-ON-CHIP

35. EPIGENETIC REGULATION OF INFLAMMATION

36. EPIGENETIC REGULATION OF INFLAMMATION
Gaps of knowledge:
How metabolic
conditions affect histone
code?

37. EPIGENETICS IN DIABETIC
ENDOTHELIAL CELLS
Reddy and Natarajan, Cardiovascular research, 2010c
Hyperglycemia and AGEs result in production of
pro-inflammatory mediators (cytokines, GF)
Pro-inflammatory mediators activate oxidant stress,
signal transduction (TK, PKC, MAPK) leading to
activation of transcription factors (NFkappaB) and
dysregulation of epigenetic mechanisms including
DNA methylation, histone modifications and miRNA
Outcome: loss of repressive chromatin marks and
gain of activation marks leading to formation of open
chromatin sate at the promoters of pathological
genes
Long-term fixation of open chromatin state works as
“metabolic memory”
Open question: why open state of chromatin is
fixed? Molecular mechanisms?
HMTs: histone methyltransferases
HDM: histone demethylases
HDAC: histone deacetylases
DNMTs: DNA methyltransferases
DeMet: DNA demethylases

38. Gaps of knowledge:
What are the key differences in
epigenetic changes in
subpopulations of macrophages in
health and disease Saeed S et al,
Science, 2014
EPIGENETICS CONTROLS
MONOCYTE TO MACROPHAGE DIFFERENTIATION

39. Our question:
which genes are controlled by
epigenetic mechanisms by metabolic conditions
in macrophages?

40. MODEL SYSTEM, MET
 Monocyte isolation from buffycoats by Biocoll and Ficoll centrifugation and
magnetic seperation of CD14+
monocytes with MACS beats
CD14+
monocytes
low glucose conditions (5 mM) high glucose conditions (25 mM)
IL-4 IL-4IFN-γ IFN-γ
 Macrophage differentitation by culuturing and stimulation for
6d
 RNA isolation and cDNA synthesis
 PCR and quantitative realtime PCR analysis

41. TLR AND DIABETES
 TLR2 knockout in diabetic mice attenuates developing chronic
inflammation or incipient diabetic nephropathy [Devaraj, 2011]
 TLR4 knockout attenuates the proinflammatory state of diabetes
[Devaraj, 2011]
 increased expression of macrophage TLR4 is reported in obesity [Olefsky,
2010]
 NF-κB signaling is more active under obese conditions or in insulin
resistance [Osborn, 2012]
 application of TNF-α has been shown to increase blood sugar levels
and insulin resistance in rodents [Könner, 2011]

42. TLR EXPRESSION
 TLR2 + 4 mRNA and protein expression in THP-1 human monocytic cell
line are increased under hyperglycemic conditions (3d) [Dasu, 2008]
 An increased TLR2 and TLR4 expression in mRNA and protein in T1D
and T2D patients’ monocytes, compared with controls, correlate with
theire HbA1C levels [Devaraj, 2008; Dasu, 2010]
 TLR2 + 4 surface expression (FACS) increased in monocytes from T1D
patients with microvascular complications compared with NC and
"healthy" T1D [Devaraj, 2011]
Does glucose alter TLR expression in macrophage
subpopulation?

43. HYPOTHESIS
 Hyperglycemia leads to an altered TLR expression in
macrophage subtypes and drive pro-inflammatory responses to
endogenous unwanted-self ligands

44. RT-PCR RESULTS
(8 INDIVIDUAL DONORS)

45. DONOR SPECIFIC
TLR2 EXPRESSION

46. DONOR SPECIFIC
TLR6 EXPRESSION

47. CHANGE OF EXPRESSION IN
HIGH GLUCOSE CONDITION
macrophage
population
number of
donors
change in TLR
expression
M1 5/ 8  TLR6
3/ 8  TLR2, 6
2/ 8
6/ 8
4/ 8
 TLR2, 6, 1,
 TLR4
 TLR2, 4
M2 6/ 8  TLR6
5/ 8  TLR2
3/ 8  TLR2, 6, 1
5/ 8  TLR4

48. TOLL-LIKE RECEPTORS
FSL-1
Pam3
CSK4

49. EDOGENOUS TLR LIGANDS
AND DIABETES
Toll-like receptor ligand
TLR2
necrotic cell products
apolipoprotein CIII
serum amyloid A
vesican
human cardiac myosin
snapin
TLR2/ CD36
oxidised phospholipids
saturated fatty acids
lipoprotein A
TLR2 or TLR4
hyaluron fragments
biglycan
oxLDL
HSP60, 70
endoplasmin
HMGB1
TLR4
surfactanc protein A
tenascin C
fibrinogen
fibronectin EDA
heparan sulphate
ß-defensin 2
amyloid ß peptid
HSP22, 72
minimally modified LDL
AGE-LDL
Shalhoub J. Inflamm. 2011
Yu J. Cell. Mol. Med. 2010
 damage-associated
molecular patterns
 endogenous activation of
TLR

50. TLR EXPRESSION IN FAT TISSUE OF
METABOLIC SYNDROME PATIENTS
qPCR results

51. CONCLUSIONS
Conclusions
higher levels of TLR2, 1,6, 4 and TLR8 expression in M1compared to M2
no increased expression of TLR4, but rather tendency to lower expression
levels of TLR4 in high glucose conditions
individual TLR expression pattern in high glucose conditions
Induction by glucose of TLR2, 1,6 in macrophage correlates with their
increase in fat tissue in metabolic syndrome patient
Next steps:
Analysis of individual TLR responses to stimulation by unwanted –self
ligands related to metabolic conditions

52. High glucose induces donor-specific
cytokine responses in human macrophages
Cytokines RT-PCR ELISA
TNFa Increased in 5 out of 6
donors after 6h in M1
Increased in 4 out of 6
donors after 6h in M1
CCL18 Decreased in 8 out of 8
donors on day 6 in M2
Decreased 8 out of 8 donors
on day 6 in M2
IL1beta Increased in 5 out of 8
donors on day 6 in M1
Increased 6 out of 8 donors
on days 1, 3 and 6 in M0,
M1 and M2
IL1ra Increased in 2 out of 8
donors on day 6 in M2
Increased in 6 out of 8
donors on day 6 in M0
Green = High glucose induces
Red = High glucose supresses

53. Sig Index for Estimate of IFNg:
High glucose vs Low Glucose
M1
M2
M0
Sig Index for Estimate of NS:
High Glucose vs Low Glucose
Sig Index for Estimate of IFNg:
High Glucose vs Low Glucose
Sig Index for Estimate of IL4:
High Glucose vs Low Glucose
M1 M0
M2
Searching for glucose regulated genes in
macrophages: Affymetrix Microarray analysis

54. EPIGENETICS IS A SENSOR TRANSLATED TO MACROPHAGES
DIFFERENTIATION AND ACTIVATION
IN METABOLIC CONDITIONS
metabolic, life style factors,
diabetic conditions
Epigenetic changes
Epigenetic memory
High glucose, ,
endogenous unwanted-self
products
Pathologic response
Pro-inflammatory
monocytes in the circulation
Changed M1/M2 balance in
the tissues
Modifications in
macrophages plasticity
Pathological
profile of cytokines
and receptors
Chronic
inflammation
Vascular
complications
Gaps of knowledge: exact
mechanisms, specific for
macrophage subtypes

55. Take home messages
 Metabolic conditions induce epigenetic changes leading to the
inflammatory responses in endothelial cell
 Histone code is major epigenetic mechanism of inflammatory
programming of macrophages in response to infection
 High glucose induces gene expression for TLRs, cytokines and other
regulatory genes not only in M1, but also and M2
 Next horizons in research: to identify epigenetic mechanism of long-
term metabolic memory in macrophages
 Translational perspective for clinic: drugs that block pathological
metabolic memory in immune cells and decrease risk of diabetic
complications.

56. Institute of Transfusion Medicine
and Immunology
Kondaiah Moganti
Bin Song
Alexandru Gudima
Shuiping Yin
Feng Li
Michael Balduff
Maria Verdiell-Lopes
Vladimir Ryabov,
Christna Schmuttermaier
Sandu Gudima
Amanda Mickley
Harald Klüter
Karen Bieback
Peter Buggert
Thank you
Financial support
DFG SFB „Immune tolerance“ Project B12
DFG GRK880 “Vascular Medicine”
Ministry of Science, Research
and Art of Baden-Württemberg
Margarete von Wrangell
Habilitationsprogramm 2003-2008
BMBF 2010-2012
BMBF BIOIN 2013-2015
FP7-ERANET
GRK1874 DIAMICOM
FP7-IMMODGEL
FP7 EULAMDIMA
Helmholtz-Zentrum, München
Elisabeth Kremmer
University of Heidelberg,
Medical Faculty
Mannheim
Oxford University
Siamon Gordon
DKFZ, Heidelberg
Berndt Arnold
University of Jena,
S. Mosig, K Rennert, Prof. H. Funke
Centre for Medical Research,
Lab for Microarray analysis
Xaolei Yu
M. Saile
C. Sticht
Department of Dermatology
Sergij Goerdt
Kai Schledzewski
Alexei Gratchev
I. Ovsij
N. Wang
S. Mamidi
L. Krusell
H. Brundiers
A. Dittmann
E. Usselmann
J. Dvoracek
H. Ahmed
A. D. Nurgazieva
A. Popova
All DIAMICOM members
UMC Groningen
Martin Harmsen
Jan-Luuk Hillebrands
EULAMDIMA consortium
University of Mexico,
Galileo Escobedo

57. OUR PROGRESS
Model system to examine
the high glucose effects on
human M0, M1 and M2
is established
Effect of high glucose on
M1 and M2 cytokines is identified
Kondaiah Moganti: glucose
affects cytokine expression
on transcriptional and post-
transcriptional levels
Affymetrix profiling: high glucose
affect transcription in M0, M1 and
M2, with strongest effect in M1
Effect of high glucose on TLR
expression profile in M1 and M2
is identified
Michael Balduff: glucose
affects TLR expression on
transcriptional level
ChIP-on-chip using
activation and repression
histone marks: effect of HG
on M1, M2, plasticity

58. Green color represents significantly upregulated
Red color represents significantly
Epigenetic regulatory factors

59. EPIGENETICS IN DIABETES
Conclusions
It is established that epigenetics plays a key role in
glucose memory and diabetic complications
development
It involves both DNA methylation and histone code
Identification of particular mechanisms requires large
clinical studies

60. EPIGENETICS IN DIABETES
Keating ST, El-Osta A. Epigenetic changes in diabetes. Clin Genet 2013
Gap of knowledge:
for how long
glucose memory
persists in vivo?

61. TLR MRNA EXPRESSION IN HU
MONOCYTE-DERIVED MACROPH
M1 M2
M1 M2

62. ENHANCERS ARE CRITICAL FOR
REGULATION OF EPIGENETIC IN MACROPHAGES
Jan Van den Bossche, Curr
Opin Lipidol, 2014
Gaps of knowledge:
What are the key
differences in
epigenetic changes in
subtypes of
macrophages

63. EPIGENETIC REGULATION OF INFLAMMATION IN
HYPOXIC CONDITIONS
HIF-1 binds to a specific site of the
promoter of histone 3 lysine 9
demethylases (JMJD1A, JMJD2A)
and activate these genes.
Brigati et al, 2010 Mediators of Inflammation

64. Jan Van den Bossche, Curr
Opin Lipidol, 2014
JMJD3: HISTONE DEMETHYLASE WITH THE MOST
COMPLEX ROLE IN MACROPHAGES POLARISATION

65. Satoh et al, Nature
Immunology, 2010
JMJD3: POTENTIAL M2-SPECFIC EPIGENETIC FACTOR?

66. HIGH RISK TLR PROFILE
BC 786-2

67. Enzymes providing histone modifications
Acetylation: HATs - CBP,p300, GCN5, ATF2, Tip 60…
Deacethylation: HDACs- class I and II
Methyaltion: Lysine: SET-domain HMTase and non-SET domain
HMTase (Dot1)
Arginine: PRMT family, CARM1
Demethylation: LSD1, JMJD3
Ubiquitination: ubiquitin conjugase Rad6/ligase Bre1 for H2B
De-Ubiquitination: SAGA-associated Ubp10

68. Macrophage activation dichotomy
(1985-1992, S.Gordon)
Th1
IFNγ
LPS
Th2IL-4,
Mannose
receptor
TNFα, IL-6,
IL-1
NO, H2O2
Classically activated macrophage
Alternatively activated macrophage
Initially alternative activation of
macrophages was defined on the
basis of IL-4- dependent induction
of macrophage mannose receptor
(MMR)