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DEPARTMENT OF IMMUNOLOGYDEPARTMENT OF IMMUNOLOGYChair of Histology and Chair of Histology and
ImmunologyImmunologyMedical University of GdańskMedical University of Gdańsk
Research groupResearch group
Head: Jolanta Myśliwska, MD, PhDHead: Jolanta Myśliwska, MD, PhDProfessor of immunologyProfessor of immunology
Joanna Więckiewicz, M. Sc.Joanna Więckiewicz, M. Sc.
Dominik Rachoń, MDDominik Rachoń, MD
Łukasz Hak, M. Sc.Łukasz Hak, M. Sc.
Monika Mlotkowska, M.Sc.Monika Mlotkowska, M.Sc.
Katarzyna Zorena, PhDKatarzyna Zorena, PhD
Hanna Suchanek, MD, PhDHanna Suchanek, MD, PhD
RESEARCH PROBLEMSRESEARCH PROBLEMS
• AGE-RELATED CHANGES OFAGE-RELATED CHANGES OF I IMMUNE MMUNE RESPONSERESPONSE
• HORMONAL MODULATION OF HORMONAL MODULATION OF IMMUNE RESPONSEIMMUNE RESPONSE
• IMMUNOLOGICAL ASPECTS OF IMMUNOLOGICAL ASPECTS OF CARDIOVASCULAR DISEASECARDIOVASCULAR DISEASE
PREVIOUS AND CURRENT
AGE-RELATED CHANGES OFAGE-RELATED CHANGES OF IIMMUNE RESPONSEMMUNE RESPONSE
ROLE OF NATURAL KILLER CELLS IN AGEING ROLE OF NATURAL KILLER CELLS IN AGEING
• We found that successful healthy ageing is We found that successful healthy ageing is associated with a shift towards high cytotoxic associated with a shift towards high cytotoxic activity of Natural Killer cells. The high values of NK-activity of Natural Killer cells. The high values of NK-associated parameters may be preserved until very associated parameters may be preserved until very old age.old age.
• Healthy elderly people have an increased number of Healthy elderly people have an increased number of the regulatory NK cells with intra-cellular IFNthe regulatory NK cells with intra-cellular IFN . .
• This status correlates with anti-viral This status correlates with anti-viral immune immune response in the elderly what we observed on the response in the elderly what we observed on the natural model of vaccination against influenza. natural model of vaccination against influenza.
• MyMyśśliwskaliwska J, et al. (1997) J, et al. (1997) Gerontology, 43, 158-167.Gerontology, 43, 158-167.
• MyMyśśliwskaliwska J, et al. (2000) J, et al. (2000) Acta Biochem. Pol., 47, 1-11.Acta Biochem. Pol., 47, 1-11.
• KmiećKmieć Z, et al. (2001) Z, et al. (2001) Gerontology, 47, 282-288.Gerontology, 47, 282-288.
• Brydak LB, Brydak LB, et al. (2003) et al. (2003) J. Clin. Immunol., 23:214-222.J. Clin. Immunol., 23:214-222.
• Myśliwska J, et al. (2003) Clin Exp. Immunol., acceptedMyśliwska J, et al. (2003) Clin Exp. Immunol., accepted
• We found that ageing process is accompanied by an We found that ageing process is accompanied by an enhanced expression of pro-inflammatory cytokines.enhanced expression of pro-inflammatory cytokines.
• Low immune response to vaccination in the elderly, who Low immune response to vaccination in the elderly, who stayed apparently healthy, is associated with an stayed apparently healthy, is associated with an increased expression of genes of such inflammatory increased expression of genes of such inflammatory cytokines as; Interleukin 1, TNFcytokines as; Interleukin 1, TNF, Interleukin 6, , Interleukin 6, Interleukin 12.Interleukin 12.
• Enhanced expression of pro-inflammatory cytokines was Enhanced expression of pro-inflammatory cytokines was associated with carrier status of latent CMV infection in associated with carrier status of latent CMV infection in the elderly low responders to influenza vaccination.the elderly low responders to influenza vaccination.
PRO-INFLAMMATORY PRO-INFLAMMATORY STATE IN THE ELDERLYSTATE IN THE ELDERLY
Myśliwska JMyśliwska J, et al. , et al. (1999) Mech. Ageing Dev. ,107: 1-14. (1999) Mech. Ageing Dev. ,107: 1-14. Myśliwska JMyśliwska J, et al. , et al. (2000) Eur. Cytokine Net. ,11:397-406. (2000) Eur. Cytokine Net. ,11:397-406.
TrzonkowskiTrzonkowski P P, , et al. (2002)et al. (2002) Mech. Ageing Dev. ,123, 1283- Mech. Ageing Dev. ,123, 1283-1293.1293.
Trzonkowski PTrzonkowski P,, et al. et al. (2003) Vaccine , 21:3826-3836.(2003) Vaccine , 21:3826-3836.
HORMONAL MODULATION OF IMMUNE HORMONAL MODULATION OF IMMUNE RESPONSERESPONSE
ESTROGENS DOWN-REGULATE PRO-ESTROGENS DOWN-REGULATE PRO-INFLAMMATORY AND UP-REGULATE ANTI-INFLAMMATORY AND UP-REGULATE ANTI-
INFLAMMATORY CYTOKINESINFLAMMATORY CYTOKINES
-actin
TNF
1 2 3 4 5 6 (-) (+)
883bp
501bp
800bp
pUC
420bp404bp
692bp
Non-stimulated ex vivo monocytes of healthy Non-stimulated ex vivo monocytes of healthy postmenopausal women express postmenopausal women express the the TNF geneTNF gene
• This may be the effect of estrogen deprivationThis may be the effect of estrogen deprivation
-actin
IL6692bp
883bp
501bp
800bp
pUC
628bp
Non-stimulated ex vivo monocytes of healthy Non-stimulated ex vivo monocytes of healthy postmenopausal women express postmenopausal women express the the IL-6 geneIL-6 gene
1 2 3 4 5 6 (-) (+)
• This may be the effect of an exaggerated TNF expression This may be the effect of an exaggerated TNF expression which in turn stimulates IL-6 gene expressionwhich in turn stimulates IL-6 gene expression
Rachoń D, Myśliwska J, et al. (2002) J Endocrinol.,172, 387-395.Rachoń D, Myśliwska J, et al. (2002) J Endocrinol.,172, 387-395.
Non-stimulated PBMC of postmenopausal women Non-stimulated PBMC of postmenopausal women produce more bioactive TNF and IL-6 produce more bioactive TNF and IL-6 comparedcompared to young to young
controlscontrols
In vitro effects of 17In vitro effects of 17-estradiol on the spontaneous -estradiol on the spontaneous TNF and IL-6 production by PBMC of postmenopausal TNF and IL-6 production by PBMC of postmenopausal
womenwomen
In vivo In vivo effects of 17effects of 17-estradiol treatment on the -estradiol treatment on the spontaneous TNF and IL-6 production by PBMC of spontaneous TNF and IL-6 production by PBMC of
postmenopausal womenpostmenopausal women
Non-stimulated PBMC of postmenopausal women Non-stimulated PBMC of postmenopausal women produce produce less IL-10 less IL-10 compared to young controlscompared to young controls
In vitro effects of 17In vitro effects of 17-estradiol on the IL-10 gene -estradiol on the IL-10 gene expression in the PBMC of postmenopausal womenexpression in the PBMC of postmenopausal women
IMMUNOLOGICAL ASPECTS OF IMMUNOLOGICAL ASPECTS OF CARDIOVASCULAR DISEASECARDIOVASCULAR DISEASE
CURRENT RESEARCHCURRENT RESEARCH
Polymorphism of the human Interleukin 6 (IL6Polymorphism of the human Interleukin 6 (IL6)) gene at gene at position –174position –174G/CG/C in coronary heart disease in coronary heart disease (CHD) (CHD)
patientspatients
YEARS FROM ONSET OF THE DISEASE TILL AN OPERATION
year
s
-202468
101214161820
CC GG GC
CONCENTRATION OF IL6 IN VITRO CULTURES
IL6
pg/m
l
0
1000
2000
3000
4000
CG and GG CC
Polymorphism of the human atrial natiuretic peptidePolymorphism of the human atrial natiuretic peptide (ANP)(ANP) gene in coronary heart disease gene in coronary heart disease (CHD) (CHD) patientspatients
• Polymorphism is analysed in CHD patients with Polymorphism is analysed in CHD patients with different cardiac arrhythmias occurring in (or different cardiac arrhythmias occurring in (or without) association with cardiac surgery.without) association with cardiac surgery.
• The A2A2 allele combination carriers constitute The A2A2 allele combination carriers constitute 55% of the whole group of CHD patients and 55% of the whole group of CHD patients and seem to be protective from atrial fibrillation.seem to be protective from atrial fibrillation.
242 bp 234 bp
213 bp
117 bp96 bp
21 bp
190 bp
147 bp
111 bp
67 bp
INFECTIONS WITH CMV AND CHLAMYDIA INFECTIONS WITH CMV AND CHLAMYDIA PNEUMONIAE IN CHD PATIENSPNEUMONIAE IN CHD PATIENS
positive control sample of mononuclear cells
185 bp
Analysis of function
of CD14+CD16+ monocytes
• detection of monocyte populations by cytophotometric technique
• tests revealing presence of intra-cytoplasmic cytokines in CD14+CD16+ monocytes
• in vitro cultures analysing an effect of Lovastatin on spontaneous and LPS-induced synthesis of intra-cytpolasmic IL6
ROLE OF ROLE OF CD14+CD16+ BLOOD MONOCYTES CD14+CD16+ BLOOD MONOCYTES IN CORONARY HEART DISEASEIN CORONARY HEART DISEASE
CD14+CD16+ CD14+CD16+ BLOODBLOOD monocytes in Cmonocytes in CORONARY ORONARY HHEART DISEASEEART DISEASE and controls and controls
Mediana 25%-75% Min.-Maks. patients
controls
2468
1012141618202224262830
% o
f C
D1
4+C
D16
+ m
onoc
ytes
_____*____
Spontaneous IL-6production
Mediana Min.-Maks. 14+16+
14+16-
0
10
20
30
40
50
60
70
80
_____*_____
LPS-induced IL-6 production
Mediana Min.-Maks. 14+16+
14+16-
0
10
20
30
40
50
60
70
80
90
____*____
CD14+ CD16+
Mediana Min.-Maks.
Control Lova LPS LPS + Lova0
1020
30
4050
60
7080
90
____*____
___*____
CD14+ CD16-
Mediana Min.-Maks.
Control Lova LPS LPS + Lova0
10
20
30
40
50
60
70
80
90
Human cytokine array systemHuman cytokine array system – expression – expression profile profile of of multiple cytokines multiple cytokines
• In serum of the patients with coronary heart disease, with CCS 3 and 4 grade of the angina pectoris, we found an expression of 18 new cytokines
positive control
positive control