DEPARTMENT OF IMMUNOLOGY Chair of Histology and Immunology Medical University of Gda ń sk DEPARTMENT OF IMMUNOLOGY Chair of Histology and Immunology Medical.

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