The role of IL-17A in postmenopausal inflammatory events,

such as in osteoporosis

1 Ildikó Molnár, MD, CSc, 2 Ilona Bohaty, MD, 1Éva Somogyiné-Vári1Immunoendocrinology and Osteoporosis Centre,

2Regional Centre of Hungarian National Blood Transfusion Service,

Debrecen, Hungary

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IL-17 cytokine

IL-17 (called as IL-17A) is characterized by:T-cell-derived cytokine secreted from Th17 cells, which are distributed from

other effector CD4+ T helper cells, such as Th1, Th2 and regulatory T (Treg) cells.

Sources of IL-17: Cells of innate and adaptive immunity (T and B cells, NK, NKT, γδ T cells, neutrophils, basophils, mast cells, monocyte-macrophages, dendritic cells). Epithelial, endothelial, vascular and stromal cells.

IL-17 initiated cytokine/chemokine productions: IL-6, TNFα, IL-1β, IL-8, CXC1, CXC2, CXC8, CCL2, MCP-1 Summit 2015, Houston

Arteriosclerosis(cardiovascular

diseases)

Increased IL-17 production-related diseases

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Local inflammation (cytokine interplay chemokine)

Autoimmunity

Allergy

Host defense mechanism(infection, transplantation)

Tissue repair

Bone loss(osteoporosis)

Immune dysfunction associated with estrogen deficiency

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Estrogen Estrogen deficiency

Th17 IL-17↑

spleen

neutrophilrecruitment

IL-6 secretion

T cell activation

peripheralmononuclear cells

osteoclastestrogen receptor

IL-1↑ TNFα ↑

osteoblast

TGFβ ↓ IFNγ ↑

IL-6 secretionBONE LOSS

RANK ligand ↑

Patients and methods

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Pre- (n=22, mean age 41 yr) and postmenopausal (n=72, mean age 65 yr) women were studied.Serum levels of IL-17A, IL-6, MCP-1 (macrophage-chemoattractant protein-1), sRANK (soluble receptor activator of NF-κB) ligand and OPG (osteoprotegerin) were measured by enzyme-linked immunosassay (ELISA).Serum levels of estradiol were measured by chemi-luminescence assay in fully automatized manner. Bone mineral density was detected by dual-energy X-ray absorptiometry (DXA) using Hologic Discoveryequipment.

We studied in pre- and postmenopausal women: The relationship among serum IL-17A levels, estrogen deficiency and postmenopausal period.

The relationships among serum IL-17A, sRANK ligand, OPG levels and bone mineral densities.

The relationship between vitamin D3 deficiency and serum IL-17A levels.

The relationship between serum IL-17A and IL-6 or MCP-1 levels.

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Synergism between bone loss and arteriosclerosis via IL-17 cytokine.

The relationship among serum IL-17A levels, estrogen deficiency

and postmenopausal period.

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Serum IL-17A levels were significantly higher in women with estrogen deficiency.

Figure 7

2,8

2,9

3

3,1

3,2

3,3

3,4

3,5Se

rum

IL-1

7A le

vels

(ng/

ml)

(mean ±SD) 3.01 3.43± 0.38 ± 0.56

P<0.001

patient groups

n=16 n=78

OR: 17.74 RR: 5.64CI95%: 3.52-79.62

estradiol 73.54±0.86 pmol/l

estradiol 304.66±234.33 pmol/l

estradiol levelsbelow 83 pmol/l

estradiol levelsin the normal range

0

1

2

3

4

Seru

m IL

-17A

leve

ls (n

g/m

l)

age-related groups

50-59 60-69 70-79 80-89n=20 n=34 n=10 n=8

ranges of age (years)

P<0.003

P<0.04

mean±SD 3.59 3.31 3.63 3.97±0.65 ±0.4 ±0.46 ±0.77

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Figure 8

Inreased serum IL-17A levels showed an age-related dependency in postmenopause.

Women with menopausal age > 10 years

Seru

m IL

-17A

lev

els

(ng/

ml) Y= 3.112 + (X*0.018)

p<0.049

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Correlation between serum IL-17A levels and the post-menopausal period of women aged 60 to 89 years.

Figure 9

3,1

3,15

3,2

3,25

3,3

3,35

3,4

3,45

3,5

3,55

3,6Se

rum

IL-1

7A le

vels

(ng/

ml)

(mean ±SD) 3.57 3.29± 0.59 ± 0.39

P<0.026

patient groups

n=54 n=18

Yes hysterectomy

No hysterectomy

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Figure 10

Inreased serum IL-17A levels showed a dependency on the history of hysterectomy in postmenopause.

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Inreased serum IL-17A levels showed a postmenopausal age-related dependency on the history of hysterectomy.

Figure 110

1

2

3

4

Seru

m IL

-17A

leve

ls (n

g/m

l)

age-related groups0-10 yr 11-20 yr 21-30 yr 31-40 yrn=17 n=3 n=20 n=5 n=14 n=7 n=3 n=3

menopausal age

mean±SD 3.54 3.49 3.46 3.25 3.62 3.13 4.3 3.55 ±0.68 ±0.6 ±0.47 ±0.26 ±0.46 ±0.28 ±1.09 ±0.56

P<0.008

Yes hysterectomy

No hysterectomy

Relationship between serum IL-17A levels and bone mineral densities (BMDs).

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Postmenopausal Premenopausal womenosteopenia osteoporosis normal in lumbar region

(-1>T-score>-2.5) (T-score<-2.5) (T-score >-1)n=31 n=41 n=22

2

2,5

3

3,5

4

4,5

5

Seru

m IL

-17A

leve

ls (n

g/m

l)

patient groups

(mean ±SD) 3.31 ± 0.08 3.65 ± 0.61 2.88 ± 0.08 ng/mlP<0.027

P<0.013 P<0.0001

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Figure 13

Increased serum IL-17A levels were associated with relevant bone loss in postmenopause compared to those in premenopause.

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Serum IL-17A levels were significantly higher in post-menopausal osteoporotic women in the lumbar and femoral total regions.

Figure 14

Fem

oral

nec

kbo

ne m

iner

al d

ensi

ty (g

/cm

2 )

Serum IL-17A levels (ng/ml)

Y=0.914 - (x*0.074)p<0.001

Fem

oral

bone

min

eral

den

sity

(g/c

m2 )

Serum IL-17A levels (ng/ml)

Y=1.122 - (x*0.086)p<0.003

Lum

bar

bone

min

eral

den

sity

(g/

cm2 )

Serum IL-17A levels (ng/ml)

Y=0.972 - (x*0.059)p<0.022

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Serum IL-17A levelscorrelated inversely with bone mineral densities in postmenopause.

Lu

mb

ar r

egio

nF

emor

al r

egio

ns

Relationship between serum IL-17A and sRANK ligand or

OPG levels.

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Premenopausal women, n=18.

Postmenopausal osteopenic women, n=31.osteoporotic women, n=41.

A

C

B

Seru

m IL

-17

leve

ls (n

g/m

l)

mean±SD 2.89±0.07 3.31±0.43 3.65±0.60

P<0.007

P<0.0001

P<0.0001

patient groups Seru

m s

RAN

K lig

and

leve

ls

(ng/

ml)

mean±SD 2.61±0.61 2.49±0.61 2.88±0.84

P<0.027

patient groups

Seru

m O

PG le

vels

(ng/

ml)

mean±SD 1.43±0.07 1.39±0.07 1.43±0.07

P<0.038

patient groups

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Figure 17

Increased serum IL-17A and sRANK ligand levels were detected in osteoporotic women.

IL-17A sRANK ligand

OPG

Serum sRANK ligand levels (ng/ml)

Seru

m IL

-17

leve

ls (n

g/m

l)

Y= 2.924 +(X*0.2)P<0.009

Ratio of serum sRANK ligand and OPG levels

Seru

m IL

-17

leve

ls (n

g/m

l)

Y= 2.971 + (X*0.26)P<0.023

Serum OPG levels (ng/ml)

Seru

m IL

-17

leve

ls (n

g/m

l)

Y= 1.294 + (X*1.55)NS

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Figure 18

Serum IL-17A levels correlated positively with sRANK ligand and did not with OPG serum levels, but with the ratio of sRANK ligand and OPG serum levels.

sRANK ligand OPG

Ratio of sRANK ligand and OPG

Vitamin D3 deficiency and serum IL-17A levels.

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ParametersVitamin D3deficiency

(n=18)<50 nmol/l

Vitamin D3insufficiency

(n=25)51-75 nmol/l

Vitamin D3sufficiency

(n=11)>75 nmol/l

P-values

Age(years)

59±17 61±13 61±12

BMI*(kg/m2)

29±5 29±6 29±6

Vitamin D3(nmol/l)

16,84±6,06 35,08±5,79 68,95±44

IL-17A(ng/ml)

13,35±2,9 o 12,07±2,61 11±1,9 o 0.033

MCP-1**(ng/ml)

17,35±2,9 16,62±2,07 16,18±1,23

IL-6(ng/ml)

26,11±12,66 26,25±12,23 22,68±13,52

Serum IL-17A levels were significantly higher in post-menopausal women with vitamin D3 deficiency.

*Body mass index (BMI) **Monocyte chemoattractant protein-1 (MCP-1)Summit 2015, Houston

Table 1

D3-

vita

min

seru

m le

vels

(nm

ol/l

)

IL-17A serum levels (ng/ml)

Y= 64,45-(X*2,34)P<0.026 r=0.3033

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Figure 20

Serum IL-17A levels correlated inversely with vitamin D3 serum levels in postmenopause.

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Relationship between serum IL-17A and IL-6 or MCP-1 levels.

IL-1

7A se

rum

leve

ls (n

g/m

l)

MCP-1 serum levels (ng/ml)

Y= (X*0,81)-1.43P<0.0001 r=0.6745

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Figure 22

Serum IL-17A levels correlated positively with serum MCP-1 levels in postmenopause.

IL-1

7A se

rum

leve

ls (n

g/m

l)

IL-6 serum levels (ng/ml)

Y= 8,47+(X*1,46)P<0.0001 r=0.6693

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Figure 23

Serum IL-17A levels correlated positively with serum IL-6 levels in postmenopause.

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Synergism between bone loss and arteriosclerosis via IL-17 cytokine.

bone loss

From Butcher M and Galkina EV , Thromb Haemost, 2011, 106(5): 787-795.

Figure 24

arteriosclerosis

Conclusions

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The high prevalence of increased serum IL-17A levels was connected to postmenopausal estrogen deficiency and showed a postmenopausal period-related dependency. Postmenopausal osteoporosis was associated with increased serum IL-17A and sRANK ligand levels, but only weakly increased serum OPG levels. Vitamin D3 deficiency was associated with higher serum IL-17A levels in postmenopause. The strong correlation between serum IL-17A and IL-6 or MCP-1 highlighted the relationship between osteoporosis and arteriosclerosis, as well as cardio- vascular diseases.

Thank youfor your attention!

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Participated in this work:Regional Centre of Hungarian National Blood Transfusion Service, Debrecen, Hungary

Ilona Bohaty MD

EndoMed, Immunoendocrinology and Osteoporosis Centre

Éva Somogyiné-Vári