Medical

Communication

Biosci. Biotech. Res. Comm. 10(3): 587-591 (2017)

Comparison of interleukin 17 and 22 in saliva of oral

lichen planus patients with healthy people

Shiva Shirazian

, Farzaneh Aghahosseini

, Eisa Salehi

, Mehdi Vatanpour

Seyede Niloofar Banijamali

and Sara Pourshahidi

Assistant Professor, Department of Oral Medicine, School of Dentistry, Tehran University of Medical

Sciences, Tehran, Iran

Full Professor, Department of Oral Medicine, School of Dentistry, Tehran University of Medical Sciences,

Tehran, Iran

Assistant Professor, Department of Immunology and Biology, School of Medicine, Tehran University of

Medical Sciences, Tehran, Iran

Assistant Professor, Department of Endodontics, Dental Branch, Islamic Azad University, Tehran, Iran

Resident of Pediatric Dentistry, Dental Branch, Islamic Azad University, Tehran, Iran

Associated Professor, Department of Oral Medicine, School of Dentistry, Tehran University of Medical

Sciences, Tehran, Iran

ABSTRACT

Lichen Planus (LP) is an in ammatory chronic mucocutaneous disease. Since the etiology of OLP is unknown, efforts to under-

stand the etiology and pathogenecity may lead to improve treatment modalities. Studies have shown the different levels of

increase or decrease of various cytokines in saliva, serum or tissue culture of patients with OLP.As saliva originates from blood

and its collection is an easy, non-invasive method, so it can be a diagnostic  uid that has logistical advantages when compared

with serum. The purpose of this study was to compare the salivary level of IL17 and IL22 in patients with and without OLP. Saliva

of 52 patients with and without OLP was collected. The saliva level of IL17 and IL 22 was measured by using ELISA. Data were

analyzed using independent T test, Mann-Whitney and Pearson Correlation test.The saliva concentration of IL 22 was signi -

cantly higher in control group than OLP but the difference was not signi cant between reticular and atrophic-erosive type (P >

0.05).The difference between mean rank of IL17 for OLP and healthy cases and between reticular and erosive-atrophic form was

not signi cant (P > 0.05). There was no correlation between the level of IL17 and IL 22 (P > 0.05). Our  ndings suggest that IL 22

may have an effect in the pathogenesis of OLP and its protective effect may be decreased in OLP patients.

KEY WORDS: INTERLEUKIN 17, INTERLEUKIN 22, ORAL LICHEN PLANUS, SALIVA

587

ARTICLE INFORMATION:

*Corresponding Author: sarapourshahidi20@hotmail.com

Received 21

July, 2017

Accepted after revision 29

Sep, 2017

BBRC Print ISSN: 0974-6455

Online ISSN: 2321-4007 CODEN: USA BBRCBA

Thomson Reuters ISI ESC and Crossref Indexed Journal

NAAS Journal Score 2017: 4.31 Cosmos IF: 4.006

reserved.

Online Contents Available at:

http//www.bbrc.in/

DOI: 10.21786/bbrc/10.3/37

588 COMPARISON OF INTERLEUKIN 17 AND 22 IN SALIVA OF ORAL LICHEN PLANUS PATIENTS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

Shiva Shirazian etal.

INTRODUCTION

Lichen Planus (LP) is an in ammatory chronic mucocu-

taneous disease that affects 1-2% of the general popula-

tion, and is the non-infected mucosal disease causing the

most referral to dental clinics (Pakfetrat et al. 2009). The

oral cavity is the most common site of the disease. Oral

Lichen Planus (OLP) usually affects middle aged women

(30-60 years), children are rarely affected.World Health

Organization (WHO) has considered OLP as a pre-cancer-

ous disorder. Although OLP is considered an immunologi-

cally mediated disease, its etiology is not known, yet. The

involvement of both speci c and non-speci c antigens

are considered to have roles in pathogenesis of OLP (van

der Meij et al. 2003 Aghahosseini et al. 2006; Pourshahidi

et al. 2011; Agha-Hosseini et al. 2015).

Alteration of the immune condition is not well

known in OLP to date, but it is clear that cytokines

exhibit immunoregulatory actions through a complex

cytokine network consisting of paracrine and autocrine

systems. Unbalanced cytokine actions are considered to

play a role in the pathogenesis of autoimmune diseases.

Numerous cytokines are secreted from oral mucosa

or blood as locally or systemically that they have an

important role in initiation and progression (courses) of

OLP (Zhang et al. 2008).

Increased production of cytokine by keratinocytes

that may be the result of stimulation by external stimu-

lants is the initial event. Then in ltration of monocytes

followed by lymphocytes could be the next step. Studies

have shown the different levels of increase or decrease

of various cytokines in saliva or serum or tissue culture

of patients with OLP (Yamamoto et al. 1990; Rhodus

et al. 2005; Zhang et al. 2008).

Interleukin (IL) 17 A, also named IL17, is the most

important IL from the IL17 family which is composed

of 6 members (IL17A-IL17F). IL 22 is a member of IL10

family and it has a role in immune reaction against

bacterial pathogens, especially in epithelium. IL17 and

22 are produced by T helper 17 and contribute in both

innate and acquired immune responses against extra

cellular pathogens (Lambiase et al. 2009). The role of

Th17 and its cytokines (IL17 and 22) have been estab-

lished in in amed tissues in various autoimmune dis-

eases such as psoriasis, multiple sclerosis, lupus ery-

thematosus, rheumatoid arthritis, in ammatory bowel

disease (IBD) and pemphigus (Miossec et al. 2009).Saliva

originates from blood. Saliva collection is an easy, non-

invasive method, and it has been shown that various

tumor markers are present in saliva. Therefore it can be

a diagnostic  uid that has logistical advantages when

compared with serum (Agha-Hosseini et al. 2015). The

purpose of this study was to compare the level of IL17

and IL22 in saliva in patients with and without OLP.

MATERIAL AND METHODS

Fifty two samples of referred patients to the Department

of Oral Medicine, Faculty of Dentistry, Tehran Univer-

sity of Medical Sciences included in this cross-sectional

study (26 in each group).this study was approved by

ethical committee and also all subjects provided writ-

ten informed consent to participate in the study. OLP

was diagnosed clinically and con rmed histopathologi-

cally according to the modi ed WHO criteria in 2003 as

below:Clinical criteria: presence of symmetric or bilat-

eral reticular or papular lesions with or without erosive–

atrophic components. Histological criteria: presence of

well-de ned band-like zones of in ammatory in ltra-

tion con ned to the super cial part of connective tis-

sue, consisting mainly of mature lymphocytes; signs of

“liquefaction degeneration” in basal cell layer, absence

of epithelial dysplasia. Exclusion criteria: application of

any local treatment for OLP lesions during the previous

month; taking any drugs during the previous 3 months;

history of allergy to foods or environmental factors,

smoking; the existence of any oral lesions except OLP,

systemic diseases (cardiovascular disease, kidney, hyper-

tension), pregnancy. We also excluded patients whose

lesion was nearby amalgam restorations. The unstimu-

lated whole saliva (UWS) was collected between 10:00

a.m. and 12:00 p.m., and at least 90 minutes after the

last intake of drink or food. All subjects were requested

to swallow  rst, tilt the head forward and then expecto-

rate at least 5cc UWS into a sterile centrifugal tube with-

out swallowing. The samples were centrifuged (2000g

for 10 min), and the supernatants stored at -20 oC until

analysis. The saliva level of IL was measured by using

Enzyme-Linked Immunosorbent Assay (ELISA) kits,

Human platiniumIL17A (BMS2017) and Human pluto-

nium IL22 (88-7522-22) (bioscience, USA).

STATISTICAL ANALYSIS

Independent T test was used to compare the mean of

IL22 between two groups; and Mann-Whitney for IL17.

Pearson Correlation test was used for assessing the cor-

relation between IL 17 and IL22 in each group. A P value

less than 0.05 was considered statistically signi cant

and primary power of analysis was set at 80%.

RESULTS AND DISCUSSION

26 patients with OLP (22 females and 4 males) and 26

sex and aged matched healthy individuals were enrolled

in this study. The mean age was 50.81±12.61 years, and

49.42±5.33 years in OLP and control groups respec-

tively. 23% (6 patients) of OLP was reticular form, and

77% (20 patients) was atrophic-erosive form. The mean

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS COMPARISON OF INTERLEUKIN 17 AND 22 IN SALIVA OF ORAL LICHEN PLANUS PATIENTS 589

Shiva Shirazian etal.

of IL22 was 184.48±4.58 pg/ml in OLP and 209.01±4.58

pg/ml in control group, there was signi cant differ-

ence between two groups (p=0.003). The mean of IL 22

was 176.5±9.40 pg/ml in reticular and 186.88±5.26 in

atrophic-erosive type but the difference was not signi -

cant (p=0.35).

The mean of IL 17 was 5.24±2.709 pg/ml in OLP and

0.526±0.164 pg/ml in control. The mean rank of IL17

was 37.33 and 33.59 for OLP and healthy cases respec-

tively. The difference was not signi cant (p=0.436). The

difference between the mean of IL17 in reticular and

erosive-atrophic form was not signi cant (p=0.25).

There was no correlation between the level of IL17

and IL22 (p=0.35, r=-0.19)in healthy patients. Although

correlation analysis revealed no signi cant correlation

between IL17 and IL 22 in OLP group but it seems to

be a reverse correlation between them according to the

statistical  ndings (p=0.054, r=- 0.049).

As the etiology of OLP is unknown and because it is

one of the most common lesion referred to dental clin-

ics, studies on the pathogenesis of the OLP are of high

importance. In this study we evaluated the relevance

of IL17 and IL22 with OLP. The saliva concentration

of Interleukin 22 was signi cantly higher in control

group than OLP but the difference was not signi cant

between reticular and atrophic- erosive type (P>0.05).

We couldn’t  nd any studies on OLP patients’ salivary

levels of IL22. But Chen et al and Shen et al studied on

IL22 in the tissue samples from OLP patients and found

it more than their control group (Chen et al. 2011; Shen

et al. 2012). More expression of IL22 in the tissue may

be due to the location of its receptors which is on the

epithelial origin cells (Ouyang et al. 2008). On the other

hand there are some researches on the role of IL22 and

psoriasis (Zheng et al. 2007; Lowes et al. 2008). Zaba et

al. (2007) in an animal study reported the higher expres-

sion of IL17 and IL22 in skin lesions of mice with pso-

riasis; but Zheng and Ma reported that de ciency and

limitation of IL22 can exaggerate skin lesion of psoriasis

in mice (Zaba et al. 2007; Zheng et al. 2007; Ma et al.

2008). This controversy reveals that more studies should

be done on the subject. The salivary level of IL17 in

OLP patients of our study was higher than the control

group but the difference was not statistically signi cant

(p=0.436). On the basis of our knowledge there is no

study on salivary level of IL17 in OLP.

Zhang et al. (2007) compared the level of IL1, IL8

and TNF in serum and saliva in 30 OLP patients and

30 healthy controls. These IL were higher signi cantly

in saliva and serum in patients than control, and the

level of IL8 was higher in saliva in erosive OLP than

other types (Zhang et al. 2008). Xie et al. (2012) assessed

Th1 and Th17 in tissue (by double immuno uorescence

staining) and in serum (by  ow cytometry) of 40 patients

with OLP (22 cases with reticular type and 18 cases with

erosive- atrophic type). Their results showed that the

Th17 and Th1 cells were similar in tissue, but both were

increased in serum. Th17 was higher in erosive- atrophic

OLP than reticular. The level of IL 17 was signi cantly

higher in serum of OLP patients than controls and in

erosive-atrophic OLP than reticular.

The role of IL17 has been assessed in other auto-

immune diseases in previous studies (Moseley et al.

2003; Nistala et al. 2009; Caproni et al. 2009). Caporni

and et al. (2009) evaluated the level of IL17 in serum

of psoriasis patients before and after treatment, and

compared with healthy controls. The level of IL17 was

higher in patients than control and was reduced to a low

level after treatment [21]. Ziolkowska et al measured the

level of IL17 in serum and synovial  uid of joints in 15

patients with rheumatoid arthritis, and compared with

healthy controls. Their results showed higher level of

IL17 in patients than control (Ziolkowska et al. 2000).

Zhao et al studied the role of Th17 in asthma sensitiv-

ity. Percentages of Th17 cells and the level of several

cytokines including IL4, IL22, IL25, IL17 and INF were

measured. Th17 cells and related cytokines had the most

increasing; additionally, the amount of IL17 and IL22

and Th17 had a positive correlation with the severity of

disease (Zhao et al. 2010).

A hypersensitivity reaction is considered in patho-

genesis of OLP when cytokines are released by activated

T cell, resulting in accumulation of in ammatory cells,

and the destruction of keratinocytes due to the cytotox-

icity of cells (Sezer et al. 2007).

The immune mechanism with speci c and non speci c

antigen are proposed in the pathogenesis of OLP (Suger-

man et al. 2005). IL17 may up regulate various cytokines

like IL1, IL8 and TNF (Hwang et al. 2004). In the study

of Zhang et al (2008) the serum and salivary level of these

cytokines was elevated in OLP patients such as other stud-

ies (Sun et al. 2005; Rhodus et al. 2006; Janardhanam et

al. 2007; Rhodus et al. 2007). Therefore, it may be pos-

tulated that IL17 probably has a role in pathogenesis of

OLP by increasing these in ammatory cytokines. In non

speci c antigen, another mechanism may be degranula-

tion of mast cells and activation of MMP (Sugerman et al.

2005). Jovanovic et al (2001) reported that IL17 acts as an

up regulator of local in ammatory factors, causing injury

of extracellular matrix through MMP (in vitro). Therefore

it may be an in uence in pathogenesis of OLP. In present

study the level of IL17 was ten times higher than the level

of healthy control (5.24 pg/ml in patients and 0.516 pg/

ml in healthy subjects), although the difference was not

signi cant statistically.

It is clear that IL22 has protection effects while IL 17

has not (Zenewicz et al. 2007; Sonnenberg et al. 2010).

Consequently, with the lower level of IL22 and higher

590 COMPARISON OF INTERLEUKIN 17 AND 22 IN SALIVA OF ORAL LICHEN PLANUS PATIENTS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

Shiva Shirazian etal.

level of other in ammatory cytokines, epithelial cells

are disposed to destroy, and OLP lesions can appear. This

hypothesis of the reverse correlation between IL17 and

IL22 in patients with OLP is strengthened, but it needs

more research.

CONCLUSION

Our  ndings suggest that IL 22 may have an effect in

the pathogenesis of OLP and its protective effect may be

decreased in OLP patients.

ACKNOWLEDGEMENT

This research has been supported by Tehran University

of Medical Sciences & health Services, Faculty of Den-

tistry grant91-01-69-16665.

REFERENCES

Agha-Hosseini F, Mirzaii-Dizgah I, Mahboobi N, Shirazian S,

Harirchi I. (2015) Serum and Saliva MMP-3 in Patients with

OLP and Oral SCC. J Contemp Dent Pract. 1; 16(2):107-11.

Aghahosseini F, rbabi-Kalati F, Fashtami LA, Fateh M, Djavid

GE. (2006) Treatment of oral lichen planus with photodynamic

therapy mediated methylene blue: a case report. Med Oral

Patol Oral Cir Bucal. (2):E126-E129.

Caproni M, Antiga E, Melani L, Volpi W, Del BE, Fabbri P.

(2009) Serum levels of IL-17 and IL-22 are reduced by etaner-

cept, but not by acitretin, in patients with psoriasis: a rand-

omized-controlled trial. J Clin Immunol. 29(2):210-214.

Chen J, Feng J, Chen X,et al. (2013) Immunoexpression of Inter-

leukin-22 and Interleukin-23 in Oral and Cutaneous Lichen

Planus Lesions: A Preliminary Study. Mediators In amm

801974. doi: 10.1155/2013/801974. Hwang SY, Kim JY, Kim

KW, Park MK, Moon Y, Kim WU,Kim YH. (2004) IL-17 induces

production of IL-6 and IL-8 in rheumatoid arthritis synovial

 broblasts via NF-kappaB-and PI3-kinase/Akt-dependent

pathways. Arthritis Res Ther . 6(2):120-128.

Janardhanam SB, Prakasam S, Swaminathan VT, Kodumudi

KN, Zunt SL, Srinivasan M. (2012) Differential expression of

TLR-2 and TLR-4 in the epithelial cells in oral lichen planus.

Arch Oral Biol. 57(5):495-502.

Jovanovic DV, Di Battista JA, Martel-Pelletier J, Reboul P, He

YULA, Jolicoeur FC, Pelletier JP. (2001) Modulation of TIMP-1

synthesis by antiin ammatory cytokines and prostaglandin E2

in interleukin 17 stimulated human monocytes/macrophages.J

Rheumatol. 28(4):712-718.

Lambiase A, Micera A, Mantelli F, et al. (2009) T-helper 17

lymphocytes in ocular cicatricial pemphigoid. Mol Vis.

15:1449-1455.

Lowes MA, Kikuchi T, Fuentes-Duculan J, et al. (2008) Pso-

riasis vulgaris lesions contain discrete populations of Th1 and

Th17 T cells. J Inv Dermatol. 128(5):1207-1211.

Ma HL, Liang S, Li J, et al. (2008) IL-22 is required for Th17

cell-mediated pathology in a mouse model of psoriasis-like

skin in ammation. J Clin Invest. 118(2):597-607.

Miossec P, Korn T, Kuchroo VK. (2009) Interleukin-17 and type

17 helper T cells. N Engl J Med . 361(9):888-898.

Moseley TA, Haudenschild DR, Rose L, Reddi AH. (2003) Inter-

leukin-17 family and IL-17 receptors. Cytokine Growth Factor

Rev. 14(2):155-174.

Nistala K, Wedderburn LR. (2009) Th17 and regulatory T cells:

rebalancing pro-and anti-in ammatory forces in autoimmune

arthritis. Rheumatology 48(6):602-606.

Ouyang W, Kolls JK, Zheng Y. (2008) The biological functions

of T helper 17 cell effector cytokines in in ammation. Immu-

nity . 28(4):454-467.

Pakfetrat A, Javadzadeh-Bolouri A, Basir-Shabestari S, Falaki

F. (2009) Oral Lichen Planus: a retrospective study of 420 Ira-

nian patients. Med Oral Patol Oral Cir Bucal . 14(7):E315-E318.

Pourshahidi S, Ebrahimi H, Andisheh Tadbir A. (2011) Eval-

uation of the Relationship between Oral Lichen Planus and

Stress. Shiraz Univ Dent J 12(1): 43-47.

Rhodus NL, Cheng B, Bowles W, Myers S, Miller L, Ondrey F.

(2006) Proin ammatory cytokine levels in saliva before and

after treatment of (erosive) oral lichen planus with dexametha-

sone. Oral Dis. 12(2):112-116.

Rhodus NL, Cheng B, Myers S, Bowles W, Ho V, Ondrey F.

(2005) A comparison of the pro-in ammatory, NF-kappaB-

dependent cytokines: TNF-alpha, IL-1-alpha, IL-6, and IL-8

in different oral  uids from oral lichen planus patients. Clin

Immunol. 114(3):278-283.

Rhodus NL, Cheng B, Ondrey F. (2007) Th1/Th2 cytokine ratio

in tissue transudates from patients with oral lichen planus.

Mediators In amm. 19854.

Sezer E, Ozugurlu F, Ozyurt H, Sahin S, Etikan I. (2007) Lipid

peroxidation and antioxidant status in lichen planus. Clin Exp

Dermatol . 32(4):430-434.

Shen Zh, Du G, Zhou Z, Liu W, Shi L, Xu H. (2016) Aber-

rant expression of interleukin-22 and its targeting microRNAs

in oral lichen planus: a preliminary study.J Oral Pathol Med

45(7):523-527.

Sonnenberg GF, Nair MG, Kirn TJ, Zaph C, Fouser LA, Artis

D. (2010) Pathological versus protective functions of IL-22

in airway in ammation are regulated by IL-17A. J Exp Med.

207(6):1293-1305.

Sugerman PB, Savage NW, Walsh LJ, Zhao ZZ, Zhou XJ, Khan

A, Seymour GJ, Bigby M. (2002) The pathogenesis of oral

lichen planus. Crit Rev Oral Biol Med. 213(4):350-365.

Sun A , Wang JT, Chia JS, Chiang CP. (2005) Serum interleu-

kin-8 level is a more sensitive marker than serum interleukin-6

level in monitoring the disease activity of oral lichen planus.

Br J Dermatol. 152(6):1187-1192.

van der Meij EH, van der Waal. (2003) Lack of clinicopatho-

logic correlation in the diagnosis of oral lichen planus based

on the presently available diagnostic criteria and suggestions

for modi cations. J Oral Pathol Med. 32(9):507-512.

Shiva Shirazian etal.

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS COMPARISON OF INTERLEUKIN 17 AND 22 IN SALIVA OF ORAL LICHEN PLANUS PATIENTS 591

Xie S, Ding L, Xiong Z, Zhu S. (2012) Implications of Th1 and

Th17 cells in pathogenesis of oral lichen planus. J Huazhong

Univ Sci Technolog Med Sci. 32(3):451-457.

Yamamoto T, Yoneda K, Ueta E, Osaki T. (1990) Cellular

immunosuppression in oral lichen planus. J Oral Pathol Med.

19(10):464-470.

Zaba LC, Cardinale I, Gilleaudeau P, et al. (2007) Amelioration

of epidermal hyperplasia by TNF inhibition is associated with

reduced Th17 responses. J Exp Med. 204(13):3183-3194.

Zenewicz LA, Yancopoulos GD, Valenzuela DM, Murphy AJ,

Karow M, Flavell RA. (2007) Interleukin-22 but not interleu-

kin-17 provides protection to hepatocytes during acute liver

in ammation. Immunity. 27(4):647-659.

Zhang Y, Lin M, Zhang S, et al. (2008) NF-kappaB-dependent

cytokines in saliva and serum from patients with oral lichen

planus: a study in an ethnic Chinese population. Cytokine.

41(2):144-149.

Zhao Y, Yang J, Gao Yd, Guo W. (2010) Th17 immunity in

patients with allergic asthma. Int Arch Allergy Immunol.

151(4):297-307.

Zheng Y, Danilenko DM, Valdez P, Kasman I, Eastham-Ander-

son J, Wu J, Ouyang W. (2007) Interleukin-22, a TH17 cytokine,

mediates IL-23-induced dermal in ammation and acanthosis.

Nature. 445(7128):648-651.

Ziolkowska M, Koc A, Luszczykiewicz G, Ksiezopolska-

Pietrzak K, Klimczak E, Chwalinska-Sadowska H, Maslin-

ski W. (2000) High levels of IL-17 in rheumatoid arthritis

patients: IL-15 triggers in vitro IL-17 production via cyclo-

sporin A-sensitive mechanism. J Immunol . 164(5):2832-

2838.