RESEARCH ARTICLE Open Access
IL28B and IL10R −1087 polymo phisms a e
p o ec i e o ch onic geno ype 1 HCV in ec ion
and p edic o s o esponse o in e e on-based
he apy in an Eas -Cen al Eu opean coho
Alajos Pá
1*†
, Gab iella Pá
1†
, Is án To nai
2
, Fe enc Szalay
3
, Dalma Vá szegi
4
, Edi F á e
2
, Má ia Papp
2
,
Gab iella Lengyel
5
, János Fehé
5
ˆ, Má a Va ga
6
, Judi Ge ain
7
, János Schulle
8
, Zsuzsanna Nemes
1
, Zol án Pé e i
1
,
Anna Tusnádi
9
, Béla Hunyady
1
, A ila Ha agh
10
, Zsol Szinku
10
, Á on Vincze
1
, László Sze eday
11
, Pé e Kis ali
12
and Béla Melegh
12
Abs ac
Backg ound: P e ious s udies ha e shown ha single nucleo ide polymo phisms (SNP) in IL28B and IL10R a e
associa ed wi h sus ained i ological esponse (SVR) in ch onic hepa i is C pa ien s ea ed wi h pegila ed in e e on
plus iba i in (P/R). The p esen s udy ex ends ou ea lie in es iga ions on a la ge Eas -Cen al Eu opean coho . The
allele equencies o IL28B and IL10R in geno ype 1 HCV in ec ion we e compa ed wi h ha o heal hy con ols o
he pu pose o examining he ela ionship be ween he polymo phisms and he SVR o P/R ea men .
Me hods: A o al o 748 ch onic HCV1 in ec ed pa ien s (365 male, 383 emale; 18–82 yea s) and 105 olun a y
blood dono s as con ols we e en olled. Fou hund ed and wen y HCV pa ien s we e ea ed wi h P/R o 24–72
weeks, ou o hem 195 (46.4%) achie ed SVR. The IL28 s12979860 SNP was de e mined using Cus om Taqman
SNP Geno yping Assays. The IL10R −1087 (also known as IL10R −1082 ( s1800896) p omo e egion SNP was
de e mined by RT-PCR and es ic ion agmen leng h polymo phism analysis.
Resul s: The IL28B CC geno ype occu ed wi h lowe equency in HCV pa ien s han in con ols (26.1% s 51.4%,
p<0.001). P/R ea ed pa ien s wi h he IL28B CC geno ype achie ed highe SVR a e, as compa ed o pa ien s wi h
CT (58.6% s 40.8%, p=0.002). The p e alence o IL10R −1087 GG geno ype was lowe in pa ien s han in con ols
(31.8 % s 52.2%, p<0.001). Among pa ien s achie ing SVR, he IL10R −1087 GG geno ype occu ed wi h highe
equency han he AA (32.0% s 17.4%, p=0.013). The IL28B T allele plus IL10R A allele combina ion was ound wi h
highe p e alence in pa ien s han in con ols (52% s 20.7%, p<0.001). The IL28B CC plus IL10R A allele
combina ion occu ed wi h highe equency among pa ien s wi h SVR han in non- esponde s (21.3% s 12.8%,
p=0.026). Bo h he IL28B CC plus IL10R GG and he IL28B CC plus IL10R A allele combina ions occu ed wi h lowe
equency in pa ien s han in con ols.
Conclusions: In ou HCV1 pa ien s, bo h he IL28B CC and IL10R GG geno ypes a e associa ed wi h clea ance o
HCV. Mo eo e , dis inc IL28B and IL10R allele combina ions appea o be p o ec i e agains ch onic HCV1 in ec ion
and p edic o s o esponse o P/R he apy.
Keywo ds: Gene ic polymo phism, Hepa i is C i us, In e e on, In e leukin-28B, In e leukin-10
* Co espondence: [email p o ec ed]
†
Equal con ibu o s
ˆDeceased
1
Fi s Depa men o Medicine, Uni e si y o Pécs, Rákóczi u. 2, 7623 Pécs,
Hunga y
Full lis o au ho in o ma ion is a ailable a he end o he a icle
© 2014 Pá e al.; licensee BioMed Cen al L d. This is an open access a icle dis ibu ed unde he e ms o he C ea i e
Commons A ibu ion License (h p://c ea i ecommons.o g/licenses/by/2.0), which pe mi s un es ic ed use, dis ibu ion, and
ep oduc ion in any medium, p o ided he o iginal wo k is p ope ly ci ed.
Pá e al. BMC Resea ch No es 2014, 7:12
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Backg ound
Hepa i is C i us (HCV) is a majo global heal h p oblem:
cu en ly 170 million subjec s a e su e ing om HCV
in ec ion wo ldwide [1,2]. The ou come o HCV in ec ion
anges om spon aneous i al clea ance o hepa i is,
ci hosis and hepa ocellula ca cinoma. Some indi iduals
ha e apidly p og essi e li e disease, while o he s emain
symp om- ee i us ca ie s. The exac causes o he di e -
en disease cou ses a e no known. Immune mechanisms,
as well as en i onmen al ac o s a e esponsible o
he a ious HCV- ela ed e en s [3]. As gene ic ac o s
con olling he hos immune esponse play a pi o al
ole in HCV in ec ion, a gene ic p edisposi ion may also
be c ucial [4-6]. Cy okines a e among he p edominan
mechanisms o hos de ense agains in ec ion; hey induce
in lamma o y esponse ha o en leads o issue inju y,
bu also se e as an i i al e ec o s. Mo eo e , cy okine
syn hesis capaci y has a signi ican gene ic componen .
This explains why di e ences exis be ween indi iduals in
hei abili y o p oduce cy okines, a ac which may also
be due o single-nucleo ide polymo phisms (SNPs) wi hin
he coding egions o cy okine genes. Cy okine genes a e
polymo phic and ce ain SNPs modi y he cy okine
p oduc ion and a ec he hos immune esponse. In
HCV in ec ion, sec e ion o an inapp op ia e amoun o
cy okines may be associa ed wi h ch onici y o esis ance
o in e e on (IFN) ea men [7].
The in e leukin-28B (IL-28B)–also e e ed o as
in e e onλ-3 (IFNλ-3) [8-12] –was shown o be
in ol ed in he con ol o HCV in ec ion. The gene ic
polymo phisms o he encoding IL28B gene egion de e -
mine he spon aneous and ea men induced clea ance o
geno ype 1 HCV (HCV1) [13-15].
IL-28B is exp essed by pe iphe al blood mononuclea
cells, dend i ic cells, and hepa ocy es upon in ec ion
wi h i uses o s imula ion wi h double-s anded RNA.
IL28B in u n ac i a es signal ansduc ion h ough he
JAK-STAT pa hway, exe s an i i al ac i i y and has an
impac on na u al clea ance o HCV. IL-28B exhibi s
ewe IFN-like ad e se e ec s because IL-28B ecep o s a e
exp essed on a limi ed numbe o cell ypes. The ecep o
o IL-28 is composed o a unique IL-28R-αchain ha pai s
wi h he IL-10R-βchain [10,11]. IL28B gene on human
ch omosome 19q was disco e ed using genomic sc eening
p ocess in which he en i e human genome was scanned
o pu a i e unc ional a ian s [8,9].
Ge e al. [12] iden i ied i s a SNP ( s12979860) loca ed
only 3 kilobases ups eam om he IL28B gene, ha
encodes IL-28B. They applied genome-wide associa ion
s udy (GWAS), e ospec i ely analysed samples o 1,137
HCV in ec ed indi iduals pa icipa ing in a clinical ial
wi h pegyla ed in e e on plus iba i in (P/R) he apy, and
ound ha he high IL-28B p oduc ion CC geno ype was
associa ed wi h app oxima ely a h ee old g ea e a e o
SVR o P/R when compa ed wi h he TT geno ype. This
inding was con i med by o he s [13,14]. A he same
ime i was also shown ha he equency o he CC
geno ype was signi ican ly lowe in HCV pa ien s
when compa ed o ma ched con ols; hus, sugges ing
ha his a ian may be linked o a highe a e o
na u al clea ance o HCV. This hypo hesis was soon
s eng hened by Thomas e al., who epo ed ha he
IL28B CC geno ype enhanced he spon aneous eso-
lu ion a e o HCV in ec ion [15]. Thus, i became e iden
ha HCV pa ien s who ha bo he CC geno ype a
s12979860 a e mo e p one o espond o P/R ea men
and o clea he i us han pa ien s who do no possess
his gene ic polymo phism.
The s anda d o ca e (SOC) ea men o HCV in ec ion
is pegin e e on plus iba i in (P/R) he apy o 24–72 mon h
du a ion, depending on HCV geno ype, p e- ea men i al
load and i al kine ic. This ype o ea men may esul in
sus ained i ological esponse (unde ec able HCV RNA
a e 24 weeks o he end o he apy) in app oxima ely
40–45% o pa ien s in ec ed by geno ype 1 HCV (HCV1)
he mos common geno ype in Eu ope. In he pas 2 yea s,
new compounds, “di ec ac ing an i i als”(DAAs)ha ealso
been de eloped such as HCV p o ease- and polyme ase
inhibi o s, ha signi ican ly enhance SVR compa ed
wi h SOC e en o 68–74% in ea men -naï e HCV1
indi iduals [16].
Since he e a e con lic ing da a on he ole o he
egula o y Th2 cy okine in e leukin-10 (IL-10) in HCV
in ec ion [17-20], he polymo phism o he IL10R
gene has also been examined. In ou p elimina y s udies,
he i s om an Eas -Cen al Eu opean coun y, we
epo ed ha no only IL28B bu also IL10R −1087
(also known as IL10R-1082) gene ic a ian s may play
a ole in HCV in ec ion [21,22].
In e leukin-10 (IL-10), p oduced by monocy es,
mac ophages and T cells, inhibi s bo h he ac i a ion
o CD4+ T-helpe cells and he unc ion o cy o oxic
CD8 + T, NK and an igen-p esen ing cells, and also
modula es hepa ic s ella e cell collagen syn hesis. IL-10
plays a egula o y ole in immune eac ion and supp esses
in lamma o y esponses by inhibi ing he p oduc ion o
p o-in lamma o y cy okines. I s e ec is media ed h ough
he IL-10 ecep o (IL-10R), which is a he e odime
ha consis s o bo h IL-10R1 equi ed o binding,
and IL-10R2 equi ed o signaling. Inc eased IL-10
p oduc ion was epo ed in associa ion wi h he -1082G,
-819C, -592GCC IL-10 p omo e (ATA) haplo ype [23,24].
I mac ophages sec e e la ge amoun o IL-10, i may
dec ease ci cula ing TFN-αand IL-6 le els, hus, educing
hei ha m ul e ec s. A highe equency o IL10 -1082 GG
geno ype was ound in olde heal hy con ols han in
pa ien s wi h myoca diac in a c ion; high IL-10 p oduc ion
was p o ec i e o longe i y [25].
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The p esen s udy ex ends ou p e ious in es iga ions
on a la ge Eas -Cen al Eu opean coho in Hunga y. The
allele equencies o IL28B and IL10R −1087 in HCV1
in ec ion we e compa ed wi h ha o heal hy con ols o
he pu pose o examining he ela ionship be ween he
polymo phisms and he esponse o PEG-IFN and iba i in
(P/R) ea men . We ind ha in HCV1-in ec ed
Eas -Cen al Eu opean pa ien s, bo h he IL28B CC and
IL10R GG geno ypes a e associa ed wi h clea ance o HCV.
Mo eo e , dis inc IL28B and IL10R allele combina ions
appea o be p o ec i e agains ch onic HCV1 in ec ion
and p edic o s o esponse o P/R he apy.
Me hods
E hics
This wo k has been ca ied ou in acco dance wi h he
guidelines o he 1975 Decla a ion o Helsinki and has
been app o ed by bo h he Na ional E hics Commi ee
(ETT TUKEB Budapes , No 490/010) and he Regional
E hics Commi ee a he Uni e si y o Pécs.
All he pa ien s ag eed o pa icipa e in he s udy, and
p o ided w i en in o med consen .
Pa ien s
A o al o 748 HCV1 in ec ed pa ien s (365 men, 383 emale,
anging in age om 18 o 82 yea s, mean 54 ± 10 yea s)
we e en olled. All subjec s we e inpa ien s in 10 Hunga ian
ci y hospi als o uni e si y clinics be ween Janua y 2007
and Decembe 2009.
The diagnosis o ch onic hepa i is C was based on
pe sis en ele a ion o se um alanine amino ans e ase
(ALT) le els >6 mon hs in he p esence o an i-HCV by
ELISA and HCV RNA by eal- ime e e se ansc ip ase-
polyme ase chain eac ion (RT-PCR). Quan i a ion o
plasma HCV RNA was ca ied ou using Cobas Amplico
2.0 (Ho mann-La Roche Inc.). HCV geno ype was de e -
mined by a geno ype speci ic p obe based assay in he 5′
un ansla ed egion (LiPA; Innogen ics, Ghen , Belgium).
(In Hunga y, >95% o HCV pa ien s ha e been in ec ed
wi h HCV geno ype 1. In Eas e n Eu opean coun ies he
p e alence o geno ype 1 anges be ween 57% and 99%
[26]). In 70% o ou pa ien s pe cu aneous li e biopsy
was pe o med.
O he 748 HCV1 pa ien s, 420 we e ea ed wi h pegy-
la ed in e e on al a 2a/2b (Pegasys, Ho mann-La Roche
Inc./Pegin on, SP Labo N.V. Belgium) 135–180 μg/1.0-
1.5 μg/kg subcu aneously pe week, and iba i in (Copegus,
Ho mann-La Roche Inc./Rebe ol, SP Labo N.V. Belgium)
1000–1200 mg o ally pe day o 24–72 weeks, hen
ollowed up o 24 weeks.
Subjec s who had unde ec able HCV RNA le els
ollowing 24 weeks o he apy we e designa ed as sus ained
i ological esponse (SVR), unde ec able se um HCV RNA
a week 4 a e s a ing ea men as apid i ological
esponse (RVR), unde ec able HCV RNA a week 12 a e
s a ing he apy as ea ly i ological esponse (EVR). In ou
pape , pa ien s who did no achie e SVR we e ega ded as
“non- esponde s”(non-SVR).
One hund ed and i e heal hy indi iduals (64 men, 41
emale, mean age 45 ± 3 yea s), who we e consecu i e
olun a y blood dono s wi h no mal li e unc ion es s
and nega i e o HBV, HCV and HIV se ology, se ed as
con ols.
Geno yping
DNA was isola ed om pe iphe al blood by a s anda d
desal ing me hod. The IL28B s12979860 SNP was
de e mined using Cus om Taqman SNP Geno yping
Assays (Applied Biosys ems, Li e Technologies, Fos e , CA,
USA). The IL10R −1087 (also known as IL10R −1082)
( s1800896) p omo e egion was equi ed o he o ma ion
o he EcoNI ecogni ion sequence 5′-AAGACAACAC
TACTAAGGCT-3′; helowe p ime was5′-TAAA
TATCCTCAAAGTTCC-3′. A e cu ing he ampli ied
p oduc (584 bp) by EcoNI, homozygo e GG was iden i ied
by wo agmen s 315 and 279 bp, while he e ozygo e AG
had 310, 280, 252 and 28 bp agmen s and homozygo e
AA had 310, 252 and 28 bp agmen s [27].
S a is ical analysis
S a is ical analysis was ca ied ou using he SPSS 16.0
o Windows (SPSS Inc., Chicago, IL, USA). The allele
and geno ype equencies we e compa ed wi h Pea son’s
χ2 es . Bina y logis ic eg ession analysis was pe o med o
obse e he indi idual con ibu ions o he polymo phisms.
A p alue o <0.05 was conside ed signi ican .
Resul s
O 420 pegin e e on and iba i in (P/R) ea ed pa ien s
195 (46.4%) achie ed SVR. Pa ien s wi h RVR (20% o
ea ed) achie ed a 86% SVR a e, while hose wi h EVR
(55% o ea ed) showed a 65% SVR a e.
As shown in Table 1, when compa ing he IL28B
geno ype equencies be ween g oups o heal hy con ols
and pa ien s, he CC geno ype occu ed wi h lowe
equency in HCV pa ien s han in con ols, hus,
sugges ing i s p o ec i e e ec agains ch onic hepa i i s C.
On he o he hand, CT he e ozygosi y and T alleles we e
mo e p e alen in pa ien s, and may, he e o e, con ey
suscep ibili y o he disease.
As shown in Table 2, subjec s ha ecei ed P/R he apy
and who had he IL28B CC geno ype achie ed highe a es
o SVR han subjec s who had CT geno ype (58.6% s
40.8%) (OR 2.057, 95% CI: 1.305-3.058, p = 0.002), o hose
ca ying he T allele (41.8%, OR 1.976, 95% CI: 1.263-3.058,
p = 0.002).
As shown in Table 3, IL10R GG geno ype occu ed
wi h lowe equency in pa ien s (31.8%) han in con ols
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(52.2%) (OR: 0.428, p < 0.001). The p e alence o IL10R
AA allele was 68.15% in pa ien s and 47.8% in heal hy
indi iduals (OR: 2.335, p < 0.001).
Among P/R ea ed pa ien s wi h SVR, he IL10R GG
geno ype occu ed wi h highe equency han he AA
geno ype, (57/178, 32.0% s 31/178, 17.4%, (OR: 1.84,
95% CI 1.13-2.98, p = 0.013).
The SVR a e in pa ien s wi h IL10R GG geno ype was
42.2% (59/125); in hose wi h GA 47.4% (88/178) and in
AA pa ien s 39.7% (31/78). HCV pa ien s wi h Aallele
(non-GG geno ype) achie ed SVR in 46.4% (119/256).
Geno ype combina ions
Heal hy con ols and HCV1 pa ien s
The p e alence o he double wild ype combina ion
o he IL28B CC plus IL10R GG was lowe (9.2%) in
HCV pa ien s han in heal hy con ols (23.9%) (OR 0.322,
p < 0.001).
The IL28B CC plus IL10R A allele combina ion
likewise occu ed wi h lowe equency in pa ien s
(16.1%) when compa ed wi h con ols (27.2,%, OR 0.515,
p = 0.009). Bo h combina ions sugges a p o ec i e e ec
agains ch onic HCV in ec ion.
The p e alence o IL28B T allele plus IL10R A allele
combina ion was highe in pa ien s (52.4%) han in heal hy
con ols (20.7%), hus, leading o a mo e han ou old isk
inc ease o he disease (OR 4.231, p < 0.001) (Table 4).
Compa ison o subjec s wi h sus ained i ologic esponse
(SVR) e sus non- esponde (non-SVR) pa ien s
The IL28B CC plus IL10R −1087 AA allele combina ion
was ound wi h highe equency among pa ien s achie ing
SVR (21.3%), han in non- esponde s (12.8%, OR 1.848,
95% CI 1.070-3.190, p = 0.026). In pa ien s wi h SVR, he
p e alence o he IL28B CC plus IL10R GG combina ion
was 12.4%, whe eas in non- esponde s i was 7.9%
(OR 1.648, NS) (Table 4).
Cumula i ely, he da a sugges ha SNPs in bo h IL28B
and he IL-10 p omo e egion, and speci ic combina ions
he eo , a e associa ed wi h p o ec ion agains HCV in ec ion
and di e en ial esponsi eness o IFN he apy.
Discussion
P e ious s udies ha e shown ha IL28B SNP a he poly-
mo phic si e s129798060 wi h he CC geno ype, and IL10
ecep o SNP a −1082 wi h he G allele a e associa ed wi h
suscep ibili y o ch onic hepa i is C i us in ec ion and
esponse o combined an i i al he apy [12-14,17,28,29]. In
ou coho o Hunga ian pa ien s, we e alua ed hese SNPs
in a o al o 748 pa ien s in ec ed wi h geno ype 1 HCV
om 10 cen e s h oughou Hunga y. To ou knowledge,
his is la ges s udy o i s ype on HCV-in ec ed subjec s
om Eas -Cen al Eu ope. Ou esul s show ha polymo -
phisms o bo h IL28B and IL10R genes play a ole in he
ou come o ch onic hepa i is C in a new coho . We we e
able o demons a e ha IL28B CC geno ype is a s ong
p edic o o SVR in pa ien s ea ed wi h P/R, and p o ec s
agains he disease. The cu en s udy on a la ge coho
con i ms he esul s o ou ea lie s udies [21,22]. This gen-
e ic a ian o IL28B may be associa ed wi h an ele a ed
p oduc ion o IL-28B (IFNλ-3) bu his has no been ound
in all s udies [12].
Table 2 Sus ained i ological esponse (SVR) a e
s a i ied by IL28B geno ypes in Hunga ian HCV pa ien s
T ea ed SVR
IL28B geno ype n Numbe o pa ien s Pe cen age
CC 116 68 58.6%
CT 228 93 40.8%
TT 76 34 44. 7%
T allele (non-CC) 304 127 41.8%
Table 1 P e alence o IL28B geno ypes in Hunga ian HCV
pa ien s and heal hy con ols
IL28B geno ype HCV1
(n = 748)
Con ols
(n = 105)
OR
CC 195 (26.1%) 54 (51.4%) 0.333 (0.22 - 0.505)
P < 0.001
CT 411 (54.9%) 39 (37.1%) 2.064 (1.354 - 3.145)
p = 0.001
C allele 606 (81.02%) 93 (88.6%) 0.551 (0.294-1.032)
p = 0.059
TT 142 (19.0%) 12 (11.4%) 1.816 (0.969 –3.404)
p = 0.059
T allele (non-CC) 553 (73.9%) 51 (48.6%) 3.003 (1.981 –4.552)
P < 0.001
Table 3 P e alence o IL10R −1087 geno ypes in HCV
pa ien s and heal hy con ols
IL10R −1087
Geno ype
HCV1
(n = 672)
Con ols
(n = 92)
OR
GG 214 (31.8) 48 (52.2%) 0.428 (0.276 - 0.665)
p < 0.001
GA 333 (49.6%) 32 (34.8%) 1.842 (1.169 –2.903)
p = 0.008
G allele 547 (81.4%) 80 (87.0%) 0.656 (0.347-1.241)
p = 0.192
AA 125 (18.6%) 12 (13.0%) 1.523 (0.806 –2.881)
p = 0.192
A allele (non-GG) 458 (68.15%) 44 (47.8%) 2.335 (1.504 –3.625)
p < 0.001
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Ou esul s a e in acco dance wi h he p e ious indings
[12-14]. The equency o IL28B CC in HCV1 pa ien s
was lowe han in heal hy con ols, his ac may mean
ha indi iduals wi h IL28B CC geno ype a e p o ec ed
agains ch onic hepa i is C. Pa ien s wi h IL28 CC geno-
ype may ha e a high likelihood o SVR and possibly need
only he s anda d o ca e P/R ea men , pa icula ly i
hey ha e o he posi i e p edic o s: low i al load, young
age, emale, sho du a ion o in ec ion, absence o ci ho-
sis and low BMI. Unde such ci cums ances, an IL28B CC
will no only p edic SVR bu also mo i a e bo h he pa-
ien and he doc o , se ing o enhance adhe ence as well
Howe e , i mus be also emphasised ha he s onges
posi i e p edic o o SVR is no he IL28B CC, bu he
RVR [30,31] as i was ound in ou cases oo, when 86% o
pa ien s wi h RVR elimina ed hei HCV1 in ec ions.
The ques ion a ises whe he HCV1 pa ien s ca ying
he IL28B TT geno ype always equi e a mo e complex
(e.g. iple) he apy wi h di ec ac ing an i i als (DAAs)?
I is concei able ha pa ien s wi h he TT homozygosi y,
who also ha e o he nega i e p edic o s (such as p e i-
ous non- esponse, high i al load, ci hosis, obesi y,
insulin esis ance), will be candida es o ea men wi h
DAAs.
Recen ly, i was ound ha bo h he iple he apy
wi h DAAs, (e.g. p o ease inhibi o s plus P/R) and he
IFN- ee an i-HCV ea men may o e come he e ec o
he un a ou able TT IL28B polymo phism. Thus, in he
u u e, he ele ance o IL28B geno yping may become
limi ed in he indi idualized ea men o HCV1 in ec ion
[32-35]. Howe e , B onowicki e al. epo ed ha 100% o
ea men -nai e HCV1 pa ien s wi h IL28B CC achie ed
SVR, e en wi h 12 weeks o a p o ease inhibi o elap e i
and P/R combina ion he apy [36]. The e o e, IL28B
geno ypingmaycon inue obeo impo ancein he
sho ening o ea men du a ion.
Thus, he indings o IL28B polymo phisms will allow
one o p edic wi h be e accu acy which pa ien s a e
likely o espond o he s anda d o ca e P/R he apy.
Howe e , i mus be s essed ha he e ec o he
a ou able a ian is no absolu e: no all ca ie s o he
“good esponse geno ype”clea he i us; no do all pa ien s
lacking i ail o bene i om he P/R ea men [30,31].
I was no ed p e iously ha IL10 -1082 GG geno ype
was equen in HCV, when mononuclea cells p oduced
wo- old g ea e quan i ies o IL-10 compa ed wi h pa ien s
wi h IL10 AA geno ype [25,26]. Vigidal e al. epo ed, ha
IL10 gene p omo e −1082 GG geno ype occu ed mo e
equen ly in HCV pa ien s han in con ols and ha he
GG homozygosi y was associa ed wi h poo esponse o
IFN. This geno ype was iden i ied in 34.6% o 80
HCV pa ien s compa ed wi h 6/33 (16.7%) o con ols
(p = 0.048). Howe e , his s udy ocused on limi ed numbe s
o bo h pa ien s and con ol subjec s [28]. Me a-analysis o
Zhang e al., also showed ha he IL10 -1082 GG geno ype
was associa ed wi h suscep ibili y o ch onic HCV in ec ion
[29]. Ye , A zal e al. epo ed ha he equency o −1082
GA geno ype was highe in heal hy indi iduals han in
HCV pa ien s, sugges ing he p o ec i e e ec o his
a ian [19]. Lio e al. no ed ha he IL10 -1082 GG
was associa ed wi h spon aneous eco e y om HCV
in ec ion [18], while acco ding o Gao e al. -1082 AA
con e ed an inc eased isk o pe sis en HCV in ec ion
[37]. Knapp e al. ound ha HCV pa ien s wi h SVR
showed highe equency o IL10 -1082 GG geno ype
han non- esponde s (26.9% s 14.0% OR 2.28, 95%
CI 1.21-4.32, p = 0.05) [17].
In his s udy we in es iga ed he IL10R p omo e gene
polymo phisms a posi ion −1087 ela i e o ansc ip ion
s a si e and p o ed ha he −1087 GG occu s a a lowe
equency in HCV pa ien s as compa ed wi h con ols;
hus, his a ian may be p o ec i e agains he disease. A
Table 4 IL28B and IL10R −1087 geno ype combina ions in he di e en s udy g oups
IL28B geno ype
CC CC T allele T allele
IL10R geno ype HCV n = 664 GG 61 (9.2%) OR:0.322 (0.186-0.556) 148 (22.3%) 0.728 (0.446-1.188)
A allele 107 (16.1%) p < 0.001 348 (52.4%) p = 0.202
Con ols n = 92 GG 22 (23.9%) OR:0.515 (0.311-0.852) 26 (28.3%) OR:4.231 (2.497-7.169)
A allele 25 (27.2%) p = 0.009 19 (20.7%) p < 0.001
SVR n = 178 GG 22 (12.4%) OR:1.648 (0.837-3.247) 37 (20.8%) OR:0.803 (0.496-1.301)
A allele 38 (21.3%) p = 0.146 81 (45.6%) p = 0.372
Non- esponde s n = 203 GG 16 (7.9%) OR: 1.848 (1.070-3.190) 50 (24.6%) OR:0.692 (0.462-1.037)
A allele 26 (12.8%) p = 0.026 (54.7%)111 p = 0.074
P e alence o IL28B CC plus IL10R GG in HCV pa ien s s. con ols OR: 0.322.
IL28B CC plus IL10R A allele OR: 0.515.
IL28B T allele plus IL10R A allele OR: 4.231.
P e alence o IL28B CC plus IL10R A allele in p s wi h SVR s. non- esponde s OR:1.848.
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he same ime, he equency o GA geno ype and A allele
was highe in he pa ien s’g oup - as a po en ial gene ic
ma ke o suscep ibili y o he HCV1 in ec ion. In pa ien s
who achie ed SVR, he IL10R GG geno ype occu ed wi h
highe (32.%) equency han AA (17.4%), and he IL28B
CC plus IL10R −1087 A allele combina ion was also ound
wi h highe a e among pa ien s achie ing SVR (21.3%),
han in non- esponde s (12.8%).
Ou indings on he ole o IL10R gene polymo phism
in HCV1 in ec ion a e a he con lic ing, and may
con adic bo h some o he p e iously published da a
[28,29] as well as he known biological e ec o IL-10.
The easons o hese disc epancies a e no clea . The
s udy popula ions may di e wi h ega d o ac o s ha
in luence SVR: i al load, obesi y, ib osis, e hnici y and
sex. Addi ional s udies a e equi ed o de ine he oles o
hese ac o s in connec ion wi h IL10 polymo phisms in
ch onic hepa i is C.
Conclusions
We demons a ed ha no only IL28B CC bu also
IL10R GG geno ype and ce ain IL28B plus IL10R
allele combina ions may be p o ec i e gene ic a ian s
in Hunga ian pa ien s wi h ch onic HCV1 in ec ion.
By p edic ing which pe sons a e mos likely o espond o
IFN-based he apy, he examina ion o hese polymo phisms
may help o manage an indi idualized app oach o ea
HCV pa ien s. Geno yping bo h IL28B and IL10R may
o e a baseline sc eening es , ha , howe e , should be
combined wi h o he well-known p e- and on- ea men
p edic o s o he ou come o he disease and choosing
he mos app op ia e ype o an i-HCV he apy. In he e a
o DAAs o he ea men o HCV in ec ion, he p ognos-
ic ole o hese gene ic es s may be o less impo ance.
Ye , in ce ain ci cums ances, he IL28B geno yping can
be use ul; e.g. in he sho ening o ea men du a ion
in P/R plus DAA- ea ed HCV pa ien s. I is e iden ,
howe e , ha as an on- ea men p edic o , apid i o-
logical esponse emains he s onges p ognos ic ac o ,
e en ac oss IL28B geno ypes [38].
Abb e ia ions
ALT: Alanine amino ans e ase; DDA: Di ec ac ing an i i al; GWAS: Genome-
wide associa ion s udy; HCV: Hepa i is C i us; IFN: In e e on; IL: In e leukin;
NK: Na u al kille ; PEG-IFN: Pegyla ed in e e on; P/R: Pegyla ed in e e on
plus iba i in; RBV: Riba i in; RVR: Rapid i ological esponse; SNP: Single
nucleo id polymo phism; SVR: Sus ained i ological esponse; Th: T helpe ;
TNF: Tumo nec osis ac o .
Compe ing in e es s
The au ho s decla e ha hey ha e no compe ing in e es s.
Au ho s’con ibu ions
AP, GP, IT, FS, BH ini ia ed and pa icipa ed in he design o he s udy, PK
ca ied ou he geno yping s udies; DV, EF, MP, GL, JF, MV, JG, JS, ZN ZP, AT,
AH, ZS, ÁV con ibu ed o acquisi ion o pa ien s and hei da a; AP, GP, LS
d a ed he manusc ip ; BM e ised he pape c i ically o in ellec ual
con en . All au ho s ead and app o ed he inal manusc ip .
Acknowledgemen s
Alajos Pá has ecei ed esea ch unding om he Hunga ian Na ional
Resea ch Fund (OTKA K81454) and om he Uni ed Eu opean
Gas oen e ology Fede a ion (UEGF) (2011 Top Abs ac P ize).
The au ho s hank Noemi Polga and Laszlo Po a o pe o ming s a is ical
analysis.
Au ho de ails
1
Fi s Depa men o Medicine, Uni e si y o Pécs, Rákóczi u. 2, 7623 Pécs,
Hunga y.
2
Second Depa men o Medicine, Uni e si y o Deb ecen, 4012
Deb ecen, Hunga y.
3
Fi s Depa men o Medicine, Semmelweis Uni e si y,
1082 Budapes , Hunga y.
4
Depa men o De ma ology, Uni e si y o Pécs,
7627 Pécs, Hunga y.
5
Second Depa men o Medicine, Semmelweis
Uni e si y, 1088 Budapes , Hunga y.
6
Ré hy Pál Hospi al, 5600 Békéscsaba,
Hunga y.
7
Szen Gyö gy Hospi al, 8000 Székes ehé á , Hunga y.
8
Uni ed
Szen Is án and Szen László Hospi al, 1097 Budapes , Hunga y.
9
He ényi
Géza Hospi al, 5004 Szolnok, Hunga y.
10
Kaposi Mó Teaching Hospi al, 7400
Kapos á , Hunga y.
11
Depa men o Medical Mic obiology and Immunology,
Uni e si y o Pécs, 7624 Pécs, Hunga y.
12
Depa men o Medical Gene ics,
Uni e si y o Pécs, 7624 Pécs, Hunga y.
Recei ed: 12 Sep embe 2013 Accep ed: 3 Janua y 2014
Published: 8 Janua y 2014
Re e ences
1. La anchy D: The global bu den o hepa i is C. Li e In 2009,
29(Suppl 1):74–81.
2. G a i z L: In oduc ion: a smoulde ing public-heal h c isis. Na u e 2011,
474:S2–S4.
3. Dus in LB, Rice CM: Flying unde he ada : he immunobiology o
hepa i is C. Annu Re Immunol 2007, 25:71–99.
4. Thimme R, Bukh J, Spangenbe g HC, Wieland S, Pembe on J, S eige C,
Go inda ajan S, Pu cell RH, Chisa i FV: Vi al and immunological
de e minan s o hepa i is C i us clea ance, pe sis ence, and disease.
P oc Na l Acad Sci U S A 2002, 99:15661–15668.
5. And ade DR: AJR: The in luence o he human genome on ch onic i al
hepa i is ou come. Re Ins Med T op Sao Paulo 2004, 46:119–126.
6. Donaldson PT: Gene ics o li e disease: immunogene ics and disease
pa hogenesis. Gu 2004, 53:599–608.
7. Thio CL: Hos gene ic ac o s and an i i al immune esponses o
hepa i is C i us. Clin Li e Dis 2008, 12:713–726. xi.
8. Ko enkoSV,Gallaghe G,Bau inVV,Lewis-An esA,ShenM,ShahNK,Lange JA,
Sheikh F, Dickenshee s H, Donnelly RP: IFN-lambdas media e an i i al
p o ec ion h ough a dis inc class II cy okine ecep o complex.
Na Immunol 2003, 4:69–77.
9. Sheppa d P, Kinds ogel W, Xu W, Hende son K, Schlu smeye S, Whi mo e TE,
Kues ne R, Ga igues U, Bi ks C, Ro aback J, Os ande C, Dong D, Shin J,
P esnell S, Fox B, Haldeman B, Coope E, Ta D, Gilbe T, G an FJ, Tacke M,
K i an W, McKnigh G, Clegg C, Fos e D, Kluche KM: IL-28, IL-29 and hei
class II cy okine ecep o IL-28R. Na Immunol 2003, 4:63–68.
10. Robek MD, Boyd BS, Chisa i FV: Lambda in e e on inhibi s hepa i is B and
C i us eplica ion. J Vi ol 2005, 79:3851–3854.
11. Uzé G, Monne on D: IL-28 and IL-29: newcome s o he in e e on amily.
Biochimie 2007, 89:729–734.
12. Ge D, Fellay J, Thompson AJ, Simon JS, Shianna KV, U ban TJ, Heinzen EL,
Qiu P, Be elsen AH, Mui AJ, Sulkowski M, McHu chison JG, Golds ein DB:
Gene ic a ia ion in IL28B p edic s hepa i is C ea men -induced i al
clea ance. Na u e 2009, 461:399–401.
13. Suppiah V, Moldo an M, Ahlens iel G, Be g T, Wel man M, Aba e ML,
Bassendine M, Spengle U, Do e GJ, Powell E, Rio dan S, She idan D,
Smedile A, F agomeli V, Mülle T, Bahlo M, S ewa GJ, Boo h DR, Geo ge J:
IL28B is associa ed wi h esponse o ch onic hepa i is C in e e on-alpha
and iba i in he apy. Na Gene 2009, 41:1100–1104.
14. Tanaka Y, Nishida N, Sugiyama M, Ku osaki M, Ma suu a K, Sakamo o N,
Nakagawa M, Ko enaga M, Hino K, Hige S, I o Y, Mi a E, Tanaka E, Mochida S,
Mu awaki Y, Honda M, Sakai A, Hiasa Y, Nishiguchi S, Koike A, Sakaida I,
Imamu a M, I o K, Yano K, Masaki N, Sugauchi F, Izumi N, Tokunaga K,
Mizokami M: Genome-wide associa ion o IL28B wi h esponse o pegyla ed
in e e on-alpha and iba i in he apy o ch onic hepa i is C. Na Gene
2009, 41:1105–1109.
Pá e al. BMC Resea ch No es 2014, 7:12 Page 6 o 7
h p://www.biomedcen al.com/1756-0500/7/12
15. Thomas DL, Thio CL, Ma in MP, Qi Y, Ge D, O’Huigin C, Kidd J, Kidd K,
Khakoo SI, Alexande G, Goede JJ, Ki k GD, Don ield SM, Rosen HR, Toble LH,
Busch MP, McHu chison JG, Golds ein DB, Ca ing on M: Gene ic a ia ion in
IL28B and spon aneous clea ance o hepa i is C i us. Na u e 2009,
461:798–801.
16. Thompson A, Pa el K, Tillamn H, McHu chison JG: Di ec ly ac ing an i i als
o he ea men o pa ien s wi h hepa i is C in ec ion. A clinical
de elopmen upda e add essing key u u e challenges. J Hepa ol 2009,
50:184–194.
17. Knapp S, Hennig BJW, F odsham AJ, Zhang L, Hellie S, W igh M, Goldin R,
Hill AVS, Thomas HC, Thu sz MR: In e leukin-10 p omo e polymo phisms
and he ou come o hepa i is C i us in ec ion. Immunogene ics 2003,
55:362–369.
18. Lio D, Ca uso C, Di S e ano R, Colonna Romano G, Fe a o D, Scola L,
C i ello A, Lica a A, Valenza LM, Cando e G, C axì A, Almasio PL: IL-10 and
TNF-alpha polymo phisms and he eco e y om HCV in ec ion.
Hum Immunol 2003, 64:674–680.
19. Selzne N, McGil ay I: Can gene ic a ia ions p edic HCV ea men
ou comes? J Hepa ol 2008, 49:494–497.
20. A zal MS, Tahi S, Salman A, Baig TA, Sha i T, Zaidi NUSS, Qad i I: Analysis o
in e leukin-10 gene polymo phisms and hepa i is C suscep ibili y in
Pakis an. J In ec De C ies 2011, 5:473–479.
21. Pa A, Kis ali P, Melegh B, Mise a A, To nai I, Papp M, Hunyady B, Vincze A,
Pa G: Gene ic polymo phisms in IL-10R, IL-28B and LTA genes in HCV
in ec ion. Do hey ha e p o ec i e ole and p edic sus ained i ological
esponse? J Hepa ol 2010, 52:S457.
22. Pá A, Kis ali P, Melegh B, To nai I, Lengyel G, Nemes Z, Pé e i Z, Hunyady B,
Vincze Á, Pá G: Cy okine (IL-10, IL-28B and LT-A) Gene Polymo phisms in
Ch onic Hepa i is C Vi us In ec ion. Clin Exp Med J 2011, 5:9–19.
23. Kuzushi a N, Hayashi N, Ka ayama K, Kan o T, Oshi a M, Hagiwa a H,
Kasaha a A, Fusamo o H, Kamada T: High le els o se um in e leukin-10
a e associa ed wi h a poo esponse o in e e on ea men in pa ien s
wi h ch onic hepa i is C. Scand J Gas oen e ol 1997, 32:169–174.
24. Yee LJ, Tang J, Gibson AW, Kimbe ly R, Van Leeuwen DJ, Kaslow RA: In e leukin
10polymo phismsasp edic o so sus ained esponse in an i i al he apy
o ch onic hepa i is C in ec ion. Hepa ology 2001, 33:708–712.
25. Lio D, Cando e G, C i ello A, Scola L, Colonna-Romano G, Ca allone L,
Ho mann E, Ca uso M, Licas o F, Calda e a CM, B anzi A, F anceschi C,
Ca uso C: Opposi e e ec s o in e leukin 10 common gene polymo phisms
in ca dio ascula diseases and in success ul ageing: gene ic backg ound o
male cen ena ians is p o ec i e agains co ona y hea disease. J Med Gene
2004, 41:790–794.
26. Ge ain J, Simon G J , Simon J: Geno ype dis ibu ion o hepa i is C i us
in he Hunga ian popula ion wi h ch onic i al hepa i is C. Eu J
Gas oen e ol Hepa ol 2003, 15:449–450.
27. Padyuko L, Hahn-Zo ic M, Lau YL, Hanson LA: Di e en allelic equencies
o se e al cy okine genes in Hong Kong Chinese and Swedish Caucasians.
Genes Immun 2001, 2:280–283.
28. Vidigal PG, Ge me JJ, Zein NN: Polymo phisms in he in e leukin-10,
umo nec osis ac o -alpha, and ans o ming g ow h ac o -be a1
genes in ch onic hepa i is C pa ien s ea ed wi h in e e on and
iba i in. J Hepa ol 2002, 36:271–277.
29. Zhang L-Z, Zhang T-C, Pan F-M, Zhang Z-H, Li X: In e leukin-10 gene
polymo phisms in associa ion wi h suscep ibili y o ch onic hepa i is C
i us in ec ion: a me a-analysis s udy. A ch Vi ol 2010, 155:1839–1842.
30. Asselah T: Gene ic polymo phism and esponse o ea men in ch onic
hepa i is C: he u u e o pe sonalized medicine. J Hepa ol 2010,
52:452–454.
31. Cla k PJ, Thompson AJ, McHu chison JG: IL28B genomic-based ea men
pa adigms o pa ien s wi h ch onic hepa i is C in ec ion: he u u e o
pe sonalized HCV he apies. Am J Gas oen e ol 2011, 106:38–45.
32. Poo dad F, B onowicki J-P, Go don SC, Zeuzem S, Jacobson IM, Sulkowski MS,
Poyna d T, Mo gan TR, Bu oughs M, Sniukiene V, Bopa ai N, B ass CA, Alb ech
JK, Bacon BR: IL28B polymo phism p edic s i ological esponse in pa ien s
wi h hepa i is CC geno ype 1 ea ed wi h Bocep e i (BOC) combina ion
he apy. JHepa ol2011, 54:S6.
33. Pol S, Ae ssens J, Zeuzem S, And eone P, Lawi z EJ, Robe s S, Younossi Z,
Fos e GR, Focaccia R, Ho ban A, Pock os PJ, Van Heeswijk RPG, De Meye S,
Luo D, Bo ield M, Beumon M, Picchio G: Limi ed impac o IL28B
geno ype on esponse a es in elap e i - ea ed pa ien s wi h p io
ea men ailu e. J Hepa ol 2013, 58:883–889.
34. Poo dad F, Lawi z E, Kowdley KV, Cohen DE, Podsadecki T, Siggelkow S,
Heckaman M, La sen L, Menon R, Koe G, T ipa hi R, Pilo -Ma ias T, Be ns ein B:
Explo a o y s udy o o al combina ion an i i al he apy o hepa i is C.
NEngJMed2013, 368:45–53.
35. King LY, Chung RT: IL28B es ing in a apidly changing wo ld: s ill
ele an ? J Hepa ol 2013, 58:847–849.
36. B onowicki J-P, Hezode C, Beng sson L, Pol S, Bou lie e M, Se a y L, De
Ledinghen V, T an A, Benhamou Y, G ange J-D, Ma hu in P, Ma cellin P,
T epo C, Za ski J-P, Seepe saud S, Kellihe K, Bo ield M, Pawlo sky J-M:
100% SVR in IL28B CC pa ien s ea ed wi h 12 weeks o Telap e i ,
Pegin e e on and Riba i in in he PROVE2 T ial. J Hepa ol 2012,
56:S430–S431.
37. Gao Q-J, Liu D-W, Zhang S-Y, Jia M, Wang L-M, Wu L-H, Wang S-Y, Tong L-X:
Polymo phisms o some cy okines and ch onic hepa i is B and C i us
in ec ion. Wo ld J Gas oen e ol 2009, 15:5610–5619.
38. F ied MW, Hadziyannis SJ, Shi man ML, Messinge D, Zeuzem S: Rapid
i ological esponse is he mos impo an p edic o o sus ained
i ological esponse ac oss geno ypes in pa ien s wi h ch onic hepa i is
C i us in ec ion. J Hepa ol 2011, 55:69–75.
doi:10.1186/1756-0500-7-12
Ci e his a icle as: Pá e al.:IL28B and IL10R −1087 polymo phisms
a e p o ec i e o ch onic geno ype 1 HCV in ec ion and p edic o s
o esponse o in e e on-based he apy in an Eas -Cen al Eu opean
coho . BMC Resea ch No es 2014 7:12.
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