Autoimmunity, January 2009; 42(1): 50–55

q Informa Healthcare USA, Inc.

ISSN 0891-6934 print/1607-842X online
DOI: 10.1080/08916930802292510

Gene polymorphisms of TNF-a2308 (G/A), IL-1021082 (G/A),

IL-62174 (G/C) and IL-1Ra (VNTR) in Egyptian cases with type 1
diabetes mellitus

AHMAD SETTIN1,2,†, AZZA ISMAIL4, MEGAHED ABO EL-MAGD3, RIZK EL-BAZ1, &
AMIRA KAZAMEL4
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1
Genetics Unit, Children Hospital, Mansoura University, Mansoura, Egypt, 2College of Medicine, Qassim University,
Buraydah, Saudi Arabia, 3Diabetes and Endocrinology Unit, Internal Medicine Specialist Hospital, Mansoura University,
Mansoura, Egypt, and 4Zoology Department, Faculty of Science, Mansoura University, Mansoura, Egypt

(Submitted 14 March 2008; accepted 9 June 2008)

Abstract
Background. Type 1 diabetes (T1D) is a genetically conditioned autoimmune disease in which cytokines play an important role.
Objectives. To check for the association of polymorphisms of cytokine genes with type 1 diabetes.
For personal use only.

Subjects. This work included 50 cases with T1D and 98 healthy individuals from the Nile Delta region of Egypt. Cases included
20 males and 30 females with a median age of 25 and range of 15–50 years.
Methods. DNA was amplified using PCR with sequence-specific primers for detection of polymorphisms related to tumor
necrosis factor (TNF)-a2308 (G/A), interleukin (IL)-1021082 (G/A), IL-62174 (G/C), and IL-1Ra (VNTR).
Results. Cases with T1D showed significant higher frequency of genotypes of TNF-a2308 AA ( p , 0.001, odds ratio
(OR) ¼ 7.91), IL-6-17CC ( p , 0.05, OR ¼ 3.36) and IL-1Ra A1A1 ( p , 0.05, OR ¼ 3.68) with significant lower frequencies
of TNF-a2308 GA, and IL-1Ra A1A2 genotypes ( p , 0.001 and ,0.05, respectively). They also showed significant higher
frequency of TNF-a2308 allele A ( p , 0.05, OR ¼ 2.0), IL-1Ra allele A1 ( p , 0.05, OR ¼ 2.98) with a significant lower
frequency of TNF-a2308 G allele and IL-1Ra A2 allele ( p , 0.05). No significant difference was detected among cases in relation
to IL-1021082 (G/A) genotypes or alleles nor in relation to age, sex, consanguinity or family history of the disease.
Conclusions. Polymorphisms related to TNF-a and IL-1Ra genes may be considered genetic markers for T1D among Egyptians
with a potential impact on family counseling and management.

Keywords: Type 1 diabetes, cytokines, gene polymorphism, Egypt

Introduction cause beta cell death by direct cytotoxic effects,

Type 1 diabetes mellitus (T1D) is an autoimmune
disease in which the body destroys insulin-producing
beta cells in the pancreas [1]. Using genome screen-
ing, candidate gene testing, in addition to studies of
human homologs of mouse susceptibility genes, at
least 20 different chromosomal regions have been
linked to T1D [2,3].
Cytokines also play an important role in T1D as
they are produced by immune cells and are thought to
mediate beta cell destruction [4,5]. Cytokines can
potentially acting through the induction of nitric oxide
production. Alternatively, cytokines may enhance T
cell-mediated beta cell destruction [6,7].
The interleukin (IL)-1 cluster on human chromo-
some 2q12– 2q14 harbors various promising candi-
date genes for inflammatory diseases [8]. One of the
important polymorphisms at the IL-1 gene cluster is
IL-1Ra (VNTR) caused by variable numbers of an
86 bp tandem repeat located in the second intron of
the gene. The most common alleles have been termed

Correspondence: A. Settin, Genetics Unit, Children Hospital, Mansoura University, Mansoura, Egypt. Fax: +20 50 223 4092.
E-mail: settin@mans.edu.eg; settin60@gmail.com
 Cytokine gene polymorphism in T1D 51

allele one (A1, four repeats), allele two (A2, two
repeats) in addition to other three alleles [9].
Tumor necrosis factor (TNF)-a play a central role
in the autoimmune destruction of pancreatic beta-
cells, whereas IL-6 inhibits TNF-a secretion, and
may have some protective effects [10]. In human,
the TNF-a gene is located within the highly
polymorphic major histocompatibility complex
region on chromosome 6p21.3 [11]. Many studies
have shown that SNP at position 2 308 G/A was
associated with various inflammatory conditions
[12]. Also, a polymorphism in the 50 flanking region
of the IL-6 gene on chromosome 7 at position 2 174
has been reported to exert an effect on its secretion
and function [13].
IL-10 is an anti-inflammatory and immunosup-
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Subjects and methods
This study included 50 cases diagnosed as T1D with
positive antibodies against insulin and glutamic acid
decarboxylase with very low C-peptide. They were
recruited from the Department of Endocrinology and
Diabetes, Internal Medicine Specialist Hospital,
Mansoura University, which is the main referral
hospital in the Nile Delta region of Egypt. They
comprised 20 (40%) males and 30 (60%) females with
a median age of 25 years and range of 15 – 50 years.
Of these cases, 21 (42%) had an age of onset of the
disease # 18 years, 33 (66%) were presented with
ketoacidosis, 13 (26%) had a positive parental
consanguinity and 19 (38%) had a positive family
history of T1D. Case genotypes were compared to 98
healthy unrelated blood donor volunteers from the

pressive substance produced within the body and
plays a role in the regulation of immune responses
[14,15]. The gene for IL-10 in chromosome region
1q31 – q32 has three biallelic single nucleotide poly-
morphisms (SNPs) within the promoter region, at
positions 2 1082, 2 819, and 2 592 from the tran-
scription initiation site [16]. The 2 1082 (G/A)
polymorphism was shown to affect the in vitro IL-10
production in peripheral blood cells from healthy
For personal use only.
same locality. They included 52 males and 46 females
with a median age of 45 years. Informed consent was
taken from all subjects and an approval was also
obtained from University Scientific and Ethical
Committees to carry out the research.
For all subjects, three SNPs were analyzed includ-
ing gene promoter sites TNF-a2308 (G/A), IL-
1021082 (G/A) and IL-62174 (G/C) in addition to
IL-1Ra gene VNTR in intron 2 as previously

individuals [17]. described [18 –21]. For SNPs identification, PCR

This work was planned in order to test the
association of polymorphisms of two pro-inflamma-
tory cytokines (TNF-a at position 2 308 and IL-6 at
position 2 174) and two anti-inflammatory cytokines
(IL-10 genes at position 2 1082 and of IL-1Ra
VNTR) with type 1 diabetes.
with sequence-specific primers was used in two
separate reactions employing a common forward and
two reverse primers, while IL-1Ra VNTR polymorph-
ism was characterized via a single PCR reaction
employing a forward and a reverse primers. All
primers, Taq polymerase, dNTP, and MgCl2 were

Figure 1. PCR amplification for studied cytokine gene polymorphisms including: Strip (I) TNF-a2308 (band size: 836 bp) showing: lane M:
DNA size marker, lanes 1, 2, 4, 6 with þ ve bands for G and A alleles, i.e. GA genotypes, lane 5 with only þ ve band for A allele, i.e. AA
genotype, lane 7 with only þ ve band for G allele, i.e. GG genotype and lane 3 with 2ve control; Strip (II) IL-1021082 (band size 179 bp)
showing: lanes 1, 3, 4 with þ ve bands for G and A alleles, i.e. GA genotypes, lane 6 with only þ ve band for A allele, i.e. AA genotype, lanes 5,
7 with only þ ve band for G allele, i.e. GG genotypes and lane 2 with 2ve control. Strip (III) IL-62174 (band size 234 bp) showing lanes 1, 6, 7
with þ ve bands for G and C alleles, i.e. GC genotypes, lanes 2 –4 with only þ ve band for G allele, i.e. GG genotypes and lane 5 with only þ ve
band for C allele, i.e. CC genotype.
 52 A. Settin et al.

for positive association using Fisher’s exact test and

OR with 95% confidence intervals (CI). A minimum
level of p , 0.05 is considered significant. It is to be
noted that the statistical power of these association
tests may be still considered weak. Therefore, we have
to recommend further bigger studies in multiplex
families using segregation and linkage analysis.

Figure 2. PCR amplification for IL-1Ra VNTR in intron 2
showing two alleles: band size 410 bp for A1 allele and band size
240 bp for A2 allele. Lane M shows DNA size marker. Lanes 1, 4, 7
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Results
Compared to controls, cases with T1D showed
significant higher frequency of TNF-a2308 AA
genotype ( p , 0.001, OR ¼ 7.91, 95% CI ¼ 3.4 –
18.3), IL-6-17CC genotype ( p , 0.05, OR ¼ 3.36,
95% CI ¼ 1.12 – 10.1) and IL-1Ra A1A1 genotype

show þ ve bands of A1 and A2 alleles, i.e. A1A2 genotype, lanes 2, ( p , 0.05, OR ¼ 3.68, 95% CI ¼ 1.56 – 8.68);
5, 6 show only one band of A1 allele giving A1A1 genotype while (Table I).
lane 3 is a negative control sample with no DNA showing failure of
Regarding allele frequencies, total cases showed
amplification.
significant higher frequency of TNF-a2308 allele A
( p , 0.05, OR ¼ 2.0, 95% CI ¼ 1.2 –3.3), IL-1Ra
purchased from QiaGene (Valencia, CA, USA). The
allele A1 ( p , 0.05, OR ¼ 2.98, 95% CI ¼ 1.33 –
assay was performed in Techne-Genius thermal-cycler
6.66) with a significant lower frequency of TNF-a2308
(Cambridge, England). We were careful to prepare
G allele and IL-1Ra allele A2 ( p , 0.05; Table II).
PCR master mixes applied for multiple cases and for
In the meantime, these cases showed significant
different polymorphisms together with positive and
lower frequency of TNF-a2308 GA and IL-1Ra A1A2
negative controls at the same sitting to obtain accurate
For personal use only.

genotypes ( p , 0.001 and , 0.05, respectively) with

subject genotyping. Detection of amplified fragments
was done through electrophoresis on 2% agarose after
staining with ethidium bromide and visualization with
UV light (Figures 1 and 2).
Statistical analysis
Data were processed and analyzed using the Statistical
Package of Social Science (SPSS, version 15).
The frequency of studied allelic polymorphisms
among cases was compared to that of controls
describing number and percent of each and tested
no significant difference in IL-1021082 (G/A) poly-
morphic genotypes or alleles ( p . 0.05).
Analysis of combined genotypes showed that cases
had higher significant frequency of combined geno-
type of IL-1Ra A1A1 and TNF-a2308 AA ( p , 0.001,
OR ¼ 14.6, 95% CI ¼ 5.1 – 42.1) followed by
IL-6 2174 C/C and TNF-a 2308 AA ( p , 0.05,
OR ¼ 5.3, 95% CI ¼ 1.09 – 28.6) followed by
IL-6 2174 CC and IL-1Ra A1A1 ( p , 0.05,
OR ¼ 3.8, 95% CI ¼ 1.2 – 13.8). Comparing differ-
ent genotype and allele frequencies in between sub-
groups with respect to age (, 18 years vs. . 18 years),

Table I. IL-62147, IL-1021082, TNF-a2308 and IL-1Ra VNTR genotype frequencies among cases with type 1 diabetes compared to controls
with their statistical significance.

Genotypes Cases, n ¼ 50 (%) Controls, n ¼ 98 (%) OR (95% CI)

IL-6-147
GG 3 (6.0) 5 (5.1) 1.19 (0.27–5.2)
GC 38 (76.0) 87 (88.8) 0.40 (0.16–0.98)
CC 9 (18.0)* 6 (6.1) 3.36 (1.12–10.1)
IL-1021082
GG 5 (10.0) 5 (5.1) 2.17 (0.67–7.51)
GA 40 (80.0) 85 (86.7) 0.61 (0.24–1.51)
AA 5 (10.0) 8 (8.2) 1.25 (0.38–4.04)
TNF-a2308
GG 6 (12.0) 6 (6.1) 2.1 (0.6–6.7)
GA 19 (38.0)** 81 (82.7) 0.1 (0.1–0.3)
AA 25 (50.0)** 11 (11.2) 7.9 (3.4–18.3)
IL-1Ra VNTR
A1A1 42 (84.0)* 57 (58.2) 3.68 (1.56–8.68)
A1A2 8 (16.0)* 41 (41.80) 0.27 (0.11–0.63)

* p , 0.05; ** p , 0.001. OR (95th CI) ¼ odds ratio (95% confidence interval)

 Cytokine gene polymorphism in T1D 53

Table II. IL-62147, IL-1021082, TNF-a2308 and IL-1Ra VNTR reported no association of IL-Ra polymorphisms with
allele frequencies among cases with type 1 diabetes compared to T1D [27] contrasting what was reported before in the
controls with their statistical significance. same population of increased frequency of the A1A1
Cases, n ¼ 100 Controls, n ¼ 196 OR genotype with T1D [28–30].
Alleles (%) (%) (95% CI) Regarding IL-6, our results were in agreement to
what was previously reported in Hungary that T1D
IL-6-147
G 44 (44.0) 97 (49.5) 0.80 (0.59 –1.30)
was significantly associated with IL-62174 CC geno-
C 56 (56.0) 99 (50.5) 1.24 (0.87 –2.02) type [31,32]. Also, English researcher found that the
IL-1021082 IL-62174 CC genotype was significantly different
G 50 (50.0) 95 (48.5) 1.16 (0.65 –1.72) considering age and sex of studied cases [10].
A 50 (50.0) 101 (51.5) 0.94 (0.58 –1.52) However, in another study among Caucasoid patients
TNF-a2308
G 31 (31.0)* 93 (47.4) 0.5 (0.3–0.8)
with T1D, homozygous IL-6 GG genotype was found
A 69 (69.0)* 103 (52.6) 2.0 (1.2–3.3) increased in the patients compared to the normal
IL-1Ra VNTR controls [33].
A1 92 (92.0)* 154 (79.4) 2.98 (1.33 –6.66) Regarding IL-10 gene polymorphism at position
A2 8 (8.0)* 42 (20.6) 0.33 (0.15 –0.74) 2 1082, we have found no significant difference of
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genotype and alleles frequencies among cases com-

* p , 0.05. OR (95% CI) ¼ odds ratio (95th confidence interval)
sex (males vs. females), consanguinity pattern, and
family history revealed no significant difference (data
not shown).
Discussion
The presence of inherited immunoregulatory dysfunc-
tion including the possibility of stimulation of certain
For personal use only.
pared to controls. These results are in agreement with
that reported by Reynier et al. [34]—in a French
study—that IL-1021082 polymorphism may contrib-
ute, but to a minor extent, to disease susceptibility in
juvenile T1D and should not be included in the
routine genetic screening of high-risk individuals.
Also, Ide et al. [35] in a study on Japanese cases
demonstrated that IL-10 gene promoter region
polymorphisms were not associated with genetic

cytokines resulting in a specific clinical behavior is well
reported in T1D. Cytokines largely influence the
immunologic function, initiation and progression of
the disease [22].
The most prominent feature from the current study is
the significant higher frequencies of TNF-a2308 AA
and IL-1Ra A1A1 genotypes with consequent higher
frequencies of TNF-a2308 A and IL-1Ra A1 alleles
among Egyptian cases of T1D. IL-62174 CC genotype
frequency was also high among cases but with no
significant increase in the frequency of C allele that
seems to be due to its association with the above-
mentioned genotypes. Homozygosity in these cases—
due to either parental consanguinity or random
relations—is obviously augmenting the effects of these
risk alleles and is a feature of the disease. On the other
hand, significant lower frequency was observed with
heterozygous genotypes of TNF-a2308 GA and IL-1Ra
A1A2 that may be considered a low risk or protective
genotypes. The same was previously reported by
Kumar et al. [23] that TNF-a2308 AA genotype was
significantly increased in North Indian patients with
T1D compared with the controls. Also, in a study
among Polish subjects, authors noted that TNF-a2308
GA genotype was found only in cases but not in controls
[24]. On the other hand, Deng et al. [25] reported that
no primary association was found between the 2308
polymorphism in TNF-a promoter region and T1D
cases from both US Caucasians and Chinese in Taiwan.
Likewise, polymorphisms in the IL-1 receptor genes
were found to have an association with T1D among
Dalmatian population [26]. However, Danish authors
susceptibility to T1D. In a previous study by the
same group in Japan, they reported that only ages
older than 18 years showed significant higher
frequency of AA genotype [36]. In addition, a minor
role was assumed to IL-10 genotypes in the
autoimmune diabetes risk in Spanish T1D patients
[37]. However, Polish authors demonstrated that
IL-1021082 polymorphism is related to T1D especially
AA genotypes [24]. Controversies may be explained
by the assumption that these genotypes are population
specific and co-segregate with the disease genes in
different forms among different ethnic groups.
Based on this study, we can conclude that cytokine
gene polymorphisms related to TNF-a and IL-1Ra
genes may be considered as genetic markers for T1D
among Egyptian cases with potential benefit regarding
family counseling and disease management.
Declaration of interest: Authors declare absolute
freedom of any conflict of interest related to this work.
Note
†
Visiting Professor, Qassim University, BO 6040, Buraydah 51432,
Saudi Arabia. Tel. 966 6 3851317.
References
[1] Mathis D, Vence L, Benoist C. Beta-cell death during
progression to diabetes. Nature 2001;414(6865):792–798.
[2] Pitkaniemi J, Moltchanova E, Haapala L, Harjutsalo V,
Tuomilehto J, Hakulinen T. Genetic random effects model for
 54 A. Settin et al.
family data with long-term survivors: Analysis of diabetic
nephropathy in type 1 diabetes. Genet Epidemiol 2007;31(7):
697–708.
[3] Soria J, Garagorri JM, Rodriguez M, Rodriguez G, Larrad L,
Elizalde M. Epidemiology and genetic risk of type 1 diabetes
among children in Aragon community, Spain. Diabetes Res
Clin Pract 2007;79(1):112–116.
[4] Berg AK, Korsgren O, Frisk G. Induction of the chemokine
interferon-gamma-inducible protein-10 in human pancreatic
islets during enterovirus infection. Diabetologia 2006;49(11):
2697–2703.
[5] Christen U. Chemokines as drug targets in type 1 diabetes.
Endocr Metab Immune Disord Drug Targets 2007;7(1):7–12.
[6] Reddy S, Young M. IL-1ß expression in islet cells of mouse and
its spatial relationship to beta cells and inducible nitric oxide
synthase. Ann N Y Acad Sci 2002;958:190–193.
[7] Kainonen E, Rautava S, Korkeamaki M, Isolauri E. Unique
cytokine secretion profile in children with both type I diabetes
and asthma distinct from that of solely diabetic or asthmatic
Autoimmunity Downloaded from informahealthcare.com by University of Virginia on 12/11/14
children. Cytokine 2006;34(3–4):198–205.
[8] Ramadas RA, Li X, Shubitowski DM, Samineni S,
Wills-Karp M, Ewart SL. IL-1 receptor antagonist as a
positional candidate gene in a murine model of allergic
asthma. Immunogenetics 2006;58(10):851 –855.
[9] Tarlow JK, Blakemore AI, Lennard A, Solari R, Hughes HN,
Steinkasserer A, Duff GW. Polymorphism in human IL-1
receptor antagonist gene intron 2 is caused by variable numbers
of an 86-bp tandem repeat. Hum Genet 1993;91:403–404.
[10] Gillespie KM, Nolsoe R, Betin VM, Kristiansen OP,
Bingley PJ, Mandrup-Poulsen T, Gale EA. Is puberty an
accelerator of type 1 diabetes in IL62174CC females? Diabetes
For personal use only.
2005;54(4):1245–1248.
[11] Wastowski IJ, Peres NT, Simões RT, Castelli EC, Simões AL,
Martinez-Rossi NM, Donadi EA. Identification of a novel
120 bp allele at the TNFd microsatellite locus. Tissue Antigens
2006;67(4):318–320.
[12] Hajeer AH, Hutchinson IV. Influence of TNFalpha gene
polymorphisms on TNFalpha production and disease. Hum
Immunol 2001;62(11):1191–1199.
[13] Fishman D, Faulds G, Jeffery R, Mohamed-Ali V, Yudkin JS,
Humphries S, Woo P. The effect of novel polymorphisms in
the interleukin-6 (IL-6) gene on IL-6 transcription and plasma
IL-6 levels, and an association with systemic-onset juvenile
chronic arthritis. J Clin Invest 1998;102(7):1369–1376.
[14] Tripp CS, Wolf SF, Unanue ER. Interleukin 12 and tumor
necrosis factor alpha are costimulators of interferon gamma
production by natural killer cells in severe combined
immunodeficiency mice with listeriosis, and interleukin 10 is
a physiologic antagonist. Proc Natl Acad Sci USA 1993;90(8):
3725–3729.
[15] Moore KW, de Waal Malefyt R, Coffman RL, O’Garra A.
Interleukin-10 and the interleukin-10 receptor. Annu Rev
Immunol 2001;19:683–765.
[16] Yu L, Yang MS, Zhao J, Shi YY, Zhao XZ, Yang JD, Liu ZJ,
Gu NF, Feng GY, He L. An association between poly-
morphisms of the interleukin-10 gene promoter and schizo-
phrenia in the Chinese population. Schizophr Res 2004;71(1):
179–183.
[17] Crawley E, Kay R, Sillibourne J, Patel P, Hutchinson I, Woo P.
Polymorphic haplotypes of the interleukin-10 50 flanking
region determine variable interleukin-10 transcription and are
associated with particular phenotypes of juvenile rheumatoid
arthritis. Arthritis Rheum 1999;42(6):1101–1108.
[18] Cavet J, Middleton PG, Segall M, Noreen H, Davies SM,
Dickinson AM. Recipient tumor necrosis factor-alpha and
interleukin-10 gene polymorphisms associate with early
mortality and acute graft-versus-host disease severity in
HLA-matched sibling bone marrow transplants. Blood 1999;
94(11):3941–3946.
[19] Sargen K, Demaine AG, Kingsnorth AN. Cytokine gene
polymorphisms in acute pancreatitis. JOP 2000;1(2):24–35.
[20] Cavet J, Dickinson AM, Norden J, Taylor PR, Jackson GH,
Middleton PG. Interferon-gamma and interleukin-6 gene
polymorphisms associate with graft-versus-host disease in
HLA-matched sibling bone marrow transplantation. Blood
2001;98(5):1594– 1600.
[21] Wilkinson RJ, Patel P, Llewelyn M, Hirsch CS, Pasvol G,
Snounou G, Davidson RN, Toossi Z. Influence of poly-
morphism in the genes for the interleukin (IL)-1 receptor
antagonist and IL-1beta on tuberculosis. J Exp Med 1999;
189(12):1863 –1874.
[22] Greenfield EA, Nguyen KA, Kuchroo VK. CD28/B7 costi-
mulation: A review. Crit Rev Immunol 1998;18(5):389–418.
[23] Kumar R, Goswami R, Agarwal S, Israni N, Singh SK, Rani R.
Association and interaction of the TNF-alpha gene with other
pro- and anti-inflammatory cytokine genes and HLA genes in
patients with type 1 diabetes from North India. Tissue
Antigens 2007;69(6):557 –567.
[24] Siekiera U, Jarosz-Chobot P, Janusz J, Koehler B. Polymorph-
ism of TNF-alpha (308 A/G), IL-10 (1082 A/G, 819 C/T 592
A/C), IL-6 (174 G/C), and IFN-gamma (874 A/T); genetically
conditioned cytokine synthesis level in children with diabetes
type 1. Endokrynol Diabetol Chor Przemiany Materii Wieku
Rozw 2002;8(1):29–34.
[25] Deng GY, Maclaren NK, Huang HS, Zhang LP, She JX. No
primary association between the 308 polymorphism in the tumor
necrosis factor alpha promoter region and insulin-dependent
diabetes mellitus. Hum Immunol 1996;45(2):137–142.
[26] Zemunik T, Skrabic V, Boraska V, Diklic D, Terzic IM,
Capkun V, Peruzovic M, Terzic J. FokI polymorphism, vitamin
D receptor, and interleukin-1 receptor haplotypes are
associated with type 1 diabetes in the Dalmatian population.
J Mol Diagn 2005;7(5):600–604.
[27] Kristiansen OP, Pociot F, Johannesen J, Bergholdt R,
Dinarello CA, Nerup J. Linkage disequilibrium testing of
four interleukin-1 gene-cluster polymorphisms in Danish
multiplex families with insulin-dependent diabetes mellitus.
Cytokine 2000;12(2):171 –175.
[28] Mandrup-Poulsen T, Pociot F, Molvig J, Shapiro L, Nilsson P,
Emdal T, Roder M, Kjems LL, Dinarello CA, Nerup J.
Monokine antagonism is reduced in patients with IDDM.
Diabetes 1994;43(10):1242–1247.
[29] Pociot F, Ronningen KS, Bergholdt R, Lorenzen T,
Johannesen J, Ye K, Dinarello CA, Nerup J. Genetic
susceptibility markers in Danish patients with type 1
(insulin-dependent) diabetes—evidence for polygenicity in
man. Danish Study Group of Diabetes in childhood.
Autoimmunity 1994;19(3):169–178.
[30] Mandrup-Poulsen T. The role of interleukin-1 in the
pathogenesis of IDDM. Diabetologia 1996;39(9):1005– 1029.
[31] Ladner Martha B, Valdes Ana M, Bottini Nunzio, Cho Eunji,
Noble Janelle A. PTPN22, CTLA4, MIF-1, IL6 and TCF7
polymorphisms and type 1 diabetes. Genetics 2004;5:1.
[32] Hermann C, Krikovszky D, Füst G, Kovács M, Körner A,
Szabó A, Vannay A, Madácsy L. Association between
interleukin-6 polymorphism and age-at-onset of type 1
diabetes. Epistatic influences of the tumor necrosis factor-
alpha and interleukin-1beta polymorphisms. Eur Cytokine
Netw 2005;16(4):277–281.
[33] Jahromi MM, Millward BA, Demaine AG. A polymorphism in
the promoter region of the gene for interleukin-6 is associated
with susceptibility to type 1 diabetes mellitus. J Interferon
Cytokine Res 2000;20(10):885 –888.
[34] Reynier F, Cazalis MA, Lecoq A, Paye M, Rosa A, Durand A,
Jhumka U, Mougin B, Miossec P, Bendelac N, Nicolino M,
Thivolet C. Lack of association of IL-10 promoter gene
variants with type 1 diabetes in a French population. Hum
Immunol 2006;67(4–5):311–317.

[35] Ide A, Kawasaki E, Abiru N, Sun F, Fukushima T, Ishii R,
Takahashi R, Kuwahara H, Fujita N, Kita A, Imaizumi M,
Oshima K, Usa T, Uotani S, Ejima E, Yamasaki H, Ashizawa K,
Yamaguchi Y, Eguchi K. Interleukin-10 gene promoter region
polymorphisms in patients with type 1 diabetes and autoimmune
thyroid disease. Ann NY Acad Sci 2003;1005:344–347.
[36] Ide A, Kawasaki E, Abiru N, Sun F, Takahashi R, Kuwahara
H, Fujita N, Kita A, Oshima K, Sakamaki H, Uotani S,
Autoimmunity Downloaded from informahealthcare.com by University of Virginia on 12/11/14
For personal use only.
Cytokine gene polymorphism in T1D 55
Yamasaki H, Yamaguchi Y, Eguchi K. Genetic association
between interleukin-10 gene promoter region polymorphisms
and type 1 diabetes age-at-onset. Hum Immunol 2002;63(8):
690– 695.
[37] Urcelay E, Santiago JL, de la Calle H, Martı́nez A, Figueredo
A, Fernández-Arquero M, de la Concha EG. Interleukin-10
polymorphisms in Spanish type 1 diabetes patients. Genes
Immun 2004;5(4):306–309.