Association of IL-1b +3954 C/T and IL-10-1082 G/A

Cytokine Gene Polymorphisms with Susceptibility to

Tuberculosis
P. Meenakshi*, S. Ramya*, T. Shruthi*, J. Lavanya*, H. H. Mohammed*, S. A. Mohammed*,
V. Vijayalakshmi* & G. Sumanlatha*
*Bhagwan Mahavir Medical Research Centre,
Hyderabad, Andhra Pradesh, India; and LEPRA
IndiaBlue Peter Public Health & Research
Centre, Cherlapally, Hyderabad, Andhra Pradesh,
India
Received 28 February 2013; Accepted in revised
form 25 March 2013
Correspondence to: Dr G. Sumanlatha, Scientist,
Immunology Department, Bhagwan Mahavir
Medical Research Centre, A.C. Guards,
Hyderabad, Andhra Pradesh 500 004, India.
E-mail: sumanlathag@yahoo.com
Abstract
Tuberculosis (TB) constitutes the major cause of death due to infectious diseases.
Cytokines play a major role in defence against Mycobacterium tuberculosis
infection. Polymorphisms in the genes encoding various cytokines have been
associated with tuberculosis susceptibility. Household contacts (HHC) are at
increased risk of developing the disease. In this study, we examined the
association of IL-1b and IL-10 cytokine gene polymorphisms with risk of
developing tuberculosis in TB patients, their HHC and healthy controls (HC)
using JavaStat and SPSS. Multifactor dimensionality reduction (MDR) analyses
were performed to explore the potential genegene interactions. The genotype
and allele frequencies of IL-1b +3954C/T polymorphism did not vary
signicantly between TB patients and HC. GG (P < 0.005, OR = 0.219 and
95% CI = 0.0590.735) and GA (P < 0.0001, OR = 2.938 and 95%
CI = 1.5265.696) genotypes of IL-10-1082 G/A polymorphism were found
to be signicantly associated with patients versus HC. HHC with CC (P < 0.03,
OR = 1.833 and 95% CI = 1.13.35) genotype in IL-1b and GA (P < 0.0001,
OR = 4.612 and 95% CI = 2.2259.702) genotype in IL-10 were at increased
risk of developing tuberculosis. MDR tests revealed high-risk genotypes in IL-1b
and IL-10 based on the association model. Our results demonstrate that the
polymorphisms of IL-1b and IL-10 genes may be valuable markers to predict the
risk for the development of TB in household contacts.
Introduction
Tuberculosis, primarily caused by Mycobacterium tuberculosis
(M.tb), is one of the leading causes of morbidity and
mortality worldwide despite the existence of National and
International control programmes [1, 2]. Recent data from
the World Health Organization show that about 8.59.2
million new cases arise annually, and eventually 1.21.5
million deaths occur every year [3]. It is estimated that one-
third of the worlds population is infected with M.tb, while
10% of those infected develop clinical disease [4]. This
suggests that besides Mycobacteria itself, the host genetic
factors may determine the differences in host susceptibility
to tuberculosis (TB) [5].
Several reports from different countries have shown that
household contacts of tuberculosis are at much higher risk
of infection that range from 3080% depending on the
intensity of tuberculosis disease transmission [69]. Iden-
tication of these high-risk individuals in recently exposed
or infected individuals is of great importance for reducing
the disease burden in the community [10].
Although environmental factors are important deter-
minants of progression to disease, there is a genetic
component underlying susceptibility to TB, the basis of
which may vary in different populations [11]. Manifesta-
tion of clinical TB depends on balance between T helper 1
(Th1) cytokines associated with resistance to infection and
Th2 cytokines with progressive disease [12]. Inuence of
cytokine response may be due to their polymorphisms
leading to modication of host immunological response in
the pathogenesis of TB [13, 14]. IL-1b a pro-inamma-
tory cytokine located on chromosome 2 is mainly
produced by monocytes, macrophages and dendritic cells.
In tuberculosis patients, IL-1b is expressed in excess [15]
at the site of the disease [16]. IL1 b +3954 C to T
(rs1143634) has been associated with periodontitis [17]
and tuberculosis [18]. IL-10 a Th2 cytokine gene mapped
to chromosome 1 is a potent inhibitor of T cell function,
92 2013 John Wiley & Sons Ltd.
HUMAN I MMUNOL OGY doi: 10.1111/sji.12055
..................................................................................................................................................................
major histocompatibility complex (MHC) class II expres-
sion, antigen specic proliferation and IFN-c synthesis
[19]. Interindividual variations in IL-10 production are
genetically contributed by polymorphisms within the
IL-10 promoter (rs1800896) [20]. The polymorphism
at position -1082 may affect the binding of this
transcriptional factor and therefore alter transcriptional
activation [21].
The aim of this study was to determine the association
of IL-1b +3954 C/T and IL-10-1082 G/A gene polymor-
phisms susceptible to tuberculosis in patients and their
household contacts.
Materials and methods
Subjects
A total of 300 subjects were included in the study which
consists of tuberculosis patients, their household contacts
(HHC) and agesex-matched healthy controls (HC) of
100 each group. Patients who attended free chest clinic
at Mahavir Hospital (PPM-DOTS) were recruited based
on radiographic examination, sputum culture for acid-
fast bacilli (AFB) and histocytological examination.
Tuberculin skin test (TST) positivity was assessed both
in patients and household contacts by administering 5
tuberculin units (TU) intradermally on the volar surface
of the left arm. An induration of >10 mm within 48
72 h was considered positive (TST+). In healthy
controls, TST was not performed. Body Mass Index
(BMI) was calculated in all the subjects. The study was
approved by the Institutional Ethics Committee, and
written informed consent was obtained from each
participant.
DNA extraction
Genomic DNA was extracted from venous blood (12 ml)
using DNA isolation kit (Flexi gene DNA isolation kit)
according to the manufacturers protocol. Quantity and
quality of DNA was conrmed by spectrophotometer
(Thermo scientic), and DNA was stored at 20 C.
Genotyping
IL-1b + 3954C/T genotyping
The IL-1b +3954 C/T was genotyped by restriction
fragment length polymorphism (RFLP) where a 249-bp
fragment of the IL-1b exon 5 was amplied using forward
primer 5-gtt gtc atc aga ctt tga cc-3 and reverse primer
5-ttc agt tca tat gga cca ga-3 in a 20ll reaction. The
mixture was amplied for three cycles of 95 C for 4 min,
then 30 cycles of 95 C for 30 s, 59 C for 30 s, 72 C for
30 s and then a nal 4 min at 72 C. The products were
digested overnight at 65 C with 2.5 U Taq 1 and run on a
2% agarose gel, generating the following patterns: single
band of 249 bp, TT homozygote; two bands at 135 and
114 bp, CC homozygote; all three bands, CT heterozygote
(Fig. 1A).
IL-10-1082 G/A genotyping
IL-10-1082 G/A polymorphism was genotyped by ampli-
cation refractory mutation system polymerase chain
reaction (ARMS-PCR) method. It was performed where a
161-bp fragment of the IL-10 was amplied using primer
sequences: a common antisense primer 5-gta agc ttc tgt
ggc tgg agtc-3 and two sense primers 5-aac act act aag gct
tct ttg ggt g -3 and 5-aac act act aag gct tct ttg ggt a-3.
The PCR was performed for 1 cycle of 94 C for 3 min,
then 30 cycles of 94 C for 30 s, 60 C for 30 s, 72 C for
45 s and then a nal 3 min at 72 C. PCR products were
run on 1.5% gel stained with ethidium bromide and
visualized by UV lightgenerating bands (Fig. 1B).
Statistical analysis
Pearsons v
2
test was used to examine differences in
characteristic variables and the distribution of genetic
polymorphisms. Odds ratio (O.R) and 95% condence
interval (CI) were calculated using JAVASTAT. All
epidemiologic variables were determined using IBM SPSS
Statistics 20 software, where students t-test is used to
evaluate continuous variables, and v
2
test, for categorical
variables. The genegene interaction for SNPs was analysed
by nonparametric multifactor dimensionality reduction
(MDR version 2.0 beta 8.4) analysis. Distribution of alleles
and deviation of genotype frequencies were tested by using
HardyWeinberg equilibrium (HWE). P < 0.05 was
considered to be statistically signicant for all the tests
except HWE. Bonferroni correction, an adjustment made
to P values, was used to reduce the chances of obtaining
false-positive results (P < 0.0005).
Results
Demographic characteristics
The demographic prole of tuberculosis cohort was
studied. The mean age of the patients (50 males and 50
females), their HHC (44 males and 56 females) and HC
(54 males and 46 females) was 27.4 13.9, 34.8 10.7
and 30 10.7, respectively. TST positivity was observed
in patients and HHC with a signicance of P < 0.0001.
Mean BMI was found to be 16.8 4.25, 22.6 6.85
and 23.7 4.09 in patients, HHC and HC, respectively,
and there was signicant difference in patients versus HHC
and patients versus HC (P < 0.001 and P < 0.0001)
(Table 1).
2013 John Wiley & Sons Ltd.
P. Meenakshi et al. IL-1b and IL-10 SNPs in TB Patients and their HHC 93
..................................................................................................................................................................
Genetic analysis
The genotype frequencies of IL-1 b (+3954 C/T) poly-
morphism did not vary signicantly between TB patients
and HC (P < 0.32, 0.395 and 0.89 for CC, CT and TT
respectively). CC genotype was found to be signicantly
associated with HHC versus HC (P < 0.03, OR = 1.833
and 95% CI = 1.13.35) while Bonferroni correction was
not signicant. Frequency of alleles did not differ signif-
icantly in all the subjects with T allele more frequently
found when compared with the C allele (Table 2). IL-1b
(+3954 C/T) was found to be in HardyWeinberg
equilibrium with P > 0.05 (v
2
= 0.08).
In IL-10-1082 G/A polymorphism, GG (P < 0.005,
OR = 0.219 and 95% CI = 0.0590.735) and GA
(P < 0.0001, OR = 2.938 and 95% CI = 1.5265.696)
genotypes were found to be signicantly associated with
patients versus HC. GA (P < 0.0001, OR = 0.194 and
95% CI = 0.0690.516) and AA (P < 0.0001,
OR = 4.612 and 95% CI = 2.2259.702) genotypes in
HHC versus HC have shown signicant association. Allele
frequency was found to be similar in all the subjects
(Table 3). IL-10-1082 G/A polymorphism deviates from
the HardyWeinberg equilibrium due to the excess
number of heterozygotes where P < 0.05.
Genotype combinations for IL-1b and IL-10 genes in
patients, HHC and HC were studied by MDR analysis. All
the genotypes of IL-1b have shown high risk with GA
genotype of IL-10 in patients versus HC and HHC versus
HC with GG and AA genotypes. In patients versus HHC,
Table 1 Epidemiological characteristics of patients, household contacts (HHC) and healthy controls (HC).
Variable Patients (n = 100) HHC (n = 100) HC (n = 100) P value
Age 27.4 13.9 34.8 10.7 30 10.7 0.15* Pts versus HHC
0.17*pts versus HC
Gender M/F 50(50%)/50(50%) 44(44%)/56(56%) 54 (54%)/46 (46%) 0.0001+
TST p/n 1.87 0.34 1.73 0.45 0.0001+
BMI (kg/m
2
) 16.8 4.25 22.6 6.85 23.74.09 0.001* Pts versus HHC 0.001* HHC
versus HC
*Mean SD, independent samples t-test, + v
2
tests.
p/n, positive/negative; TST, tuberculin skin test; BMI, body mass index.
1 2 3 4 5 6 7 8 9 10 11 12 13
Lane 1- 100bp ladder; Lane 2 TT genotype; Lane 3-6,12 CC genotype;
Lanes 7-11, 13 CT genotype
100bp
1000bp
249bp
135bp
114bp
1 2 3 4 5 6 7 8 9 10 11 12 13
Lane 1 100bp Ladder; Lane 4&5, 6&7,10&11 GA genotype :Lane 2 &3 to 8& 9
AA genotype; Lane 12&13 GG genotype
161bp
1000bp
100bp
A
B
Figure 1 (A) Agarose gel picture showing
amplied RFLP-PCR products of IL-1b
+3953C/T Polymorphism. (B) Agarose gel
picture showing amplied ARMS PCR
products of IL-10 -1082 G/A polymorphism.
Scandinavian Journal of Immunology, 2013, 78, 9297
94 IL-1b and IL-10 SNPs in TB Patients and their HHC P. Meenakshi et al.
..................................................................................................................................................................
high risk was observed between CC and CT genotypes of
IL-1 b and GA genotype of IL-10 (Fig. 2).
Discussion
Host genetic factors may be important determinants of
susceptibility to tuberculosis, and several candidate gene
polymorphisms have shown variable associations with
severity of tuberculosis disease in different populations
[22, 23]. IL-1b participates in aberrant immune responses
in lung diseases but controls M.tb infection [24]. It
regulates inammatory reaction and immune response
through promoting other cytokine expressions, such as
IL-6 and IL-12.
In the present study, IL-1b +3954 C/T polymorphism
was not found to be associated with tuberculosis suscep-
tibility. The distribution of their genotypes and alleles did
not signicantly differ between the patients and healthy
controls in concordance with studies in London on
idiopathic pulmonary brosis patients [25], in Gambian
population [26] and in Gujarat Asians in east London with
tuberculosis [27]. Studies in other diseases like hypo-
gammaglobulinaemia, autoimmunity, cancers [28] and
asthma [29] have shown similar results, whereas in contrast
to our study IL-1b +3954 C/T polymorphism have shown
an association with extrapulmonary tuberculosis in
American population [30], in Gambian population with
malaria [31] and in Turkish population with behcets
disease [32].
IL-10 considered as a key mediator of immunosup-
pression, and tolerance appears to be primarily produced
by monocytes and T regulatory lymphocytes. It converts
human dendritic cells into macrophage-like cells with
increased antimycobacterial activity. Modulation of T cell
responses by IL-10 inuences the host susceptibility to
TB [33].
Our study reported the association of IL-10-1082 G/A
polymorphism with tuberculosis. Earlier studies in the
Hong Kong, Chinese [34], Colombian [35], Spanish,
Turkish and Cambodian populations [36] have also shown
the same. The GG genotype was signicantly associated
with the present study and also in Colombian population,
whereas in the Tunisian[37], Iranian [38], West African
[39], Macedonian [40] Gambian [18], Spanish [41] and
Korean population [42], it was not associated. The
frequency of GA genotype which is 81% in our study
was found to be similar in Iranian population (82.5%).
Signicant difference was not observed with the allele
Table 2 Analysis of IL-1 b (+3953C/T) genotype and allele frequencies in Tuberculosis patients, their household contacts (HHC) and healthy controls
(HC).
HC Patients HHC
n = 100 n = 100 Frequency (%) OR P value 95% CI n = 100 Frequency (%) OR P value 95% CI
Genotype
CC 40 47 47 1.33 0.32 0.7312.424 55 55 1.833 0.03* 1.13.35
CT 50 44 44 0.786 0.395 0.4331.424 37 37 0.587 0.06 0.3211.073
TT 10 9 9 0.89 0.89 0.3142.51 8 8 0.783 0.62 0.2672.272
Allele
C 130 138 69 1.2 0.395 0.7731.86 147 73.5 1.493 0.06 0.9522.344
T 70 62 31 0.834 0.395 0.5381.294 53 26.5 0.67 0.06 0.4271.05
*P value <0.05 statistically signicant.
OR, odds ratio; 95% CI, condence interval.
Table 3 Analysis of IL-10 (+1082G/A) candidate gene polymorphisms in Tuberculosis patients, their household contacts (HHC) and healthy controls
(HC).
HC Patients HHC
n = 100 n = 100 Frequency(%) OR P value 95% CI n = 100 Frequency(%) OR P value 95% CI
Genotype
GG 16 4 4 0.219 0.005* 0.0590.735 7 7 0.395 0.04 0.1441.044
GA 59 81 81 2.938 <0.0001* 1.5265.696 87 87 4.612 <0.0001* 2.2259.702
AA 25 15 15 0.535 0.07 0.2551.112 6 6 0.194 <0.0001* 0.0690.516
Allele
G 91 89 44.5 0.957 0.816 0.6471.415 101 50.5 1.217 0.301 0.8241.798
A 109 111 55.5 1.045 0.816 0.7071.546 99 49.5 0.822 0.301 0.5561.214
*P < 0.05 Statistically signicant.
OR, odds ratio; 95% CI, condence interval.
2013 John Wiley & Sons Ltd.
P. Meenakshi et al. IL-1b and IL-10 SNPs in TB Patients and their HHC 95
..................................................................................................................................................................
frequency in our population similar to the Tunisian
population. In contrast to our results, other recent reports
by Mosaad et al. [43] and Akgunes et al. [44] reported
signicant association with TB susceptibility. However, A
allele was associated with Italian (Sicilian) population [45].
These contradictory ndings may be due to ethnical
differences in various populations.
In our study, household contacts with IL-1b CC
genotype and IL-10 GA genotype have shown signicant
association with tuberculosis. To our knowledge, this is the
rst study to report association of these genotypes in
household contacts.
Based on MDR analysis, high-risk combination between
IL-1b and IL-10 genes suggests that these SNPs interact
synergistically affecting signalling impairment, and hence,
effector mechanisms signicantly leading to pathogenesis
of tuberculosis.
Our study illustrates that IL-1b CC and IL-10 GG
genotypes may be useful for early detection of the disease in
high-risk individuals, that is, household contacts. How-
ever, there is a need to evaluate the data in large sample
size.
Acknowledgment
We thank Bhagwan Mahavir Trust and staff of the free
chest clinic Mahavir PPMDOTS, Tuberculosis Unit (TU).
Financial support was provided by DBT-RGYI (Sanction
no: 102/IFD/PR/2029/2007-2008 dated 18/01/2008) and
COE (Sanction No: BT/01/COE/07/02, dated 30/12/08).
References
1 Rahman S, Gudetta B, Fink J et al. Compartmentalization of immune
responses in human tuberculosis. A J Pathol 2009;174:221124.
2 Dolin PJ, Raviglione MC, Kochi A. Global tuberculosis and mortality
during 19902000. Bull World Health Organ 1994;72:21320.
3 World Health Organization. The World Health Report 2011changing
history. Geneva: World Health Organization, 2011.
4 Schluger NW. Recent advances in our understanding of human host
responses to tuberculosis. Respir Res 2001;2:15763.
5 Bellamy R. Susceptibility to Mycobacterial infections:the importance
of host genetics. Genes Immun 2003;4:411.
6 Akhtar S, Carpenter TE, Rathi SK. A chain binomial model for intra
household spread of Mycobacterium tuberculosis in a low socio-economic
setting in Pakistan. Epidemiol Infect 2007;135:2733.
7 Dhingra VK, Rajpal S, Aggarwal N, Taneja DK. Tuberculosis trend
among household contacts of TB patients. Indian J Commun Med
2004;29:13.
8 Guwatudde D, Nakakeeto M, Jones-Lopez EC et al. Tuberculosis in
household contacts of infectious cases in Kampala,Uganda. Am J
Epidemiol 2003;158:88798.
9 Lemos AC, Matos ED, Pedral-Sampaio DB, Netto EM. Risk of
tuberculosis among household contacts in Salvador, Bahia. Braz J
Infect Dis 2004;8:42430.
10 Ansari A, Talat N, Jamil B et al. Cytokine gene polymorphisms across
tuberculosis clinical spectrum in Pakistani patients. PLoS ONE
2009;4:e4778.
A B
C
Figure 2 The distribution of high risk & low
risk genotypes of IL-1b & IL-10 genes in
Patients, their Household Contacts (HHC) &
Healthy Controls (HC). Dark boxes represent
high risk combinations of genotypes. Light
shaded boxes represent low-risk combinations.
White boxes represent unclassied data. Bars
represent (A) patients(left) versus healthy
controls(right); (B) patients(left) versus
household contacts(right) & (C) household
contacts(left) versus ealthy controls (right).
Above the boxes 0,1,2 represents CC,CT,TT
genotypes of IL-1b & GG,GA,AA genotypes
of IL-10 respectively.
Scandinavian Journal of Immunology, 2013, 78, 9297
96 IL-1b and IL-10 SNPs in TB Patients and their HHC P. Meenakshi et al.
..................................................................................................................................................................
11 Britton WJ, Fernando SL, Saunders BM, Sluyter R, Wiley JS. The
genetic control of susceptibility to Mycobacterium tuberculosis. Novartis
Found Symp 2007;281:7989.
12 Wallis RS, Ellner JJ. Cytokines and tuberculosis. J Leukoc Biol
1994;55:67681.
13 Pravica V, Asderakis A, Perrey C, Hajeer A, Sinnott PJ, Hutchinson
IV. In vitro production of IFN-c correlates with CA repeat
polymorphism in the human IFN-c gene. Eur J Immunogenet
1999;28:13.
14 Sher A, Coffman RL. Regulation of immunity to parasites by T cells
and T cell -derived cytokines. Annu Rev Immunol 1992;10:385409.
15 Schauf V, Rom WN, Smith KA et al. Cytokine gene activation and
modied responsiveness to interleukin-2 in the blood of tuberculosis
patients. J Infect Dis 1993;168:10569.
16 Law K, Weiden M, Harkin T, Tchou-Wong K, Chi C, Rom WN.
Increased release of interleukin-1 beta, interleukin-6, and tumor
necrosis factor-alpha by bronchoalveolar cells lavaged from involved
sites in pulmonary tuberculosis. Am J Respir Crit Care Med
1996;153:799804.
17 McDevitt MJ, Wang HY, Knobelman C et al. Interleukin-1 genetic
association with periodontitis in clinical practice. J Periodontol
2000;71:15663.
18 Bellamy R, Ruwende C, Corrah T, McAdam KP, Whittle HC, Hill
AV. Assessment of the interleukin 1 gene cluster and other candidate
gene polymorphisms in host susceptibility to tuberculosis. Tuberc
Lung Dis 1998;79:839.
19 Howard M, Garra AO. Biological properties of IL-10. Immunol Today
1992;13:19880.
20 Kim JM, Brannan CI, Copeland NG, Jenkins NA, Khan TA, Moore
KW. Structure of the mouse IL-10 gene and chromosomal localization
of the mouse and human genes. J Immunol 1992;148:361823.
21 Moller M, de Wit E, Hoal EG. Past, present and future directions in
human genetic susceptibility to tuberculosis. FEMS Immunol Med
Microbiol 2010;58:326.
22 Van Helden PD, Moller M, Babb C et al. TB epidemiology and
human genetics. Novartis Found Symp 2006;279:1731.
23 Yim JJ, Selveraj P. Genetic susceptibility in tuberculosis. Respirology
2010;15:24156.
24 Kline JN, Schwartz DA, Monick MM, Floerchinger CS, Hunninghake
GW. Relative release of interleukin-1 beta and interleukin-1 receptor
antagonist by alveolar macrophages. A study in asbestos-induced lung
disease, sarcoidosis, and idiopathic pulmonary brosis. Chest
1993;104:4753.
25 Hutyrova B, Pantelidis P, Drabek J. Interleukin-1 gene cluster
polymorphisms in sarcoidosis and idiopathic pulmonary brosis. Am J
Respir Crit Care Med 2002;165:14851.
26 Bellamy RC, Ruwende T, Corrah KP et al. Assessment of the
interleukin 1 gene cluster and other candidate gene polymorphisms
in host susceptibility to tuberculosis. Tuberc Lung Dis 1998;79:
8390.
27 Wilkinson RJ, Patel P, Llewelyn M. Inuence of Polymorphism in
the genes for the interleukin (IL)-1 receptor antagonist and IL-1 b on
tuberculosis. J Exp Med 1999;189:1863187.
28 Rezaei N, Amirzargar AA, Shakiba Y, Mahmoudi M, Moradiand B,
Aghamohammadi A. Proinammatory cytokine gene single
nucleotide polymorphisms in common variable immunodeciency.
Clin Exp Immunol 2009;155:217.
29 Trajkov D, Mirkovska-Stojkovikj J, Arsov T et al. Association of
cytokine gene polymorphisms with bronchial asthma in Macedonians.
Iran J Allergy Asthma Immunol 2008;7:14356.
30 Motsinger-Reif AA, Antas PR, Oki NO, Levy S, Holland SM,
Sterling TR. Polymorphisms in IL-1b, vitamin D receptor Fok1, and
Toll-like receptor 2 are associated with extrapulmonary tuberculosis.
BMC Med Genet 2010;11:3741.
31 Walley AJ, Aucan C, Kwiatkowski D, Hill AV. Interleukin-1 gene
cluster polymorphisms and susceptibility to clinical malaria in a
Gambian casecontrol study. Eur J Hum Genet 2003;10:17.
32 Coskun M, Bacanli A, Sallakci N, Alpsoy E, Yavuzer U, Yegin O.
Specic interleukin-1 gene polymorphisms in Turkish patients with
Behc ets disease. Exp Dermatol 2005;14:1249.
33 Fortsch D, Rollinghoff M, Stenger S. IL-10 converts human dendritic
cells into macrophage-like cells with increased antibacterial activity
against virulent Mycobacterium tuberculosis. J Immunol 2000;165:97887.
34 Tso HW, Ip WK, Chong WP, Tam CM, Chiang AK, Lau YL.
Association of interferon gamma and interleukin 10 genes with
tuberculosis in Hong Kong Chinese. Genes Immun 2005;6:35863.
35 Henao MI, Montes C, Pars SC, Garca LF. Cytokine gene polymor-
phisms in Colombian patients with different clinical presentations of
tuberculosis. Tuberculosis 2006;86:119.
36 Delgado JC, Baena A, Thim S, Goldfeld AE. Ethnic-specic genetic
associations with pulmonary tuberculosis. J Infect Dis 2002;186:
14638.
37 Ben-Selma W, Harizi H, Boukadida J. Association of TNF-a and IL-
10 polymorphisms with tuberculosis in Tunisian populations.
Microbes Infect 2011;04:914.
38 Amirzargar AA, Danesh AA, Khosravi F et al. Cytokines genes
polymorphisms in Iranian patients with pulmonary tuberculosis.
Indian J Immunol 2004;1:1259.
39 Thye T, Browne EN, Chinbuah MA et al. IL10 haplotype associated
with tuberculin skin test response but not with pulmonary TB. PLoS
ONE 2009;4:e5420.
40 Trajkov D, Trajchevska M, Arsov T et al. Association of 22 cytokine
gene polymorphisms with tuberculosis in Macedonians. Indian J
Tuberc 2009;56:11731.
41 Lopez-Maderuelo D, Arnalich F, Serantes R et al. Interferon gamma
and interleukin-10 gene polymorphisms in pulmonary tuberculosis.
Am J Respir Crit Care Med 2003;167:9705.
42 Shin HD, Park BL, Kim YH, Cheong HS, Lee IH, Park SK. Common
interleukin 10 polymorphism associated with decreased risk of
tuberculosis. Exp Mol Med 2005;37:12832.
43 Mosaad YM, Soliman OE, Tawhid ZE, Sherif DM. Interferongamma
+874 T/A and Interleukin-10 -1082 A/G single nucleotide polymor-
phism in Egyptian children with tuberculosis. Scand J Immunol
2010;72:35864.
44 Akgunes A, Coban AY, Durupinar B. Human leucocyte antigens and
cytokine gene polymorphisms and tuberculosis. Indian J Med Microbiol
2011;29:2832.
45 Scola L, Crivello A, Marino V, Gioia V, Serauto A et al. IL-10 and
TNF-alpha polymorphisms in a sample of Sicilian patients affected by
tuberculosis implication for ageing and life span expectancy. Mech
Ageing Dev 2003;124:56972.
2013 John Wiley & Sons Ltd.
P. Meenakshi et al. IL-1b and IL-10 SNPs in TB Patients and their HHC 97
..................................................................................................................................................................