ISSN: 0975 -8542
Journal of Global Pharma Technology
www.jgpt.co.in Available Online at
RESEARCH ARTICLE
©2009-2018, JGPT. All Rights Reserved 318
Polymorphism and Expression of TLR2 Gene in Iraqi Pulmonary
Tuberculosis Patients
Lara Ghali A. Ali
1*
, Nawal Mohammed Utba
1
, Ahmed Asmer Mankhi
2
1. Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq.
2. National Reference Laboratory of Tuberculosis, Ministry of Health, Baghdad, Iraq.
*Corresponding Author: Email: laragale12@gmail.com
Abstract
Objective The innate immune system has evolved as the first line of defense against microorganisms,
which involves specific pathogen recognition receptors such as toll-like receptors (TLR). The aim of the
present study was to investigate whether the TLR2 gene polymorphism could influence the susceptibility
to pulmonary tuberculosis (TB) and its expression level as well as its concentration in patients’ sera.
Methods A total of 120 subjects, including 80 patients with TB including 40 multiple drug resistance
(MDR), 20 recently diagnosed (RD) and 20 old cases (OC) of TB patients, in addition to 40 healthy
control, were examined for TLR2 polymorphism over locus -100 microsatellite guanine-thymine repeats,
24969 (T>C) and 5305 (C>T). TLR2 gene expression was assayed using a real time PCR technique;
furthermore, sera from 68 TB patients and 20 healthy controls were assayed for TLR2 level using ELISA
technique. Results we found that the TC genotypes showed a significantly increased frequency in MDR
patients than control (35% vs.12.5%, respectively; **P = 0.034), and the associated OR value 3.77 with
EF value of 0.25. That may regard TLR-2 rs3804100 TC as TB susceptibility genotype in the investigated
Iraqi patients .Whereas the TLR-2 serum levels varied significantly (*p<0.05) between studied groups in
regard to TT genotypes. For TLR-2 rs7696323, all TB patients and control groups have CC genotypes
(Wild type). In respect to (GT)n microsatellite repeat; GT repeat alleles had insignificant differences (p >
0.05) between MDR and OC than control groups, while RD group was significantly higher [**P = 0.006]
than control group in GT(24) repeat allele, with OR value of 5.2 and EF value of 0.33. Furthermore, this
GT repeat allele associated with susceptibility to TB. For TLR-2 gene expression, the results revealed a
significant decrease (*p< 0.05) in TB patients in comparison to control groups. A novel TLR-2 rs3804100
SNPs were detected in Iraqi Arabs at g.24870 in 15%, g.24864 in 1.25% and g.25212 in 32.5% of patients.
Nevertheless, in control the novel SNPs were g.24887 in 2.5%, g.25212 in 7.5% and g. 25216 in 7.5%. On
the other hand, a novel TLR-2 rs7696323 SNPs were determined in Iraq Arabs patients and control at
g.23934 in 10%, g.23564 in 1.25%, g.23460 in 6.25%, g.23445 in 11.25%, g. 23452 in 10% and g. 23477 in
10% of TB patients. While in control group, g.23957 in 7.5%, g.23438 in 7.5%, g.23445 in 7.5%, g.23452 in
32.5% and g.23443 in 2.5% of control.ConclusionTLR2 genetic polymorphisms influence susceptibility to
pulmonary TB. TLR2 variants play a role in the development of TB disease, probably by affecting TLR2
expression level. Moreover, the present study showed that during anti-tuberculosis treatment, which
were seems likely responsible for controlling infection and excess inflammation.
Keywords: Tuberculosis, TLR2, Polymorphism, TLR2 expression, SNPs, PCR, qPCR, MTB, ELISA,
microsatellite repeat.
Introduction
Mycobacterium tuberculosis is the main
causative agent of TB in human. A cough for
three weeks or longer, weight loss, loss of
appetite, fever, night sweat and general
fatigue are the most common symptoms of
TB [1]. Statistics indicate that the one-third
of the world population having a latent TB
infection, approximately 1.5 million deaths
occurring annually from the active disease.
TB occupies the second rank of the leading
cause of death after human
immunodeficiency virus (HIV) [2]. In Iraq TB
is a serious problem, it threatens all society
sectors .Iraq rank 43
rd
in the list of countries
with an incidence of 40 cases per 100,000
person or greater [3]. The occurrence of TB at
different rates among particular races,
ethnicities, and families indicates a genetic