Int. J. Chem. & LifeSci.

ISSN: 2234-8638
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Original Research Article OPEN ACCESS

HLA DQA1-0501 and DQB1-0201 Alleles in Rheumatoid Arthritis

Varun Chaithanya Gurram1*, Seema Kapoor2, Sunil Kumar Polipalli2, Vijay Kumar Karra3, Rajesh Ruttala3,
Veena Manogna Poludasu1, Appa Rao4, Ashok Kumar5
1Department of Human Genetics, Andhra University, Visakhapatnam, Andhra Pradesh, India.
2Pediatrics Research & Genetic Lab, Department of Pediatrics, MAMC & Associated Lok Nayak Hospital, New Delhi,

India.
3Department of Medicine, MAMC, New Delhi.
4Department of Microbiology, AMC, Visakhapatnam.
5Department of Orthopedics, AMC, Visakhapatnam.

Received For Publication: November 11, 2013; Accepted: December 22, 2013

Abstract: Human Leukocyte Antigens (HLA) system is the loci of genes that encode for major
histocompatibility complex (MHC) in humans. The expression of HLA Class II molecules on most cell types in
later stages of Rheumatoid Arthritis (RA) is both a hallmark of activation and indicates the significance of these
molecules in the onset and progression of RA. In the current study an attempt is made to check any possible
association between HLA DQA1-0501 and DQB1-0201 alleles and progress of RA in South Indian population.
The study was conducted on 154 subjects of which 84 are RA patients and 70 age and sex matched controls.
HLA DQA1-0501 and DQB1-0201 alleles were amplified using sequence specific primers. The amplified
product for different samples were separated by a 1.5% agarose gel stained with ethidium bromide and then
photographed. All statistical analyses were carried out using SYSTAT 12 software. Fifty seven RA patients
expressed HLA DQA1-0501 allele and 66 expressed HLA DQB1-0201. In controls, of the total seventy, 42
expressed HLA DQA1-0501 and 53 expressed HLA DQB1-0201 allele. The results of X2 analysis suggests no
significant heterogeneity between patients and controls. Odds ratio was also calculated for the two alleles
which suggests no significant association between HLA DQA1-0501 & HLA DQB1-0201 alleles and RA. It is
concluded that HLA DQA1-0501 & HLA DQB1-0201 alleles are not associated with progression of RA.

Keywords: Rheumatoid arthritis, Human Leukocyte Antigens, Major histocompatibility complex

Introduction
Rheumatoid Arthritis (RA) is an inflammatory
disease characterized by pain, swelling, stiffness,
and reduced function of the joints [1]. Onset is most
frequent during middle age, but people of any age
can be affected [2]. The exact cause of RA is still
unknown. Several factors are responsible. Genes that
play a role in the immune system seem to be
involved in the development of RA, but they are not
the only causative factor. The expression of HLA
Class II molecules on most cell types in later stages
of RA is both a hallmark of activation and indicates
the significance of these molecules in the onset and
progression of the disease [3].
Human Leukocyte Antigens (HLA): This
contains three classes (I, II and III). Class I and II
gene products have similar overall structure. There
are no functional or structural similarities between
class III gene products and class I and II gene
products [4]. The HLA class I gene is located on
short arm of chromosome 6 (6p21.31) telomeric to
HLA class II region [5].
Studies on extended HLA haplotypes in RA
have associated certain HLA class I antigens with it
[6]. On the other hand these associations are mainly
due to linkage disequilibrium between the two HLA
Class I and Class II loci [7]. The HLA class II region
is also located on short arm of chromosome 6. This
contains three major independent gene clusters
designated DP, DQ and DR and spans around 800
kilo bases (kb). The extra cellular region of HLA
class II molecule consists of a peptide binding region
and an immunoglobulin like region. The peptide
binding regions of HLA class II are polymorphic
and the immunoglobulin regions are non
polymorphic. HLA class II antigens are primarily
expressed on bone marrow derived cells, which
serve as specialized Antigen Presenting Cells
(APCs). Synthesis and expression of HLA molecules
are highly regulated at the gene transcription level
by cell type specific factors, inflammatory factors
and cytokines such as interferon γ which can up
regulate the expression of both HLA class I and II
molecules. Epidemiological studies indicate that
HLA genes are non-randomly associated with RA [8,

*Corresponding Author:
Dr. Varun Chaithanya Gurram,
Department of Human Genetics,
Andhra University, Visakhapatnam,
Andhra Pradesh, India
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Varun Chaithanya et al., Int. J. Chem. & LifeSci., 2014, 3 (01), 1265-1268

9]. These outcomes are not always consistent and

differences exist between different racial and ethnic
groups in the specific HLA alleles involved [10]. The
effect of HLA class II region genes is more
complicated than any of the existing hypotheses can
explain [11].
In the present study, HLA DQA1-0501 and
DQB1-0201 alleles were chosen because these are
class II loci genes and are involved in the
presentation of the foreign or self-antigens to the T-
Lymphocytes and their polymorphisms can affect
the antigens that are presented.
Materials and Methods
This study is carried out on 190 subjects of
which 90 (65 females and 25 males) are RA patients
and 100 age and sex matched controls. 5 ml of
intravenous blood samples were collected from
patients and controls into an EDTA vacutainer in
aseptic conditions after obtaining informed consent
from subjects. The laboratory work has been
performed at Department of Human Genetics,
Andhra University and Pediatric Research & Genetic
Lab, Maulana Azad Medical College, New Delhi,
India. All patients fulfilled the American College of
Rheumatology (ACR) 1987 revised criteria for
classification of RA [12] and had a disease history of
minimum 3 years.
PCR Amplification: The Genomic DNA is
isolated by Salting out method [13] and quantified
by using a Spectrophotometer. The absorbance ratio
of 1.8:2.0 or higher is considered and the final
solution is stored at -40c. The set of sequence specific
primers demonstrated in previous studies [14] were
used to amplify the target DNA in HLA region as
shown in the Figure 1.
DQB1*0201-F 5'-GTGCGTCTTGTGAGCAGAAG-3’
DQB1*0201-R 5'-GCAAGGTCGTGCGGAGCT-3’
DRB1*IPC-F 5’-TGC CAA GTG GAG CAC CCA A-3’
DRB1*IPC-R 5'-GCA TCT TGC TCT GTG CAG AT-3’
Figure 1: List of HLA Primers
Cycling File Parameters: Hot start period for about 5
minutes at 950 c, followed by
950 C - 30 seconds
610C - 35 seconds 35 cycles
720C - 60 seconds
720C - 5 minutes
An amplification product of 185-bp for HLA
DQA1-0501 allele and 205-bp for HLA DQB1-0201
were detected and separated by using a 1.5%
agarose gel, stained with ethidium bromide and
photographed [Figure 2].
Statistical analysis:
Frequencies of patients and controls
expressing HLA DQA1-0501 & HLA DQB1-0201
alleles were analyzed using 2 x 2 contingency table
test to test the goodness of fit for any association of
the said alleles with rheumatoid arthritis. A
probability value of <0.05 was considered
statistically significant.
Results and Discussion
HLA DQA1-0501 and HLA DQB1-0201: The
frequencies of patients and controls expressing HLA
DQA1-0501 and HLA DQB1-0201 alleles are
presented in Table 1. Out of 90 patients, only 84
could be successfully analyzed for the two alleles.
Fifty-seven (67.87) RA patients expressed HLA
DQA1-0501 allele and 66 (78.57) HLA DQB1-0201
(percentage in parenthesis). In controls, of the total
seventy, 42 (60.0) expressed HLA DQA1-0501 and 53
(76.0) expressed HLA DQB1-0201 allele (percentage
in parenthesis). The results of X2 analysis suggests
no significant heterogeneity between patients and
controls (p values = 0.090 and 0.671). Odds ratio was
also calculated for the two alleles which suggests no
significant association between HLA DQA1-0501 &
HLA DQB1-0201 alleles and RA.

Table 1: Frequencies of Patients and Controls expressing HLA DQA1-0501 and HLA DQB1-0201 alleles
RA Patients 84 Controls = 70 CI for OR
Allele X2 P-value OR
n % n % Lower Upper
HLA DQA1-0501 57 67.87 42 60.0 2.85 0.090 0.55 0.28 1.10

HLA DQB1-0201 66 78.57 53 76.0 0.18 0.671 0.85 0.40 1.80

n = number of individuals; p value =probability value of the statistical test, X2 =chi square, OR=Odds Ratio, S= Significant,
NS= Not Significant.

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Varun Chaithanya et al.,
Our study showed no significant association
between these alleles and RA. Similar to our
findings are reports from other studies. Ali et al.,
(2006) [15] demonstrated that HLA-DQB1- 0201 is
not associated with RA. Parthibhan et al., (2002) [16]
stated that HLA DQA1and HLA DQB1 alleles are
not associated with disease progression in RA.
Castro et al., (2001) [17], revealed that HLA DQA1-
0501 and HLA DQB1-0201 are not significantly
associated with RA and Taneja et al., (1996) [18]
reported that DQB locus is not associated with RA.
When other autoimmune diseases are considered
the results are also inconsistent. Maciel et al., (2001)
[19] and Yanagawa et al., (1994) [20] reported that
HLA DQA1-0501 is associated with Graves disease.
Philippou et al., (2001) [21] and Cuddihy et al., (1996)
[22] stated that HLA DQA1-0501 is not an
independent marker for Graves disease. HLA
DQB1-0201 is associated with allergy and asthma
(Movahedi et al., 2008) [23].
Conclusion
In Comparison with the previous studies and
results obtained from this study it is concluded that
HLA DQA1-0501 & HLA DQB1-0201 alleles are not
associated with RA. The study has few limitations.
This does not include patients with intermediate and
mild severity and also the number of subjects
studied is small. By resolving these limitations more
encouraging results can be obtained.
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Source of support: Nil,
Conflict of interest: None Declared
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