J. Biomedical Science and Engineering, 2011, 4, 511-515
doi:10.4236/jbise.2011.47065 Published Online July 2011 (http://www.SciRP.org/journal/jbise/
JBiSE
).
Published Online July 2011 in SciRes. http://www.scirp.org/journal/JBiSE
HLA allele frequencies in Iranian opticospinal multiple sclero-
sis patients
——HLA in Opticospinal MS
Hossein Kalanie1, Malihe Kamgooyan2, Yadollah Kholghie3, Ali Amini Harandi4*, Zahra Hos seinzadeh5
1MS Research Center, Mehr and Loghman Hospital Shahid Beheshti, University of Medical Sciences,Tehran, Iran;
2Bahar Medical Laboratory, Department of Immunology and Transplantation, Tehran, Iran;
3Loghman Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
4Loghman Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
5Mehr Hospital, Zartosht Street Tehran, Iran.
Received 4 May 2011; revised 26 May 2011; accepted 12 June 2011.
ABSTRACT
Background: In Iranian patients with opticospinal
multiple sclerosis (OSMS), a paucity of brain lesions
and short spinal cord lesions extending less than
three spinal segments are characteristic findings on
magnetic resonance imaging (MRI). It also shows a
relatively benign course with negative CSF oligo-
coonal bands. Objective: We aimed to clarify the pos-
sible relationship between clinical phenotype and
MRI features of OSMS and human leucocyte antigen
(HLA) system in Iran. Methods: Genotyping of HLA
class II allele frequencies in 20 patients with OSMS
were done, using polymerase chain reaction se-
quence-specific primer amplification method. Blood
samples were extracted and typed for HLA-DRB,
DQA, and DQB loci and compared with 100 controls.
Results: Significant positive association was observed
in DRB1*03, DQA1*0201, DQA1*03, DQB1*0201,
and DQB1*0611, while DQB1*0602 was ab sent in our
patients. Conclusion: These finding suggest that
HLA-DRB association pattern in OSMS is different
from conventional MS in Iran which is mostly asso-
ciated with DRB1*1501 and from similar Japanese
OSMS who are negative for brain lesions fulfilling
the Barkhof criteria and negative for the presence of
longitudinally extensive spinal cord lesions who car-
ries the DRB*0405 allele. OSMS is immunogeneti-
cally heterogeneous. Also absence of DQB1*0602 al-
lele may negatively be associated with the absence of
Barkhof brain lesion.
Keywords: Epidemiology, HLA-Class II Allele
Frequency, Opticospinal Multiple Sclerosis;
Neuromyelitis Optica; NMO-IgG Antibody; Devic’s
Neuromyelitis Optica; Iranian; Demyelinating Disease
1. INTRODUCTION
The etiology of multiple sclerosis (MS) is unknown, and
pathogenesis of the disease encompasses multiple in-
flammatory as well as apoptotic process in the central
nervous system [1-3]. Epidemiological studies indicate
both genetic and environmental component in MS sus-
ceptibility [4]. The human leucocyte antigen (HLA) sys-
tem provides a set of genetic loci which lend themselves
to systematic study. In fact both linkage and whole ge-
nome association screens revealed a prominent role for
alleles of the major histocompatibility complex class II
gene HLA-DRB1 [5]. Of interest, in northern Euro-
pean-descended populations, association with this gene
was identified only within families that carried the HLA
DRB1*1501 allele [6]. This observation demonstrates
that only a portion of familial MS is associated with al-
lelic variation in this gene. Therefore other genes must
contribute prominently to MS susceptibility of DRB1.
The fact that familial MS has at least 2 forms, the variety
associated with HLA-DRB1 and the form without this
association, onclusively demonstrates that MS is immu-
nogenetically heterogenesis.
In Asia, the diversity of MS has been a topic of de-
bates for years and there are wide variations in the clini-
cal presentation [7], and the prominence of Opticospinal
MS (OSMS) in Asian populations has long been recog-
nized. Many authors have divided MS into opticospinal
and conventional forms, base on their differential clini-
cal phenotypes. Kira et al. [8] considered these to be
differential entities and have also shown immunogenetic
differences between the types. They found an association
between conventional MS and DRB1*1501 and

H. Kalanie et al. / J. Biomedical Science and Engineering 4 (2011) 511-515 512
DRB5*0501 was found to play an important role in the
development of MS in general but not in OSMS [9,10].
On the basis of the presence or absence of brain lesions
fulfilling the Barkhof criteria and the longitudinally ex-
tensive spinal cord lesions (LESCLs) recently Matsuoka
et al. [11] have shown that OSMS in Japan is heteroge-
neous. One third of Japanese OSMS have Barkhof (-)
LESCLs (-) vairant of the disease and interestingly
enough this type of presentation is so far from the only
known presentation of the disease in Iran [12].
Although, OSMS is reported to be common in Japan
and some other Asian countries [13-20], it is an uncom-
mon presentation of MS in Iran [12,21]. The aim of this
study is to clarify the possible relationship between clini-
cal phenotype and magnetic resonance imaging (MRI)
features of OSMS and HLA system in Iran.
2. MATERIALS AND METHOD
HLA-Class II allele frequencies were studied in 20 pa-
tients with OSMS whose demographic data, clinical
course, MRI findings and cerebro-spinal fluid (CSF)
changes have been recently published [22] and were
compared with 100 healthy controls. There were 14
women and 6 men with gender ratio of 2.3:1. The age of
onset ranged from 10 to 50 years with the mean of 24 ±
8.2 and the mean disease duration of 8 ± 4.4 years. The
annual relapse rate was 0.66 ± 0.84 and the EDSS
ranged from 0 to 6 with the mean of 2.5 ± 1.3 with the
number of exacerbation of 5.25 ± 5.1. All 20 patients had
1 to 5 hemispheric T2 lesion in brain MRI and spinal
cord lesions that extending below three vertebral seg-
ments (short lesions) in sagittal planes with peripheral
white matter location and central sparing on the axial
plane. CSF contained normal cell count and protein level
with normal IgG index and negative oligocolonal bands.
Genomic DNA was extracted from EDTA blood sam-
ple by QIA DNA Mini Kits (QIA Gen) and DNA extrac-
tion kits of protrans. HLA-DRB, DQA and DQB typing
were performed using PCR-SSP method. Medium Reso-
lution Kit used for typing these loci was supplied by
Protrans and CTS of Heidelberg University and Taq
DNA polymerase enzyme by Roche Company (Basle,
Switzerland). The PCR reaction was carried out in 10 ul
volumes by thermal cyclers (Techne-Genius). Amplifi-
cation was followed after initial denaturation with 94
cycles (CS) for 2 minutes (including 10 CS for 15 sec-
onds, 65 CS for 60 seconds, 20 CS for 15 seconds, 61
CS for 50 seconds, and 72 CS for 30 seconds). After
running PCR products on agarose gel and observation of
specific bands using UV transilluminator, the results
were interpreted and alleles assigned [23,24]. The same
processes were performed for 100 healthy persons as
control group who were volunteers of organ transplanta-
tion.
Statistical analysis was performed using Chi-Square
and Fisher’s Exact Test. A p value less than 0.05 was
considered statistically significant and analysis was done
by SPSS version 11.0 software (SPSS Inc., Chicago, IL).
3. RESULTS
The HLA class II allele frequencies for DRB, DQA and
DQB in 20 OSMS patients comparing to 100 healthy
subjects are shown in Table 1, 2 and 3, respectively.
There were strong positive association between DRB1*03
(70% vs. 16%; P-value = 0.009), DQA1*0201 (35% vs.
10%; P-value = 0.01), DQA1*03 (45% vs. 10%; P-value
= 0.009), DQB1*0201 (90% vs. 19%; P-value = 0.009),
and DQB1*0611(30% vs. 7%; P-value = 0.009) and
OSMS patients, while DQB1*0602 (0% vs. 10%;
P-value = 0.210) and DRB1*11 (20% vs. 46%; P-value
= 0.045) haplotype were negatively associated with our
patients. Interestingly enough, no significant association
was found between OSMS and HLA- DRB1*1501
which is the most common HLA class II [5,7,9,22] in
conventional MS patients in Iran and other countries.
The HLA-DRB1*0103, DRB1*09, DRB1*12,
DRB1*14 were not observed in our OSMS patients, so
was for HLA-DQA1*03011 and DQA1*04011.
4. DISCUSSION
While OSMS is a common clinical presentation of MS
in oriental countries including Japan [13-20] with dis-
tinct reported features [8] and immunogenetic differ-
ences with conventional MS (CNMS), it is an uncom-
mon clinical presentation of the disease in Iran [12,21].
Heterogeneity in the clinical course and MRI findings
[12,25,26] may indicate to the presence of possible ge-
netic contributing factors and in fact previously it has
been shown that HLA-DRB1*1501 which plays an im-
portant role in the development of MS in general are not
observed in OSMS. On the basis of presence or absence
of brain lesions fulfilling the Barkhof criteria ± LESCLs,
have shown that Barkhof (-) patients had remarkably
lower frequency of HLA-DRB1*0901, despite this being
the most common allele in Japanese [27]. This allele also
was not presented in our patients, however it is not a
common allele in Iranian population. In contrast our
cases showed negative association with DQB1*0602
allele (0.0% vs. 10%). Therefore, absence of this allele
along with paucity of DRB1*1501 may negatively be
associated with the absence of Barkhof brain lesions in
our patients. In Matsouka’s study [27] the Barkhof (-)
LESCLs (-) patients which are in close similarity to our
study group, as a unique subtype of OSMS in Asian
showed a significant increase in the frequency of
DRB1*0405 compared with controls. But in our patients
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Table 1. HLA-DRB allele in 20 Iranian Opticospinal MS and 100 normal population.
HLA frequency No. of pts No. of controls P value Significancy
DRB1*0101 1 (5%) 15 (15%) 0.305 NS
DRB1*0103 0 0 − −
DRB1*03 14 (70%) 16 (16%) 0.009 Ext.sig
DRB1*04 7 (35%) 28 (28%) 0.719 NS
DRB1*07 8 (40%) 27 (27%) 0.369 NS
DRB1*08 1 (5%) 2 (2%) 0.424 NS
DRB1*09 0 2 (2%) 1.0 NS
DRB1*10
1 (5%) 6 (6%)
1.0 NS
DRB1*11 4 (20%) 46 (46%) 0.045 Ext.sig
DRB1*12 0 2 (2%) 1.0 NS
DRB1*13 2 (10%) 20 (20%) −
DRB1*14 0 9 (9%) 0.362 NS
DRB1*15 4 (20%) 20 (20%) 1.0 NS
DRB1*16 2 (10%) 10 (10%) 1.0 NS
DRB 303 16 (80%) 92 (92%) 0.114 NS
DRB 40404 15 (75%) 58 (58%) 0.242 NS
DRB 50101 6 (30%) 29 (29%) 1.0 NS
No of Pts: number of patients. NS: Not significant. Ext.sig: Extremely significant.
Table 2. HLA-DQA allele in 20 Iranian Opticospinal MS and 100 normal population.
HLA frequency No. of pts No. of controls P value Significancy
DQA1*01 7 (35%) 45 (%) 0.564 NS
DQA1*0101 1 (5%) 10 (%) 0.689 NS
DQA1*0102 4 (20%) 14 (%) 0.499 NS
DQA1*0103 1 (5%) 12 (%) 0.693 NS
DQA1*0104 1 (5%) 9 (%) 1.0 NS
DQA1*0201 7 (35%) 10 (%) 0.010 Ext.sig
DQA1*03 9 (45%) 10 (%) 0.009 Ext.sig
DQA1*03011 0 0 − −
DQA1*04 0 2 (%) 1.0 NS
DQA1*04011 0 0 −
DQA1*05 10 (50%) 33 (%) 0.233 NS
DQA1*06 0 0 − −
No of Pts: number of patients. NS: Not significant. Ext.sig: Extremely significant.
Table 3. HLA-DQB allele in 20 Iranian Opticospinal MS and 100 normal population.
HLA frequency No. of pts No. of controls P value Significancy
DQB1*0201 18 (90%) 19 (19%) 0.009 Ext.sig
DQB1*0203 0 1 (1%) 1.0 NS
DQB1*0301 5 (25%) 29 (29%) 0.928 NS
DQB1*0302 2 (10%) 5 (5%) 0.330 NS
DQB1*0303 1 (5%) 4 (4%) 0.342 NS
DQB1*0305 2 (10%) 1 (1%) 0.072 NS
DQB1*0306 0 1 (1%) 1.0 NS
DQB1*0401 2 (10%) 2 (2%) 1.0 NS
DQB1*0501 3 (15%) 18 (18%) 1.0 NS
DQB1*05051 1 (5%) 0 1.0 NS
DQB1*0531 0 3 (3%) 1.0 NS
DQB1*0611 6 (30%) 7 (7%) 0.009 Ext.sig
DQB1*0602 0 10 (10%) 0.210 NS
DQB1*0604 0 3 (3%) 1.0 NS
No of Pts: number of patients. NS: Not significant. Ext.sig: Extremely significant.
HLA-DRB1*04 is the third common HLA-gene in this
respect, while significant over-representation was seen in
the frequency of DRB1*03 in our group comparing with
controls. Interestingly, over-representation of this allele
recently has been reported in neuromyelitis optica
(NMO) from Brazil [28]. Our previous study [12]
showed that although there are some demographic simi-
larities between a subpopulation of OSMS in Japan with
Barkhof (-), LESCLs (-) and Iranian OSMS, still there
are genetic dissimilarities between two groups which
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H. Kalanie et al. / J. Biomedical Science and Engineering 4 (2011) 511-515 514
pinpoints to further heterogeneity of the disease in this
respect. Nowadays, with the availability of NMO im-
munoglobulin G (NMO-IgG) which targets the water
channel protein aquaporin-4, NMO spectrum of disorder
is wider than previously known and covers some cases
of Asian OSMS [29-31]. At present, probably the type of
OSMS in Iran [12] and a subpopulation of OSMS in
Japan according to Matsuoka et al. [11,27] with Barkhof
(-), LESCLs (-) MRI features, lower female to male ratio,
low annual relapse rate and lower number of exacerba-
tion and lower EDSS scores is the best candidate to be
nominated as pure opticospinal MS (POSMS).
5. CONCLUSIONS
Since evaluation of NMO-IgG was not feasible in our
country, it is our main study limitation. We wish to em-
phasize that the present study is a preliminary one as a
result of small number of patients involved due to low
prevalence of the disease in Iran. Nonetheless the find-
ings are encouraging enough to warrant further study of
these patients on a larger scale both in Iran and other
Asian countries.
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