HLA allele frequencies in Iranian opticospinal multiple sclerosis patients<br>——HLA in Opticospinal MS

doi:10.4236/jbise.2011.47065

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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.

Email: *[email protected]

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

H. Kalanie et al. / J. Biomedical Science and Engineering 4 (2011) 511-515 513

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

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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