VOLUME 16 NUMBER
6 November/December 2003
Distribution of C282Y and H63D mutations
in the HFE gene in healthy Asian Indians and patients with thalassaemia
Gurvinder Kaur, C. Chen Rapthap, M. Xavier,
R. Saxena, V. P. Choudhary, S. K. Reuben,
N. K. Mehra
Background. Mutations in the HFE gene have been shown to be strongly
associated with hereditary haemochromatosis, an autosomal recessive
disease of iron overloading. The majority of patients with
hereditary haemochromatosis possess a homo-zygous mutation
C282Y that disrupts the binding of the HFE gene with b2 microglobulin
and prevents its surface expression. Another HFE mutation H63D
is known to increase the relative risk of developing hereditary
haemochromatosis. This disease is rare in India although secondary
haemochromatosis is commonly seen among children suffering
from thalassaemia major. The status of HFE mutations has not
been explored among Indians, particularly in patients with
thalassaemia major. It is also possible that in India clinical
haemochromatosis could be masked by iron deficiency.
Methods. We examined a cohort of 59 unrelated, healthy individuals
from north India, 57 from south India and 75 thalassaemia major
patients from north India for HFE mutations (C282Y and H63D)
in cis/trans by the polymerase chain reaction sequence-specific
Results. The C282Y and H63D mutations in the HFE gene were rare
among Indians. Although the HFE mutations were increased among
patients of thalassaemia their effect on iron burden or disease
pathogenesis remains unclear.
Conclusions. Hereditary haemochromatosis is rarely observed among
Indians and so are the C282Y and H63D mutations in the HFE gene.
Long term follow up studies would be required to determine whether
the relatively higher frequency of these mutations among patients
of thalassaemia has any influence on iron accumulation.
Natl Med J India 2003;16:309–10
Hereditary haemochromatosis is an autosomal recessive disorder
of iron overload affecting approximately 5 per 1000 Caucasians
while 1 in every 12 Caucasians in northwestern Europe is a carrier.1 It is a multigenic disorder with the possible involvement of
several genetic loci that include HFE-1 (C282Y, H63D and S65C
mutations in HFE at 6p21),2 HFE-2 (1q in juvenile haemochromatosis),
HFE-3 (Y250X and other mutations in TFR-2 at 7q22),3 NRAMP-2
(duodenal metal transporter, DMT-1),4 SFT (stimulator of iron
transport)5 and other unknown factors.
The HFE gene, originally called human histocompatibility leucocyte-H
(HLA-H), is a non-peptide binding major histocompatibility complex
(MHC) class I homologue that binds with b2 microglobulin and
modulates iron transport by binding the transferrin receptor
(TfR)6 in a pH-dependent manner. Two mutations in the HFE gene,
namely (i) C282Y that results from a change in cysteine (C) at
position 282 to tyrosine (Y), and (ii) H63D that results from
a change in histidine (H) at position 63 to aspartate (D) disrupt
cell surface expression of the mutant HFE gene, and alter binding
with TfR, respectively. These mutations are strongly associated
with predisposition to hereditary haemochromatosis and are also
implicated in other disorders such as rheumatoid arthritis,7 type 2 diabetes mellitus,8 porphyria cutanea tarda9 and coronary
Considerable ethnic variation is observed in the frequency distribution
of HFE mutations. Hereditary haemochromatosis is rare in India11 and the status of HFE mutations is virtually unexplored. On the
other hand, iron overload is frequently observed among patients
with thalassaemia major, a widely prevalent haemoglobinopathy.12 We investigated the prevalence of HFE mutations in two healthy
populations from north and south India and in patients with thalassaemia
major from north India.
This study was done on 116 healthy, unrelated subjects (59 from
north India [from the states of Punjab, Delhi, Haryana and Uttar
Pradesh] and 57 from south India [from the states of Kerala,
Andhra Pradesh, Karnataka and Tamil Nadu]) and 75 patients with
thalassaemia major from north India referred to the All India
Institute of Medical Sciences, New Delhi. All the patients with
thalassaemia major had received multiple blood transfusions and
iron chelation therapy. Blood samples were collected after informed
consent and processed for HFE genotyping using the polymerase
chain reaction-sequence specific primer method and primer sequences
as previously published.13 Four possible HFE genotypes, namely
63H-282C, 63H-282Y, 63D-282C, 63D-282Y, were analysed and their
The relative frequencies of HFE mutations in the healthy subjects
and the thalassaemia major patients were compared and the significance
of association was tested by 2×2 contingency c2 analysis
and Fischer exact text. Serum ferritin levels were also measured
in 27 of the 75 thalassaemia patients using the standard biochemical
Genotyping of C282Y and H63D mutations
The C282Y mutation was seen in only 1 of the 59 healthy subjects
from north India (1.7%), but in none of those from south India.
On the other hand, the allele frequency of H63D was 4 of 59 (6.8%)
in subjects from north India, and 6 of 57 (10.5%) in those from
south India. The C282Y and H63D mutations were observed in 3
(4%) and 9 (12%) patients with thalassaemia major, respectively.
An analysis of the genotypes of these mutations revealed that
the frequency of genotype CY was higher among patients with thalassaemia
major (6 of 75) than healthy controls (1 of 116; p=0.03). There
was no significant difference in the prevalence of HH, HD or
DD genotypes between thalassaemia patients and healthy controls.
We did not encounter any compound heterozygotes among the subjects
tested (Table I).
Analysis of ferritin levels
High ferritin levels (2050–11 000 mg/L) were observed in
all but 1 (96.3%) of these patients. However, there was no correlation
between C282Y and H63D mutations and serum ferritin levels.
|Table I. HFE genotypes in healthy Indians
and patients with thalassaemia major
|Genotypes HD and DD represent the presence
of mutation (H to D) at amino acid residue 63 in heterozygous
and homozygous conditions, respectively. The genotypes CY
and YY represent presence of mutation (C to Y) at amino acid
residue 282 in the heterozygous and homozygous conditions,
We describe the occurrence of HFE gene mutations, C282Y and H63D,
in two healthy populations from north and south India as well
as thalassaemia major patients from north India. Generally
these mutations occur at lower frequencies in Indians as compared
with healthy Europeans.1 For example, the C282Y mutation occurs
with a frequency of 6.4% in the British population,1 5.6% in
the Hungarian,15 3.1% in the Polish16 and 3.9% in the Germans,
but is rare in Greeks (1.4%) and is absent in the Finnish,
African, Chinese and Turkish populations. Similarly, the frequency
of H63D mutation is 12.8% in the British, 10.4% in Russians,
13.6% in Europeans, but only 2.8% in Chinese, and is absent
in Papua New Guineans and Australian Aboriginals.
Homozygosity of the C282Y mutation has been reported in over
90% of patients with hereditary haemochromatosis in the UK by
the UK Haemochromatosis Consortium, 82.3% in the USA,17 92.4%
in Britain18 and in 100% of Australian patients with hereditary
haemochromatosis and a positive family history.19 Thus, the C282Y
mutation is a very good marker for hereditary haemochromatosis,
given the fact that HFE is not the only gene responsible for
this condition since the role of other factors (genetic, dietary
and environmental) cannot be ruled out. Hereditary haemochromatosis
is rarely seen in the Asia–Pacific region20 including India,
Japan,21 China22 and Korea.23 The observed low frequencies of
the C282Y and H63D mutations in these populations correlates
with this observation.
Our results suggest that the relatively infrequent occurrence
of the C282Y and H63D mutations among Indians could explain the
absence or rare occurrence of hereditary haemochromatosis. These
HFE mutations might be involved in the process of secondary iron
overload observed in some patients with thalassaemia major.
We thank the Department of Biotechnology, Ministry of Science
and Technology and the Indian Council of Medical Research for
granting financial support for the study.
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All India Institute of Medical Sciences, Ansari Nagar, New Delhi
Gurvinder Kaur, C. Chen Rapthap, M. Xavier,
N. K. Mehra Department of Transplant Immunology and Immunogenetics
R. Saxena, V. P. Choudhary Department of Haematology
Lady Doak College, Madurai, Tamil Nadu, India
S. K. Reuben Department of Biology and Research Centre
Correspondence to N. K. MEHRA; email@example.com