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First report of Rh null individuals in the Indian population and characterization of the underlying molecular mechanisms
Author(s) -
Kulkarni Swati S.,
Vasantha Kasiviswanathan,
Gogri Harita,
Parchure Disha,
Madkaikar Manisha,
Férec Claude,
Fichou Yann
Publication year - 2017
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.14150
Subject(s) - biology , null allele , genetics , gene , sanger sequencing , microbiology and biotechnology , multiplex ligation dependent probe amplification , multiplex , exon , phenotype , rh blood group system , alternative splicing , mutation , antibody
BACKGROUND Rh null phenotype is an extremely rare condition characterized by no expression of Rh antigens at the surface of red blood cells. Although rare, genetic bases of this phenotype are well known and include mutations within either the RH ( RHD and RHCE ) genes or the RHAG gene. So far Rh null has been reported in individuals of Caucasian, African, and Asian origin. Here, we report individuals from two families of Indian origin representing such a rare phenotype. STUDY DESIGN AND METHODS Serologic analysis was carried out by testing with anti‐D, ‐C, ‐c, ‐E, and ‐e in Rh null individuals and their family members. RH genes were analyzed by standard molecular approaches, including Sanger sequencing and quantitative multiplex polymerase chain reaction (PCR) of short fluorescent fragments. RHAG gene was investigated by exon‐specific PCR amplification and Sanger sequencing. RESULTS In one family, RHAG gene was found to be deleted at the homozygous state in the propositus, suggesting Rh null of the regulator type. In the other family, a novel splice site variant in RHCE in cis with whole RHD gene deletion was identified at the homozygous state. Further functional analysis by minigene splicing assay showed that this variation, that is, c.801 + 1G>A, completely impairs normal splicing, then inactivating the expression of RhCE protein. Contrary to the former case, these data suggest Rh null of the amorph type. CONCLUSION Overall, we report for the first time the molecular mechanisms responsible for Rh null phenotype in individuals of Indian origin. This study contributes to extend the molecular spectrum of variations in Rh null individuals.