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Expression of the CTL 2 transcript variants in human peripheral blood cells and human tissues
Author(s) -
Flesch Brigitte K.,
Wesche Jan,
Berthold Tom,
Goldmann Torsten,
Hundt Matthias,
Greinacher Andreas,
Bux Jürgen
Publication year - 2013
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.12160
Subject(s) - antibody , peripheral blood mononuclear cell , microbiology and biotechnology , complementary dna , antigen , messenger rna , gene , rna , chemistry , biology , immunology , in vitro , genetics
Background The HNA ‐3a antigen is an important antibody target in the pathophysiology of transfusion‐related acute lung injury ( TRALI ). It is encoded by the choline transporter‐like protein 2 ( CTL 2) gene, which exists in the two transcript variants TV 1 and TV 2, differing in the upstream promoter and coding region. Only TV 1 has been demonstrated to enable choline transport across the cell membrane. Study Design and Methods The aim of this study was to determine the CTL 2 transcript pattern in human peripheral blood cells and tissues and its capacity to bind HNA ‐3a antibodies. RNA was isolated from human whole blood, isolated neutrophils, mononuclear blood cells, leukoreduced platelets, human lung, liver, and colon. After reverse transcription, the single‐stranded cDNA was amplified using primer combinations specific for the respective transcript. Plasmids containing the entire CTL 2 coding cDNA of the transcript variant TV 1 or TV 2 served as controls. HEK 293 T cells expressing both variants were used to determine the binding of HNA ‐3a antibodies. Results The shorter TV 2 transcript was demonstrated in each RNA sample derived from human peripheral blood tested so far, as well as in human lung and liver, whereas the longer TV 1 transcript was only detected in human lung and colon. TV 1 and TV 2 had the same binding capacity to HNA ‐3a antibodies. Conclusion The expression of TV 1 and TV 2 is tissue and cell specific, with peripheral blood cells expressing only TV 2. This does not affect binding of HNA ‐3a antibodies. Whether the unequal expression might be relevant in the pathogenesis of TRALI remains to be investigated.