A new lymphoblastoid cell line defective in class II HLA expression.
A lymphoblastoid cell line, HAJ, was derived by in vitro transformation with Epstein-Barr virus of peripheral blood lymphocytes (PBL) from a patient with renal insufficiency awaiting kidney graft. Cell surface expression of class I and class II HLA molecules was determined by flow cytofluorimetry using monoclonal antibodies and compared with that of cell line PAJ similarly derived from a healthy donor. HAJ cells expressed class I antigens at levels comparable with PAJ cells. In contrast, class II antigens were absent from the cell surface of HAJ cells while they were abundant on PAJ cells. Permeabilization and fixation of cells with acetone/formaldehyde solution revealed intracellular Ki-67 antigen but not class II HLA molecules. The genes for HLA-DR beta, DQ alpha and DP alpha were present in the HAJ genome as detected with polymerase chain reaction (PCR) using locus-specific primers amplifying a second exon. In RT (reverse transcriptase)-PCR, transcripts of DQA1 and DPA1 genes were easily detectable in PAJ (positive control) but not in HAJ cells. These results suggest a defect in HAJ cells of transcription of genes for all class II antigens. The cell line HAJ may prove to be an interesting model for in vitro studies of molecular mechanisms of the regulation of class II expression.
Laboratory of Immunogenetics, Polish Academy of Sciences, Wroclaw, Poland.
M
M
Mańczak
J
J
Dubis
A
A
Kość
K
K
Baldy-Chudzik
M
M
Machnicki
R
R
Pacholczyk
M
M
Prussak
B
B
Nowakowska
P
P
Kuśnierczyk
D001692:Biological Transport / Q000276:immunology; D002461:Cell Line, Transformed; D006681:HLA-D Antigens / Q000032:analysis* / Q000096:biosynthesis*; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D017384:Sequence Deletion / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
D006681:HLA-D Antigens