Establishment of cell lines stably expressing HNA-1a, -1b, and -2a antigen with low background reactivity in flow cytometric analysis.
BACKGROUND
Antibodies to neutrophil antigens have been implicated in neonatal alloimmune neutropenia, autoimmune neutropenia, and transfusion-related acute lung injury. Most often, neutrophil-specific antibodies are directed toward human neutrophil antigen (HNA)-1 (Fcgamma receptor 3b) and HNA-2a (CD177) in these disorders.


METHODS
To detect the alloantibodies in the serum samples, a panel of cell lines was established in which the HNA-1a, HNA-1b (polymorphisms of HNA-1), or HNA-2a gene was transduced with a retrovirus vector to confer stable transgene expression in K562 cells that exhibited low background reactivity to human serum samples obtained from healthy donors in flow cytometric analysis.


RESULTS
It was shown that several well-characterized human serum samples containing antibodies against HNA-1a, -1b, and -2a were unambiguously identified by the established panel cell lines and observed a lower background reactivity and longer shelf life of the K562 panel cell lines compared with isolated neutrophils, which have been used for the cell panel to identify antibodies against HNA in human serum samples.


CONCLUSIONS
These results indicate that the K562 panel cell lines provide a good panel for detecting HNA-reactive neutrophil antibodies in human serum samples.
Yasui
Kazuta
K
Japanese Red Cross Osaka Blood Center, and the Kansai Medical University, Osaka, Japan. yasui-kazuta@osaka.bc.jrc.or.jp
Miyazaki
Toru
T
Matsuyama
Nobuki
N
Kojima
Yoshitaka
Y
Furuta
Rika A
RA
Fujisawa
Jun-Ichi
J
Tani
Yoshihiko
Y
Shibata
Hirotoshi
H
Sato
Shin-Ichirou
S
Kato
Toshiaki
T
Ikeda
Hisami
H
Hirayama
Fumiya
F
D016475:3T3 Cells; D000818:Animals; D001773:Blood Cells / Q000276:immunology / Q000378:metabolism; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D003429:Cross Reactions; D005434:Flow Cytometry / Q000379:methods*; D058851:GPI-Linked Proteins; D006367:HeLa Cells; D006801:Humans; D007519:Isoantigens / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D019169:Jurkat Cells; D020014:K562 Cells; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014162:Transfection
C407242:CD177 protein, human; D058851:GPI-Linked Proteins; D007519:Isoantigens; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; C067704:neutrophil-specific antigen NA1, human