The origin of multiple molecular forms in urine of HNL/NGAL.
OBJECTIVE
Several molecular forms of human neutrophil lipocalin/neutrophil gelatinase-associated lipocalin (HNL/NGAL), a novel biomarker for acute kidney injury (AKI), have been found in urine. The origin of these different forms and the effect of antibody configuration on assay performances were investigated in this report.


METHODS
The molecular forms of HNL/NGAL from human neutrophils and present in urine obtained from cardiac surgery patients and patients with urinary tract infection (UTI), as well as secreted from HK-2 cells, were studied by Western blotting. The levels of HNL/NGAL in urine were measured by ELISAs. Kidney injury was simulated by incubation of HK-2 cells under stressful conditions.


RESULTS
The major molecular form of HNL/NGAL secreted by neutrophils is dimeric, whereas the major form secreted by HK-2 cells is monomeric. This was reflected by a predominance of the monomeric form in urine from patients with AKI and the dimeric form in patients with UTIs. The epitope specificities of the antibody used in the ELISAs had a profound effect on assay performance and paralleled differences of the antibodies to identify the different forms of urine HNL/NGAL.


CONCLUSIONS
The monomeric form is the predominant form secreted by tubular epithelial cells, and the dimeric form is the predominant form secreted by neutrophils. The development of molecular form-specific assays for HNL/NGAL may be a means to identify the origin of HNL/NGAL in urine and construct more specific tools for the diagnosis of AKI.
Cai
Linjun
L
Department of Preventive Veterinary Medicine, Molecular Immunology and Pathogenic Microorganism, Jilin University, Changchun, People's Republic of China.
Rubin
Jenny
J
Han
Wenyu
W
Venge
Per
P
Xu
Shengyuan
S
D058186:Acute Kidney Injury / Q000175:diagnosis / Q000652:urine; D000209:Acute-Phase Proteins / Q000737:chemistry / Q000652:urine*; D015153:Blotting, Western; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000378:metabolism; D000071068:Lipocalin-2; D054834:Lipocalins / Q000737:chemistry / Q000652:urine*; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000652:urine*
D000209:Acute-Phase Proteins; C489506:LCN2 protein, human; D000071068:Lipocalin-2; D054834:Lipocalins; D011518:Proto-Oncogene Proteins