PMID- 19497055
OWN - NLM
STAT- MEDLINE
DCOM- 20090923
LR  - 20101118
IS  - 1537-2995 (Electronic)
IS  - 0041-1132 (Linking)
VI  - 49
IP  - 9
DP  - 2009 Sep
TI  - A novel enzyme-linked immunosorbent assay method for the detection of human 
      neutrophil antigen-2a antibodies.
PG  - 1819-24
LID - 10.1111/j.1537-2995.2009.02229.x [doi]
AB  - BACKGROUND: Antibodies to human neutrophil antigen (HNA)-2a are responsible for a 
      number of immune-mediated neutropenia disorders. Although several methods exist 
      for the identification of anti-HNA-2a, all these methods have several 
      limitations. In this study, a solid-phase enzyme-linked immunosorbent assay 
      (ELISA) using recombinant HNA-2a antigen (rHNA-2a) allowing rapid detection of 
      HNA-2a antibodies was developed. STUDY DESIGN AND METHODS: Soluble rHNA-2 was 
      generated by transfection of insect cells with CD177 vector. Purified rHNA-2a was 
      immobilized on microtiter wells coated with anti-CD177 and was applied to analyze 
      10 sera containing HNA-2a antibodies. For the evaluation of the ELISA method, 
      results were compared with the standard assay, MAIGA (monoclonal antibody antigen 
      capture assay) for detection of neutrophil antibodies. RESULTS: The specificity 
      of HNA-2a antibodies in all sera was confirmed by immunoblotting. Sera were then 
      tested simultaneously in ELISA and MAIGA assays. Nine of 10 sera showed positive 
      reactions in ELISA, whereas only 9 of 10 sera reacted in the standard MAIGA 
      assay. All HNA-2a antibodies were detectable in MAIGA when diluted sera were 
      applied. No reaction was observed with different sera containing 
      neutrophil-reactive antibodies (6 anti-HNA-1a, 4 anti-HNA-1b, and 20 anti-HLA 
      Class I and II) in ELISA. All HNA-2a antibodies were detectable in MAIGA when 
      diluted sera were applied. Notably, sera containing anti-proteinase 3 (PR3) from 
      patients with Wegener's granulomatosis reacted in MAIGA. In contrast, this 
      antibody showed no reaction in ELISA with purified rHNA-2a. CONCLUSIONS: These 
      results demonstrated that ELISA with rHNA-2a provides a good method for detecting 
      HNA-2a antibodies in human serum. This assay enables to exclude the presence of 
      autoantibody against PR3 in patient's sera, which cannot be differentiated from 
      anti HNA-2a with current serologic methods.
FAU - Bayat, Behnaz
AU  - Bayat B
AD  - From the Institute for Clinical Immunology and Transfusion Medicine, Justus 
      Liebig University, Giessen, Germany.
FAU - Werth, Silke
AU  - Werth S
FAU - Sachs, Ulrich J H
AU  - Sachs UJ
FAU - Santoso, Sentot
AU  - Santoso S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090527
PL  - United States
TA  - Transfusion
JT  - Transfusion
JID - 0417360
RN  - 0 (Antibodies)
RN  - 0 (CD177 protein, human)
RN  - 0 (GPI-Linked Proteins)
RN  - 0 (Isoantigens)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (Receptors, Cell Surface)
RN  - 0 (neutrophil-specific antigen NA1, human)
RN  - 0 (neutrophil-specific antigen NA2)
RN  - EC 3.4.21.76 (Myeloblastin)
SB  - IM
MH  - Antibodies/*immunology
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - GPI-Linked Proteins
MH  - Humans
MH  - Isoantigens/*immunology
MH  - Membrane Glycoproteins/immunology
MH  - Myeloblastin/immunology
MH  - Neutrophils/immunology
MH  - Receptors, Cell Surface/immunology
EDAT- 2009/06/06 09:00
MHDA- 2009/09/24 06:00
CRDT- 2009/06/06 09:00
PHST- 2009/06/06 09:00 [entrez]
PHST- 2009/06/06 09:00 [pubmed]
PHST- 2009/09/24 06:00 [medline]
AID - TRF02229 [pii]
AID - 10.1111/j.1537-2995.2009.02229.x [doi]
PST - ppublish
SO  - Transfusion. 2009 Sep;49(9):1819-24. doi: 10.1111/j.1537-2995.2009.02229.x. Epub 
      2009 May 27.