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Geno- and Phenotyping of Human Neutrophil Antigens

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Molecular Typing of Blood Cell Antigens

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1310))

Abstract

For typing of human neutrophil antigens (HNA) usually genotyping techniques are used, except for HNA-2, which—due to a gene expression defect—requires phenotyping. For genotyping, several techniques have been described. Most reference laboratories use variations of the polymerase chain reaction (PCR) for antigen typing which showed good results in international quality assessment exercises. The granulocyte immunofluorescence test has been the gold standard technique for phenotyping for all HNA antigens except for HNA-3a and -3b phenotyping. The expression of the latter antigens on neutrophils is often better shown by the use of the granulocyte agglutination test.

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References

  1. Greinacher A, Wesche J, Hammer E, Fürll B, Völker U, Reil A, Bux J (2010) Characterization of the human neutrophil alloantigen (HNA) 3a. Nat Med 16:45–48

    CAS  PubMed  Google Scholar 

  2. Kissel K, Scheffler S, Kerowgan M, Bux J (2000) Molecular basis of NB1 (HNA-2a, CD177) deficiency. Blood 99:4231–4233

    Article  Google Scholar 

  3. Lucas G, Porcelijn L, Fung YL, Green F, Reil A, Hopkins M, Green A, de Haas M, Bux J (2013) External quality assessment of human neutrophil antigen (HNA)-specific antibody detection and HNA genotyping from 2000 to 2012. Vox Sang 105:259–269

    CAS  PubMed  Google Scholar 

  4. Bux J, Chapman J (1997) Report on the second international granulocyte serology workshop. Transfusion 37:977–983

    Article  CAS  PubMed  Google Scholar 

  5. Böyum A (1968) A one-stage procedure for isolation of granulocytes and lymphocytes from human blood. Scand J Clin Lab Invest Suppl 97:51–76

    PubMed  Google Scholar 

  6. Verheugt FW, von dem Borne AE, Décary F, Engelfriet CP (1977) The detection of granulocyte alloantibodies with an indirect immunofluorescence test. Br J Haematol 36:533–544

    CAS  PubMed  Google Scholar 

  7. Jiang A, Lalezari P (1975) A micro-technique for detection of leukocyte agglutinins. J Immunol Methods 7:103–108

    CAS  PubMed  Google Scholar 

  8. Bux J, Kober B, Kiefel V, Mueller-Eckhardt C (1993) Analysis of granulocyte-reactive antibodies using an immunoassay based upon monoclonal antibody-specific immobilization of granulocyte antigens (MAIGA). Transfus Med 3:157–162

    CAS  PubMed  Google Scholar 

  9. Bux J, Stein EL, Santoso S, Mueller-Eckhardt C (1995) NA gene frequencies in the German population, determined by polymerase chain reaction with sequence-specific primers. Transfusion 35:54–57

    Article  CAS  PubMed  Google Scholar 

  10. Bux J, Stein EL, Bierling P, Fromont P, Clay M, Stroncek D, Santoso S (1997) Characterization of a new alloantigen (SH) on the human neutrophil Fc gamma receptor IIIb. Blood 89:1027–1034

    CAS  PubMed  Google Scholar 

  11. Steffensen R, Gülen T, Varming K, Jersild C (1999) FcgammaRIIIB polymorphism: evidence that NA1/NA2 and SH are located in two closely linked loci and that the SH allele is linked to the NA1 allele in the Danish population. Transfusion 39:593–598

    Article  CAS  PubMed  Google Scholar 

  12. Sachs UJ, Chavakis T, Fung L, Lohrenz A, Bux J, Reil A, Ruf A, Santoso S (2004) Human alloantibody anti-Mart interferes with Mac-1-dependent leukocyte adhesion. Blood 104:727–734

    Article  CAS  PubMed  Google Scholar 

  13. Sachs UJ, Reil A, Bauer C, Bux J, Bein G, Santoso S (2005) Genotyping of human neutrophil antigen-5a (Ond). Transfus Med 15:115–117

    CAS  PubMed  Google Scholar 

  14. Reil A, Wesche J, Greinacher A, Bux J (2011) Geno- and phenotyping and immunogenicity of HNA-3. Transfusion 51:18–24

    Article  CAS  PubMed  Google Scholar 

  15. Reil A, Sachs UJ, Siahanidou T, Flesch BK, Bux J (2013) HNA-1d: a new human neutrophil antigen located on Fcγ receptor IIIb associated with neonatal immune neutropenia. Transfusion 53:2145–2151

    CAS  PubMed  Google Scholar 

  16. Flesch BK, Reil A, Bux J (2011) Genetic variation of the HNA-3a encoding gene. Transfusion 51:2391–2397

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Angelika Reil .

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Reil, A., Bux, J. (2015). Geno- and Phenotyping of Human Neutrophil Antigens. In: Bugert, P. (eds) Molecular Typing of Blood Cell Antigens. Methods in Molecular Biology, vol 1310. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2690-9_16

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  • DOI: https://doi.org/10.1007/978-1-4939-2690-9_16

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2689-3

  • Online ISBN: 978-1-4939-2690-9

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