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Immunohistochemical detection of intra-neuronal VZV proteins in snap-frozen human ganglia is confounded by antibodies directed against blood group A1-associated antigens.

Abstract
Varicella-zoster virus (VZV) causes chickenpox, establishes latency in trigeminal (TG) and dorsal root ganglia (DRG), and can lead to herpes zoster upon reactivation. The VZV proteome expressed during latency remains ill-defined, and previous studies have shown discordant data on the spectrum and expression pattern of VZV proteins and transcripts in latently infected human ganglia. Recently, Zerboni and colleagues have provided new insight into this discrepancy (Zerboni et al. in J Virol 86:578-583, 2012). They showed that VZV-specific ascites-derived monoclonal antibody (mAb) preparations contain endogenous antibodies directed against blood group A1 proteins, resulting in false-positive intra-neuronal VZV staining in formalin-fixed human DRG. The aim of the present study was to confirm and extend this phenomenon to snap-frozen TG (n=30) and DRG (n=9) specimens of blood group genotyped donors (n=30). The number of immunohistochemically stained neurons was higher with mAb directed to immediate early protein 62 (IE62) compared with IE63. The IE63 mAb-positive neurons always co-stained for IE62 but not vice versa. The mAb staining was confined to distinct large intra-neuronal vacuoles and restricted to A1(POS) donors. Anti-VZV mAb staining in neurons, but not in VZV-infected cell monolayers, was obliterated after mAb adsorption against blood group A1 erythrocytes. The data presented demonstrate that neuronal VZV protein expression detected by ascites-derived mAb in snap-frozen TG and DRG of blood group A1(POS) donors can be misinterpreted due to the presence of endogenous antibodies directed against blood group A1-associated antigens present in ascites-derived VZV-specific mAb preparations.
AuthorsWerner J D Ouwendijk, Sarah E Flowerdew, Desiree Wick, Anja K E Horn, Inga Sinicina, Michael Strupp, Albert D M E Osterhaus, Georges M G M Verjans, Katharina Hüfner
JournalJournal of neurovirology (J Neurovirol) Vol. 18 Issue 3 Pg. 172-80 (Jun 2012) ISSN: 1538-2443 [Electronic] United States
PMID22544677 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • A-1 antigen
  • ABO Blood-Group System
  • Antibodies, Monoclonal
  • IE62 protein, Human herpesvirus 3
  • Immediate-Early Proteins
  • Membrane Proteins
  • Trans-Activators
  • Viral Envelope Proteins
  • immediate early protein 63, Human herpesvirus 3
Topics
  • ABO Blood-Group System
  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Antibodies, Monoclonal (immunology)
  • Erythrocytes (immunology)
  • False Positive Reactions
  • Female
  • Freezing
  • Ganglia, Sensory (chemistry, immunology, virology)
  • Herpes Zoster (diagnosis, immunology, virology)
  • Herpesvirus 3, Human (genetics, metabolism)
  • Humans
  • Immediate-Early Proteins (analysis, genetics, immunology)
  • Immunohistochemistry
  • Male
  • Membrane Proteins (immunology)
  • Middle Aged
  • Neurons (chemistry, immunology, virology)
  • Trans-Activators (analysis, genetics, immunology)
  • Viral Envelope Proteins (analysis, genetics, immunology)
  • Virus Latency

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