title: 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.
authors.affiliation: Department of Virology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Werner J D
authors.initials: WJ
authors.lastname: Ouwendijk
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Flowerdew
authors.forename: Desiree
authors.initials: D
authors.lastname: Wick
authors.forename: Anja K E
authors.initials: AK
authors.lastname: Horn
authors.forename: Inga
authors.initials: I
authors.lastname: Sinicina
authors.forename: Michael
authors.initials: M
authors.lastname: Strupp
authors.forename: Albert D M E
authors.initials: AD
authors.lastname: Osterhaus
authors.forename: Georges M G M
authors.initials: GM
authors.lastname: Verjans
authors.forename: Katharina
authors.initials: K
authors.lastname: Hüfner
mesh_terms: D000017:ABO Blood-Group System; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal / Q000276:immunology; D004912:Erythrocytes / Q000276:immunology; D005189:False Positive Reactions; D005260:Female; D005615:Freezing; D017950:Ganglia, Sensory / Q000737:chemistry* / Q000276:immunology / Q000821:virology; D006562:Herpes Zoster / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000276:immunology; D008875:Middle Aged; D009474:Neurons / Q000737:chemistry* / Q000276:immunology / Q000821:virology; D015534:Trans-Activators / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D017735:Virus Latency
chemical_list: C085329:A-1 antigen; D000017:ABO Blood-Group System; D000911:Antibodies, Monoclonal; C074221:IE62 protein, Human herpesvirus 3; D017874:Immediate-Early Proteins; D008565:Membrane Proteins; D015534:Trans-Activators; D014759:Viral Envelope Proteins; C074551:immediate early protein 63, Human herpesvirus 3