Multiple Antibody Targets on Herpes B Glycoproteins B and D Identified by Screening Sera of Infected Rhesus Macaques with Peptide Microarrays

Abstract

Herpes B virus (or Herpesvirus simiae or Macacine herpesvirus 1) is endemic in many populations of macaques, both in the wild and in captivity. The virus elicits only mild clinical symptoms (if any) in monkeys, but can be transmitted by various routes, most commonly via bites, to humans where it causes viral encephalitis with a high mortality rate. Hence, herpes B constitutes a considerable occupational hazard for animal caretakers, veterinarians and laboratory personnel. Efforts are therefore being made to reduce the risk of zoonotic infection and to improve prognosis after accidental exposure. Among the measures envisaged are serological surveillance of monkey colonies and specific diagnosis of herpes B zoonosis against a background of antibodies recognizing the closely related human herpes simplex virus (HSV). 422 pentadecapeptides covering, in an overlapping fashion, the entire amino acid sequences of herpes B proteins gB and gD were synthesized and immobilized on glass slides. Antibodies present in monkey sera that bind to subsets of the peptide collection were detected by microserological techniques. With 42 different rhesus macaque sera, 114 individual responses to 18 different antibody target regions (ATRs) were recorded, 17 of which had not been described earlier. This finding may pave the way for a peptide-based, herpes B specific serological diagnostic test.

The Kaposi's sarcoma-associated herpesvirus (KSHV) non-structural membrane protein K15 is required for viral lytic replication and may represent a therapeutic target

“…The production of the rat anti-KSHV K15 monoclonal antibody (clone 18E5) was described before [ 49 ]. Here, the K15 epitope necessary for antibody binding for both clones used in western blot (clone 10A6) and IFA (clone 18E5) was mapped by using a microarray of 44 overlapping synthetic peptides from the K15 cytoplasmic tail sequence with 3 amino acid shifts as described before [ 99 ]. In brief, the peptides were synthesized using the SPOT method on a soluble cellulose matrix [ 100 ].…”

Diagnosing Zika virus infection against a background of other flaviviruses: Studies in high resolution serological analysis

“…In the context of the overlapping peptides method, the term ‘antibody target’ denotes a stretch of polypeptide chain of an antigen encompassing either a single linear B cell epitope or a locally clustered set thereof. In the latter case, it is called an antibody target region (ATR) 25 . Since a spots series as such often does not allow distinguishing between single epitope and ATR, the latter term is occasionally also used as a synonym of the generic name “antibody target” with epitopes regarded as extreme cases within that class.…”

“…dengue, yellow fever, tick-borne encephalitis, and West Nile virus). As shown earlier 25 , the necessary deconvolution of complex B cell responses to the level of contributions made by individual short stretches of polypeptide chain can be achieved experimentally by peptide microarray techniques. Comparing ATRs resulting from inspection of ZIKV and non-ZIKV flaviviral antisera will reduce the former to the ZIKV-specific subset.…”

“…Each serum pool was tested in triplicate (three individual chips, corresponding to nine arrays total). For unknown reasons, all four sera pools produced atypically high levels of background which precluded application of the previously developed, automated spot calling procedure 25 . Therefore, array patterns were visually inspected and positive responses were identified in two steps – with the burden of safeguarding against false positives laid predominantly on stringent selection in step 1: (i) Individual signals were called positive if they showed bright red fluorescence spread uniformly across the entire spot area (Fig.…”

Species-Specific Conservation of Linear Antigenic Sites on Vaccinia Virus A27 Protein Homologs of Orthopoxviruses

“…In order to obtain equal antibody concentrations of 2 µg/µL per chamber, protein concentrations were measured using a NanoDrop ND-1000 Spectrophotometer. The screening procedure with the microarray chip was performed as previously reported [75].…”