An Immunofluorescence Technique with Counterstain on Fixed Cells for the Detection of Antibodies to Human Herpesviruses; Antibody Patterns in Patients with Hodgkin’s Disease and Nasopharyngeal Carcinoma

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Abstract
An indirect immunofluorescence (IF)1 test on fixed cells with Evans’ blue counterstain is described for all four human herpesviruses, i.e., herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2), varicella-zoster virus (VZV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Comparison with immunodiffusion (ID) for HSV-2 and with ID and complement fixation (CF) for VZV and CMV demonstrated the specificity and high sensitivity of the IF test. Also introduced is a modification of the anti-complement immunofluorescence (ACIF) test for EBV-determined nuclear antigen (EBNA), permitting simultaneous titration of antibodies to this nuclear antigen and of the anti-nuclear factor (ANF). Seroepidemiological studies of these viruses in patients with Hodgkin’s disease (HD) in the Netherlands revealed the following pattern: (1) in nodular sclerosing (NS) HD there is a 4-fold (significant) elevation in antibody titer to EBV-VCA, but no elevation to EBV-EA and EBNA; (2) in mixed cellularity (MC) HD a 10-fold (significant) elevation to both EBV-VCA and EA, but no elevation to EBNA is found compared to the control groups. These patterns in NS and MC HD are different from the pattern in nasopharyngeal carcinoma (NPC) which manifests elevations in antibody titers to EBV-VCA and EA as well as to EBNA. Antibody titers to HSV, VZV and CMV are not significantly elevated in either HD or NPC.