Antibody class capture assays for varicella-zoster virus.

Pooled monoclonal antibodies to varicella-zoster virus (VZV) were used as "detector" antibodies in a four-phase enzyme immunofluorescence assay for determination of immunoglobulin M (IgM), IgA, and IgG antibodies to VZV. Polyclonal antisera specific for heavy chains of human IgM, IgA, and IgG were employed as "capture" antibodies on the solid phase. The antibody class capture assay (ACCA) for VZV IgM antibody detected high titers of virus-specific IgM in all patients with varicella and in 5 of 10 zoster patients. VZV IgM antibody was not detected in patients with primary herpes simplex virus infections or in other individuals without active VZV infection, with one exception, a patient with encephalitis who had other serological findings compatible with a reactivated VZV infection. VZV-specific IgA and IgG antibody titers demonstrable by ACCA were compared with those measured by solid-phase indirect enzyme immunofluorescence assay (EIFA). VZV IgA antibody titers detected in patients with varicella and zoster were variable and could not be considered to be reliable markers of active VZV infection. IgA antibody titers detected by ACCA tended to be higher than those demonstrated by solid-phase indirect EIFA in varicella and zoster patients. VZV IgG antibody titers detected by ACCA in patients with varicella, and to a lesser extent in zoster patients, were as high as or higher than those demonstrated by solid-phase indirect EIFA. However, ACCA was totally insensitive in detecting VZV IgG antibody in individuals with past infections with VZV and would not be a suitable approach for determination of immunity status to VZV.

Use of monoclonal antibodies to human immunoglobulin M in "capture" assays for measles and rubella immunoglobulin M.

Monoclonal antibodies to human immunoglobulin M (IgM) were used in a four-phase enzyme immunofluorescence "capture" assay for determination of IgM antibodies to measles and rubella viruses. Little or no background reactivity was seen in the test system, and interfering effects of rheumatoid factor were avoided by preabsorption of test sera with aggregated human IgG. Virus-specific IgM antibody was demonstrable in 23 of 24 patients with serological evidence of measles virus infections and in 36 of 36 patients with serological evidence of postnatal rubella infection. A few of the rubella patients did not show IgM antibody until 5 days after onset of illness. The enzyme immunofluorescence assay was able to demonstrate rubella IgM antibody in congenitally infected newborns, whereas indirect immunofluorescence results for virus-specific IgM were negative. Viral IgM antibody was not detected in persons with past infections with the test viruses, in young children without evidence of past infection, or in patients infected with heterotypic viruses, rickettsiae, chlamydiae, or mycoplasmas.

Production of monoclonal antibodies to human IgM for assay of viral IgM antibodies.

Monoclonal antibodies to human IgM were produced by fusing the Sp 2/0-Ag 14 line of mouse myeloma cells with spleen cells from BALB/c mice immunized with purified human IgM. From 6 clones which secreted antibody to human IgM, the one which produced the highest levels of antibody and grew relatively rapidly was selected for expansion and production of immune reagents for viral IgM antibody assays. Mouse ascitic fluids produced with this clone of cells had antibody titers for human IgM of 1 X 10(-10) by indirect enzyme immunofluorescence assay (EIFA). The monoclonal antibodies were found to belong to subclass 1 of murine IgG, and their specificity was shown to be directed against the Fab portion of the mu chain of human IgM. Antibodies from murine ascitic fluid conjugated with horseradish peroxidase were shown to be suitable for assay of measles IgM antibody by indirect immunoperoxidase staining. Antibodies conjugated with alkaline phosphatase could be used in an indirect EIFA for determination of measles IgM antibodies; use of monoclonal conjugates in this system eliminated the nonspecific activity observed in tests utilizing polyclonal anti-mu reagents. Further, the monoclonal antibodies were highly satisfactory for use in a 'capture' system for viral IgM antibody assays. The availability of monoclonal antibodies to human IgM overcomes problems with specificity, consistency and supply which have previously hindered development and standardization of viral IgM antibody assays.

Serological reactivity of some monoclonal antibodies to varicella-zoster virus.

Monoclonal antibodies to varicella-zoster virus, free from host cell reactivity, were produced by cell fusion technics. Antibodies from four different clones showed diverse activities in neutralization immunofluorescence and complement fixation assays. The antibodies provide useful reagents for viral diagnosis and viral antigenic characterization.