Comparison of immunoglobulin A (IgA), IgG, and IgM enzyme-linked immunosorbent assays, plaque reduction neutralization assay, and complement fixation in detecting seroresponses to rotavirus vaccine candidates

Abstract

In a phase 1 study to evaluate human-rhesus rotavirus reassortant vaccines, 116 infants 1 to 5 months of age received one of the following five preparations: the serotype 1 reassortant, the serotype 2 reassortant, rhesus rotavirus (serotype 3), a bivalent preparation (serotypes 1 and 3), or a placebo. Seroresponses to the different vaccines were measured by plaque reduction neutralization assay (PRNA); rotavirus-specific immunoglobulin A (IgA), IgG, and IgM enzyme-linked immunosorbent assays (ELISAs); and complement fixation (CF). The seroresponse rate, calculated by using a fourfold or greater antibody rise by any assay, was similar in the four vaccine groups (83 to 96%). When the data from all the vaccinees were pooled, IgA ELISA, IgG ELISA, and PRNA were comparable in detecting seroresponses (67, 62, and 70%, respectively) and more efficient than IgM ELISA (53%) and CF (44%). When the vaccinees were analyzed by age, the overall seroresponse rates were the same for infants 1 to 2 and 3 to 5 months old (90%). The IgA ELISA and PRNA were the most efficient for detecting antibody rises in both age groups. IgG ELISA was among the least efficient methods for detecting antibody rises in the younger age group but among the most efficient in the older age group (44 versus 78%). CF was among the least efficient methods in both age groups but was significantly better in the older age group than in the younger age group (54 versus 21%). Our findings show that ELISA, in particular rotavirus-specific IgA ELISA, is a sensitive indicator of vaccine takes in 1-to 5-month-old infants, the target population for vaccination. The ELISA should also be very useful in demonstrating natural rotavirus infections in field studies in which a stool specimen from a diarrheal episode is not always available. The ELISA has the advantages of being easier and quicker and requiring less serum than PRNA, but it does not give serotype-specific information about the immune response. * Corresponding author. tant human rotavirus serotypes are represented by the reassortants (serotypes 1, 2, and 4) and RRV (serotype 3). An integral part of early trials with these different rotavirus vaccine candidates is the evaluation of their immunogenicity. In this study, serologic responses in 1to 5-month-old

Rotaviruses: Immunological Determinants of Protection Against Infection and Disease

“…To further enhance the response against human G types, single gene reassortant viruses were constructed that expressed vp7 from human G type 1, 2, or 4 (Midthun et al, , 1986. Oral inoculation of infants with RRV x human reassortant strains induced rotavirus-specific IgA and RRV-specific neutralizing antibodies in serum Midthun et al, 1989;Perez-Schael et al, 1990b;Wright et al, 1991;Dagan et al, 1992). However, neutralizing antibody responses directed against G type 1, 2, or 4 after immunization with reassortant viruses containing these G types were not dramatically greater than heterotypic neutralizing antibody responses obtained after immunization with RRV alone Perez-Schael et al, 1990b;Dagan et al, 1992).…”

Clinical evaluation of a single oral dose of human‐bovine (UK) reassortant rotavirus vaccines Wa x UK (P1A[8],G6) and Wa x (DS‐1 x UK) (P1A[8],G2)

“…Preinoculation and postinoculation sera were tested for rotavirus-speci®c IgA and IgG antibodies by ELISA, using rhesus rotavirus (P5B[3], G3) as antigen as previously described [Midthun et al, 1989;Clements-Mann et al, 1999]. The amount of antibody determined by this method re¯ects the quantity of antibody speci®c for shared common antigens such as the major inner capsid protein VP6 and not those speci®c for different rotavirus serotypes.…”

“…Such shared antigenicity can be demonstrated, for example, by cross-reactivity of antisera to various strains of rotaviruses in a complement ®xation test [Kapikian et al, 1976]. Paired sera were also tested by plaque reduction neutralization (PRN) antibody assay as previously described [Midthun et al, 1989]. Viruses used in the PRN assays included: G4), and AU32 (P1A[8], G9) human rotavirus strains; Wa x UK (P1A[8], G6) and Wa x (DS-1 x UK) (P1A[8], G2) reassortant vaccine strains; and the UK (Compton, P7[5], G6) bovine rotavirus strain.…”

Take of rhesus‐human reassortant tetravalent rotavirus vaccine in breast‐fed infants

“…The IgA rotavirus ELISA antibody test on paired sera has been described in detail previously (22). Briefly, microtiter plates were coated with hyperimmune goat serum and subsequently with supernatant of RRV infected or uninfected MA104 cell culture.…”