Group and Strain-Specific Neutralization Sites of Infectious Bursal Disease Virus Defined with Monoclonal Antibodies

Abstract

Two somatic cell hybridizations were performed utilizing splenocytes from mice immunized with one or more strains of infectious bursal disease virus (IBDV). Supernatants from hybridoma cell lines were initially screened by the enzyme-linked immunosorbent assay (ELISA) against multiple strains of IBDV. Cell lines that secreted antibodies with ELISA reactivity patterns of interest were cloned, and their monoclonal antibodies (MCAs) were subsequently tested in cross-virus-neutralization tests. Two of the nine MCAs selected exhibited strong neutralizing activity and precipitated IBDV antigens in agar gel precipitin tests as well. MCA B69 significantly neutralized only the cloned D78 strain of IBDV, whereas MCA R63 neutralized all IBDV strains (representing both serotype I and II viruses) against which it was tested. Results of competitive ELISAs that used the R63 and B69 MCAs showed that the two neutralization sites on the D78 strain were not overlapping.

A comparison of the sequences of segment A of four infectious bursal disease virus strains and identification of a variable region in VP2

“…VP3 has been used to induce antibodies but these were only very weakly neutralizing (Fahey et aL, 1985b). Monoclonal antibodies (MAbs) have been raised against VP2 and VP3, but only those reacting to VP2 have the ability to neutralize the virus (Azad et al, 1987;Becht et al, 1988;Snyder et al, 1988). One neutralizing MAb recognition site has been mapped to a specific region of VP2 between amino acids 206 and 350 (Azad et al, 1987).…”

Anin vivoexperimental model to determine antigenic variations among infectious bursal disease viruses

“…c Number of chickens in the corresponding serum dilution group that showed the appropriate VN titre. (Snyder et al, 1988a(Snyder et al, , 1988b(Snyder et al, , 1992Snyder, 1990) and their subsequent use in an antigen-capture enzyme-linked immunosorbent assay was an important step for antigenic characterization of field isolates (van der Marel et al, 1990) and served as an effective surveillance tool. More rapid techniques were developed and the use of RT-PCR followed by restriction enzyme digestion (RFLP) of the amplified cDNA fragment (Jackwood & Jackwood, 1994) became mainstream for typing IBDV.…”

“…Surprisingly, one isolate caused 70 to 80% mortality, typical for vvIBDV, while the other isolate caused only 10% mortality (Hoque et al, 2001). Various antigenic strains of IBDV have been described in the USA based on their reactivity with mAbs: E/Dellike IBDV reacted with mAbs R63 and 67, GLS-like IBDV reacted with mAbs 10 and 57, and classical IBDV reacted with mAbs 10, R63, and B69 (Snyder et al, 1988a(Snyder et al, , 1988b(Snyder et al, , 1992Snyder, 1990). In addition, an IBDV isolated in Belgium reacted with mAbs 10, R63, 67, and B69, a combination previously not described (Letzel et al, 2007).…”

“…IBDV strains described in the early 1960s from the USA belong to what is now referenced as the classical subtype. IBDV strains isolated in the USA in the mid-1980s were described as variant strains due to change in their pathogenic phenotype and were later determined to be antigenically different from earlier viruses (Saif, 1984;Rosenberger et al, 1985;Snyder et al, 1988aSnyder et al, , 1988bSnyder, 1990). All three pathotypes of IBDV induce immunosuppression, leading to an increase in the opportunity for secondary pathogens to invade the host and cause multisystemic diseases (Cho, 1970;Fadly et al, 1976;Rosenberger & Gelb, 1978;Pejkovski et al, 1979;Yuasa et al, 1980;Moradian et al, 1990).…”

Restriction fragment length polymorphism analysis of isolates of infectious bursal disease viruses from Turkey

“…Because antigenic typing of IBDV strains using cross virus neutralization requires adaptation of the virus to grow in cell culture and production of antiserum to the new strain, new methods for characterization of these viruses were sought (Jackwood et al, 2001). Monoclonal antibodies have also been used to characterize the antigenic differences among IBDV strains (Snyder et al, 1988a). The antigen-capture enzyme linked immunosorbent assay (AC-ELISA) was used to differentiate IBDV strains (Snyder et al, 1988b).…”