Characterization of Schistosoma mansoni monoclonal antibodies which block in‐vitro killing: failure to demonstrate blockage of immunity in vivo

Abstract

We have characterized various anti-schistosomular surface MoAbs previously shown to partially block in-vitro killing of schistosomula by human sera and eosinophils (Dunne et al. 1987). Immunodiffusion analysis showed that four IgM and one IgG3 MoAbs recognized periodate sensitive epitopes on the same molecular species present in schistosomular antigen but their patterns of reactivity with soluble egg antigen demonstrated that at least three distinct epitopes were involved. SDS-PAGE analysis showed the IgMs to react with 125I-labelled surface antigens of Mr 35,000-38,000 and Mr 20,000, and the IgG3 to react with an Mr 38,000 antigen. In spite of their effect in vitro, transfer of the IgM MoAbs at the time of challenge of mice vaccinated with irradiated cercariae or of mice injected with an unrelated (anti-Mr 16,000) protective MoAb failed to produce in-vivo blocking. Similarly, injection of Schistosoma mansoni eggs, prior infection with worms of one sex, or passive transfer of serum from single-sex infected mice failed to influence the resistance conferred by vaccination with irradiated cercariae.

Structural characterization of N-glycans from the freshwater snail Biomphalaria glabrata cross-reacting with Schistosoma mansoni glycoconjugates

“…The hemolymph was collected by cardiac puncture of the snails, shortly centrifuged after sampling, and stored at 220 8C. The supernatant from M2D3H hybridoma cell culture (Bickle and Andrews 1988), producing a monoclonal antibody (mAb M2D3H) recognizing the Fuc(a1-3)GalNAc-epitope , was used 10-fold concentrated for ELISA and western blot analysis. Rabbit hyperimmune serum raised against SEAs from S. mansoni [anti-SEA (Hamilton et al 1999)] was used for immunoaffinity chromatography as well as for ELISA and western blot analyses.…”

Induction of immunity against Schistosoma mansoni by drug (Roll‐3128)‐terminated infections: analysis of surface antigen recognition

“…The monoclonal antibodies (MoAbs) M7B3A, M22H12C and M22CIC against the M, 16000, M, 32 000 and M, 38 000 larval surface antigens respectively have been described (Bickle et al 1986, Bickle & Andrews 1988. The indirect fluorescent antibody tests (IFAT) were carried out at 37°C using a 1/50 dilution of MoAb ascites fluid and a 1/60 dilution of fluorescein-conjugated rabbit anti-mouse whole immunoglobulin serum (Nordic).…”

Identification and Characterization of Keyhole Limpet Hemocyanin N-Glycans Mediating Cross-reactivity with Schistosoma mansoni

“…Anti-SEA antibodies, coupled to NHS-activated Sepharose, were provided by T. Lehr, Institute of Biochemistry, University of Giessen. The monoclonal antibody M2D3H was produced and kindly provided by Q. Bickle, London School of Hygiene and Tropical Medicine, London University, UK (25).…”

“…Incubation with 100 l per well of the primary antibody in TTBS-10 (TBS 1:10 diluted, 0.05% Tween 20) containing 0.25% bovine serum albumin was performed for 1 h at 37°C. Primary antibodies used were: monoclonal antibody M2D3H (ascites fluid, dilution 1:100,000 (25)), murine S. mansoni infection serum (dilution 1:2,000), an anti-KLH rabbit hyperimmune serum (␣KLH, dilution 1:100,000) (22)), and an anti-soluble egg antigen rabbit hyperimmune serum (␣SEA, dilution 1:100,000) (24). After multiple washes with TTBS-10, alkaline phosphatase-conjugated goat anti-mouse Ig (Dako Diagnostics, Hamburg, Germany, diluted 1:1,000) or goat anti-rabbit Ig (Sigma, diluted 1:1,000) in TTBS-10 containing 0.25% bovine serum albumin were applied.…”