ALTEX :: Alternatives to Animal Experiments

1998, Supplement

A Comparison of Commercially Available Adjuvants in Balb/C-Mice Immunised with a Weekly Immunogenic Peptide

Christine Zwerger, Roland Plesker, Kyriakos Papadopulos, Klaus Cussler, and Joachim Hartinger

Paul-Ehrlich-Institut, D-Langen

SUMMARY

The antibody-stimulating activities and side effects of commercially available adjuvants were compared in BALB/c mice immunised with the immunosuppressive (ISU) peptide of HIV I. Clinical and pathological-histological parameters as well as behavioral changes, were used to assess the distress and pain caused to the animals. Complete Freund's Adjuvant (FAk) was used as the positive control and PBS as the negative control. The oil-based adjuvants Montanide ISA 51® (M 51), Specol®, and Hunter's TiterMax Gold® (HTMG) were used with the ISU-peptide conjugated to KLH. The water-soluble Gerbu Adjuvant® was administered together with the antigen KLH conjugate and also with the ISU-peptide conjugated to cholera toxin B subunit (ChTxB). The Dutch "Code of Practice" was used as a guideline for all immunisations. No changes in animal activity or behavior was observed in any of the groups. All the oil-based adjuvants gave rise to swellings and encapsulations, which were most pronounced in the HTMG-group. Although body weight increased throughout the study, no increase was seen in any adjuvant group for a short period after each booster immunisation, nor after the first immunisation in the HTMG group. FAk induced a light fever after all immunisations. FAk, Specol, and M 51 as well as Gerbu-ChTxB induced antibody titres which were detectable in the ELISA, but no detectable antibody the water-soluble adjuvants or following administration of HTMG applied with the ISU-peptide-KLH conjugate.

Keywords: adjuvant, animal welfare, antibody titer, Cholera toxin B subunit, ELISA, mice, monoclonal antibody