Comparison of adjuvants for immune potentiating properties and side effects in mice

doi:10.1016/0165-2427(94)05413-M

Veterinary Immunology and Immunopathology

Volume 48, Issues 1–2, September 1995, Pages 123-138

https://doi.org/10.1016/0165-2427(94)05413-M Get rights and content

Abstract

Four types of adjuvants were evaluated as alternatives to the use of Freund's complete adjuvant in mice. The adjuvants evaluated included a water-in-oil emulsion (Specol), a microorganism (Lactobacillus), preformed immune-stimulating complexes (ISCOM) containing rabies virus glycoprotein and a saponin, Quil A. The adjuvants and saline were combined with three weak immunogens (a synthetic peptide, a self antigen and a particulate antigen) and given by three different routes (intra-peritoneal, subcutaneous and dorsal in the foot). The evaluation was based on clinical observations, behavioural studies, pathological lesions and capacity to support immunological responses to weak immunogens. Lesions were most severe after injection of antigen combined with Freund's adjuvant or Quil A, mild to moderate with Specol and minimal with Lactobacillus, iscom conjugates or saline. Despite pathological changes, no signs of prolonged pain or distress could be demonstrated based on clinical observations and behavioural studies. Minimal immunological responses were found after injection of antigen in combination with saline or Lactobacillus. T-cell activation and high antibody responses were found after injection of antigen-iscom conjugates or antigen in Freund's adjuvant emulsions. After Specol/antigen immunisations T-cell activation was demonstrated and high antibody titres were found except for Specol/self antigen immunisations. Presented data suggest that Specol is a possible alternative to Freund's complete adjuvant for the induction of an immune response against weak immunogens except possibly self antigens, for which preformed iscoms seem very suitable.

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The protein was combined with an oily adjuvant (Specol) to improve the immune response, because many recombinant proteins are usually poor as immunogens. Some authors have used Specol as an alternative to Freund's Adjuvant, since its administration causes fewer adverse effects [26,27]. Based on the results obtained by Western blot in Fig. 3b (line 1), we concluded that the mice inoculated with the F0 protein produced antibodies that recognized the proteins homonymous to CDV.
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Saponin based adjuvants such as Quil A elicit the formation of immune stimulating complexes (ISCOMs). The use of ISCOMs as adjuvants for experimental polyclonal antibody production is limited by their complicated and labor-intensive preparation [11]. Mineral-based adjuvants such as aluminum salt adjuvants are known to increase responses against weak immunogens.
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These therapeutic aids need hyperimmunisation of animals. Since these are subject to a certain number of interventions such as injections and blood samplings, animal welfare prescriptions have to be taken into account.
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Typically, Specol mainly induced antibodies of the IgG1 isotype1. Previous study in mice7 concluded that Specol might be an acceptable alternative to Freund's complete adjuvant (FCA). Specol is generally regarded as safe for use in animals1,2, showing fewer adverse effects than other wateroil adjuvants like FCA and had demonstrated strong immunostimulating properties.
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This chapter discusses antibodies as critical reagents in a wide variety of research methodologies. The antibodies used are either polyclonal or monoclonal, with each having distinct advantages and disadvantages. Polyclonal antibodies are heterogeneous and bind to several different antigen epitopes. While decreasing the specificity and potentially increasing nonspecific reactions, the polyclonal antibody is also more likely to successfully bind to the specific antigen in a variety of different test conditions. The production of polyclonal antibodies remains a common research procedure involving vertebrate animal use. Rabbits are the most commonly used laboratory animals for the production of polyclonal antibodies. The rabbit offers many advantages over other species. The adequate body size of the rabbit and the ready accessibility of the marginal ear vein and central auricular artery afford a technically easy procedure for the collection of large blood samples. The excellent responsiveness of the rabbit to a wide variety of antigens, the presence of a single primary Immunoglobulin G isotype, the vast amount of information available on the production, and the reavailability of secondary reagents further enhance the reputation of the rabbit in polyclonal antibody production. Polyclonal antibody production is essential in research activity and rabbits continue to serve as one of the primary species used in polyclonal antibody production. The schedules and methodologies used to immunize rabbits and produce polyclonal antibodies continue to vary depending on the immunogen, the adjuvant, and the end purpose of the antibody.
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Comparison of adjuvants for immune potentiating properties and side effects in mice

Abstract

Vet. Immunol. Immunopathol.

J. Immunol. Methods

Vaccine