A method to induce hen egg lysozyme-specific humoral immune tolerance in mice by pre-exposition with the protein's oligomers

Abstract

During treatment with protein therapeutics, such as monoclonal antibodies, the development of anti-drug antibodies is a serious side-effect of modern pharmacology. Anti-drug antibodies are produced as the number and exposure to therapeutic proteins increase. In this context, less immunogenic responses could diminish these noxious effects. Biophysical characterization of antigens, that is size, chemical composition, physical form, and degrability, are known to influence the outcome of immune responses. Here, using chemical modification, we have prepared oligomers of hen egg lysozyme (HEL), 3- to 5-mer, as a typical antigen in immunology and evaluated the efficacy as a tolerogen in HEL-specific antibody responses. Our results clearly demonstrated that pre-exposed the HEL-oligomers into mice effectively suppressed HEL-specific IgG responses regardless of the cross-linking mode. Therefore, the oligomerization is a method to induce tolerogenicity of proteins and may emerge as a promising strategy to control the production of undesirable anti-protein drug antibodies.

Relationship between protein conformational stability and its immunogenicity when administering antigens to mice using adjuvants—Analysis employed the CH2 domain in human antibodies

“…The immunization schedule we propose, with 1-week intervals, may appear shorter when contrasted with the conventional method that typically utilizes a 2-week interval for protein vaccine immunization. The immunization protocol employed in this study was developed following methodologies previously established in our laboratory [ 17 , 18 , 21 , 36 , 38 ]. Blood samples were drawn every week under anesthesia using sevoflurane to remove pain.…”

Modulation of Aggregation and Immunogenicity of a Protein: Based on the Study of Hen Lysozyme

“…. HEL monomer (thick line) and HEL oligomers (dotted line) at a concentration of 0.05 mg/mL were derived from a pre-exposed solution used in the experiments shown in Fig.8Aof ref 74)…”