Enhanced and long term immunogenicity of a Her-2/neu multi-epitope vaccine conjugated to the carrier CRM197 in conjunction with the adjuvant Montanide

Autor: Christoph C. Zielinski, Joshua Tobias, Ursula Wiedermann, Nicholas J. Ede, Michael Kundi, Karin Baier, Joanna Jasinska
Rok vydání: 2017
Předmět:
0301 basic medicine
Cancer Research
Antibodies
Neoplasm
Receptor
ErbB-2
medicine.medical_treatment
Epitope
Mice
0302 clinical medicine
Adjuvant
Mice
Inbred BALB C
biology
Immunogenicity
Antibody titer
Humoral and cellular response
Her-2/neu
medicine.anatomical_structure
Oncology
030220 oncology & carcinogenesis
Vaccines
Subunit
Female
Antibody
Research Article
Recombinant Fusion Proteins
T cell
Antineoplastic Agents
Cancer Vaccines
03 medical and health sciences
Adjuvants
Immunologic
Bacterial Proteins
Antigens
Neoplasm
Cell Line
Tumor
Genetics
medicine
Animals
Humans
Cell Proliferation
business.industry
Th1-deriving response
Virology
030104 developmental biology
Polyclonal antibodies
Immunoglobulin G
Peptide vaccine
biology.protein
Hybrid peptide
Drug Screening Assays
Antitumor
business
Epitope Mapping
Neoplasm Transplantation
Zdroj: BMC Cancer
ISSN: 1471-2407
Popis: Background We previously identified three short single peptides (P4, P6 and P7) representing different B-cell epitopes on the extracellular domain of Her-2/neu for a vaccine that was tested in a phase-I clinical trial. Here we describe the improvement of the multi peptide vaccine by fusing the single peptides to a hybrid peptide P467. Methods After coupling to either virosomes or to diphtheria toxoid CRM197 (CRM), the hybrid peptide was tested in different concentrations in combination with either Montanide or Aluminium hydroxide (Alum) in preclinical studies. Results Already low amount (10 μg) of P467 conjugated to CRM led to faster onset of high antibody levels compared to the P467-virosome. The formulation P467-CRM-Montanide induced higher serum IgG antibody titers, compared with P467-CRM-Alum, as examined by ELISA using recombinant Her-2/neu or Her-2/neu natively expressed on the tumor cell line SK-BR-3. Compared to P467-CRM-Alum, higher in vitro production of IL-2 and IFNγ in the Montanide-immunized mice was induced after re-stimulation of splenocytes with CRM but also with P467, indicating a clear Th1-biased response. In contrast to the single B cell peptides, the hybrid peptide led to T cell proliferation and cytokine production as CD4 T cell epitopes were generated in the fusion region of the single peptides P4 and P6 or P6 and P7. Additionally, a significantly higher proportion IFNγ-producing CD8+ T cells was found in the P467-CRM-Montanide immunized mice, probably by Montanide-driven bystander activation. Importantly, anti-P467 IgG antibodies exhibited anti-tumor properties and the combination of anti-P467 specific IgG with Herceptin® was found to inhibit the proliferation of Her-2/neu-overexpressing cell line SK-BR-3 in a significantly higher capacity than Herceptin® alone. Conclusions Fusion of the B cell peptides has led to additional generation of CD4 T cell epitopes, and this P467-multi epitope vaccine was found to induce polyclonal antibody responses with anti-proliferative capacity against Her-2/neu. The hybrid vaccine together with Montanide induced higher and long-lasting antibody levels, Th1-biased cellular responses being superior to vaccination with the single B cell peptides. This vaccine formulation is now planned to be evaluated in a phase Ib/II study in Her-2/neu overexpressing cancer patients.
Databáze: OpenAIRE
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