Immunogenicity of a multi-antigen vaccine made from a lysate of tumor malignant cells or from primary solid tumor tissue

Abstract

Introduction. Personalized vaccination with tumor antigens is one of the most promising trends in immunotherapy for cancer patients. Various technological platforms are used to create a therapeutic vaccine. The most progressive are vaccines based on synthetic peptides that copy mutant neoantigens, as well as vaccines that are RNA- or DNA-vectors encoding tumor neoantigens. The mentioned technological approaches are very complex and expensive, and require considerable time for their implementation. We are developing an extremely simple, low-cost method for preparing a personalized antitumor vaccine for its wide use in the complex treatment of cancer patients. Our approach is to use tumor tissue as a source of tumor antigens for a personalized vaccine. Vaccines prepared from tumor tissue or cancer malignant cells we define as personalized multi-antigen cancer vaccines.

Aim – to study the immunogenicity of two variants of a multiantigen antitumor vaccine prepared from a tissue lysate of primary solid 4T1 carcinoma or from a pure culture of malignant 4T1 carcinoma cells.

Material and methods. The multi-antigenic vaccines studied in this work were prepared from solid 4T1 mammary carcinoma tissue of BALB/c mice or from a pure population of malignant 4T1 carcinoma cells grown under in vitro cell culture conditions. The homogenate (lysate) of tumor tissue or 4T1 cells was supplemented with molecular immunoadjuvants from the class of PRR-agonists. The resulting compositions were used to immunize BALB/c mice. Immunogens were administered intraperitoneally, four times, with intervals of 2 weeks between injections. Systemic immune responses against 4T1 carcinoma antigens used in the studied immunogens were determined according to the numbers of antitumor T cells in the spleen and the levels of tumor-specific antibodies in the blood serum of mice. Antigen-reactive CD4 and CD8 T memory cells, and T effector cells secreting interferon-γ were analyzed by ELISPOT. Antibodies to 4T1 carcinoma cell surface antigens were analyzed by flow cytometry. Antibo- dies to intracellular antigens of 4T1 carcinoma were studied by enzyme immunoassay.

Results. Multi-antigenic antitumor vaccines based on 4T1 tumor tissue homogenate (Multivac-1) or on 4T1 malignant cell lysate (Multivac-4) demonstrated high immunogenicity. After several injections of these immunogens, BALB/c mice developed intense T cell and antibody responses against 4T1 carcinoma antigens. The total response of CD4 T cells (effector cells + memory cells) in the spleen was about 3000 T cells (per 1 million T cells) after immunization with the Multivac-1 and about 8000 T cells after immunization with the Multivac-4. The total response of CD8 T cells (effector cells + memory cells) was about 3000 T cells after immunization with multivac-1 and about 5000 T cells after immunization with Multivac-4. Both vaccine preparations induced the production of antitumor antibodies. Significantly more antitumor antibodies were produced in response to the multivac-4 vaccine than in response to the Multivac-1 vaccine.

Conclusion. The use of solid tumor tissue or tumor malignant cells as a source of antigens seems to be a promising technological approach for the preparation of a personalized therapeutic antitumor vaccine. Immunogens prepared from tissue lysate of 4T1 primary solid carcinoma or from a pure culture of malignant 4T1 carcinoma cells, enhanced with molecular adjuvants from the class of PRR-agonists, induce strong antitumor adaptive immune responses such as the generation of CD4 and CD8 effector T cells and memory T cells, and also production of specific antibodies to 4T1 carcinoma antigens.

Keywords:therapeutic cancer vaccine; T cell responses; antibodies against tumor antigens

For citation: Ushakova E.I., Fedorova A.A., Lebedeva E.S., Pichugin A.V., Ataullakhanov R.I. Immunogenicity of a multi-antigen vaccine made from a lysate of tumor malignant cells. Immunologiya. 2022; 43 (4): 389–400. DOI: https://doi.org/10.33029/0206-4952-2022-43-4-389-400 (in Russian)

Funding. The study was supported by a grant from the Russian Science Foundation No. 20-15-00391.

Conflict of interests. The authors declare no conflict of interests.

Authors` contribution. Conducting experiments, statistical processing, editing the article – Ushakova E.I.; conduc- ting experiments, editing the article – Fedorova A.A.; vaccine design, data analysis, article editing – Lebedeva E.S.; planning and conductiong experiments, data analysis – Pichugin A.V.; concept, design of experiments, data analysis, article writing – Ataullakhanov R.I.

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EDITOR-IN-CHIEF
EDITOR-IN-CHIEF
Musa R. Khaitov

Corresponding member of Russian Academy of Sciences, MD, Professor, Director of the NRC Institute of Immunology FMBA of Russia

Вскрытие
Medicine today

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