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.

References

1. Lebedeva E.S., Ataullakhanov R.I., Khaitov R.M. Vaccines for the treatment of malignant neoplasms. Immunologiya. 2019; 40 (4): 64–76. DOI: https://doi.org/10.24411/0206-4952-2019-14008 (in Russia)

2. Sahin U., Derhovanessian E., Miller M., Kloke B.P., et al. Persona- lized RNA mutanome vaccines mobilize poly-specific therapeutic immunity against cancer. Nature. 2017; 547: 222–6. DOI: https://doi.org/10.1038/nature2300

3. Sahin U., Oehm P., Derhovanessian E., et al. An RNA vaccine drives immunity in checkpoint-inhibitor-treated melanoma. Nature. 2020; 585 (7823): 107–12. DOI: https://doi.org/10.1038/s41586-020-2537-9

4. Blass E., Ott P.A. Advances in the development of personalized neoantigen-based therapeutic cancer vaccines. Nat Rev Clin Oncol. 2021; 18 (4): 215–29. DOI: https://doi.org/10.1038/s41571-020-00460-2

5. Ott P.A., Hu Z., Keskin D.B., et al. An immunogenic personal neoantigen vaccine for patients with melanoma [published correction appears in Nature. 2018 Mar 14;555(7696):402]. Nature. 2017; 547 (7662): 217–21. DOI: https://doi.org/10.1038/nature22991

6. Keskin D.B., Anandappa A.J., Sun J., et al. Neoantigen vaccine generates intratumoral T cell responses in phase Ib glioblastoma trial. Nature. 2019; 565 (7738): 234–9. DOI: https://doi.org/10.1038/s41586-018-0792-9

7. Lang F., Schrörs B., Löwer M., Türeci Ö., Sahin U. Identification of neoantigens for individualized therapeutic cancer vaccines. Nat Rev Drug Discov. 2022; 21 (4): 261–82. DOI: https://doi.org/10.1038/s41573-021-00387-y

8. Ushakova E.I., Lebedeva E.S., Bagaev A.V., Pichugin A.V., Ataullakhanov R.I. Combined immunotherapy of metastatic carcinoma by resection of the primary tumor and subsequent reprogramming of macrophages and dendritic cells using a TLR4 agonist in laboratory mice. Immunologiya. 2021; 42 (5): 490–501. DOI: https://doi.org/10.33029/0206-4952-2021-42-5-490-501 (in Russian)

9. Bagaev A.V., Rybinets A.S., Fedorova A.A., Ushakova E.I., Lebe- deva E.S., Pichugin A.V., Ataullakhanov R.I. Synergism of TLR3 and TLR4 agonists during reprogramming of macrophages to antitumor state. Immunologiya. 2021; 42 (6): 615–30. DOI: https://doi.org/10.33029/0206-4952-2021-42-6-615-630 (in Russian)

10. Shah N.J., Najibi A.J., Shih T.Y., et al. A biomaterial-based vaccine eliciting durable tumour-specific responses against acute myeloid leukaemia. Nat Biomed Eng. 2020; 4 (1): 40–51. DOI: https://doi.org/10.1038/s41551-019-0503-3

11. Shih T.Y., Blacklow S.O., Li A.W., et al. Injectable, Tough Alginate Cryogels as Cancer Vaccines. Adv Healthc Mater. 2018; 7 (10): e1701469. DOI: https://doi.org/10.1002/adhm.201701469

12. Wang H., Najibi A.J., Sobral M.C., et al. Biomaterial-based scaffold for in situ chemo-immunotherapy to treat poorly immunogenic tumors. Nat Commun. 2020; 11 (1): 5696. DOI: https://doi.org/10.1038/s41467-020-19540-z

13. Langellotto F., Dellacherie M.O., Yeager C., et al. A Modular Biomaterial Scaffold-Based Vaccine Elicits Durable Adaptive Immunity to Subunit SARS-CoV-2 Antigens. Adv Healthc Mater. 2021; 10 (22): e2101370. DOI: https://doi.org/10.1002/adhm.202101370

14. Lebedeva E.S., Bagaev A.V., Garaeva A.Y., Chulkina M.M., Pichugin A.V., Ataullakhanov R.I. The cooperative interaction of TLR4-, TLR9- and NOD2-signaling pathways in mouse macrophages. Immunologiya. 2018; 39 (1): 4–11. DOI: http://dx.doi.org/10.18821/0206-4952-2018-39-1-4-11 (in Russian)

15. Pichugin A.V., Bagaev A.V., Lebedeva E.S., Chulkina M.M., Ataullakhanov R.I. Combined activation with agonists of TLR4, TLR9 AND NOD2 receptors synergistically increases production of cytokine-proteins in mouse macrophages. Immunologiya. 2018; 39 (4): 172–7. DOI: http://dx.doi.org/10.18821/0206-4952-2018-39-4-172-177 (in Russian)

16. Chulkina M.M., Bagaev A.V., Lebedeva E.S., Garaeva A.Ya., Pichugin A.V., Ataullakhanov R.I. Synergistic activation of inos, ifn-β, il12p40, il6, tnf-α genes transcription in macrophages under simultaneous influence with agonists of TLR4, TLR9 and NOD2 receptors. Immunologiya. 2018; 39 (4): 178–85. DOI: http://dx.doi.org/10.18821/0206-4952-2018-39-4-178-185 (in Russian)

17. Lebedeva E.S., Dzharullaeva A.Sh., Bagaev A.V., Erokhova A.S., Chulkina M.M., Tukhvatulin A.I., Pichugin A.V., Logunov D.Yu., Ataullakhanov R.I. Combined stimulation of receptors of TLR4, TLR9 and NOD2 synergistically increases protection of laboratory mice in lethal Salmonella enterica infection model. Immunologiya. 2018; 39 (5-6): 252–7. DOI: http://dx.doi.org/10.18821/0206-4952-2018-39-5-6-252-257 (in Russian)

18. Lebedeva E., Bagaev A., Pichugin A., Chulkina M., Ataullakhanov R., Lysenko A., Tutykhina I., Shmarov M., Logunov D., Naroditsky B. The differences in immunoadjuvant mechanisms of TLR3 and TLR4 agonists on the level of antigen-presenting cells during immunization with recombinant adenovirus vector. BMC Immunology. 2018; 19 (1): 26. DOI: https://doi.org/10.1186/s12865-018-0264-x

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