A synthetic peptide CTL vaccine targeting nucleocapsid confers protection from SARS-CoV-2 challenge in rhesus macaques

Harris, Paul E.; Brasel, Trevor; Massey, Christopher; Herst, C.v.; Burkholz, Scott; Lloyd, Peter; Blankenberg, Tikoes; Bey, Thomas; Carback, Richard T.; Hodge, Thomas; Ciotlos, Serban; Wang, Lu; Comer, Jason E.; Rubsamen, Reid M.

doi:10.1101/2021.04.24.441228

Cited by 5 publications

(1 citation statement)

References 79 publications

(128 reference statements)

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“…We have previously demonstrated the delivery system to immunize C57BL/6 mice against the Ebola virus nucleocapsid protein with prevention of mortality and morbidity after a single intraperitoneal injection (25). An additional study showed that rhesus macaques receiving pre-exposure prophylaxis with adjuvanted microspheres from the same platform described here, loaded with peptides directed against the SARS-CoV-2 nucleocapsid protein, did not have radiographic evidence of pulmonary infiltrates characteristic of COVID-19 seen in unvaccinated controls (33). As we show here, peptides delivered using this adjuvanted microsphere technology can create a TAA-specific immune response capable of reducing tumor growth rate using the aggressive 4T1 triple negative murine breast cancer model.…”

Section: Discussionmentioning

confidence: 87%

Survivin (BIRC5) peptide vaccine in the 4T1 murine mammary tumor model: a potential neoadjuvant T-cell immunotherapy for triple negative breast cancer

Burkholz

Herst

Carback

et al. 2022

Preprint

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Background Triple negative breast cancer (TNBC) cells are, by definition, estrogen and progesterone receptor negative and do not produce high levels of the HER2 protein. Checkpoint inhibitor immunotherapy has been approved for patients with TNBC, but the prognosis for these patients is poor compared with patients presenting with other common forms of breast cancer. Survivin (BIRC5) is a tumor-associated antigen (TAA) previously described to be overexpressed in triple negative breast cancer, but not expressed, or expressed at very low levels, in normal tissue, representing targets for cytotoxic T lymphocyte (CTL) immunotherapy. Study Design A BALB/c TNBC mouse model using the 4T1 cell line was used to explore the effect of an adjuvanted microsphere synthetic vaccine containing survivin peptides on tumor growth rate. After conducting a 4T1 tumor cell inoculation dose ranging study, adjuvanted peptide vaccine microspheres were studied for their effect on tumor growth using 250 4T1 cells in one group and 500 4T1 cells in the other. The vaccine was administered via intraperitoneal injection at study start with a second dose given 14 days later. An orthotopic injection of 4T1 cells into mammary tissue was performed on the same day as the administration of the second vaccine dose. The mice were followed for up to 41 days with subcutaneous measurements of tumor volume made every three days (Figure 1). Results Vaccination with survivin peptide antigens resulted in a statistically significant diminution of tumor-takes and a deceleration of primary tumor growth volume in BALB/c mice challenged with 4T1 cells versus control. This effect was seen in both the 250 4T1 cell and the 500 4T1 cell challenge groups. The mice produced an ELISpot immune response to one of the survivin peptide antigens used in the vaccine and this response was only seen after the adjuvanted microsphere vaccine was administered. Conclusion The synthetic, adjuvanted microsphere peptide vaccine given 14 days before challenge, and on the day of challenge, was able to diminish tumor-takes and decelerate tumor growth in BALB/c mice challenged with 250 or 500 4T1 cells injected into mammary tissue. This model suggests that T-cell immunotherapy specifically targeting survivin might be an applicable neoadjuvant immunotherapy therapy for triple negative breast cancer. More pre-clinical studies and clinical trials will be needed to explore this concept further.

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Section: Discussionmentioning

confidence: 87%

Survivin (BIRC5) peptide vaccine in the 4T1 murine mammary tumor model: a potential neoadjuvant T-cell immunotherapy for triple negative breast cancer

Burkholz

Herst

Carback

et al. 2022

Preprint

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Transient Expression in HEK-293 Cells in Suspension Culture as a Rapid and Powerful Tool: SARS-CoV-2 N and Chimeric SARS-CoV-2N-CD154 Proteins as a Case Study

Lao,

Farnos,

Bueno

et al. 2023

Biomedicines

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In a previous work, we proposed a vaccine chimeric antigen based on the fusion of the SARS-CoV-2 N protein to the extracellular domain of the human CD40 ligand (CD154). This vaccine antigen was named N-CD protein and its expression was carried out in HEK-293 stably transfected cells, grown in adherent conditions and serum-supplemented medium. The chimeric protein obtained in these conditions presented a consistent pattern of degradation. The immunization of mice and monkeys with this chimeric protein was able to induce a high N-specific IgG response with only two doses in pre-clinical experiments. In order to explore ways to diminish protein degradation, in the present work, the N and N-CD proteins were produced in suspension cultures and serum-free media following transient transfection of the HEK-293 clone 3F6, at different scales, including stirred-tank controlled bioreactors. The results showed negligible or no degradation of the target proteins. Further, clones stably expressing N-CD were obtained and adapted to suspension culture, obtaining similar results to those observed in the transient expression experiments in HEK-293-3F6. The evidence supports transient protein expression in suspension cultures and serum-free media as a powerful tool to produce in a short period of time high levels of complex proteins susceptible to degradation, such as the SARS-CoV-2 N protein.

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Survivin (BIRC5) Peptide Vaccine in the 4T1 Murine Mammary Tumor Model: A Potential Neoadjuvant T Cell Immunotherapy for Triple Negative Breast Cancer: A Preliminary Study

et al. 2023

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A triple negative breast cancer model using the murine 4T1 tumor cell line was used to explore the efficacy of an adjuvanted survivin peptide microparticle vaccine using tumor growth as the outcome metric. We first performed tumor cell dose titration studies to determine a tumor cell dose that resulted in sufficient tumor takes but allowed multiple serial measurements of tumor volumes, yet with minimal morbidity/mortality within the study period. Later, in a second cohort of mice, the survivin peptide microparticle vaccine was administered via intraperitoneal injection at the study start with a second dose given 14 days later. An orthotopic injection of 4T1 cells into the mammary tissue was performed on the same day as the administration of the second vaccine dose. The mice were followed for up to 41 days with subcutaneous measurements of tumor volume made every 3–4 days. Vaccination with survivin peptides was associated with a peptide antigen-specific gamma interferon enzyme-linked immunosorbent spot response in the murine splenocyte population but was absent from the control microparticle group. At the end of the study, we found that vaccination with adjuvanted survivin peptide microparticles resulted in statistically significant slower primary tumor growth rates in BALB/c mice challenged with 4T1 cells relative to the control peptideless vaccination group. These studies suggest that T cell immunotherapy specifically targeting survivin might be an applicable neoadjuvant immunotherapy therapy for triple negative breast cancer. More preclinical studies and clinical trials are needed to explore this concept further.

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A synthetic peptide CTL vaccine targeting nucleocapsid confers protection from SARS-CoV-2 challenge in rhesus macaques

Cited by 5 publications

References 79 publications

Survivin (BIRC5) peptide vaccine in the 4T1 murine mammary tumor model: a potential neoadjuvant T-cell immunotherapy for triple negative breast cancer

Survivin (BIRC5) peptide vaccine in the 4T1 murine mammary tumor model: a potential neoadjuvant T-cell immunotherapy for triple negative breast cancer

Transient Expression in HEK-293 Cells in Suspension Culture as a Rapid and Powerful Tool: SARS-CoV-2 N and Chimeric SARS-CoV-2N-CD154 Proteins as a Case Study

Survivin (BIRC5) Peptide Vaccine in the 4T1 Murine Mammary Tumor Model: A Potential Neoadjuvant T Cell Immunotherapy for Triple Negative Breast Cancer: A Preliminary Study

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