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. 2020 Mar 24;14(3):2994-3003.
doi: 10.1021/acsnano.9b07865. Epub 2020 Mar 5.

Antibody Response against Cowpea Mosaic Viral Nanoparticles Improves In Situ Vaccine Efficacy in Ovarian Cancer

Antibody Response against Cowpea Mosaic Viral Nanoparticles Improves In Situ Vaccine Efficacy in Ovarian Cancer (V体育2025版)

Sourabh Shukla et al. ACS Nano. .

VSports在线直播 - Abstract

Cancer immunotherapies are designed to facilitate recognition and elimination of transformed cells by the immune system. We have established the immunotherapeutic efficacy of the plant virus cowpea mosaic virus (CPMV) as an in situ vaccine in several syngeneic tumor mouse models as well as in companion dogs with metastatic melanoma. Intratumoral injection of CPMV modulates the local tumor microenvironment to relieve immunosuppression and potentiate antitumor immunity VSports手机版. The viral nucleocapsid that drives this antitumor immunity, however, also is a potent immunogen itself, and thus immune response in the form of anti-CPMV antibodies is expected during the treatment based on repeat administrations. Moreover, being part of the food chain, pre-existing antibodies to plant viruses may be prevalent. The presence of such pre-existing anti-CPMV immunity could potentially impact immunotherapeutic efficacy of the in situ vaccine and could have translational implications. To address such concerns, this study evaluated the efficacy of CPMV in situ vaccine in the presence of pre-existing antibodies in a syngeneic mouse model of ovarian cancer. Our results indicate that prior exposure to CPMV had no negative impact on the efficacy of CPMV in situ vaccine. Strikingly, an improved efficacy of CPMV in situ vaccine was observed. This study therefore presents an important milestone in the translational development of plant viral-based in situ vaccines and alleviates concerns about the presence of anti-CPMV antibodies, which are developed during the course of treatment but have no impact on immunotherapeutic efficacy. .

Keywords: CPMV; antiviral antibodies; cancer immunotherapy; in situ vaccine; ovarian cancer; plant viruses V体育安卓版. .

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Conflict of interest statement (V体育安卓版)

The authors declare no competing financial interest.

Figures

Figure 1.
Figure 1.
Bioconjugation and characterization of CPMV and eCPMV particles. CPMV and eCPMV were propagated in plants. Postpurification, particle integrity was determined using size exclusion chromatography (A); N-hydroxysuccinimide (NHS) chemistry was used to conjugate fluorescent dye Sulfo-Cy5 and/or PEG2000 to the (e) CPMV coat proteins via the lysine residues to obtain CPMV-Cy5, eCPMV-Cy5, PEG-eCPMV, and PEG-eCPMV-Cy5 (B). SDS-PAGE (4–12% NuPAGE gel) gels were imaged on a FluorChem R imager, first using the MultiFluor Red channel under 607 nm excitation (fluorescence image) to verify dye conjugation and then stained with GelCode Blue Safe protein stain and visualized under white light (stained image). Stained gel image was used to verify and quantify covalent conjugation of PEG molecules to the small (S) and large (L) coat proteins (CP) of eCPMV.
Figure 2.
Figure 2.
Immunogenicity of CPMV, eCPMV, and PEG-eCPMV leads to generation of anti-CPMV antibodies. Immunogenicity of intraperitoneal (i.p.) administered CPMV, eCPMV, and PEG-eCPMV was evaluated in CPMV naive (SET 1) or preimmunized (SET 2) C57BL6 female mice; mice received 6 weekly i.p. doses to mimic the typical administration schedule for i.p. in situ vaccines (A, B). Anti-CPMV antibody titers (C, D) and isotypes (E) were measured using ELISAs performed on CPMV-coated plates. Statistical analysis was performed by ordinary one-way ANOVA and Tukey’s multiple comparisons test using GraphPad Prism software.
Figure 3.
Figure 3.
Pre-exposure to CPMV improves efficacy of CPMV in situ vaccine in ID8-Defb29/Vegf-a ovarian tumors. Efficacy of CPMV in situ vaccination (100 μg in 200 μL of PBS) against i.p. established ID8-Defb29/Vegf-a ovarian tumors in female C57BL6 mice was tested in CPMV-naive vs CPMV-pre-exposed mice (A, B). Tumor progression was monitored by measuring gains in circumference (C) and body weight (D). Survival was monitored for 125 days (E). Kaplan–Meier plots were used to compare survival between groups. Statistical analysis on the survival curves was performed using the log-rank (Mantel–Cox) test using GraphPad Prism 8 software.
Figure 4.
Figure 4.
Anti-CPMV antibodies enhance the phagocytic uptake of CPMV, eCPMV, and PEG-eCPMV by bone-marrow-derived macrophages (BMDMs). BMDMs cultured for 10 days in the presence of M-CSF were gated as CD11b+ F4/80+ cells. Phagocytic uptake of Cy5-labeled CPMV, eCPMV, and PEG-eCPMV was compared using flow cytometry in the absence and presence of day 49 sera (immunized sera) from CPMV-pre-exposed mice and sera from naive mice (naive sera) (B, C, and D). Particles (1 μg) were pre-exposed to sera (1:200 dilutions) for 20 min and subsequently incubated with 200 000 BMDMs for 30 min. Cell uptake was analyzed using flow cytometry. Statistical analysis was performed using ordinary one-way ANOVA–Tukey’s multiple comparisons test (****p < 0.0001; **p = 0.0048).
Figure 5.
Figure 5.
eCPMV and PEG-eCPMV in situ vaccines display improved efficacy against ID8-Defb29/Vegf-a-Luc ovarian tumors in CPMV pre-exposed mice. The efficacy of eCPMV and PEG-eCPMV was tested in CPMV-naive and CPMV-pre-exposed female C57BL6 mice challenged i.p. with bioluminescent ID8-Defb29/Vegf-a-Luc ovarian cancer cells (A). Tumor progression was monitored using bioluminescence imaging (B), body weight (C), and survival (D); color scheme in C and D: PBS, green; naive eCPMV, red; naive PEG-eCPMV, purple; pre-exposed/eCPMV, blue; pre-exposed/PEG-eCPMV, orange. Statistical analysis was performed using unpaired t test for weights (d61: eCPMV naive vs pre-exposed **p = 0.006; PEG-eCPMV naive vs pre-exposed **p = 0.03; d78: eCPMV naive vs pre-exposed * p = 0.02); survival curves were compared by log-rank (Mantel–Cox) test using GraphPad Prism 8 software.

References

    1. Rosenberg SA Decade in Review-Cancer Immunotherapy: Entering the Mainstream of Cancer Treatment. Nat. Rev. Clin. Oncol 2014, 11, 630–632. - PMC - PubMed
    1. Joyce JA; Fearon DT T Cell Exclusion, Immune Privilege, and the Tumor Microenvironment. Science 2015, 348, 74–80. - "VSports注册入口" PubMed
    1. Goto T Radiation as an In Situ Auto-Vaccination: Current Perspectives and Challenges. Vaccines (Basel, Switz.) 2019, 7, 100. - PMC - PubMed
    1. Le QV; Suh J; Choi JJ; Park GT; Lee JW; Shim G; Oh YK In Situ Nanoadjuvant-Assembled Tumor Vaccine for Preventing Long-Term Recurrence. ACS Nano 2019, 13, 7442–7462. - PubMed
    1. Bommareddy PK; Kaufman HL Unleashing the Therapeutic Potential of Oncolytic Viruses. J. Clin. Invest 2018, 128, 1258–1260. - VSports最新版本 - PMC - PubMed

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