ovalbumin and Neoplasm-Metastasis

The targeted delivery of messenger RNA (mRNA) to desired organs remains a great challenge for in vivo applications of mRNA technology. For mRNA vaccines, the targeted delivery to the lymph node (LN) is predicted to reduce side effects and increase the immune response. In this study, we explored an endogenously LN-targeting lipid nanoparticle (LNP) without the modification of any active targeting ligands for developing an mRNA cancer vaccine. The LNP named 113-O12B showed increased and specific expression in the LN compared with LNP formulated with ALC-0315, a synthetic lipid used in the COVID-19 vaccine Comirnaty. The targeted delivery of mRNA to the LN increased the CD8

Topics: Amino Alcohols; Animals; Antigens; Cancer Vaccines; CD8-Positive T-Lymphocytes; Decanoates; Immunologic Memory; Liposomes; Lymph Nodes; Mice; mRNA Vaccines; Nanoparticles; Neoplasm Metastasis; Neoplasms; Ovalbumin

Myeloid derived suppressor cells (MDSC) are important regulators of immune responses. We evaluated the mechanistic role of MDSC depletion on antigen presenting cell (APC), NK, T cell activities and therapeutic vaccination responses in murine models of lung cancer.. Individual antibody mediated depletion of MDSC (anti-Gr1 or anti-Ly6G) enhanced the antitumor activity against lung cancer. In comparison to controls, MDSC depletion enhanced the APC activity and increased the frequency and activity of the NK and T cell effectors in the tumor. Compared to controls, the anti-Gr1 or anti-Ly6G treatment led to increased: (i) CD8 T cells, (ii) NK cells, (iii) CD8 T or NK intracytoplasmic expression of IFNγ, perforin and granzyme (iv) CD3 T cells expressing the activation marker CD107a and CXCR3, (v) reduced CD8 T cell IL-10 production in the tumors (vi) reduced tumor angiogenic (VEGF, CXCL2, CXCL5, and Angiopoietin1&2) but enhanced anti-angiogenic (CXCL9 and CXCL10) expression and (vii) reduced tumor staining of endothelial marker Meca 32. Immunocytochemistry of tumor sections showed reduced Gr1 expressing cells with increased CD3 T cell infiltrates in the anti-Gr1 or anti-Ly6G groups. MDSC depletion led to a marked inhibition in tumor growth, enhanced tumor cell apoptosis and reduced migration of the tumors from the primary site to the lung compared to controls. Therapeutic vaccination responses were enhanced in vivo following MDSC depletion with 50% of treated mice completely eradicating established tumors. Treated mice that rejected their primary tumors acquired immunological memory against a secondary tumor challenge. The remaining 50% of mice in this group had 20 fold reductions in tumor burden compared to controls.. Our data demonstrate that targeting MDSC can improve antitumor immune responses suggesting a broad applicability of combined immune based approaches against cancer. This multifaceted approach may prove useful against tumors where MDSC play a role in tumor immune evasion.

Topics: Angiogenesis Inhibitors; Animals; Antigen-Presenting Cells; Antineoplastic Agents; Apoptosis; Biomarkers, Tumor; Bone Marrow Cells; Carcinoma, Lewis Lung; Cell Adhesion; Cell Proliferation; Cytotoxicity, Immunologic; Disease Models, Animal; Killer Cells, Natural; Mice; Mice, Inbred C57BL; Myeloid Cells; Neoplasm Metastasis; Ovalbumin; Spleen; T-Lymphocytes; Treatment Outcome; Tumor Burden; Vaccination

To develop a more economic and applicable substitute for modification of dendritic cells (DCs) by capped mRNA in induction of anti-tumor immunity.. Four DNA plasmids as templates of mRNA in vitro transcription were constructed: pmRNA luciferase (Luc), pmRNA internal ribosomal entry site (IRES)-Luc, pmRNA ovalbumin (OVA), and pmRNA IRES-OVA. The translational efficiency of uncapped-Luc, capped-Luc, or IRES-Luc mRNA in murine DCs was detected via a Luc reporter system. Nine C57BL/6 mice were divided into 3 equal groups to be injected intraperitoneally with DCs transfected with IRES-OVA or capped-OVA mRNA and untreated DCs respectively, 1 weeks later the mice were injected with splenocytes wrapped with the target peptide or control peptide labeled by CFSE, and 4 hours later the mice were killed and suspension of splenocytes was made to test the activity of cytotoxic lymphocytes (CTLs). Another 30 mice were divided into 3 equal groups to undergo immunization by DCs as mentioned above, 1 week later mice melanoma cells of the line MO5 stably expressing OVA were injected the caudal vein, and 3 weeks later the mice were killed and their lungs were taken out to observe the metastasis of tumor, using OVA as a target antigen.. Insertion of IRES into upstream of gene of interest in mRNA transcriptional templates didn't affect the yield of mRNA in vitro transcription. The level of Luc activity expressed by IRES-Luc mRNA in the DCs was 20 times higher than that by expressed by Uncapped Luc mRNA, and one time higher than that by Capped-Luc mRNA 8 h after transfection of DC. Expression of Luc could be detected up to 96 h after the transfection with IRES-Luc and Capped-Luc mRNA. Four hours after the injection of peptides the CTL activity of the mice immunized with DCs pulsed with Capped-OVA mRNA was (28 +/- 3)%, not significantly different from that of the mice immunized with DCs pulsed with IRES-OVA mRNA [(32 +/- 4)%, P > 0.05]. Mata static melanoma nodules could be seen in all control mice, only one mouse of the Capped-OVA mRNA group, and none of the mice of the IRES-OVA mRNA group.. IRES-containing tumor-associated antigen (TAA) mRNA as a more economic and applicable substitute fully replaces Capped-TAA mRNA for mRNA-based DC vaccines and DCs pulsed with IRES-containing TAA mRNA induces the same effective antigen-specific cellular response as the DCs pulsed with Capped-TAA mRNA.

Topics: Animals; Binding Sites; Cell Line, Tumor; Dendritic Cells; Female; Immunotherapy, Adoptive; Luciferases; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Ovalbumin; Plasmids; Protein Biosynthesis; Ribosomes; RNA, Messenger; Spleen; T-Lymphocytes, Cytotoxic; Transcription, Genetic; Transfection

This study was undertaken to test the effect of immunization against luteinizing hormone-releasing hormone (LHRH) fusion proteins on the development and progression of prostate cancer in the transgenic adenocarcinoma mouse prostate (TRAMP) model. Two LHRH fusion proteins, ovalbumin with seven LHRH peptides (OV-LHRH-7), and thioredoxin with seven LHRH peptides (TH-LHRH-7) were used in a cocktail vaccine. Two groups of male TRAMP mice were immunized with the cocktail. Primary immunizations were at either 4 or 8 weeks of age. LHRH immunized mice (n=19) were compared with castrated (n=19) and intact mice (n=18) for testosterone concentration, tumor weight, and lifespan. Immunization against LHRH in the TRAMP mice resulted in significant production of antibodies to LHRH compared with surgically castrated and intact control mice. Testicular weight was significantly reduced in the LHRH immunized groups compared with intact control mice. Serum testosterone was reduced (P<0.05) in the immunized mice compared with intact control mice and was not different from that of castrated mice (P>0.05). Tumor weight was variable and inconsistent throughout all treatment groups. Lifespan was not increased by immunization against LHRH or castration. Intact control mice (lived the longest (227+/-11 days), whereas immunized mice lived 206+/-11 days and castrated mice lived 213+/-13 days. Tumors from immunized TRAMP mice appeared more aggressive than tumors of castrated and intact mice, as demonstrated by 35% expression of gross lung tumors in the immunized mice whereas none were observed in the castrated or intact TRAMP mice. Prostate cancer is initially dependent upon androgens for growth and development, but cells have the ability to escape androgen dependence and progress to an androgen independent state, which was evident in this study. The TRAMP mouse model immunized against LHRH may have utility in future studies and treatments of the androgen independent prostate cancer.

Topics: Adenocarcinoma; Animals; Disease Models, Animal; Disease Progression; Gonadotropin-Releasing Hormone; Immunization; Male; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Orchiectomy; Organ Size; Ovalbumin; Prostatic Neoplasms; Recombinant Fusion Proteins; Testis; Testosterone; Thioredoxins

Archaeal ether glycerolipid vesicles (archaeosomes) efficiently deliver exogenous antigen for induction of humoral and cell-mediated immunity. Because induction of CD8 cytotoxic T cells is critical for protective vaccination against tumors, we compared the ability of various archaeosome lipid compositions to evoke a strong CD8 CTL response to entrapped antigen. Subcutaneous immunization of mice with ovalbumin (OVA) entrapped in all archaeosome lipid compositions evoked a primary (day 10) splenic CTL response indicating processing for MHC class I presentation. Interestingly, several polar lipid compositions from halophilic archaea were very potent to adjuvant this early CTL response. Despite this, the lytic units reduced substantially by weeks 6-7. More importantly, at >50 weeks, only Methanobrevibacter smithii and Thermoplasma acidophilum both rich in bipolar membrane-spanning caldarchaeols, demonstrated recall memory CTLs. Immunization of mice with OVA entrapped in M. smithii, Halobacterium salinarum or T. acidophilum vesicles provided prophylactic protection against challenge with OVA-expressing solid tumors at 6 weeks. Even a dose of 3 microg OVA in archaeosomes significantly delayed tumor growth. Tumor protection was also noted in a therapeutic design wherein OVA-archaeosomes were injected concurrent with the tumor challenge. Interestingly, antigen-free T. acidophilum but not antigen-free H. salinarum archaeosomes provided innate therapeutic protection. Vaccination with a CTL peptide epitope from the melanoma differentiation antigen, tyrosinase-related protein 2, in archaeosomes induced a protective CD8 response against B16OVA metastasis, indicating potential for targeting self, tumor antigens. Thus, lipid structural properties of archaea may differentially modulate primary, long-term and/or innate immunity, impacting adjuvant choice for vaccine design.

Topics: Animals; Archaea; Cancer Vaccines; Cell Line, Tumor; Drug Carriers; Female; Immunity, Innate; Lipids; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasms; Ovalbumin; T-Lymphocytes, Cytotoxic

Vaccination with peptides isolated from tumor cells circumvents the need for identifying specific tumor rejection antigens and extends the use of active immunotherapy to the majority of cancers where specific tumor antigens have not yet been identified. In this study, we examined the efficacy of tumor vaccines composed of unfractionated tumor peptides presented by antigen-presenting cells (APC) to induce cytotoxic T lymphocyte (CTL) responses and tumor immunity. RMA-S cells pulsed with peptides isolated from ovalbumin (OVA)-expressing tumor cells were highly effective at inducing primary, OVA-specific CTL responses in vitro and priming CTL responses in vivo. In addition, tumor peptide-pulsed RMA-S cells induced protective immunity in mice when challenged with OVA-expressing tumor cells. To enhance the clinical relevance of these studies, cells pulsed with tumor peptides were evaluated in the poorly immunogenic, B16/F10.9 melanoma post-surgical metastasis model. Treatment of tumor-bearing mice with peptide-pulsed RMA-S cells or with adherent splenocytes (enriched for professional APC) caused a significant reduction in lung metastases. The antimetastatic effect of peptide-pulsed splenocytes could be further enhanced by pretreating the cells with antisense oligonucleotides directed against the TAP-2 gene which was previously shown to increase the density of specific peptide/MHC class I complexes and thereby improve the APC function of the treated cells (Nair et el., J. Immunol. 1996. 156: 1772). This study suggests that APC loaded with unfractionated peptides derived from poorly immunogenic, highly metastatic tumor cells may represent a potent form of tumor vaccine.

Topics: Animals; Antigen-Presenting Cells; Antigens, Neoplasm; Cancer Vaccines; Histocompatibility Antigens Class I; Immunity, Cellular; Immunotherapy; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasm Proteins; Ovalbumin; T-Lymphocytes, Cytotoxic

Topics: Animals; Biotin; Breast Neoplasms; Drug Synergism; Female; Humans; Laryngeal Neoplasms; Male; Neoplasm Metastasis; Neoplasms; Ovalbumin; Pharyngeal Neoplasms; Swine

Topics: Animals; Anti-Bacterial Agents; Antibody Formation; Cattle; Chemotherapy, Cancer, Regional Perfusion; Cyclophosphamide; Diphtheria Toxoid; Fibrosarcoma; Geriatrics; Immunity; Immunosuppressive Agents; Mechlorethamine; Melanoma; Neoplasm Metastasis; Neoplasms; Ovalbumin; Pharmacology; Rabbits; Rats; Research; Sarcoma, Yoshida; Serum Albumin; Serum Albumin, Bovine; Toxicology