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Listeria-based compositions comprising a peptide minigene expression system and methods of use thereof

A technology based on Listeria and genome, applied in the direction of microorganism-based methods, translation products of oncogenes, biochemical equipment and methods, etc., can solve the problem of loss of antigen presentation

Pending Publication Date: 2017-11-28
ADVAXIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This can lead to loss of antigen presentation

Method used

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  • Listeria-based compositions comprising a peptide minigene expression system and methods of use thereof
  • Listeria-based compositions comprising a peptide minigene expression system and methods of use thereof
  • Listeria-based compositions comprising a peptide minigene expression system and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0369] (B) Example 1: LLO-antigen fusions induce anti-tumor immunity

[0370] cell line

[0371] C57BL / 6 syngeneic TC-1 tumors were immortalized with HPV-16 E6 and E7 and transformed with the c-Ha-ras oncogene. TC-1 provided by T.C. Wu (Johns Hopkins University School of Medicine, Baltimore, MD) is a highly tumorigenic lung epithelial cell that expresses low levels of HPV-16 E6 and E7 and is infected by the c-Ha-ras oncogene transform. Keep TC-1 at 37°C and 10% CO 2 Grow in RPMI 1640, 10% FCS, 2mM L-glutamine, 100U / ml penicillin, 100μg / ml streptomycin, 100μM non-essential amino acids, 1mM sodium pyruvate, 50 micromolar (mcM) 2-ME, 400 Micrograms (mcg) / ml G418 and 10% National Collection Type Culture-109 medium (National Collection TypeCulture-109 medium). C3 are mouse embryonic cells from C57BL / 6 mice immortalized with the complete HPV 16 genome and transformed with pEJ-ras. EL-4 / E7 is thymoma EL-4 transduced with E7 retrovirus.

[0372] Listeria monocytogenes strains ...

example 2

[0394] Example 2: LM-LLO-E7 Treatment Causes Proliferation of TC-1 Specific Splenocytes

[0395] To measure the induction of T cells by Lm-E7 with Lm-LLO-E7, E7-specific proliferative responses (a measure of antigen-specific immune activity) were measured in immunized mice. Splenocytes from mice immunized with Lm-LLO-E7 were exposed to irradiated TC-1 cells at splenocyte:TC-1 ratios of 20:1, 40:1, 80:1, and 160:1 (as Proliferation occurs when the source of E7) ( Figure 4 ). In contrast, splenocytes from mice immunized with Lm-E7 and rLm controls showed only background levels of proliferation.

example 3

[0396] Example 3: ActA-E7 and PEST-E7 fusions confer anti-tumor immunity

[0397] Materials and Experimental Methods

[0398] Construction of Lm-ActA-E7

[0399] Lm-ActA-E7 is a recombinant strain of LM containing a plasmid expressing the E7 protein fused to a truncated form of the actA protein. Lm-actA-E7 was generated by introducing the plasmid vector pDD-1 constructed by modifying pDP-2028 into Listeria. pDD-1 contains an expression cassette expressing a copy of the 310bp hly promoter and the hly signal sequence (ss), which drives the expression and secretion of ActA-E7; the 1170bp actA gene comprising four PEST sequences (the truncated ActA polypeptide is composed of The first 390 AAs of the molecule), the 300bp HPV E7 gene, the 1019bp prfA gene (controlling the expression of virulence genes) and the CAT gene (chloramphenicol resistance gene) were used to select transformed bacterial clones (Sewell et al. (2004 ), Arch. Otolaryngol. Head Neck Surg., 130:92-97).

[...

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Abstract

This disclosure provides compositions, including Listeria delivery vectors comprising minigene expression constructs, and methods of using the same for inducing an immune response against an antigen-expressing tumor and for treating the same, and vaccinating against the same in subjects bearing the tumors.

Description

technical field [0001] Disclosed herein are compositions comprising a Listeria delivery vector comprising a minigene expression construct, and methods of using the compositions for inducing an immune response against and for treating antigen-expressing tumors or cancers , and a method of vaccinating against a tumor or cancer in a subject having said tumor or cancer. Background technique [0002] Listeria monocytogenes is an intracellular pathogen that primarily infects antigen-presenting cells and has adapted to live in the cytoplasm of these cells. Host cells such as macrophages, active phagocytic L. monocytogenes and most bacteria are degraded in phagolysosomes. Some bacteria escape into the host cytosol by piercing the lysosomal membrane of the phage through the action of the hemolysin, Listeriatolysin O (LLO). Once in the cytosol, L. monocytogenes can polymerize host actin and pass directly from cell to cell, further invading the host immune system and producing neglig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74A61K35/74A61K48/00A61P35/00C12R1/01
CPCA61K35/74A61K48/005C07K14/195C12N9/10C12N9/90C12N15/74A61K39/001106A61K38/00C07K14/00C12P21/02C07K2319/036C12N2710/20034A61K2039/522A61K2039/523A61K39/12A61K2039/572A61K2039/585C12N2710/20071A61P35/00C07K14/82C07K2319/02A61K48/00
Inventor R·珀蒂麦可·F·宾矽欧塔
Owner ADVAXIS