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Anti-tumor vaccine derived from normal cells

a normal cell, anti-tumor technology, applied in the direction of cancer antigen ingredients, biochemistry apparatus and processes, blood/immune system cells, etc., can solve the problems of difficult production and standardization of antigenic mixtures, no fully effective treatment methods suggested, and inability to meet the needs of patients

Inactive Publication Date: 2018-02-08
CYTOVAC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This approach allows for the generation of long-lasting cytotoxic T lymphocytes with memory cell characteristics, capable of recognizing a wide range of tumor cells, facilitating effective anti-tumor immune responses without the use of foreign agents, thus simplifying vaccine production and regulatory approval.

Problems solved by technology

Advanced cancers represent one of the major causes of human death, but no fully effective methods of treatment have been suggested so far.
Nevertheless, due to high heterogeneity in the expression of separate CTA members, this group of antigens is not popular as immunological target compared to differentiation or over-expressed antigens.
The major limitation of the polyvalent vaccine approach is difficulty in production and standardization of the antigenic mixture.
Direct use of original biopsy material lacks standardization, and also as well as such a material is often not available.
Use of standard cell lines has not shown high clinical efficiency so far (Palucka et al., 2006; Salcedo et al., 2005).
One of the reasons for low efficiency of using whole tumor cell material is low efficiency of cross-presentation.
The major limiting factor of RNA transfection is lack of formation of complexes of antigenic peptides and MHC class II molecules that is required for induction of T-helper response.
In addition, under conditions of tumor growth, formation of CD8+ memory response may be corrupted due to the presence of regulatory cells (Klebanoff et al., 2006).
However a major issue in this procedure is the use of foreign agents such as pokeweed mitogen (PWM) and phytohemagglutinin (PHA) for activation the cells.

Method used

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  • Anti-tumor vaccine derived from normal cells
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  • Anti-tumor vaccine derived from normal cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

Generation of Proliferating Cultures of Lymphocytes Enriched in CD4+ Cells

[0072]In order to induce expression of CTA in antigen presenting cells by chemical treatment (for example, with Aza-5-CdR), this culture should be proliferating. Professional antigen presenting cells, dendritic cells, do not proliferate under normal conditions, and therefore the inventors were looking for another type of antigen presenting cells. There are several types of antigen-presenting cells that may stimulate antigen-specific activation of CD8+ T lymphocytes. In particular, it has been shown that activated CD4+ helper T cells may be employed as antigen-presenting cells (Naota et al., 2006; Kennedy et al., 2005). Additional advantage of CD4+ cells as antigen presenting cells is that they are able to stimulate formation of memory CD8+ cells (Kennedy et al., 2005). The present inventors therefore first decided to optimize the preparation of proliferating cultures of CD4+-enriched lymphocytes. In the exampl...

example 2

Induction of Expression of Cancer / Testis Antigens in Proliferating Lymphocytes

[0076]In preliminary experiments on kinetics and Aza-5-CdR concentration dependence, variable induction of expression of CTA in lymphocyte cultures was observed. Most easily inducible were MAGE-A1, MAGE-A4 and MAGE-A12 (not shown). In order to standardize the conditions of induction, induction of CTA in lymphocytes taken from log phase of cell growth (usually at day 7 or 8, when cell concentration was about 106 / ml) was compared with lymphocytes taken from “plateau” (day 9 or 10, when lymphocyte concentration exceeds 2×106 / ml). Lymphocyte population expanded from peripheral blood in the presence of autologous mature dendritic cells was harvested, washed, re-suspended at 0.5×106 / ml in lymphocyte culture medium containing 10 μm of 5-aza-2′-deoxycytidine (Sigma) and 150 IU / ml of IL-2 and placed in the wells of 24-well tissue culture plate in 2 ml per well. Cells were incubated for 3 days, after which they were...

example 3

Induction of Cytotoxic T Cells Response Against 5-Aza-Cdr-Treated CD4-Enriched Proliferating Autologous Lymphocytes

[0082]After induction of CTA in normal lymphocytes after 5-Aza-CdR treatment, these lymphocytes were employed for the generation of tumor-specific immune response in vitro. 5-Aza-CdR-treated cultures were harvested, counted, and seeded in 96-well plate at 20×103 per well together with 20-40×103 of thawed non-adherent autologous lymphocytes in 0.1 ml of lymphocyte medium. After 2 days of incubation, 0.05 ml of lymphocyte medium supplemented with 75 IU / ml of IL-2 was added into each well. Addition of 0.05 ml of lymphocyte medium alone was repeated at days 5 and 7. At day 9, cultures were harvested and counted and used for analysis of immune response. Phenotyping of the cells were done by staining with the direct conjugated antibodies, CD4-fluorescein isothiocyanate (FITC), CD8-phycoerythrin (PE), CD56-PE, CD3-FITC and CD62L-phycoerythrin-cyanide 5 (PC5) (BD Biosciences, D...

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Abstract

A composition for inducing an immune response in a mammal, comprises lymphoid cells in which expression of tumor antigens has been chemically induced. The tumor antigens are induced in proliferating normal lymphoid cells, especially during the log phase of proliferation. The proliferation of the normal lymphoid cells is stimulated by normal mature dendritic cells. Most conveniently, the lymphoid cells are lymphocytes, especially peripheral blood lymphocytes. The tumor antigens are typically cancer / testis antigens, which may be chemically induced by DNA demethylation. Cancer / testis antigens are expressed in a wide range of tumors, so the composition is able to raise an immune response that is effective against a wide range of tumors, despite the fact that it is derived from normal cells. The composition may be used for preparation of an anti-tumor vaccine for prophylactic or therapeutic use. The composition may also be used for ex vivo activation of cytotoxic T lymphocytes, followed by expansion of the cytotoxic T lymphocyte population by normal dendritic cells, for cancer treatment by adoptive T cell immunotherapy.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]The present application is a continuation of U.S. patent application Ser. No. 12 / 522,048, filed on Dec. 3, 2009, which is the National Stage of International Application No. PCT / EP2008 / 050050, filed on Jan. 3, 2008, which claims the benefit of GB0700058.1, filed on Jan. 3, 2007. The content of all of these applications is hereby incorporated by reference in its entirety.BACKGROUNDField of the Invention[0002]The invention relates primarily to compositions suitable for inducing immune responses against malignancies; therefore the compositions can be used as a vaccine or for generating cytotoxic cells for adoptive immunotherapy. More particularly, the invention pertains to preparation of a cellular anti-tumor vaccine based on use of only normal cells of the immune system.Background of the Invention[0003]Advanced cancers represent one of the major causes of human death, but no fully effective methods of treatment have been suggested so far. C...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/00C12N5/0783
CPCA61K39/0011C12N5/0637A61K2039/5158C12N2502/1121A61K2039/572C12N2501/06C12N2501/23C12N5/0636A61P35/00A61K39/464484A61K39/4622A61K39/4611
Inventor KIRKIN, ALEXEIDZHANDZHUGAZYAN, KARINE
Owner CYTOVAC