JP2002541811A - Telomerase-specific cancer vaccine - Google Patents

Abstract

  (57) [Summary] A telomerase-specific T cell antigen is provided. This is useful for generating a T cell response to telomerase. Formulations of telomerase antigens as vaccines are useful for treating and preventing cancer using in vivo or ex vivo techniques.

Description

DETAILED DESCRIPTION OF THE INVENTION

BACKGROUND OF THE INVENTION Telomeres, DNAs at the chromosomal ends, consist of simple tandem repeats. In most somatic cells, telomere sequences are lost during DNA replication due to the need for a DNA-dependent DNA polymerase for the RNA primer annealed to the template strand. Since RNA primers cannot anneal beyond the 5 'end of the DNA strand, some telomere DNA is lost from generation to generation as cell DNA is replicated. Cells that exhibit such telomere shortening enter senescence after a certain number of population doublings, and aging is directly associated with eroding the telomere to a critical minimum length.

[0002] However, not all cells undergo such loss. Normal human somatic cells lose telomere repeats during the cell division cycle, while human germline and, importantly, cancer cells maintain a certain number of telomere repeats. Telomere length is maintained in these cells by the action of telomerase, a ribonucleoprotein enzyme that uses short endogenous RNA as a template for telomere addition. In fact, cancer cells express high levels of telomerase, while somatic cells express little telomerase.

[0003] Normal somatic cells do not appear to express or require telomerase, but because cancer cells express high levels of telomerase, telomerase enzymes represent an attractive therapeutic target. However, due to the fact that telomerase is a normal "self" antigen, conventional vaccination strategies are not available. Thus, the focus of telomerase-based therapeutics has been on various types of enzyme inhibitors, rather than on a vaccine-based approach.

[0004] Therefore, new therapeutic approaches based on telomerase are needed. This need extends to vaccines based on telomerase antigens.

SUMMARY OF THE INVENTION It is, therefore, an object of the present invention to provide an antigen useful for generating an immune response to telomerase. According to an object of the present invention, there is provided a telomerase antigen capable of inducing an immune system against a telomerase-expressing cell. In one embodiment, a telomerase antigen is provided based on the peptide sequence of the protein portion of telomerase. In another embodiment, the telomerase antigen is provided as a nucleic acid that can be used to express a peptide-based telomerase antigen.

[0006] Another object of the present invention is to provide a vaccine composition comprising at least one telomerase antigen. Accordingly, in one aspect, the invention provides a vaccine composition comprising a telomerase peptide antigen. In another aspect, a polynucleotide is provided that encodes a protein-based telomerase antigen.

[0007] Yet another object of the present invention is to provide a method for treating or preventing cancer. To this end, the telomerase antigens of the invention can be administered directly to the patient in a beneficial amount. According to this purpose, the present telomerase antigen encoded by the nucleic acid can be administered to a patient. This object is also achieved by an in vitro method comprising contacting the cell with a telomerase antigen and administering the cell to a patient. In one embodiment, the contacted cells can be antigen presenting cells, which can be used to produce sensitized T cells in vitro, and then administering the sensitized T cells to a patient. obtain.

DETAILED DESCRIPTION The present invention relates to telomerase-specific vaccines useful for producing T cells that are activated for telomerase. More specifically, the vaccines of the present invention provide antigen-specific major histocompatibility (MH) for telomerase presented by antigen presenting cells.
C) Useful for generating a limited T cell response. It is believed that the vaccines of the present invention act to reverse tolerance or no response induced through self-tolerance to telomerase in normal individuals. Telomerase is, in one embodiment,
Because they present cancer-specific therapeutic targets, the vaccines of the invention are useful for treating or preventing a variety of cancers.

[A. Definitions As used herein, "activated T cells" are those that are in the following phases of the cell cycle. G ₁ phase, S phase, G ₂ phase or M (mitosis) phase. Thus, an “activated T cell” has undergone mitosis and / or cell division. The activated T cells can be T helper (T _H ) cells or cytotoxic T cells (cytotoxic T lymphocytes (CTL or T _c )). Activation of untreated T cells will
To antigen presenting cells (APCs), including antigen / MHC complexes, and to I
It can be initiated by exposure to molecules such as L-1, IL-2, IL-12, IL-13, γ-IFN and similar lymphokines. The antigen / MHC complex interacts with a receptor on the surface of T cells (T cell receptor (TCR)). Golub et al., Eds., Immunology; Synthesis, Chapter 2: "T Cell Receptors" (1991).

[0010] As used herein, "priming" refers to exposing an animal (including a human) or a cultured cell to an antigen to provide activation and / or memory. Used for And CD8 ^{- -} CD4 to target antigens the production of the T cell responses is generally due by natural infection or deliberate immunization depends on sensitization in vivo.

As used herein, a “naive” T cell is one that has not been exposed to a foreign (non-self) antigen or that has not been exposed to a potential self antigen. It is. "Untreated" T cells are sometimes referred to as "unsensitized" T cells. One of ordinary skill in the art, "resting (resting)" cells, located in the G ₀ phase of the cell cycle, and therefore, will recognize that you have not received not division or mitosis. Those skilled in the art also
"Anergic" T cells will recognize that they cannot function properly, ie, cells that lack the ability to mediate a normal immune response. T cells from a patient suffering from a disease can include T cells that are sensitized but anergic.

As used herein, “memory T”, also known as “memory phenotype” T cells
`` Cells '' have previously encountered peptide antigens, but are now dormant,
Used to refer to one type of T cell that can be activated. Memory T cells
It is a T cell that has been exposed to an antigen and subsequently survives in the body for a long time in the absence of a stimulating antigen. However, these memory T cells respond to "recall" antigens. In general, memory T cells are more responsive to "recall" antigens compared to untreated T cell responses to peptide antigens. Memory cells can be recognized by the presence of certain cell surface antigens, such as CD45R0, CD58, CD11α, CD29, CD44 and CD26, which are markers of differentiated T cells.

As used herein, a “telomerase-specific” T cell response is a telomerase antigen stimulation, eg, a T cell response to a peptide (eg, proliferation, cytotoxicity and / or cytokine secretion), which It is not evident with other stimuli, such as peptides with different amino acid sequences (control peptides). T cell responsiveness is measured by assessing the appearance of cell surface molecules characteristic of T cell activation, including but not limited to CD25 and CD69. Such assays are known in the art.

In the context of the present invention, the term “treating”, used in various grammatical forms, refers to the prevention of the deleterious effects of a disease state, the progression of a disease, the causative agent or other abnormal conditions. Means healing, reversing, attenuating, reducing, minimizing, inhibiting or discontinuing.

[B. Telomerase antigen] [1. Generally useful antigens] The telomerase antigens according to the present invention share a characteristic ability to generate a specific T cell response. This response may be class I or class II specific. In one embodiment, the response is MHC class I specific, antigen-restricted and MH
C Including limited cytotoxicity. Class I molecules are HLA-A, HLA-B and HLA.
LA-C. Thus, preferred antigens are class I molecules, such as HLA-A1
Binds to HLA-A2, HLA-A3 or HLA-A11. More preferred antigens bind to all class I molecules. In contrast, where a class II-specific (eg, helper function) response is desired, a class II binding antigen is used. Class II molecules include HLA-DR, HLA-DQ and HLA-DP. Useful antigens can be determined as set forth below.

[0016] Telomerase antigens are typically described in US Pat. No. 5,837,857 (1998).
) And GenBank Accession Nos. AF015950 and AF018167 from the sequence of the protein portion of telomerase, which sequences are hereby incorporated by reference. They can be produced, for example, by proteolytic digestion of telomerase proteins and / or by recombinant DNA means. Generally, relatively short peptide variants are prepared by synthetic means.

The telomerase antigens according to the invention are not limited by size, they may be part or even all of the telomerase protein, but they are usually small peptide antigens. Small size is preferred because of its ease of manufacture and high specificity. Thus, unless they are multimeric (ie, multiple copies of the same epitope), most telomere antigens are about 50 amino acids or less. Preferred antigens are
Other antigens that are less than about 25 amino acids in length, with other preferred antigens being between about 8 and about 12 amino acids in length, are also contemplated as short as 6 or 7 amino acids. 9mer is typical for class I antigens. Because they usually retain the necessary functional characteristics. They are Ile-Leu-Ala-Lys-Phe-
Leu-His-Trp-Leu (ILAKFLHWL) is included as a preferred species.

[0018] Variants of the telomerase antigen are also contemplated. Importantly, any variant is a functional feature of the telomerase antigen (1) MHC molecules, such as the protein HLA-
All that is required is to retain the ability to bind to A2 and (2) the ability to induce telomerase-specific T cell responses. Amino acid substitutions, i.e., "conservative substitutions" that result in "conservative variants" include, for example, the polarity, charge, solubility, hydrophobicity, hydrophilicity,
And / or based on similarities in the amphipathic nature.

For example, (a) non-polar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; and (b) polar neutral amino acids are glycine, serine, threonine. , Cysteine, tyrosine, asparagine, and glutamine; (c) positively charged (basic) amino acids include arginine, lysine, and histidine; and (
d) Negatively charged (acidic) amino acids include aspartic acid and glutamic acid.
Substitutions can typically be made within groups (a)-(d). In addition, glycine and proline can be exchanged based on their relatively small size and lack of side chains.

Similarly, certain amino acids, such as alanine, cysteine, leucine, methionine, glutamic acid, glutamine, histidine and lysine, have an α-helix,
While more commonly found, valine, isoleucine, phenylalanine, tyrosine, tryptophan and threonine are more commonly found in β-plate sheets. Glycine, serine, aspartic acid, asparagine, and proline are commonly found in turns. Of course, the importance of structure-based substituents increases with the length of the antigen.

Some preferred conservative substitutions may be made in the following groups: (I) S and T, (ii) P and G, and (iii) A, V, L and I. Since the genetic code, and recombination and synthesis techniques are known, those skilled in the art can readily produce DNA encoding conservative amino acid variants. Of course, smaller variants can also be synthesized. One such conservative HLA-A2 binding mutant species has the following structure:
A / V / L / I) (A / V / L / I) (A / V / L / I) (A / V / L / I) K
F (A / V / L / I) HW (A / V / L / I) peptide. Accordingly, some preferred conservative variants are LLAKFLHWL, ILAKFLHWI, ILA
IAKFLHWL, IIAKFLHWI, ILARFLHWL, and ILVK
Includes FLHWL and permutations as long as the required functional features are retained.

In addition, one or more amino acids of the HLA-A2 binding peptide may be replaced with glycine. Parker et al. Immunol. 149: 3580-87
(1992) discloses that up to six amino acids in the 9mer can be replaced with glycine, and thus GLFGGGGGGV can bind to HLA-A2. 9me
The only true conservation observed for the rHLA-A2 binding peptide was Ile or Leu at approximately position 2 (counting from N-terminal to C-terminal) and Val or Leu at approximately position 9. Therefore, some simple mutants have been placed in the presence of the required functional features, GLAKFLHWL, ILAGFLHWL, ILAKGLHWL
, ILAKFGHWL, ILAKFLGWL and ILAKFLHGL.

An important source for guidance on the design of class I and class II antigens and the implementation of conservative substitutions is Ramensee et al., Immunogenic.
s41: 178-228 (1995), which is hereby incorporated by reference. As shown in the Rammensee reference, both class I and class II molecule motifs have certain "maintenances" that retain a high degree of conservation.
It has an "anchor" residue. For example, HLA-A0201, a molecule designed (actually binds) to the telomerase peptide ILAKFLHWL (
HLA-A2 molecule) has anchor residues at positions 2 and 9 corresponding to the above conserved positions. This molecule also has an "auxiliary" position at position 6, whose relative conservation is important but not as important as the anchor residue. Thus, using Rammensee's general guidelines, one of skill in the art would recognize that while the anchor and auxiliary residues are relatively conserved in HLA binding, the rest of the antigen may vary widely, possibly Will be involved in the specific antigenic characteristics of telomerase, ie, it will distinguish telomerase from non-telomerase.

[0024] Other substitutions involve replacing L-amino acids with the corresponding D-amino acids.
Further, this theory can be combined with the conservative substitution theory described above. For example,
D-leucine can be replaced with L-isoleucine. Furthermore, compared to the native sequence,
These D-amino acid-containing peptides having the reverse sequence can be prepared. Therefore, I
LAKFLHWL becomes LWHLFKALI. Such "reverse sequence" peptides are expected to have improved properties, such as increased half-life in vivo. this is,
Translate into smaller and more economically viable production.

[0025] Some embodiments contemplate multimers of the peptide. Multimers may contain multiple copies of the same peptide, or they may be mixed and matched. Multimers can be direct tandem repeats and can include a spacer sequence of amino acids such as Gly and / or Pro (eg, 2-5 residues), or other suitable spacer. . Multimers can be of any length, but typically are no more than about 100 amino acids. Preferred multimers are less than about 60 amino acids and have about 2-5 copies of the peptide, about 8 to about 12 amino acids in length. Multimers may also contain several different telomerase antigens.

The telomerase antigen may be glycosylated or partially glycosylated according to methods known in the art. They can be modified with large molecular weight polymers such as polyethylene glycol. Furthermore, lipid modification is preferred. Because they can facilitate the encapsulation or interaction of the derivative with the liposome. Useful Exemplary lipid moieties for this purpose are, palmitoyl, myristoyl, stearoyl and decanoyl groups or, more generally, the saturation of the C ₂ -C _{3 0,} including unsaturated or polyunsaturated fatty acyl group which It is not limited to.

For convenience in performing chemical modifications, it is sometimes useful to include in the telomerase antigen one or more amino acids with side chains that can undergo modification. A preferred amino acid is lysine, which can be easily modified with an ε-amino group. The side chain carboxyl of aspartate and glutamate is
It is easily modified like serine, threonine and tyrosine hydroxyl groups, cysteine sulfhydryl groups and histidine amino groups. The introduction of two cysteine residues at spaced locations in the peptide antigen may help to create structural constraints via disulfide bridges that can improve binding to MHC molecules.

Also, examples of telomerase antigens within the present invention include non-peptide “mimetics”
) ", Ie, compounds that mimic one or more functional characteristics of the telomerase antigen. Mimetics are generally water-soluble, resistant to proteolysis, and non-immunogenic. Conformationally constrained cyclic organic peptides that mimic telomerase antigens are described, for example, in Saragovi et al., Science 253: 7.
92 (1991).

The telomerase antigen may also be configured as a hybrid with immunostimulatory molecules such as cytokines and adjuvants (and / or as described below,
(Can be formulated together as separate molecules in liposomes). Interleukin-2 (
IL-2) is one such cytokine. Other cytokines are GM
-Includes CSF, IL-12 and flt-3 ligand. Telomerase antigen
As fusion proteins with IL-2, for example, they can be made by recombinant DNA or chemical synthetic means, or they can be made as chemical conjugates using bifunctional chemical linkers. Such a chimeric protein is expected to have an increased ability to produce a T cell-specific response to telomerase. Adjuvants include monophosphoryl lipid A (MPLA), and derivatives thereof, which can also be attached to telomerase antigen by conventional linkers.
Other conventional immunostimulatory molecules are keyhole limpet hemocyanin (keyhole li
mpet hemocyanin (KLH).

[2. Identification of Other Useful Antigens] It is interesting to identify additional, especially small, telomerase antigens that are expected to produce a more specific response, eg, associated with a particular epitope. In addition, these small antigens can be produced more economically.

It is advantageous to identify additional telomerase antigens and further purify the telomerase T cell antigenicity to the epitope level. One classic method involves proteolytic processing of large antigens to obtain smaller antigens. In addition, fragments of a protein antigen can be produced by recombinant DNA technology and assayed to identify a particular epitope. In addition, small peptides can be produced and assayed by in vitro synthesis methods.

As an alternative to a randomized approach to producing portions of the intact antigen and assaying them, a more biologically relevant approach is possible. In particular, MHC Class I
One example approach is to isolate the MHC molecules themselves, and then their associated peptides, since antigen fragments that bind to Class II molecules, especially Class I molecules, are of particular interest. That is. For a description of such general methods, see PCT / US98 / 09288; Agrawal et al., In.
t'l Immunol. 10: 1907-16 (1998); and Agra.
wal et al., Cancer Res. 55: 5151-56 (1998).

In a typical method, a primary tumor cell or cell line expressing the antigen of interest is provided. Further, it is recognized that phagocytic antigen presenting cells (or any APC), such as macrophages, can phagocytize large antigens (or portions thereof) and thus act as starting materials for these methods. MHC class I or class II
Molecules can be isolated from these starting cells using known methods such as antibody affinity (MHC-specific antibodies) and chromatography techniques.

[0034] The isolated MHC molecule is then processed to release the binding peptide. This can be achieved by treatment with substances that disrupt the interaction between the binding peptide and the MHC molecule, such as detergents, urea, guanidinium chloride, divalent cations, various salts and extreme pH. The released peptide can be further purified using conventional chromatography and antibody affinity (using antigen-specific antibodies) methods. The purified peptide may then be subjected to sequence determination and structure determination using, for example, peptide sequencing, gas chromatography and / or mass spectrometry.

Thus, the sequence structure of most broad peptide epitopes associated with class I and / or class II molecules can be determined. Given this sequence / structure information, determined sequence substitutions can be made as detailed above and assayed using known T cell assays. Rammensee et al., Supra, provide extensive methods and guidance related to the recognition of both class I and class II motifs.

Yet another method of producing a telomerase antigen may use algorithms known in the art for predicting binding sequences. Publicly available comparison programs for comparing known peptide sequences to different HLA binding motifs using these algorithms are described in, for example, http: // www-bimas. dcr
t. nih. gov / hla_bind. The different class I and class II binding motifs can be found at that site or in Rammensee et al.
arcer et al., both can be found in such publications as described above.

C. Vaccine Compositions In general, any telomerase-specific antigen described above is useful in formulating a telomerase-specific vaccine. Preferred antigens will typically be associated with the lipid by direct lipid modification of the antigen and / or by liposome association, as described below. The antigen may be administered as a peptide or peptidomimetic, or in a nucleic acid form.

[1. Liposomal formulation] In one embodiment of the present invention, the telomerase antigen is associated with the liposome. Techniques for preparing liposomes and formulating (eg, encapsulating or complexing) liposomes with various molecules, including peptides, are known to those of skill in the art. Liposomes are small vesicles composed of one or more lipid bilayers surrounding an aqueous compartment. See generally, Baker-Woodenberg et al., Eur. J. Clin.
Microbiol. Infect. Dis. 12 (Supplement 1): S61 (199
3) and Kim, Drugs 46: 618 (1993). Liposomes are
Due to their similar composition to cell membranes, they are generally safe to administer and biodegradable.

Depending on the method of preparation, the liposomes may be unilamellar or multilamellar, and may vary in diameter size from 0.02 μm to 10 μm or more. Various substances can be encapsulated in the liposome. Hydrophobic material partitions into bilayer, hydrophilic material inside aqueous space (s)
Distribute to See, eg, Machy et al., Cell Biology and Pharmacology Liposomes (J.
ohn Libbyey 1987), and Ostro et al. (1989) Ameri.
can J. Can. Hosp. Pharm. 46: 1576.

Liposomes are able to absorb virtually any type of cell and then release the encapsulating material. Alternatively, the liposome fuses with the target cell, where the contents of the liposome enter and empty the target cell. Alternatively, the adsorbed liposomes may undergo endocytosis by phagocytic cells. After endocytosis, lysosomal degradation of liposomal lipids and release of the encapsulating material occur. Scherpho
f, et al. (1985) Ann. N. Y. Acad. Sci. 446: 368.

[0041] Anionic liposome vectors have also been studied. These are disrupted or fused with the endosomal membrane after endocytosis and endosomal acidification, pH
Contains sensitive liposomes. However, among liposome vectors, cationic liposomes have been best studied because of their efficacy in mediating mammalian cell transfection in vitro.

[0042] Cationic lipids are not found in nature and can be cytotoxic. Because these complexes appear to be incompatible with the in vivo physiological environment rich in anionic molecules. Liposomes are selectively phagocytosed by the reticuloendothelial system. However, the reticuloendothelial system can be circumvented by several methods, including saturation with large amounts of liposome particles, or selective inactivation of macrophages by pharmacological means. Cla
ssen et al. (1984), Biochim. Biophys. Acta 802
: 428. In addition, glycolipids or polyethylene glycol derivatized (deri
vatised) Incorporation of phospholipids into liposome membranes has been shown to significantly reduce uptake by the reticuloendothelial system. Allen et al. (1991) Biochi.
m. Biophys. Acta 1068: 133; Allen et al. (1993)
Biochim. Biophys. Acta 1150: 9.

[0043] Cationic liposome preparations can be prepared by conventional methods. For example, F
elgner et al., Proc. Nat'l. Acad. Sci. USA 84:74
13 (1987); Schreier, J. et al. of Liposome Res.
2: 145 (1992); Chang et al. (1988) supra. Commercial preparations,
For example, Lipofectin (registered trademark) (Life Technologies, Gaithersburg, MD, USA) is also available. The amount of liposomes and the amount of DNA can be optimized for each cell type based on a dose response curve. Felgner et al., Supra. For some recent reviews on the methods used, see Wassef et al., Immunomethods 4: 217-22.
2 (1994) and Weiner, A .; L. Immunomethods 4
: 217-222 (1994).

[0044] Other suitable liposomes for use in the methods of the invention include multilamellar vesicles (MLV
), Oligolamellar vesicles (OLV), unilamellar vesicles (UV), small unilamellar vesicles (SUV), medium-sized unilamellar vesicles (MUV), large unilamellar vesicles (L
UV), giant unilamellar vesicles (GUV), multi vesicular vesicles (MVV), unilamellar or lamella vesicles produced by reverse phase evaporation (REV)
), Multilamellar vesicles (MLV-REV) produced by reverse phase evaporation, stable multilamellar vesicles (SPLV), frozen and thawed MLV (FATMLV), vesicles prepared by extrusion (VET) ), Vesicles prepared by French press (F
PV), vesicles prepared by fusion (FUV), dehydrated-rehydrated vesicles (DRV)
, And bubblesomes (BSV). One skilled in the art will recognize that these liposome preparation techniques are known in the art. Colloidal drug delivery system, No. 6
Volume 6 (edited by J. Kreuter, Marcel Delkker, Inc. 199)
See 4).

[2. Appropriate Adjuvants and Excipients] The vaccine formulations of the invention (liposomes or otherwise) can be advantageously formulated with several types of adjuvants. As is conventionally known in the art, an adjuvant is a substance that works with a specific antigenic stimulus to enhance a specific response to an antigen. For example, MPLA has been shown to serve as an effective adjuvant in causing increased presentation of liposomal antigens by APCs to specific T lymphocytes. Alving, C.I. R. 1993, Imm
unobiol. 187: 430-446. Further, those skilled in the art will recognize Detox, alum, QS21, complete and / or incomplete Freund's adjuvant, MD
It will be appreciated that other adjuvants such as P, lipid A and their derivatives are also suitable.

Another class of adjuvants includes stimulatory cytokines, such as IL-2.
Thus, the vaccines of the present invention can be formulated with IL-2 or
Can be administered separately for optimal antigenic response. IL-2 is advantageously formulated with liposomes.

[0047] The vaccine may also be formulated with pharmaceutically acceptable excipients. Such additives are known in the art, but typically should be physiologically acceptable, inert, or enhance with respect to the vaccine properties of the compositions of the present invention. Examples include liquid vehicles such as sterile saline. When using additives, it is
It may be added at any point in the formulation of the vaccine or mixed with the completed vaccine composition.

[0048] Vaccines may be formulated for multiple routes of administration. Particularly preferred routes include:
Intramuscular, transdermal, subcutaneous or intradermal injection, aerosol, oral or a combination of these routes, in single or multiple unit doses. The administration of the vaccine is known and ultimately depends on the specific formulation and the judgment of the attending physician.

A vaccine formulation can be maintained as a suspension or lyophilized and then hydrated to make a usable vaccine.

[D. Targeting Antigens and Vaccines of the Invention The compositions of the invention may act to provide higher specificity, and thus reduce the risk of toxicity or other undesirable effects during in vivo administration. It is advantageous to target to cells designed for, ie, antigen presenting cells. This can be achieved using conventional targeting techniques. One exemplary form of targeting using an antibody or similar specifically binding molecule is one that is associated in some manner with the antigen and / or vaccine composition.

[0051] The targeting molecule has characteristics that allow it to distinguish target APCs from background non-APCs to some extent. Targeting molecules include mannose that targets antigen presenting cells and the Fc portion of an antibody. The targeting molecule can be directly associated with the telomerase antigen, for example, in the case of a protein-based targeting sequence, by chemical conjugate, or by fusion protein production.

[0052] Antibodies and antibody derivatives are preferred targeting molecules because of their simplicity and well-known in the art. Antibodies and derivatives thereof are polyclonal antibodies,
Monoclonal antibodies (mAbs), humanized or chimeric antibodies, single chain Fv (scF
v) single chain antibodies, including fragments, Fab fragments, F (ab ') ₂ fragments, fragments produced by Fab expression libraries, epitope binding fragments, and humanized forms of any of the above. . Of course, smaller forms of these molecules are preferred based on the fact that they more easily target APCs.

In general, the techniques for preparing polyclonal and monoclonal antibodies and hybridomas that are capable of producing the desired antibodies are known in the art (Campbell).
, A. M. Monoclonal antibody technology: experimental techniques in biochemistry and molecular biology, El
Sever Science Publishers, Amsterdam, The Netherlands (1984); St. Groth et al. Immunol. Methods
35: 1 to 21 (1980); Kohler and Milstein, Nat.
ure 256: 495-497 (1975)), trioma technology, human B cell hybridoma technology (Kozbor et al., Immunology Today 4).
: 72 (1983); Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan.
R. Liss, Inc. (1985) 77-96). The affinity of antisera for antigens is described, for example, in Fisher, Chapter 42, Manual of Clinical Immunology: Second Edition, Rose and Friedman eds., Amer. Soc. For Micro
biology, Washington D.C. C. (1980) can be determined by generating a competitive binding curve.

Antibody fragments or derivatives include any portion of an antibody that can bind to an APC target molecule, typically a surface antigen. Antibody fragments include, among others, F (ab ′) ₂ , Fab, Fab
'And Fv fragments. These can be made from any class of antibody, but are typically made from IgG or IgM. They are conventional recombinant DNA
It can be made by proteolytic digestion with papain or pepsin, using techniques or using classical methods. Current Protocols in Immunology, Chapter 2,
See Coligan et al. (John Wiley & Sons 1991-92).

F (ab ′) ₂ fragments are typically about 110 kDa (IgG) or about 150 kDa.
2 kDa (IgM), hinged by disulfide bond (s)
It contains two antigen binding regions. Substantially all, if not all, of the Fc are absent from these fragments. Fab 'fragments are typically about 55 kDa (IgG) or about 75 kDa (IgM) and can be formed, for example, by reduction of the disulfide bond (s) of the F (ab') ₂ fragment. The resulting free sulfhydryl group (s) can be used to conveniently conjugate the Fab 'fragment to another molecule, such as a telomerase antigen or an adjuvant molecule.

Fab fragments are unit price and are usually around 50 kDa (from any source). F
ab fragments include antigen-binding portion of an antibody, the light chain (L) and heavy (H), the variable (V _L and V _H, respectively) and constant (respectively C _L C _H) region. The H and L moieties are linked by one or more intramolecular disulfide bridges.

[0057] Fv fragments are typically about 25 kDa (regardless of origin), including the variable region of both light and heavy chains (V _L and V _H, respectively). Normally, the _VL and _VH chains are held together only by non-covalent interactions, and thus they dissociate easily. However, they have the advantage of being small in size and retaining the same binding properties as larger Fab fragments. Thus, methods have been developed to crosslink _VL and _VH chains using, for example, glutaraldehyde (or other chemical crosslinking agents), intermolecular disulfide bonds (by incorporation of cysteines), and peptide linkers. The resulting Fv is currently single-chain (ie, scFv).

Other antibody derivatives include single chain antibodies (US Pat. No. 4,946,778; Bi).
rd, Science 242: 423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85: 5879-5883
(1988); and Ward et al., Nature 334: 544-546 (1
989)). Single chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single chain FV (scFV).

Derivatives also include “chimeric antibodies” (Morrison et al., Proc. Na.
tl. Acad. Sci. 81: 6851-6855 (1984); Neube.
rger et al., Nature, 312: 604-608 (1984); Taked.
a, Nature, 314: 452-454 (1985)). These chimeras are made by splicing DNA encoding a mouse antibody molecule with the appropriate specificity, for example, with a DNA encoding a human antibody molecule with the appropriate specificity. Thus, a chimeric antibody is a molecule in which different portions are derived from different animal species, for example, a molecule having a variable region derived from a murine mAb and a human immunoglobulin constant region. Human framework regions and murine complementarity determining regions (C
Recombinant molecules having DR) are also made using known techniques. These are also
Sometimes known as "humanized" antibodies, they and chimeric antibodies or antibody fragments offer the additional advantage of at least partial shielding from the human immune system. As such, they are particularly useful for in vivo therapeutic applications.

[E. Nucleic Acid-Based Vaccines] In recent years, interest in polynucleotide-based vaccines has increased, and such applications will be considered here. These vaccines generally comprise a DNA vector encoding the antigen of interest under the operable control of transcription and translation signals, or an RNA vector encoding the antigen of interest under the operable control of translation signals. Depends on. It is believed that upon administration of these vaccines, they are taken up by surrounding cells and then express the target antigen. The expressed antigen becomes apparently associated with the major histocompatibility (MHC) antigen of the cell and is therefore localized on the surface of the cell and presented to immune cells. See, for example, Corr et al. Exp. Med
. 184: 1555-60 (1996). Such vaccines include naked DNA (
Or the DNA may be associated with or captured by liposomes (Gregoridis et al., FEBS Lett. 402: 107-10 (1).
997)).

[0061] These so-called "naked DNA" vaccines or vaccines containing RNA are widely applicable. They include, for example, inter alia, anti-cancer vaccines (Scheur)
s et al., Cancer Res. 58: 2509-14 (1998); Hurpi.
N et al., Vaccine 16: 208-15 (1998)) and antiviral vaccines (Bohm et al., Vaccine 16: 949-54 (1998); Le.
kutis et al. Immunol. 158: 4471-77 (1997)). Naked DNA vaccines are class I (Scheurs et al., Supra;
Bohm et al., Supra; Hurpin et al., Supra) and Class II restricted response (Lek).
utis et al., supra; Leukoc. Biol. 61:
125-32 (1997)).

Accordingly, any of the foregoing telomerase antigens may be administered as a naked DNA vaccine. These vaccines include a nucleic acid vector that encodes a telomerase antigen under the control of transcription and translation signals operable in a mammal, preferably a human. They may be administered in association with or encapsulated with (usually cationic) liposomes as detailed above, or they may be administered in any other physiologically tolerable excipient.

[F. Method of the Invention] According to one aspect of the present invention, the telomerase antigen (or vaccine composition) described above can be used directly as a simple vaccine to induce an immune response to telomerase. However, in some cases, the antigen may be associated with the liposome, as indicated above, for a therapeutically or prophylactically appropriate enhanced T cell response.

Because telomerase is expressed at high levels on cancer cells, the antigens and vaccines of the invention are particularly suitable for methods of treating and / or preventing cancer. An exemplary method is to provide a cancer patient with an effective amount of one or more of the telomerase antigens (
This can be formulated as a vaccine). Again, smaller peptide antigens are preferred.

In addition, the telomerase antigens and vaccines of the invention are believed to be particularly useful for in vitro techniques. In general, these techniques involve isolating cells from a patient, contacting them with a telomerase antigen (or vaccine, including a nucleic acid vaccine), and administering the contacted cells back to the patient. Some cases (
For example, in an MHC-compatible subject), cells may be harvested from one patient for administration to another patient.

In one embodiment, autologous or compatible antigen presenting cells (usually dendritic cells or peripheral blood lymphocytes) are sensitized in vitro with a telomerase antigen. Then
These "telomerase-sensitized" cells can be introduced in beneficial amounts into patients in need of treatment or prevention. As in all aspects of the invention, the in vitro sensitization step comprises lipid-
And / or using small peptide antigens associated with liposomes.

[0067] Yet another adoptive approach is conceivable, in which antigen presenting cells are produced as described above and used to produce self or compatible T cell effectors in vitro. The T cells thus generated can be recruited and introduced into a patient in need thereof in a beneficial amount. For a description of the art-recognized techniques of adoptive T cell transfer therapy employed, see Bartels et al., Annals of Sur.
See Gical Oncology, 3 (1): 67 (1996), which is hereby incorporated by reference.

Co-treatment with the other immunostimulants listed above is also conceivable. IL-2, GM-
Molecules such as CSF, IL-12, flt-3 ligand, CD40, etc., are expected to be very useful. For example, IL-2 can be administered simultaneously, separately or in a combined formulation, or can be administered in a separate mode of administration using a telomerase antigen or vaccine. In one method, IL-2 is formulated with a liposome.

In a further embodiment of the present invention, any of the aforementioned antigens or vaccines may be used with known anti-cancer agents. One example includes a therapeutic agent based on MUC-1. Many additional examples of these are known in the art. Conventional chemotherapeutics are
Including alkylating agents, antimetabolites, various natural products (eg, vinca alkaloids, epipodophyllotoxins, antibiotics, and amino acid depleting enzymes), hormones and hormone antagonists. Specific substance classes are: nitrogen mustard, alkylsulfonates, nitrosoureas, triazenes, folic acid analogs, pyrimidine analogs, purine analogs, platinum complexes, adrenal cortex inhibitors, corticosteroids,
Includes progestins, estrogens, antiestrogens and androgens. Some exemplary compounds include cyclophosphamide, chlorambucil, methotrexate, fluorouracil, cytarabine, thioguanine, vinblastine, vincristine, doxorubicin, daunorubicin, mitomycin, cisplatin, hydroxyurea, prednisone, hydroxyprogesterone caproate, medroxyprogesterone, Includes megestrol acetate, diethylsilvestol, ethinylestradiol, tomoxifen, testosterone propionate and fluoxymesterone.

[0070] A therapeutically or prophylactically beneficial or effective amount is an amount sufficient to induce a clinically relevant telomerase-specific T cell response, as defined above. Clinical matters can be determined by the clinician.

[0071] Administration can be by any number of routes, including parenteral and oral. Cell-based vaccines are advantageously administered intravenously. Other vaccines are typically administered intramuscularly, intradermally, subcutaneously or orally. One skilled in the art will recognize that the route of administration will vary depending on the nature of the vaccine formulation. Determination of the optimal route for a vaccine can be determined empirically and is within the level of ordinary skill in the art.

The nucleic acid vaccine may also be administered by various routes, the optimal route being determined empirically. For example, some antigens have been found to elicit superior cytotoxic responses when administered intravenously. Hurpin et al., Supra. For superior immune response to oral administration, it may be beneficial to co-administer the vaccine with a mucosal adjuvant such as cholera toxin or cationic lipid. Ethchart et al. Ge
n. Virol. 78: 1577-80 (1997). Intramuscular, intradermal and subcutaneous administration are also preferred.

EXAMPLES Example 1 This example illustrates the use of a telomerase-specific peptide to generate an antigen-specific cytotoxic T cell response.

[A. Materials and Methods] In general, PCT / US98 / 09288; Agrawal et al., Int'l I
mmunol. 10: 1907-16 (1998); and Agrawal et al.,
Cancer Res. 55: 5151-56 (1998) provide suitable methods, the disclosures of which are hereby incorporated by reference.

[Peptide] The peptide was prepared according to http: // www-bimas. dcrt. nih. gov
HLA peptide search program. HLA-A2 specificity
Selected using default parameters. Three of the top 20 scores were synthesized and tested. These peptides have the sequences, RLVDDFLLV, ELLRSFFYV and ILAKFLHWL.

Preparation of Liposomes The bulk liquid composition of the liposomes consists of dipalmitoyl phosphatidylcholine (DPPC), cholesterol (Chol) and dimyristoyl phosphatidylglycerol (DMPG) in a ratio of 3: 1: 0.25, 1% of
w / w). Synthetic telomerase peptides are present in the aqueous layer at a concentration of 0.7 mg / ml in the liposome formulation, and approximately 28% of the input peptide.
% Were captured within the liposome structure. The formulated product contained 2 mg of bulk lipid, 20 μg of lipid A and about 20 μg of peptide per 100 μl injection dose.

The bulk lipid and lipid A were dissolved in chloroform / methanol (methanol was used first to solubilize DMPG). For each 4 ml liposome preparation, the lipid mixture was 64 mg DPPC, 11 mg in 12 ml chloroform / methanol at a final lipid concentration of 30 mM, at a molar ratio of 3: 1: 0.25.
Chol, 5 mg DMPG, 0.8 mg lipid A. Each 12
The lipid mixture of ml was dried to a film by rotary evaporation at 53 ° C. in a 250 ml round bottom flask and the residual solvent was removed under high vacuum. The lipid film was hydrated by adding 4 ml of PBS containing the peptide, gently rotating the flask at 53 ° C, then vortexing for 5 cycles and warming to 53 ° C.

The liposome structure was further improved by a series of five freeze / thaw cycles consisting of freezing in a dry ice bath, thawing, warming to 41 ° C., and vortexing before the next cycle began. Reformed into uniform size. The liposomes were then collected by ultracentrifugation at 1500,000 × g at 4 ° C. for 20 minutes, washed with two additions of PBS, and ultracentrifuged again. The liposomes were finally reconstituted to the desired volume.

Cytokines To promote CTL production, a human recombinant cytokine, IL-12 (R & D
Systems, Minneapolis, MN), IL-7 (Intermedico, M
arkham, Ontario) was diluted in serum-free AIM-V medium (Life Technologies) immediately before use.

[General procedure for adding APC together with liposome-encapsulated peptide] Human peripheral blood lymphocytes (PBL) were prepared using Ficoll Hypaque (Pharmacia,
Purified from heparinized blood by centrifugation in Uppsala, Sweden). The interface layer between ficoll and blood obtained by centrifugation was collected and washed twice with RPMI before use.

Briefly, one dose of liposomes containing the peptide preparation was added to 2-10 × 10 ⁶ PBLs in 0.9 ml of AIM-V medium and the PBLs were incubated overnight at 37 ° C.
O and incubated at ₂ supplemented incubator. After incubation, PBL
Were treated with mitomycin C or gamma irradiation (3000 rad) and then AIM-
Washed with V medium.

Cytotoxic T Lymphocyte Assay Three (HLA.A2 ⁺ ) normal donor PBLs were used for the CTL assay. T cells were expanded for 5 weeks in bulk culture as described above. At the end of the two weeks, live T cells were collected from the flask and counted. The target was a mutant T2 cell. Houbier et al., Eur. J. Immunol. 2
3: 2072-2077 (1993); Stauss et al., Proc. Natl.
Acad. Sci U. S. A. 89: 7871-7875 (1992). T2
HLA. Telomerase peptide-mediated up-regulation of A2 expression was determined by the HLA-A2-associated peptide ILAKFLHWL (BP1-187), RLV
DDFLLV (BP1-190) and ELLRSFFYV (BP1-191)
) Using known methods. Townsend et al., Nature
346: 476 (1989). T2 cells were incubated overnight at 37 ° C. in 7% CO ₂ for 2 hours.
00 μM of various telomerase synthetic peptides were added in the presence of 8 μg of exogenous β2 microglobulin. Houbier et al., Supra; Stauss et al., Supra.
Peptide-loaded T2 target cells received ⁵¹ Cr (using NaCrO ₄ )
Added for 0 minutes and washed. CTL assays were performed as described above. Agra
Wal et al. Immunol. 156: 2089 (1996). Percent specific kill was calculated as experimental release-spontaneous release / maximum release-spontaneous release x 100. The effector to target ratios used were 50: 1, 25: 1, 10: 1 and 5:
It was one. Each group was set up in replicates of 4 and the average% specific kill was calculated.

[Cell Surface Immunofluorescent Staining] For detection of cell surface antigen, T2 cells that had been fed with a peptide were once treated with 1% BS
Wash in cold PBS containing A, then 1 μg of anti-A2 monoclonal antibody, MA
2.1 or control antibody was added and incubated on ice for 45 minutes. Next, the cells were washed, and a second antibody, goat anti-mouse IgG (H + L) -FITC label (Southern Biotech) was added for 30 minutes on ice.

[B. Results] The cytotoxic activity of T cells stimulated with autologous APCs pulsed with liposomal telomerase peptide was determined using the method described above. The T cell source consists of three HL
A. PBL from A2 ⁺ donor. The telomerase peptide described above was added to target T2 cells (HLA.A2 ⁺ ).

The negative control was T2 cells and the positive control was 10 m
er flu peptide FLPSDYFPSV, which is HLA on T2 cells.
. A2 expression was strongly up-regulated. These data confirm that the method can be used to generate specific, biologically important T cell responses to telomerase, such as cytotoxicity.

FIG. 1 shows FACS analysis of cells treated according to the above method, Panel A shows
A negative control is shown. Panel B is a positive control, showing a 227% increase in median channel intensity of A2 expression compared to the negative control. Panel C is an experiment using BP1-187, showing a 396% increase in median channel intensity of A2 expression compared to the negative control. Panel D
And E show the results with BP1-190 and BP1-191, respectively, resulting in 0% and 6% increases over the negative control, respectively.

Table 1 shows the specific killing of targets by (BP1-187) cytotoxic T cells sensitized with telomerase antigen. The negative control was SIINFEKL.

[0088] These data show that, contrary to expectations, the immune response can be produced against telomerase, a self-antigen.

[0089]

[Table 1]

The foregoing detailed description and examples have been presented for the purpose of illustration only,
It is not meant to be limiting. Further embodiments of the present invention will be readily apparent to one of ordinary skill in the art in light of the present disclosure.

[Brief description of the drawings]

FIG. 1 shows a fluorescence activated cell sorter (FACS) analysis of T cells activated by a telomerase-specific antigen. Panels A and B are a negative control and a positive control, respectively, and panels C, D and E are antigen candidates.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C07K 7/06 C12N 15/00 ZNAA 4H045 7/08 5/00 B C12N 5/10 A61K 37/02 (81 ) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF) , CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SL, SZ, TZ, UG, ZW) ), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR , BY, CA, CH, CN, CR, CU, CZ, DE, DK, DM, DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT , RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (72) Inventor Rongenecker , Brian Michael, Tea 6 Earl, Alberta, Canada 1 Sea 8, Edmonton, Rooney Clement 440 F-term (reference) 4B024 AA01 BA31 CA04 DA02 EA04 GA11 4B065 AA90X AA93Y AB01 BA02 CA25 CA45 4C076 AA19 CC27 FF67 A034A03 BA03 CA56 DC50 MA24 NA14 ZB2 62 4C085 AA03 BB11 CC12 DD23 DD32 DD33 EE01 4H045 AA10 AA11 AA30 CA42 DA86 DA89

Claims (41)

[Claims] 1. A peptide comprising up to about 60 amino acids of a native telomerase protein sequence optionally having one or more conservative substitutions, wherein the peptide is capable of binding at least one human leukocyte antigen (HLA). 2. The peptide according to claim 1, wherein said HLA molecule is a class I molecule. 3. The peptide according to claim 1, which is about 8 to about 12 amino acids in length. 4. The peptide according to claim 1, which comprises the sequence ILAKFLHWL, or a conservative variant thereof. 5. A vaccine comprising at least a portion of a native telomerase protein sequence optionally having one or more conservative amino acid substitutions, or a polynucleotide encoding said telomerase portion, and a lipid, wherein said telomerase is a telomerase. A vaccine whose portion can bind to at least one human leukocyte antigen (HLA). 6. The vaccine according to claim 5, wherein said telomerase moiety is a peptide having a length of about 60 amino acids or less. 7. The vaccine according to claim 5, wherein the telomerase moiety is a peptide having a length of about 8 to about 12 amino acids. 8. The vaccine according to claim 5, wherein the lipid is part of a liposome. 9. A method for treating or preventing cancer, comprising: administering to a patient in need thereof:
Administering an effective amount of a composition comprising at least a portion of a native telomerase protein sequence optionally having one or more conservative amino acid substitutions, or a polynucleotide encoding said telomerase portion, wherein said portion comprises: A method for treating or preventing cancer characterized by being able to bind to at least one human leukocyte antigen (HLA). 10. The method of claim 9, wherein said composition further comprises a lipid. 11. The method according to claim 9, wherein the telomerase moiety is a peptide having a length of about 60 amino acids or less. 12. The method according to claim 9, wherein the telomerase moiety is a peptide having a length of about 8 to about 12 amino acids. 13. The method according to claim 10, wherein the lipid is part of a liposome. 14. A method for treating or preventing cancer comprising administering to a patient in need thereof an antigen-presenting cell sensitized with telomerase. 15. Use of a composition wherein the antigen presenting cell comprises at least a portion of a native telomerase protein sequence optionally having one or more conservative amino acid substitutions, or a polynucleotide encoding the telomerase portion. 15. The method of claim 14, wherein the telomerase moiety is capable of binding to at least one human leukocyte antigen (HLA). 16. The method of claim 15, wherein said telomerase moiety is a peptide of about 60 amino acids or less. 17. The method of claim 16, wherein said telomerase moiety is a peptide of about 8 to about 12 amino acids in length. 18. The method according to claim 15, wherein the composition further comprises a lipid. 19. The method of claim 18, wherein said lipid is in a liposome. 20. The method according to any of claims 14 to 19, further comprising the step of administering an effective amount of interleukin-2 (IL-2) to said patient. 21. An isolated polynucleotide encoding a telomerase-specific antigen, wherein said polynucleotide is no more than about 180 nucleotides in length. 22. The method of claim 21, which is about 24 to about 36 nucleotides in length.
3. The polynucleotide according to item 1. 23. The antigen according to claim 21, wherein the antigen is a class I-specific antigen.
3. The polynucleotide according to 2. 24. The polynucleotide according to any one of claims 21 to 23, which encodes a peptide comprising the sequence ILAKFLHWL or a conservative variant thereof. 25. A method of producing a telomerase-sensitized antigen presenting cell, comprising: providing the antigen presenting cell with at least a portion of a native telomerase protein sequence optionally having one or more conservative amino acid substitutions. Or a telomerase-sensitized antigen presenting cell comprising contacting with a composition comprising a polynucleotide encoding said telomerase moiety, wherein said telomerase moiety is capable of binding to at least one human leukocyte antigen (HLA). A method for producing 26. The method of claim 25, wherein said telomerase moiety is a peptide of about 60 amino acids or less. 27. An antigen-presenting cell sensitized with telomerase produced by the method according to claim 25 or 26. 28. The peptide of claim 2, which is no more than about 25 amino acids in length. 29. The polynucleotide of claim 21, which is 75 or fewer nucleotides in length. 30. At least a portion of a native telomerase protein sequence optionally having one or more conservative amino acid substitutions in the treatment or prevention of cancer.
Or use of a composition comprising a polynucleotide encoding said telomerase moiety, wherein said telomerase moiety comprises at least one human leukocyte antigen (HLA)
Use of a composition characterized in that it can be bound to. 31. The use according to claim 30, wherein the composition further comprises a lipid. 32. The use according to claim 31, wherein said lipid is part of a liposome. 33. The use according to any one of claims 30 to 32, wherein the telomerase moiety is a peptide of up to about 60 amino acids in length. 34. The use according to any of claims 30 to 33, wherein said telomerase moiety is a peptide of about 8 to about 12 amino acids in length. 35. Use of a telomerase-sensitized antigen-presenting cell for treating or preventing cancer. 36. Use of a composition wherein said antigen presenting cell comprises at least a portion of a native telomerase protein sequence optionally having one or more conservative amino acid substitutions, or a polynucleotide encoding said telomerase portion. Use according to claim 35, characterized in that the telomerase moiety is capable of binding to at least one human leukocyte antigen (HLA). 37. The use according to claim 36, wherein said telomerase moiety is a peptide of about 60 amino acids or less. 38. Use according to claim 36 or 37, wherein the telomerase moiety is a peptide of about 8 to about 12 amino acids in length. 39. The use according to any one of claims 36 to 38, wherein the composition further comprises a lipid. 40. The use according to claim 39, wherein said lipid is in a liposome. 41. Use of a composition comprising telomerase-sensitized antigen-presenting cells according to any of claims 15 to 19 in the treatment or prevention of cancer.
Use of a composition characterized in that the composition further comprises interleukin-2 (IL-2).