Fusion protein and its use

Abstract

A fusion protein comprising a first peptide and a second peptide is provided. The first peptide enables the fusion protein to bind to a receptor expressed on a cell, and the second peptide having a cleavage site enables the fusion protein to kill the cell. Therefore, the fusion protein can be used to prevent or treat an infection caused by a pathogen. Also provided are nucleic acids encoding the fusion protein and methods for preparing and using the fusion protein.

Description

Technical Field

[0001] Fusion protein relates to a fusion protein comprising a first peptide and a second peptide. The first peptide enables the fusion protein to bind to a receptor expressed on a cell, which can then be internalized, and the second peptide contains a cleavage site with an optimized selection property, which can therefore be referred to as an optimized or selective cleavage site, which enables the fusion protein to kill the cell. Therefore, the fusion protein can be used to prevent or treat an infection caused by a pathogen (such as a virus), wherein CMV is an example of this virus. Nucleic acids encoding the fusion protein and methods for preparing and using the fusion protein are also provided. Background Art

[0002] Immunotoxins are proteins with a targeting moiety attached to the toxin. Immunotoxins are examples of fusion proteins. Immunotoxins can be used to treat infections caused by a range of pathogens. For example, toxins attached to antibodies have previously been used to target CMV-infected cells, but with limited success.

[0003] Human cytomegalovirus (HCMV) is a species-specific herpesvirus and a notable pathogen particularly in immunocompromised individuals, neonates and patients undergoing transplantation.

[0004] Currently, all drugs used in clinical treatment of HCMV infection are associated with considerable adverse side effects. In addition, the emergence of drug resistance often leads to therapy failure.

[0005] In addition to the pharmacological challenges and poor cell-targeting selectivity of existing immunotoxins, another reason for therapeutic failure is often insufficient internalization, resulting in inadequate killing of infected cells.

[0006] These limitations support the importance of developing novel drug treatments for pathogen-induced infections such as HCMV.

[0007] WO 08 / 003327 discloses immunotoxins that can be used to treat diseases associated with CMV infection. The immunotoxins disclosed therein comprise a variant of the chemokine domain of human CX3CL1 and a toxin. Summary of the invention

[0008] The invention is as defined in the claims.

[0009] Provided herein is a fusion protein comprising a first peptide that provides target selectivity and a second peptide that provides a toxin and also provides internal cleavage of the toxin due to a cleavage site and effector toxicity produced in a targeted cell. The fusion proteins provided herein can be used to treat pathogen-induced infections, including treatment of latent infections.

[0010] The inventor has produced a fusion protein that effectively targets and kills infected cells (such as CMV infected cells, including latent infected cells). The fusion protein comprises a first peptide that can be combined with at least one receptor (which can be a receptor such as US28 encoded by a virus) and a second peptide containing an optimized cleavage site and a toxin (such as the selected domain (such as domain II and III) of exotoxin A). When the target is combined, the receptor is preferably internalized, and the second peptide is at least partially cut at the cleavage site, thereby releasing the toxin part, and then it turns to kill infected cells. Surprisingly, the inventor found that the specific cleavage site motif (ArgX1X2Arg, wherein X2 is alkaline such as Arg) corresponding to the mutation in the natural furin cleavage site of exotoxin A increases the selectivity of the cell expressing the receptor (such as US28) encoded by the pathogen. This discovery allows improvement in targeting and killing of infected cells such as CMV infected cells. Therefore, the optimized cleavage site described herein can be used for fusion proteins, and particularly immunotoxins, to increase toxicity / cell killing efficacy and particularly increase selectivity.

[0011] In one aspect, the present invention provides a fusion protein comprising

[0012] a) a first peptide that binds to at least one receptor expressed on a cell; and

[0013] b) a second peptide comprising a cleavage site having the amino acid sequence ArgX1X2Arg,

[0014] wherein X2 is Arg or Lys, and wherein the second peptide comprises a toxin.

[0015] In another aspect, the present invention provides an isolated nucleic acid molecule encoding a fusion protein according to the above aspects.

[0016] In one aspect, the present invention provides a vector comprising a nucleic acid molecule according to the above aspects.

[0017] In another aspect, the present invention provides a recombinant host cell comprising the nucleic acid molecule or vector according to the above aspects.

[0018] In one aspect, the present invention provides a pharmaceutical composition comprising the fusion protein, nucleic acid, vector or recombinant host cell according to the above aspects and a pharmaceutically acceptable carrier, diluent and / or excipient.

[0019] In another aspect, the present invention provides a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition according to the above aspects for use as a medicament.

[0020] In one aspect, the present invention provides a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition according to the above aspects, which is used to prevent or treat infection caused by pathogens and / or pathogen-related disorders.

[0021] In another aspect, the present invention provides a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition according to the above aspects, which is used for preventing or treating CMV infection and / or CMV-related disorders.

[0022] In one aspect, the present invention provides a method for treating or preventing an infection caused by a pathogen, such as a CMV infection, in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition according to any of the above aspects.

[0023] In another aspect, the present invention provides an ex vivo use of the fusion protein or pharmaceutical composition according to any one of the above aspects for treating solid organs for transplantation and / or hematopoietic stem cells for transplantation.

[0024] In one aspect, the present invention provides a fusion protein, nucleic acid, vector, host cell or pharmaceutical composition according to the above aspects, which is used for the manufacture of a medicament for preventing or treating infection caused by a pathogen and / or a pathogen-related disorder, such as for preventing or treating CMV infection and / or CMV-related disorders.

[0025] In another aspect, the present invention provides a method for producing a fusion protein according to any of the above aspects, the method comprising culturing a host cell as defined in any of the above aspects under conditions allowing expression of the encoded fusion protein. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 : Domain structures of exemplary first peptides of human CX3CL1, Pseudomonas exotoxin A, and fusion protein SYN002.

[0027] A. Schematic diagram of the following domain structures: human CX3CL1 (S = signal sequence, CX3CL1 = chemokine domain, handle = mucin-like handle, M = transmembrane portion and C = cytoplasmic domain), Pseudomonas aeruginosa exotoxin A (S = signal sequence, domain I = receptor binding domain, domain II = translocation domain, Ib = domain Ib with unknown function and domain III = ADP-ribosylation domain), with the amino acid number of the precursor protein given above each protein. Disulfide bridges are shown below each protein in square brackets and the number of the amino acid involved. B. Schematic diagram of SYN002. Mutations in a given domain are written with the single letter code of the amino acid involved and its number. For example, P303R means that the proline at position number 303 has been replaced by arginine. The single letter code is also used at the N and C termini of the construct and between domains. The dotted line between two domains indicates that the amino acids are connected. Furin cleaves between amino acids 304 and 305 of domain II of exotoxin A.

[0028] Figure 2 : Selectivity due to the F49A mutation.

[0029] Substituting a single amino acid in the chemokine portion of SYNx results in selectivity for the US28 receptor (A) versus the CXCR1 receptor (B). SYN000 is the wild-type human chemokine (C-X3-C motif) ligand 1 (CX3CL1) (the native chemokine sequence), while SYN001 has a single mutation (F49A) in the receptor binding portion of the protein.

[0030] Figure 3 : Introduction of a furin cleavage site (in this case, the full-length translocation domain of exotoxin A) increased potency.

[0031] Addition of the full-length translocation domain of exotoxin A containing a furin cleavage site to the SYN001 construct generated SYN016, which increased potency against both endogenous CX3CR1 receptor-expressing cells and virally encoded US28 receptor-expressing cells.

[0032] Figure 4 : In vitro cleavage by furin.

[0033] Compared to the fusion protein construct SYN016 with the native furin cleavage site sequence (RQPR), furin did not improve the in vitro cleavage of the SYN002 construct with the optimized cleavage site (RQRR). In vitro furin digestion of SYNx. Lane 1: Mark 12 protein standard, lane 2: SYN000, lane 3: SYN017, lane 4: SYN014, lane 5: SYN001, lane 6: SYN016 and lane 7: SYN002. SYN000, SYN017 and SYN014 have the native CX3CL1 chemokine sequence, while SYN001, SYN016 and SYN002 have the F49A mutation in the chemokine portion. SYN000, SYN017, SYN001, and SYN016 have a native furin cleavage site (RQPR), while SYN014 and SYN002 have an optimized cleavage site (RQRR).

[0034] Figure 5 : SYN002 is cleaved by furin over a wide pH range.

[0035] SYN002 is cleaved in vitro over a wide pH range. Lane 1: Mark 12 protein standard, Lane 2: no furin added, Lane 3: +8 mm HCl (about pH 5.2), Lane 4: +6 mm HCl (about pH 6.3), Lane 5: +4 mm HCl (about pH 6.6), Lane 6: +2 mm HCl (about pH 7.0), Lane 7: no titrant added (about pH 7.4), Lane 8: 2 mm NaOH (about pH 7.7), and Lane 9: +4 mm NaOH (about pH 9.0).

[0036] Figure 6 : Optimized cleavage site increases selectivity

[0037] When the optimized cleavage site ArgX1X2Arg is introduced into the second peptide, in this particular case, the optimized cleavage site ArgGlnArgArg is introduced into SYN016 to obtain SYN002, thereby obtaining an increase in selectivity for virally encoded receptors. Compared with SYN016 with a cleavage site ArgGlnProArg (therefore, X2 = Pro), the cell killing efficacy of SYN002 for cells expressing virally encoded receptors such as US28 remains approximately unchanged, but the efficacy of cells expressing endogenous / human receptors such as CX3CR1 is reduced compared to SYN016. When compared with a second peptide that does not contain a furin cleavage site (e.g., SYN000 (not shown in the figure)), both selectivity and efficacy are increased. DETAILED DESCRIPTION

[0038] The present disclosure relates to a fusion protein comprising a first peptide that provides target recognition and target selectivity and a second peptide that provides a toxin and also provides intracellular cleavage and release of the toxin and effector toxicity produced in the targeted cells. The fusion proteins provided herein can be used to treat pathogen-induced infections, including treatment of latent infections.

[0039] The present inventors have generated fusion proteins that effectively target and kill infected cells (such as CMV-infected cells, including latently infected cells) that can advantageously be constitutively internalized upon binding to a receptor, such as a virally encoded receptor such as US28.

[0040] US28 is a G protein coupled receptor encoded by the human cytomegalovirus open reading frame US28. US28 is a constitutive internalization receptor. Therefore, chemokines or other compounds in combination with US28 are internalized into cells expressing the receptor. US28 is expressed on CMV infected cells (including cells latently infected by CMV).

[0041] The fusion protein comprises a first peptide capable of binding to a receptor, particularly a receptor encoded by a pathogen, such as US28, and a second peptide containing an optimized cleavage site and a selected domain of a toxin, such as exotoxin A. After mutation in the cleavage site, the selectivity of the fusion protein for a receptor encoded by a pathogen, such as US28, is surprisingly enhanced.

[0042] The optimized cleavage sites described herein can be used in various fusion proteins, and in particular immunotoxins, to increase cell killing selectivity by altering cell killing potency in a target receptor-selective manner.

[0043] In another aspect, the present invention provides a fusion protein comprising

[0044] a) a first peptide that binds to at least one receptor expressed on a cell; and

[0045] b) a second peptide comprising a cleavage site having the amino acid sequence ArgX1X2Arg,

[0046] wherein X2 is Arg or Lys, and wherein the second peptide comprises a toxin.

[0047] The term "amino acid" as used herein includes the standard twenty genetically encoded amino acids and their corresponding stereoisomers in the "D" form (compared to the natural "L" form), ω-amino acids and other naturally occurring amino acids, unconventional amino acids (e.g., α, α-disubstituted amino acids, N-alkyl amino acids, etc.), and chemically derivatized amino acids. Unless otherwise expressly stated, when specifically enumerating an amino acid such as "alanine" or "Ala" or "A", the term refers to both l-alanine and d-alanine.

[0048] In one embodiment of the invention, the fusion protein consists only of naturally occurring amino acids.

[0049] Cleavage site

[0050] The second peptide of the present disclosure comprises a cleavage site having the following amino acid sequence

[0051] ArgX1X2Arg

[0052] X1 can be any amino acid. In one embodiment, X1 is selected from Gln, Ser, Thr and Asn. In one embodiment, X1 is Gln.

[0053] X2 is selected from Arg and Lys. In an advantageous embodiment, X2 is Arg. In one embodiment, X2 is Lys. By introducing a basic amino acid such as Arg at amino acid position X2, cell killing selectivity is enhanced. Thus, for example, cells infected with a virus expressing a receptor encoded by the virus and binding to the first peptide are killed more effectively than cells expressing an endogenous receptor that can also bind to the first peptide.

[0054] In a preferred embodiment, the cleavage site comprises or consists of the amino acid sequence ArgGlnArgArg(RQRR). In one embodiment, the cleavage site comprises or consists of the amino acid sequence ArgGlnLysArg. In one embodiment, the cleavage site comprises or consists of the amino acid sequence ArgSerLysArg. In one embodiment, the cleavage site comprises or consists of the amino acid sequence ArgSerArgArg. In one embodiment, the cleavage site comprises or consists of the amino acid sequence ArgThrLysArg. In one embodiment, the cleavage site comprises or consists of the amino acid sequence ArgThrArgArg. In one embodiment, the cleavage site comprises or consists of the amino acid sequence ArgAsnLysArg. In one embodiment, the cleavage site comprises or consists of the amino acid sequence ArgAsnArgArg.

[0055] In one embodiment, cleavage occurs at the C-terminus of the cleavage site.

[0056] The fusion protein can be cleaved with furin, but other enzymes or proteases may also be involved in cleavage of the cleavage site.

[0057] Thus, in one embodiment, the cleavage site is an enzymatic cleavage site.

[0058] In one embodiment, the enzymatic cleavage site is a furin cleavage site.

[0059] Receptors

[0060] In one embodiment, the first peptide binds to at least one receptor expressed on a cell. In one embodiment, the first peptide binds to at least two receptors expressed on a cell.

[0061] In one embodiment, one receptor that binds to the first peptide is a receptor encoded by a pathogen, and another receptor that binds to the first peptide is a human-encoded receptor and / or an endogenous receptor for the first peptide or a variant thereof.

[0062] In one embodiment, the receptor is a G protein coupled receptor (GPCR), such as US28 of SEQ ID NO:10.

[0063] In one embodiment, the receptor is a chemokine receptor. In one embodiment, the chemokine receptor is a CC chemokine receptor and / or a CX3C chemokine receptor.

[0064] In one embodiment, the CC chemokine receptor is US28. In one embodiment, the CX3C chemokine receptor is CX3CR1.

[0065] In one embodiment, the receptor is encoded by a pathogen. In one embodiment, the pathogen is a bacterium. In some embodiments, the pathogen is a virus. In some embodiments, the virus is a DNA virus. In some embodiments, the virus is an RNA virus. In one embodiment, the virus is a herpes virus. In one embodiment, the virus is a cytomegalovirus.

[0066] In one embodiment, the receptor is capable of internalization. In one embodiment, the receptor is internalized after binding to the first peptide. In one embodiment, the receptor is constitutively internalized. Receptors encoded by pathogens are advantageously constitutively internalized because this will ensure that the fusion protein is efficiently absorbed by cells infected by pathogens, thereby causing death of infected cells and having minimal undesirable toxicity and side effects. "Internalization" refers to the process by which a receptor is moved into a cell expressing it. For example, the receptor can enter a cell by endocytosis or phagocytosis.

[0067] In some embodiments, the invention provides a fusion protein that targets a specific cell by binding to a receptor in a selective manner, thereby causing internalization of the receptor. The toxin portion of the fusion protein is then cleaved, killing the cell.

[0068] toxin

[0069] In one embodiment, the fusion protein is an immunotoxin. An "immunotoxin" refers to a protein consisting of a targeting moiety attached to a toxin. The targeting moiety may also be referred to as a ligand.

[0070] The toxins can be different types of toxins. In one embodiment, the second peptide comprises a toxin selected from Pseudomonas exotoxin A, gelonin, bouganin, saporin, ricin, ricin A chain, brentux, restrictocin, diphtheria toxin, diphtheria toxin A chain and variants and fragments thereof. The toxin portion of the fusion protein enables cell killing. The toxin is cytotoxic.

[0071] In one embodiment, the second peptide comprises one or more domains of Pseudomonas exotoxin A. In one embodiment, the second peptide comprises at least a portion of exotoxin A of SEQ ID NO:9.

[0072] Fusion Protein

[0073] The first peptide is a targeting moiety which allows binding to a receptor expressed on a cell.

[0074] In one embodiment, the first peptide and the second peptide are operably linked. Generally, "operably linked" means that the linked sequences are contiguous and / or in a functional relationship with each other, such as covalently linked.

[0075] In one embodiment, the first peptide comprises or consists of

[0076] a. the amino acid sequence of SEQ ID NO: 1;

[0077] b. a variant of SEQ ID NO: 1, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 1, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 1, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 1;

[0078] c. A fragment of SEQ ID NO: 1 having a length of more than 50 amino acids, such as more than 60, 70 or 75 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 1, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 1, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 1.

[0079] In one embodiment, the first peptide comprises or consists of a variant of SEQ ID NO: 1, the variant comprising or consisting of an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises or consists of a variant of SEQ ID NO: 1, the variant comprising or consisting of an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises or consists of a variant of SEQ ID NO: 1, the variant comprising or consisting of an amino acid sequence having at least 96% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises or consists of a variant of SEQ ID NO: 1, the variant comprising or consisting of an amino acid sequence having at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises or consists of a variant of SEQ ID NO: 1, the variant comprising or consisting of an amino acid sequence having at least 98% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises or consists of a variant of SEQ ID NO:1, said variant comprising or consisting of an amino acid sequence having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:1.

[0080] In one embodiment, the first peptide comprises or consists of a fragment of SEQ ID NO: 1 having a length of more than 50 amino acids. In one embodiment, the first peptide comprises or consists of a fragment of SEQ ID NO: 1 having a length of more than 60 amino acids. In one embodiment, the first peptide comprises or consists of a fragment of SEQ ID NO: 1 having a length of more than 70 amino acids. In one embodiment, the first peptide comprises or consists of a fragment of SEQ ID NO: 1 having a length of more than 75 amino acids.

[0081] In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 96% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 96% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 98% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 98% sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the first peptide comprises a fragment of SEQ ID NO: 1 or a variant thereof having at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 1, or consists of a fragment of SEQ ID NO: 1 or a variant thereof having at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 1.

[0082] In one embodiment, the amino acid residue at position 49 of SEQ ID NO: 1 is mutated to Ala, Lys or Asp, preferably Ala. These substitutions can result in increased selectivity for cell killing of cells expressing a pathogen-encoded receptor that binds the first peptide, compared to a human or endogenously encoded receptor that also binds the first peptide.

[0083] In one embodiment, the first peptide further comprises a methionine (M) at the N-terminus.

[0084] In one embodiment, the first peptide comprises or consists of one or more additional amino acids inserted at the N- and / or C-terminus and / or within the amino acid sequence of SEQ ID NO: 1.

[0085] In one embodiment, one or more amino acids are deleted at the N- and / or C-terminus and / or within the amino acid sequence of SEQ ID NO: 1.

[0086] In some embodiments, the first peptide is less than 100 amino acids in length.

[0087] In some embodiments, the first peptide is less than 90 amino acids in length.

[0088] In some embodiments, the first peptide is less than 85 amino acids in length.

[0089] In some embodiments, the first peptide is less than 80 amino acids in length.

[0090] In one embodiment, the second peptide comprises domain A, which may be a translocation domain, such as domain A having an amino acid sequence according to SEQ ID NO:3, or a fragment or variant thereof; and / or domain B, such as domain B having an amino acid sequence according to SEQ ID NO:4, or a fragment or variant thereof; and / or domain C, which may be a cytotoxic domain, such as domain C having an amino acid sequence according to SEQ ID NO:5, or a fragment or variant thereof.

[0091] In one embodiment, the second peptide comprises domain A, such as domain A having an amino acid sequence according to SEQ ID NO: 3, or a fragment or variant thereof. In one embodiment, the second peptide comprises domain A, such as domain A having an amino acid sequence according to SEQ ID NO: 3, or a fragment or variant thereof; and domain B, such as domain B having an amino acid sequence according to SEQ ID NO: 4, or a fragment or variant thereof. In one embodiment, the second peptide comprises domain A, such as domain A having an amino acid sequence according to SEQ ID NO: 3, or a fragment or variant thereof; and domain B, such as domain B having an amino acid sequence according to SEQ ID NO: 4, or a fragment or variant thereof; and domain C, such as domain C having an amino acid sequence according to SEQ ID NO: 5, or a fragment or variant thereof. In one embodiment, the second peptide comprises domain B having an amino acid sequence according to SEQ ID NO: 4, or a fragment or variant thereof. In one embodiment, the second peptide comprises domain C, such as domain C having an amino acid sequence according to SEQ ID NO: 5, or a fragment or variant thereof.

[0092] In one embodiment, the second peptide comprises

[0093] a. the amino acid sequence of SEQ ID NO: 3;

[0094] b. a variant of SEQ ID NO: 3, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 3, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 3;

[0095] c. A fragment of SEQ ID NO:3 having a length of more than 80 amino acids, such as more than 90, 100 or 110 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:3, more preferably at least 85% or 90% sequence identity to SEQ ID NO:3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:3.

[0096] In one embodiment, the second peptide comprises

[0097] a. the amino acid sequence of SEQ ID NO: 4;

[0098] b. a variant of SEQ ID NO: 4, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 4, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 4;

[0099] c. A fragment of SEQ ID NO:4 having a length of more than 6 amino acids, such as a length of more than 8, 10 or 12 amino acids, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:4, more preferably at least 85% or 90% sequence identity to SEQ ID NO:4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:4.

[0100] In one embodiment, the second peptide comprises

[0101] a. the amino acid sequence of SEQ ID NO: 5;

[0102] b. a variant of SEQ ID NO: 5, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 5, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 5;

[0103] c. A fragment of SEQ ID NO:5 having a length of more than 180 amino acids, such as more than 190, 200 or 210 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:5, more preferably at least 85% or 90% sequence identity to SEQ ID NO:5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:5.

[0104] In one embodiment, the second peptide comprises

[0105] a. the amino acid sequence of SEQ ID NO: 3;

[0106] b. a variant of SEQ ID NO: 3, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 3, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 3;

[0107] c. a fragment of SEQ ID NO: 3 having a length of more than 80 amino acids, such as more than 90, 100 or 110 amino acids, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 3, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 3; and

[0108] d. the amino acid sequence of SEQ ID NO: 4;

[0109] e. a variant of SEQ ID NO: 4, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 4, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 4;

[0110] f. A fragment of SEQ ID NO:4 having a length of more than 6 amino acids, such as a length of more than 8, 10 or 12 amino acids, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:4, more preferably at least 85% or 90% sequence identity to SEQ ID NO:4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:4.

[0111] In one embodiment, the second peptide comprises

[0112] a. the amino acid sequence of SEQ ID NO: 3;

[0113] b. a variant of SEQ ID NO: 3, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 3, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 3;

[0114] c. a fragment of SEQ ID NO: 3 having a length of more than 80 amino acids, such as more than 90, 100 or 110 amino acids, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 3, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 3; and

[0115] d. the amino acid sequence of SEQ ID NO: 4;

[0116] e. a variant of SEQ ID NO: 4, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 4, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 4;

[0117] f. a fragment of SEQ ID NO: 4 having a length of more than 6 amino acids, such as a length of more than 8, 10 or 12 amino acids, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 4, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 4; and

[0118] g. the amino acid sequence of SEQ ID NO: 5;

[0119] h. a variant of SEQ ID NO:5, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:5, more preferably at least 85% or 90% sequence identity to SEQ ID NO:5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:5;

[0120] i. A fragment of SEQ ID NO:5 having a length of more than 180 amino acids, such as more than 190, 200 or 210 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:5, more preferably at least 85% or 90% sequence identity to SEQ ID NO:5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:5.

[0121] In one embodiment, the second peptide comprises

[0122] a. the amino acid sequence of SEQ ID NO: 3;

[0123] b. a variant of SEQ ID NO: 3, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 3, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 3;

[0124] c. a fragment of SEQ ID NO: 3 having a length of more than 80 amino acids, such as more than 90, 100 or 110 amino acids, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 3, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 3, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 3; and

[0125] d. the amino acid sequence of SEQ ID NO: 5;

[0126] e. a variant of SEQ ID NO: 5, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 5, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 5;

[0127] f. A fragment of SEQ ID NO:5 having a length of more than 180 amino acids, such as more than 190, 200 or 210 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:5, more preferably at least 85% or 90% sequence identity to SEQ ID NO:5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:5.

[0128] In one embodiment, the second peptide comprises

[0129] a. the amino acid sequence of SEQ ID NO: 4;

[0130] b. a variant of SEQ ID NO: 4, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 4, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 4;

[0131] c. a fragment of SEQ ID NO: 4 having a length of more than 6 amino acids, such as a length of more than 8, 10 or 12 amino acids, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 4, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 4, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 4; and

[0132] d. the amino acid sequence of SEQ ID NO: 5;

[0133] e. a variant of SEQ ID NO: 5, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 5, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 5;

[0134] f. A fragment of SEQ ID NO:5 having a length of more than 180 amino acids, such as more than 190, 200 or 210 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:5, more preferably at least 85% or 90% sequence identity to SEQ ID NO:5, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:5.

[0135] In one embodiment, domain A is a translocation domain and domain C is a cytotoxicity domain, such as an ADP-ribosylation domain.

[0136] In one embodiment, the second peptide comprises the amino acid sequence KDEL of SEQ ID NO:8 in the C-terminus. In one embodiment, the last 5 amino acids of the second peptide are replaced with the amino acid sequence of SEQ ID NO:8.

[0137] In one embodiment, the second peptide comprises or consists of

[0138] a. the amino acid sequence of SEQ ID NO: 2;

[0139] b. a variant of SEQ ID NO: 2, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 2, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 2, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2;

[0140] c. A fragment of SEQ ID NO:2 having a length of more than 300 amino acids, such as more than 310, 330 or 340 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:2, more preferably at least 85% or 90% sequence identity to SEQ ID NO:2, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2.

[0141] In one embodiment, the second peptide comprises or consists of a variant of SEQ ID NO:2, the variant comprising or consisting of an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO:2. In one embodiment, the second peptide comprises or consists of a variant of SEQ ID NO:2, the variant comprising or consisting of an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO:2. In one embodiment, the second peptide comprises or consists of a variant of SEQ ID NO:2, the variant comprising or consisting of an amino acid sequence having at least 96% sequence identity with the amino acid sequence of SEQ ID NO:2. In one embodiment, the second peptide comprises or consists of a variant of SEQ ID NO:2, the variant comprising or consisting of an amino acid sequence having at least 97% sequence identity with the amino acid sequence of SEQ ID NO:2. In one embodiment, the second peptide comprises or consists of a variant of SEQ ID NO:2, the variant comprising or consisting of an amino acid sequence having at least 98% sequence identity with the amino acid sequence of SEQ ID NO:2. In one embodiment, the second peptide comprises or consists of a variant of SEQ ID NO:2, said variant comprising or consisting of an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:2.

[0142] In one embodiment, the second peptide comprises or consists of a fragment of SEQ ID NO:2 having a length of more than 300 amino acids. In one embodiment, the second peptide comprises or consists of a fragment of SEQ ID NO:2 having a length of more than 310 amino acids. In one embodiment, the second peptide comprises or consists of a fragment of SEQ ID NO:2 having a length of more than 330 amino acids. In one embodiment, the second peptide comprises or consists of a fragment of SEQ ID NO:2 having a length of more than 340 amino acids.

[0143] In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO: 2. In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 2. In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 2. In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 2. In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 96% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 96% sequence identity with the amino acid sequence of SEQ ID NO: 2. In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 2. In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 98% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 98% sequence identity with the amino acid sequence of SEQ ID NO: 2. In one embodiment, the second peptide comprises a fragment of SEQ ID NO: 2 or a variant thereof having at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 2, or consists of a fragment of SEQ ID NO: 2 or a variant thereof having at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 2.

[0144] In one embodiment, the second peptide is less than 400 amino acids in length, such as less than 380, 370, 360, 350 or 345 amino acids in length.

[0145] In some embodiments, the second peptide is less than 400 amino acids in length.

[0146] In some embodiments, the second peptide is less than 380 amino acids in length.

[0147] In some embodiments, the second peptide is less than 370 amino acids in length.

[0148] In some embodiments, the second peptide is less than 360 amino acids in length.

[0149] In some embodiments, the second peptide is less than 350 amino acids in length.

[0150] In some embodiments, the second peptide is less than 345 amino acids in length.

[0151] In one embodiment, the fusion protein comprises or consists of

[0152] a. the amino acid sequence of SEQ ID NO: 6;

[0153] b. a variant of SEQ ID NO: 6, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 6, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 6, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 6;

[0154] c. A fragment of SEQ ID NO:6 having a length of more than 360 amino acids, such as more than 380, 400 or 420 amino acids in length, or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO:6, more preferably at least 85% or 90% sequence identity with SEQ ID NO:6, and most preferably at least 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO:6.

[0155] In one embodiment, the fusion protein comprises or consists of a variant of SEQ ID NO:6, the variant comprising or consisting of an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO:6. In one embodiment, the fusion protein comprises or consists of a variant of SEQ ID NO:6, the variant comprising or consisting of an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO:6. In one embodiment, the fusion protein comprises or consists of a variant of SEQ ID NO:6, the variant comprising or consisting of an amino acid sequence having at least 96% sequence identity with the amino acid sequence of SEQ ID NO:6. In one embodiment, the fusion protein comprises or consists of a variant of SEQ ID NO:6, the variant comprising or consisting of an amino acid sequence having at least 97% sequence identity with the amino acid sequence of SEQ ID NO:6. In one embodiment, the fusion protein comprises or consists of a variant of SEQ ID NO:6, the variant comprising or consisting of an amino acid sequence having at least 98% sequence identity with the amino acid sequence of SEQ ID NO:6. In one embodiment, the fusion protein comprises or consists of a variant of SEQ ID NO:6, wherein the variant comprises or consists of an amino acid sequence having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:6.

[0156] In one embodiment, the fusion protein comprises or consists of a fragment of SEQ ID NO:6 having a length of more than 360 amino acids. In one embodiment, the fusion protein comprises or consists of a fragment of SEQ ID NO:6 having a length of more than 380 amino acids. In one embodiment, the fusion protein comprises or consists of a fragment of SEQ ID NO:6 having a length of more than 400 amino acids. In one embodiment, the fusion protein comprises or consists of a fragment of SEQ ID NO:6 having a length of more than 420 amino acids.

[0157] In one embodiment, the fusion protein comprises a fragment of SEQ ID NO: 6 or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO: 6, or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO: 6. In one embodiment, the fusion protein comprises a fragment of SEQ ID NO: 6 or a variant thereof having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 6, or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 6. In one embodiment, the fusion protein comprises a fragment of SEQ ID NO: 6 or a variant thereof having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 6, or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 6. In one embodiment, the fusion protein comprises a fragment of SEQ ID NO: 6 or a variant thereof having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 6, or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 6. In one embodiment, the fusion protein comprises a fragment of SEQ ID NO: 6 or a variant thereof having at least 96% sequence identity with the amino acid sequence of SEQ ID NO: 6, or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 96% sequence identity with the amino acid sequence of SEQ ID NO: 6. In one embodiment, the fusion protein comprises a fragment of SEQ ID NO: 6 or a variant thereof having at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 6, or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 6. In one embodiment, the fusion protein comprises or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 98% sequence identity with the amino acid sequence of SEQ ID NO: 6. In one embodiment, the fusion protein comprises or consists of a fragment of SEQ ID NO: 6 or a variant thereof having at least 98% sequence identity with the amino acid sequence of SEQ ID NO: 6.

[0158] In one embodiment, the fusion protein is less than 500 amino acids in length, such as less than 490, 480, 470, 460, 450, 440, 430, 425 or fewer amino acids in length.

[0159] In some embodiments, the fusion protein is less than 500 amino acids in length. In some embodiments, the fusion protein is less than 490 amino acids in length. In some embodiments, the fusion protein is less than 480 amino acids in length. In some embodiments, the fusion protein is less than 470 amino acids in length. In some embodiments, the fusion protein is less than 460 amino acids in length. In some embodiments, the fusion protein is less than 450 amino acids in length. In one embodiment, the fusion protein is less than 440 amino acids in length. In some embodiments, the fusion protein is less than 430 amino acids in length. In some embodiments, the fusion protein is less than 420 amino acids in length.

[0160] In one embodiment, the fusion protein kills cells infected with a pathogen, such as cells latently infected with a pathogen.

[0161] In one embodiment, the pathogen is a virus such as a DNA virus, such as a herpes virus, such as a cytomegalovirus; or an RNA virus.

[0162] In one embodiment, the fusion protein induces cell death of a cell expressing the receptor.

[0163] In one embodiment, the cell death is selected from apoptosis, necrosis, autophagic cell death and mitosis-related cell death. The terms "apoptosis" and "apoptotic activity" are used in a broad sense and refer to an orderly or controlled form of cell death in mammals that is usually accompanied by one or more characteristic cell changes, including condensation of cytoplasm, loss of plasma membrane microvilli, nuclear fragmentation, degradation of chromosomal DNA, or loss of mitochondrial function.

[0164] In one embodiment, the fusion protein induces a direct or indirect effect on the pathogen, resulting in inhibition of pathogen growth, replication, genomic stability, maturation, packaging, latency, reactivation, dissemination and / or immunosuppression.

[0165] In one embodiment, the fusion protein selectively kills cells expressing a receptor encoded by the pathogen.

[0166] In one embodiment, the fusion protein kills cells expressing US28.

[0167] In one embodiment, the fusion protein has an IC50 value for a virally encoded receptor such as US28 of less than 10 nM, such as less than 5 nM, for example less than 1 nM, such as less than 0,5 nM, for example less than 0,1 nM or less than 0,001 nM.

[0168] In one embodiment, compared to the effectiveness of cells expressing endogenous receptors or human encoded receptors such as CX3CR1, the fusion protein has an increased effectiveness, such as at least 100 times the effectiveness of cells expressing virally encoded receptors such as US28, such as at least 300 times the effectiveness of cells expressing CX3CR1, such as at least 400 times the effectiveness of cells expressing CX3CR1, such as at least 500 times the effectiveness of cells expressing US28. In one embodiment, compared to the effectiveness of cells expressing CX3CR1, the fusion protein has an increased effectiveness, such as at least 100 times the effectiveness of cells expressing US28. In one embodiment, compared to the effectiveness of cells expressing CX3CR1, the fusion protein has an increased effectiveness, such as at least 300 times the effectiveness of cells expressing US28. In one embodiment, compared to the effectiveness of cells expressing CX3CR1, the fusion protein has an increased effectiveness, such as at least 400 times the effectiveness of cells expressing US28. In one embodiment, compared to the effectiveness of cells expressing CX3CR1, the fusion protein has an increased effectiveness, such as at least 500 times the effectiveness of cells expressing US28.

[0169] In one embodiment, the fusion protein has an increased affinity for a virally encoded receptor such as US28, such as at least a 50-fold increased affinity, such as at least a 100-fold increased affinity, compared to the affinity for an endogenous receptor or a human-encoded receptor such as CX3CR1. In one embodiment, the fusion protein has an increased affinity for US28, such as at least a 100-fold increased affinity, compared to the affinity for CX3CR1.

[0170] In one embodiment, the selectivity ratio US28 / CX3CR1 of the fusion protein of the present invention is at least 750, such as at least 800, such as at least 850, such as at least 900. In one embodiment, the selectivity ratio US28 / CX3CR1 of the fusion protein of the present invention is at least 880. In one embodiment, the selectivity ratio US28 / CX3CR1 of the fusion protein of the present invention is at least 750. In one embodiment, the selectivity ratio US28 / CX3CR1 of the fusion protein of the present invention is at least 800. In one embodiment, the selectivity ratio US28 / CX3CR1 of the fusion protein of the present invention is at least 850. In one embodiment, the selectivity ratio US28 / CX3CR1 of the fusion protein of the present invention is at least 900.

[0171] Nucleic Acids

[0172] In one aspect, the invention provides an isolated nucleic acid molecule encoding a fusion protein as described herein.

[0173] "Nucleic acid molecules" include DNA (eg, genomic DNA or complementary DNA) and mRNA molecules, which may be single-stranded or double-stranded. "Isolated" means that the nucleic acid molecule is not located or otherwise provided within a cell.

[0174] Carrier

[0175] In one aspect, the invention provides a vector comprising a nucleic acid molecule as described herein.

[0176] In one embodiment, the vector is an expression vector.

[0177] Host cells

[0178] In one aspect, the invention provides a recombinant host cell comprising a nucleic acid molecule or a vector as described herein.

[0179] In one embodiment, the host cell is a bacterial cell, a yeast cell, a mammalian cell such as a human cell, or an insect cell.

[0180] Pharmaceutical composition

[0181] In one aspect, the present invention provides a pharmaceutical composition comprising a fusion protein, nucleic acid, vector or recombinant host cell as described herein and a pharmaceutically acceptable carrier, diluent and / or excipient.

[0182] In one embodiment, the pharmaceutical composition further comprises one or more pharmaceutical agents. In one embodiment, the pharmaceutical agent is selected from an immunosuppressant, an antiviral agent, and an immunotherapy.

[0183] In one embodiment, the antiviral agent is selected from valganciclovir, ganciclovir, cidofovir, leflunomide, letermovir, maribavir, and cidofovir.

[0184] In one embodiment, the immunotherapy is a cell-based therapy. In one embodiment, the immunotherapy is a T cell therapy.

[0185] disease

[0186] In one aspect, the present invention provides a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition as described herein for use as a medicament.

[0187] In another aspect, the present invention provides a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition as described herein, for use in preventing or treating an infection caused by a pathogen and / or a pathogen-related disorder.

[0188] In one embodiment, the pathogen is a bacterium or a virus. In one embodiment, the virus is a DNA virus, such as a herpes virus, such as a cytomegalovirus; or an RNA virus.

[0189] In one aspect, the present invention provides a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition as described herein, which is used to prevent or treat CMV infection and / or CMV-related disorders.

[0190] In one embodiment, the CMV infection is a latent and / or lytic CMV infection.

[0191] In one embodiment, CMV infection can be detected in:

[0192] i) one or more tissues selected from the group consisting of retina, cornea, heart, liver, kidney, lung, gastrointestinal tissue, thymus, spleen, skin and muscle, and / or

[0193] ii) a body fluid selected from one or more of saliva, blood, urine, semen and breast milk.

[0194] In one embodiment, the CMV infection is an infection in an immunocompromised patient selected from HIV patients, neonates and immunosuppressed patients, bone marrow transplant patients, solid organ transplant patients, immunotherapy patients, cancer patients, intensive care patients, trauma patients, stem cell patients, gene therapy patients, cell therapy patients, elderly patients, and patients with multiple diseases.

[0195] In one embodiment, the CMV infection is an infection in a patient suffering from coronary artery disease and / or vascular disease.

[0196] In one embodiment, the fusion protein, the nucleic acid, the vector, the host cell, or the composition is administered intravenously, intratumorally, intraperitoneally, intrathecally, and / or intralymphatically.

[0197] In one embodiment, the fusion protein, the nucleic acid, the vector, the host cell or the composition is administered to the individual one or more times.

[0198] In one embodiment, the individual is a human.

[0199] In one embodiment, the human is an immunocompromised patient. An immunocompromised patient is one whose immune system is damaged or weakened, resulting in the patient's inability to respond to infection. For example, an immunocompromised patient may be diagnosed with a disease that affects the immune system, such as diabetes and HIV. An immunocompromised patient may have a suppressed immune response after treatment such as chemotherapy.

[0200] In one embodiment, the human is a child. In one embodiment, the human is an adult.

[0201] In one embodiment, the human is in need of a solid organ transplant and / or a hematopoietic stem cell transplant. Hematopoietic stem cell transplantation is a transplantation of pluripotent hematopoietic stem cells, usually derived from a bone marrow, peripheral blood or umbilical cord blood. The transplant can be autologous (stem cells are separated from the same patient) or allogeneic (stem cells are separated from different patients).

[0202] In one embodiment, the CMV-associated disorder is selected from cytomegalovirus pneumonia, cytomegalovirus hepatitis, cytomegalovirus pancreatitis, cytomegalovirus mononucleosis, CMV polyradiculomyelopathy, cytomegalovirus inclusion disease, cytomegalovirus colitis, cytomegalovirus esophagitis, cytomegalovirus retinitis, Guillain-Barré syndrome, mucoepidermoid carcinoma and ulcerative colitis, graft-versus-host disease (GVHD), solid organ graft-versus-host disease (SOT-GVHD).

[0203] In one aspect, the present invention provides a fusion protein, nucleic acid, vector, host cell or pharmaceutical composition as described herein, for use in the manufacture of a medicament for preventing or treating an infection caused by a pathogen and / or a pathogen-associated disorder, such as for preventing or treating a CMV infection and / or a CMV-associated disorder.

[0204] In another aspect, the present invention provides an ex vivo use of the fusion protein or pharmaceutical composition as described herein for treating solid organs for transplantation and / or hematopoietic stem cells for transplantation.

[0205] As used herein, the terms "treating", "treatment" and "therapy" refer to curative, preventive, ameliorative and palliative therapy. Preferably, the treatment is curative.

[0206] Treatment In one aspect, the present invention provides a method for treating or preventing an infection caused by a pathogen, such as a CMV infection, in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of a fusion protein, nucleic acid, vector, recombinant host cell or pharmaceutical composition as described herein.

[0207] An "effective amount" or "therapeutically effective amount" of a compound is an amount of the compound sufficient to provide a beneficial effect to a subject to which the compound is administered. As used herein, the phrase "therapeutically effective amount" may refer to an amount of the fusion protein sufficient or effective to treat an infection (delay or prevent the onset of an infection, prevent the progression of an infection, inhibit, reduce or reverse an infection).

[0208] Generation method In one aspect, the present invention provides a method for producing a fusion protein as described herein, the method comprising culturing a host cell as defined herein under conditions allowing expression of the encoded fusion protein.

[0209] sequence

[0210] SEQ ID NO: 1 (Exemplary first peptide)

[0211] QHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLFCADPKEQWVKDAMQHLDRQAAALTRNG

[0212] SEQ ID NO: 2 (Exemplary second peptide)

[0213] GGSLAALTAHQACHLPLETFTRHRQRRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVG YHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPP

[0214] SEQ ID NO:3 (Domain A of Exotoxin A)

[0215] GGSLAALTAHQACHLPLETFTRHRQRRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAAS

[0216] SEQ ID NO:4 (Domain B of Exotoxin A)

[0217] GPADSGDALLERNYP

[0218] SEQ ID NO:5 (Domain C of Exotoxin A)

[0219] TGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPP

[0220] SEQ ID NO:6 (SYN002)

[0221] MQHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLACADPKEQWVKDAMQHLDRQAAALTRNGGGSLAALTAHQACHLPLETFTRHRQRRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPKDEL

[0222] SEQ ID NO:7(human CX3CL1)MAPISSLSWLLRLATFCHLTVLLAGQHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLFCADPKEQWVKDAMQHL DRQAAALTRNGGTFEKQIGEVKPRTTPAAGGMDESVVLEPEATGESSSLEPTPSSQEAQRALGTSPELPTGVTGSSGTRLPPTPKAQDGGPVGTELFRVPPVS TAATWQSSAPHQPGPSLWAEAKTSEAPSTQDPSTQASTASSPAPEENAPSEGQRVWGQGQSPRPENSLEREEMGPVPAHTDAFQDWGPGSMAHVSVVPVSSEGTPSREPVASGSWTPKAEEPIHATMDPQRLGVLITPVPDAQAATRRQAVGLLAFLGLLFCLGVAMFTYQSLQGCPRKMAGEMAEGLRYIPRSCGSNSYVLVPV

[0223] SEQ ID NO: 8 (sequence ensuring binding to the KDEL receptor for retrograde transport inside the cell) KDEL

[0224] SEQ ID NO:9 (full length exotoxin A)

[0225] MHLTPHWIPLVASLGLLAGGSFASAAEEAFDLWNECAKACVLDLKDGVRSSRMSVDPAIADTNGQGVLHYSMVLEGGNDALKLAIDNALSITSDGLTIRLEGGVEPNKPVRYSYTRQARGSWSLNWLVPIGHEKPSNIKVFIHELNAGNQLSHMSPIYTIEMGDELLAKLARDATFFVRAHESNEMQPTLAISHAGVSVVMAQAQPRREKRWSEWASGKVLCLLDPLDGVYNYLAQQRCNLDDTWEGKIYRVLAGNPAKHDLDIKPTVISHRLHFPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASADVVSLTCPVAAGECAGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPREDLK

[0226] SEQ ID NO:10(US28)

[0227] MTPTTTTAELTTEFDYDEAATPCVFTDVLNQSKPVTLFLYGVVFLFGSIGNFLVIFTITWRRRIQCSGDVYFINLAAADLLFVCTLPLWMQYLLDHNSLASVPCTLLTACFYVAMFASLCFITEIALDRYYAIVYMRYRPVKQACLFSIFWWIFAVIIAIPHFMVVTKKDNQCMTDYDYLEVSYPIILNVELMLGAFVIPLSVISYCYYRISRIVAVSQSRHKGRIVRVLIAVVLVFIIFWLPYHLTLFVDTLKLLKWISSSCEFERSLKRALILTESLAFCHCCLNPLLYVFVGTKFRQELHCLLAEFRQRLFSRDVSWYHSMSFSRRSSPSRRETSSDTLSDEVCRVSQIIP

[0228] SEQ ID NO:11(SYN000)

[0229] MQHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLFCADPKEQWVKDAMQHLDRQAAALTRNRQPRGWEQLEQSGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPKDEL

[0230] SEQ ID NO:12(SYN001)

[0231] MQHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLACADPKEQWVKDAMQHLDRQAAALTRNRQPRGWEQLEQSGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPKDE

[0232] SEQ ID NO:13(SYN014)

[0233] MQHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLFCADPKEQWVKDAMQHLDRQAAALTRNGGGSLAALTAHQACHLPLETFTRHRQRRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPKDEL

[0234] SEQ ID NO:14(SYN016)

[0235] MQHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLACADPKEQWVKDAMQHLDRQAAALTRNGGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPKDEL

[0236] SEQ ID NO:15(SYN017)

[0237] MQHHGVTKCNITCSKMTSKIPVALLIHYQQNQASCGKRAIILETRQHRLFCADPKEQWVKDAMQHLDRQAAALTRNGGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPKDEL

[0238] Item

[0239] 1. A fusion protein, comprising:

[0240] a) a first peptide that binds to at least one receptor expressed on a cell; and

[0241] b) a second peptide comprising a cleavage site having the amino acid sequence ArgX1X2Arg,

[0242] wherein X2 is Arg or Lys, and wherein the second peptide comprises a toxin.

[0243] 2. The fusion protein according to item 1, wherein the fusion protein is an immunotoxin.

[0244] 3. The fusion protein according to any one of the preceding items, wherein the cleavage site comprises or consists of the amino acid sequence ArgGlnArgArg.

[0245] 4. The fusion protein according to any one of the preceding items, wherein the first peptide binds to at least two different receptors expressed on cells.

[0246] 5. The fusion protein according to any one of the preceding items, wherein one receptor binding to the first peptide is a receptor encoded by a pathogen, and another receptor binding to the first peptide is a human-encoded receptor and / or an endogenous receptor of the first peptide or a variant thereof.

[0247] 6. The fusion protein according to any one of the preceding items, wherein the receptor is a G protein coupled receptor (GPCR), such as US28 of SEQ ID NO: 10.

[0248] 7. The fusion protein according to any one of the preceding items, wherein the second peptide comprises a toxin selected from Pseudomonas exotoxin A, gelonin, bouganin, saporin, ricin, ricin A chain, brentux, restrictocin, diphtheria toxin, diphtheria toxin A chain, and variants and fragments thereof.

[0249] 8. The fusion protein according to any one of the preceding items, wherein the receptor is internalized upon binding to the first peptide.

[0250] 9. The fusion protein according to any one of the preceding items, wherein the receptor is constitutively internalized.

[0251] 10. The fusion protein according to any one of the preceding items, wherein the first peptide comprises or consists of the following items

[0252] a. the amino acid sequence of SEQ ID NO: 1;

[0253] b. a variant of SEQ ID NO: 1, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 1, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 1, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 1;

[0254] c. A fragment of SEQ ID NO: 1 having a length of more than 50 amino acids, such as more than 60, 70 or 75 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 1, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 1, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 1.

[0255] 11. A fusion protein according to any one of the preceding items, wherein the second peptide comprises or consists of the following

[0256] a. the amino acid sequence of SEQ ID NO: 2;

[0257] b. a variant of SEQ ID NO: 2, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 2, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 2, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2;

[0258] c. A fragment of SEQ ID NO:2 having a length of more than 300 amino acids, such as more than 310, 330 or 340 amino acids in length, or a variant thereof having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:2, more preferably at least 85% or 90% sequence identity to SEQ ID NO:2, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2.

[0259] 12. The fusion protein according to any one of the preceding items, wherein the fusion protein comprises or consists of the following items

[0260] a. the amino acid sequence of SEQ ID NO: 6;

[0261] b. a variant of SEQ ID NO: 6, comprising or consisting of an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 6, more preferably at least 85% or 90% sequence identity to SEQ ID NO: 6, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 6;

[0262] c. A fragment of SEQ ID NO:6 having a length of more than 360 amino acids, such as more than 380, 400 or 420 amino acids in length, or a variant thereof having at least 80% sequence identity with the amino acid sequence of SEQ ID NO:6, more preferably at least 85% or 90% sequence identity with SEQ ID NO:6, and most preferably at least 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO:6.

[0263] 13. The fusion protein according to any one of the preceding items, wherein the fusion protein kills cells infected by a pathogen, such as cells latently infected by a pathogen.

[0264] 14. A fusion protein according to any of the preceding items, wherein the fusion protein has increased potency, such as at least 300-fold increased potency, such as at least 400-fold increased potency, such as at least 500-fold increased potency, such as at least 700-fold increased potency, against cells expressing a virally encoded receptor such as US28 compared to the potency against cells expressing an endogenous receptor or a human-encoded receptor such as CX3CR1.

[0265] 15. The fusion protein according to any one of items 1 to 14, for use in preventing or treating infections caused by pathogens and / or pathogen-related disorders.

[0266] 16. The fusion protein according to any one of items 1 to 14, for use in preventing or treating CMV infection and / or CMV-related disorders.

[0267] Example

[0268] Example 1: Mutations in the first peptide of the fusion protein SYNx lead to selectivity in cell killing.

[0269] Materials and Methods

[0270] Inducible US28 or CX3CR1-expressing HEK293 cells

[0271] For binding and cell killing assays, e.g. GM et al., 2013, described in constructing cells. In brief, stable and inducible clones of HA-CX3CR1 expressing cells and HA-US28 expressing cells are produced by co-transfection of Flp-In T-Rex-293 cells with Flp recombinase expression vector pOG44 and any one of pcDNA5 / FRT / TO receptor constructs. This targeted cloning at the FRT site puts the receptor gene under the control of the tetracycline repressor / operator gene system. The expression of the receptor is confirmed by Western blotting. For culturing cells, one or more cells are grown in a humidified incubator (37°C, 5% CO2) in 1x DMEM (with 9% (v / v) FBS, 180U / mL penicillin and 45μg / mL streptomycin, 13.5μg / mL blasticidin and 1.32mg / mL hygromycin B).

[0272] Cell killing assay

[0273] 80 μ L of 20 μ g / mL poly-D-lysine (PDL) in 1x PBS was added to each well of a white 96-well plate and incubated at room temperature for 30 min. Inducible US28 or CX3CR1 expression HEK293 cells were washed in 10 mL 1x PBS and released with 2 mL 0.05% (w / v) trypsin-EDTA for 2 min. Cells were resuspended in 10 mL growth medium (1xDMEM, 9% (v / v) FBS, 180 U / mL penicillin and 45 μ g / mL streptomycin) and then cell density was determined. PDL solution was aspirated from the plate and each well was washed with 100 μ L 1x PBS. After aspirating the wash buffer, 96-well plates were inoculated with 2,000 to 10,000 cells / well in 100 μ L and placed in a CO2 incubator (37 ° C, 10% CO2) overnight.

[0274] For receptor expression, 10 μL of tetracycline, 1.375 μg / mL US28, and 5.5 μg / mL CX3CR1 in growth medium without selection were added to each well of the plate and returned to the CO2 incubator (37°C, 10% CO2) overnight.

[0275] The fusion toxin protein aliquots were thawed on ice and used to prepare a dilution series in 1 mM acetic acid and 5 g / L BSA. The receptor expression growth medium was aspirated and replaced with 100 μL fresh growth medium. 5 μL of fusion toxin protein dilution series was added to each well, and 5 μL of 1 g / L cycloheximide was added to one well as a positive control. The plate was returned to a CO2 incubator (37°C, 10% CO2) overnight.

[0276] Preparation measures the 1:10 dilution of AlamarBlue in the growth medium. The solution is covered with tin foil and heated in a 37°C water bath for 10 min, then filtered through a 0.2 μm filter. Aspirate the well solution, and add 100 μL of AlamarBlue solution to each well, and place the plate back in a CO2 incubator (37°C, 10% CO2) for 4 hours. Use 540nm excitation and 585nm emission to collect fluorescence data on FlexStation 3.

[0277] result

[0278] The cell killing efficiency of SYNx was determined using tetracycline-inducible HEK293 cells expressing either a virally encoded receptor (e.g., US28) or an endogenous receptor (e.g., CX3CR1) as well as non-inducible cells without receptor expression (negative control). Data were normalized to the maximum number of viable cells. Substitution of a single amino acid in the first peptide of SYNx (here, the chemokine moiety) resulted in selectivity for the virally encoded receptor US28, e.g. Figure 2 Both constructs bind to US28 with similar selectivity ( Figure 2 A), while SYN001, which has a single mutation (F49A, corresponding to F73A using precursor numbering) in the first peptide (receptor binding portion) of the protein, is less selective for the endogenous receptor (here CXC3R1) ( Figure 2 B).

[0279] in conclusion

[0280] A single amino acid F49A substitution in the first peptide (eg, chemokine portion) of the fusion protein construct induces selectivity of the fusion protein (SYNx) for the virally encoded receptor US28.

[0281] Example 2: Increased potency is obtained by adding a furin cleavage site to the full-length translocation domain of exotoxin A.

[0282] Materials and Methods

[0283] See Example 1.

[0284] result

[0285] Tetracycline-inducible HEK 293 cells expressing virally encoded receptor US28 or endogenous receptor CX3CR1 and non-inducible cells without receptor expression (negative control) were used to determine the cell killing efficiency of SYNx. Data were normalized to the maximum number of viable cells. Adding a furin cleavage site (such as a full-length translocation domain) to the SYN001 construct produced SYN016, which increased the potency against cells expressing endogenous / human CX3CR1 receptors and pathogen-encoded US28 receptors, such as Figure 3 And as shown in Table 1.

[0286] in conclusion

[0287] Domain II of exotoxin A has been identified as the translocation domain responsible for transferring the catalytic domain, domain III, from the endocytic vesicle to the cytosol (Hwang J et al., 1987; Siegall CB et al., 1989). Adding the translocation domain, domain II of exotoxin A, containing a furin cleavage site, to SYN001 produced SYN016, thereby producing a fusion protein with increased potency for both the endogenous receptor CX3CR1 and the virally encoded receptor US28.

[0288] Table 1. IC50 values ​​of different fusion protein constructs

[0289]

[0290] Example 3: In vitro cleavage of SYNx by human furin.

[0291] Materials and Methods

[0292] In vitro cleavage of SYNx by human furin

[0293] Thaw the purified SYNx construct on the lab bench. Once thawed, the sample was then centrifuged and precipitated. Measure the concentration of the sample and digest 50 μL of 20 μM SYNx in 1x PBS containing 5 mM CaCl2 in a new tube. Remove 5 μg for SDS-PAGE analysis. Add 1 μL of human furin (NEB, 2 units / μL) to the mixture and place in a water bath at 37°C. After 1.5 hours, remove 5 μg of SYNx for SDS-PAGE analysis. Adjust the pH of the reaction mixture with 1M HCL or 1M NaOH before adding SYNx.

[0294] Reducing SDS-PAGE analysis

[0295] Samples were prepared and analyzed on NuPAGE Bis-Tris 4%-12% gels according to the manufacturer's protocol along with protein marker standards using 1xMES running buffer containing 2 mM DTT. The gel was run at a constant voltage of 125 V for 75 min. TM The gel was stained with SafeStain Manual (Novex).

[0296] result

[0297] Purified SYNx constructs were treated with purified human furin in vitro to determine the cleavage efficiency of furin on different SYNx constructs. Other cleavage mechanisms other than furin may occur. Results were analyzed by SDS-PAGE. Adding a complete translocation domain (such as in this case, a complete ExoA translocation domain) to SYN000 or SYN001 produced SYN017 and SYN016, respectively, which improved the in vitro cleavage of furin. Optimization of the furin cleavage site in SYN017 and SYN016 produced SYN014 and SYN002, respectively, which did not show further improvement in the in vitro cleavage of furin, as shown in Figure 2. Figure 4 shown.

[0298] Purified SYN002 was treated with purified human furin in vitro at different pH values ​​to determine the cleavage efficiency, and the results were analyzed by SDS-PAGE. SYN002 was cleaved by furin over a wide pH range, such as Figure 5 shown.

[0299] in conclusion

[0300] Compared to constructs with native sequences (ArgGlnProArg) as in SYN016, adding an optimized cleavage site (in this case, ArgGlnArgArg) to the SYNx construct (where the SYN002 construct also contains a translocation domain) did not result in an improvement in in vitro cleavage of furin. SYN002 can be cleaved in vitro over a wide pH range, suggesting that cleavage of furin can occur at any cellular location, not just in endosomes.

[0301] Example 4: Cell killing selectivity between endogenous receptor expressing cells and virally encoded receptor expressing cells is also enhanced and mediated by optimized cleavage sites.

[0302] Materials and Methods

[0303] See Example 1.

[0304] result

[0305] The cell killing efficiency of SYNx was determined using tetracycline-induced HEK 293 cells expressing US28 or CX3CR1 as well as non-induced cells without receptor expression (negative control). Data were normalized to the maximum number of viable cells.

[0306] When the optimized cleavage site ArgX1X2Arg was introduced into the second peptide, in this specific case, into SYN016 to give SYN002, a surprising increase in selectivity was obtained. The cell killing potency of SYN002 against cells expressing virally encoded receptors such as US28 remained approximately unchanged compared to SYN016 with the cleavage site ArgGlnProArg (thus, X2 = Pro) ( Figure 6 A, Table 1). However, the potency is reduced (about 0.9 log(EC50)) on cells expressing endogenous receptors such as CX3CR1 (see Figure 6 B), thus producing an effect that can support a more effective drug with fewer side effects. Both selectivity and potency were increased when compared to a second peptide that did not contain the optimized cleavage site.

[0307] in conclusion

[0308] Surprising effects on cell killing selectivity were obtained by optimizing the cleavage site. Without being bound by any theory, this may be due to differences in internalization and intracellular trafficking between virally encoded receptors (such as the US28 receptor) and endogenous receptors (such as the CX3CR1 receptor).

[0309] References

[0310] Hwang J et al.,Cell.1987Jan 16;48(1):129-36;

[0311] Siegall CB et al., J Biol Chem. 1989Aug 25;264(24):14256-61). Sequence Listing <110> Sinclair Noel <120> Fusion protein and its use <130> P5162EP00 <140> 19195122.7 <141> 2019-09-03 <160> twenty three <170> PatentIn Version 3.5 <210> 1 <211> 76 <212> PRT <213> Homo sapiens <400> 1 Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met Thr 1 5 10 15 Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln Ala 20 25 30 Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg Leu 35 40 45 Phe Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln His 50 55 60 Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Gly 65 70 75 <210> 2 <211> 340 <212> PRT <213> Pseudomonas aeruginosa <400> 2 Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro 1 5 10 15 Leu Glu Thr Phe Thr Arg His Arg Gln Arg Arg Gly Trp Glu Gln Leu 20 25 30 Glu Gln Cys Gly Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala 35 40 45 Ala Arg Leu Ser Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu 50 55 60 Ala Ser Pro Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln 65 70 75 80 Pro Glu Gln Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu 85 90 95 Arg Phe Val Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser 100 105 110 Gly Pro Ala Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr 115 120 125 Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg 130 135 140 Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln 145 150 155 160 Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu 165 170 175 Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln 180 185 190 Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala 195 200 205 Leu Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg 210 215 220 Ile Arg Asn Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu 225 230 235 240 Pro Gly Phe Tyr Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala 245 250 255 Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp 260 265 270 Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu 275 280 285 Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro 290 295 300 Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro 305 310 315 320 Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro 325 330 335 Gly Lys Pro Pro 340 <210> 3 <211> 112 <212> PRT <213> Pseudomonas aeruginosa <400> 3 Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro 1 5 10 15 Leu Glu Thr Phe Thr Arg His Arg Gln Arg Arg Gly Trp Glu Gln Leu 20 25 30 Glu Gln Cys Gly Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala 35 40 45 Ala Arg Leu Ser Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu 50 55 60 Ala Ser Pro Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln 65 70 75 80 Pro Glu Gln Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu 85 90 95 Arg Phe Val Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser 100 105 110 <210> 4 <211> 15 <212> PRT <213> Pseudomonas aeruginosa <400> 4 Gly Pro Ala Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro 1 5 10 15 <210> 5 <211> 213 <212> PRT <213> Pseudomonas aeruginosa <400> 5 Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr 1 5 10 15 Arg Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg 20 25 30 Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe 35 40 45 Leu Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser 50 55 60 Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro 65 70 75 80 Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly 85 90 95 Arg Ile Arg Asn Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser 100 105 110 Leu Pro Gly Phe Tyr Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala 115 120 125 Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu 130 135 140 Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile 145 150 155 160 Leu Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile 165 170 175 Pro Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile 180 185 190 Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln 195 200 205 Pro Gly Lys Pro Pro 210 <210> 6 <211> 421 <212> PRT <213> Artificial Sequence <220> <223> Fusion Protein SYN002 <400> 6 Met Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met 1 5 10 15 Thr Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln 20 25 30 Ala Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg 35 40 45 Leu Ala Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln 50 55 60 His Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Gly Gly Gly Ser 65 70 75 80 Leu Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Thr 85 90 95 Phe Thr Arg His Arg Gln Arg Arg Gly Trp Glu Gln Leu Glu Gln Cys 100 105 110 Gly Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu 115 120 125 Ser Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro 130 135 140 Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln 145 150 155 160 Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val 165 170 175 Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Gly Pro Ala 180 185 190 Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu 195 200 205 Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln 210 215 220 Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu 225 230 235 240 Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala 245 250 255 Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp 260 265 270 Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr 275 280 285 Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn 290 295 300 Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe 305 310 315 320 Tyr Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val 325 330 335 Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr 340 345 350 Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro 355 360 365 Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro 370 375 380 Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu 385 390 395 400 Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro 405 410 415 Pro Lys Asp Glu Leu 420 <210> 7 <211> 397 <212> PRT <213> Homo sapiens <400> 7 Met Ala Pro Ile Ser Leu Ser Trp Leu Leu Arg Leu Ala Thr Phe Cys 1 5 10 15 His Leu Thr Val Leu Leu Ala Gly Gln His His Gly Val Thr Lys Cys 20 25 30 Asn Ile Thr Cys Ser Lys Met Thr Ser Lys Ile Pro Val Ala Leu Leu 35 40 45 Ile His Tyr Gln Gln Asn Gln Ala Ser Cys Gly Lys Arg Ala Ile Ile 50 55 60 Leu Glu Thr Arg Gln His Arg Leu Phe Cys Ala Asp Pro Lys Glu Gln 65 70 75 80 Trp Val Lys Asp Ala Met Gln His Leu Asp Arg Gln Ala Ala Ala Leu 85 90 95 Thr Arg Asn Gly Gly Thr Phe Glu Lys Gln Ile Gly Glu Val Lys Pro 100 105 110 Arg Thr Thr Pro Ala Ala Gly Gly Met Asp Glu Ser Val Val Leu Glu 115 120 125 Pro Glu Ala Thr Gly Glu Ser Ser Ser Leu Glu Pro Thr Pro Ser Ser 130 135 140 Gln Glu Ala Gln Arg Ala Leu Gly Thr Ser Pro Glu Leu Pro Thr Gly 145 150 155 160 Val Thr Gly Ser Ser Gly Thr Arg Leu Pro Pro Thr Pro Lys Ala Gln 165 170 175 Asp Gly Gly Pro Val Gly Thr Glu Leu Phe Arg Val Pro Pro Val Ser 180 185 190 Thr Ala Ala Thr Trp Gln Ser Ser Ala Pro His Gln Pro Gly Pro Ser 195 200 205 Leu Trp Ala Glu Ala Lys Thr Ser Glu Ala Pro Ser Thr Gln Asp Pro 210 215 220 Ser Thr Gln Ala Ser Thr Ala Ser Ser Pro Ala Pro Glu Glu Asn Ala 225 230 235 240 Pro Ser Glu Gly Gln Arg Val Trp Gly Gln Gly Gln Ser Pro Arg Pro 245 250 255 Glu Asn Ser Leu Glu Arg Glu Glu Met Gly Pro Val Pro Ala His Thr 260 265 270 Asp Ala Phe Gln Asp Trp Gly Pro Gly Ser Met Ala His Val Ser Val 275 280 285 Val Pro Val Ser Ser Glu Gly Thr Pro Ser Arg Glu Pro Val Ala Ser 290 295 300 Gly Ser Trp Thr Pro Lys Ala Glu Glu Pro Ile His Ala Thr Met Asp 305 310 315 320 Pro Gln Arg Leu Gly Val Leu Ile Thr Pro Val Pro Asp Ala Gln Ala 325 330 335 Ala Thr Arg Arg Gln Ala Val Gly Leu Leu Ala Phe Leu Gly Leu Leu 340 345 350 Phe Cys Leu Gly Val Ala Met Phe Thr Tyr Gln Ser Leu Gln Gly Cys 355 360 365 Pro Arg Lys Met Ala Gly Glu Met Ala Glu Gly Leu Arg Tyr Ile Pro 370 375 380 Arg Ser Cys Gly Ser Asn Ser Tyr Val Leu Val Pro Val 385 390 395 <210> 8 <211> 4 <212> PRT <213> Homo sapiens <400> 8 Lys Asp Glu Leu 1 <210> 9 <211> 638 <212> PRT <213> Pseudomonas aeruginosa <400> 9 Met His Leu Thr Pro His Trp Ile Pro Leu Val Ala Ser Leu Gly Leu 1 5 10 15 Leu Ala Gly Gly Ser Phe Ala Ser Ala Ala Glu Glu Ala Phe Asp Leu 20 25 30 Trp Asn Glu Cys Ala Lys Ala Cys Val Leu Asp Leu Lys Asp Gly Val 35 40 45 Arg Ser Ser Arg Met Ser Val Asp Pro Ala Ile Ala Asp Thr Asn Gly 50 55 60 Gln Gly Val Leu His Tyr Ser Met Val Leu Glu Gly Gly Asn Asp Ala 65 70 75 80 Leu Lys Leu Ala Ile Asp Asn Ala Leu Ser Ile Thr Ser Asp Gly Leu 85 90 95 Thr Ile Arg Leu Glu Gly Gly Val Glu Pro Asn Lys Pro Val Arg Tyr 100 105 110 Ser Tyr Thr Arg Gln Ala Arg Gly Ser Trp Ser Leu Asn Trp Leu Val 115 120 125 Pro Ile Gly His Glu Lys Pro Ser Asn Ile Lys Val Phe Ile His Glu 130 135 140 Leu Asn Ala Gly Asn Gln Leu Ser His Met Ser Pro Ile Tyr Thr Ile 145 150 155 160 Glu Met Gly Asp Glu Leu Leu Ala Lys Leu Ala Arg Asp Ala Thr Phe 165 170 175 Phe Val Arg Ala His Glu Ser Asn Glu Met Gln Pro Thr Leu Ala Ile 180 185 190 Ser His Ala Gly Val Ser Val Val Met Ala Gln Ala Gln Pro Arg Arg 195 200 205 Glu Lys Arg Trp Ser Glu Trp Ala Ser Gly Lys Val Leu Cys Leu Leu 210 215 220 Asp Pro Leu Asp Gly Val Tyr Asn Tyr Leu Ala Gln Gln Arg Cys Asn 225 230 235 240 Leu Asp Asp Thr Trp Glu Gly Lys Ile Tyr Arg Val Leu Ala Gly Asn 245 250 255 Pro Ala Lys His Asp Leu Asp Ile Lys Pro Thr Val Ile Ser His Arg 260 265 270 Leu His Phe Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln 275 280 285 Ala Cys His Leu Pro Leu Glu Thr Phe Thr Arg His Arg Gln Pro Arg 290 295 300 Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr Pro Val Gln Arg Leu Val 305 310 315 320 Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp Asn Gln Val Asp Gln Val 325 330 335 Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser Gly Gly Asp Leu Gly Glu 340 345 350 Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg Leu Ala Leu Thr Leu Ala 355 360 365 Ala Ala Glu Ser Glu Arg Phe Val Arg Gln Gly Thr Gly Asn Asp Glu 370 375 380 Ala Gly Ala Ala Ser Ala Asp Val Val Ser Leu Thr Cys Pro Val Ala 385 390 395 400 Ala Gly Glu Cys Ala Gly Pro Ala Asp Ser Gly Asp Ala Leu Leu Glu 405 410 415 Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile 420 425 430 Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu 435 440 445 Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr 450 455 460 His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val 465 470 475 480 Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile 485 490 495 Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro 500 505 510 Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg Val Tyr Val 515 520 525 Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg Thr Gly Leu Thr Leu Ala 530 535 540 Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu 545 550 555 560 Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg 565 570 575 Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile 580 585 590 Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp 595 600 605 Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp 610 615 620 Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg Glu Asp Leu Lys 625 630 635 <210> 10 <211> 354 <212> PRT <213> Cell virus <400> 10 Met Thr Pro Thr Thr Thr Thr Ala Glu Leu Thr Thr Glu Phe Asp Tyr 1 5 10 15 Asp Glu Ala Ala Thr Pro Cys Val Phe Thr Asp Val Leu Asn Gln Ser 20 25 30 Lys Pro Val Thr Leu Phe Leu Tyr Gly Val Val Phe Leu Phe Gly Ser 35 40 45 Ile Gly Asn Phe Leu Val Ile Phe Thr Ile Thr Trp Arg Arg Arg Ile 50 55 60 Gln Cys Ser Gly Asp Val Tyr Phe Ile Asn Leu Ala Ala Ala Asp Leu 65 70 75 80 Leu Phe Val Cys Thr Leu Pro Leu Trp Met Gln Tyr Leu Leu Asp His 85 90 95 Asn Ser Leu Ala Ser Val Pro Cys Thr Leu Leu Thr Ala Cys Phe Tyr 100 105 110 Val Ala Met Phe Ala Ser Leu Cys Phe Ile Thr Glu Ile Ala Leu Asp 115 120 125 Arg Tyr Tyr Ala Ile Val Tyr Met Arg Tyr Arg Pro Val Lys Gln Ala 130 135 140 Cys Leu Phe Ser There Is There Trp Trp There Is No Val There Is There Is There 145 150 155 160 Pro His Phe Met Val Val Thr Lys Lys Asp Asn Gln Cys Met Thr Asp 165 170 175 Tyr Asp Tyr Leo Glu Will Ser Tyr Pro Ile Leo Asn Val Leo Glu 180 185 190 Met Leu Gly Ala Phe Val Ile Pro Leu Ser Val Ile Ser Tyr Cys Tyr 195 200 205 Tyr Arg With Arg Is Not Gln Served With Arg His Lys Gly 210 215 220 Arg Ile Val Arg Val Leu Ile Ala Val Val Leu Val Phe Ile Phe 225 230 235 240 Trp Leu Pro Tyr His Leu Thr Leu Phe Val Asp Thr Leu Lys Leu Leu 245 250 255 Lys Trp Ile Ser Ser Cys Glu Phe Glu Arg Ser Leu Lys Arg Ala 260 265 270 Leu Ile Leu Thr Glu Ser Leu Ala Phe Cys His Cys Cys Leu Asn Pro 275 280 285 Leu Leu Tyr Val Phe Val Gly Thr Lys Phe Arg Gln Glu Leu His Cys 290 295 300 Leu Leu Ala Glu Phe Arg Gln Arg Leu Phe Ser Arg Asp Val Ser Trp 305 310 315 320 Tyr His Ser Met Ser Phe Ser Arg Arg Ser Ser Pro Ser Arg Arg Glu 325 330 335 Thr Ser Ser Asp Thr Leu Ser Asp Glu Val Cys Arg Val Ser Gln Ile 340 345 350 Ile Pro <210> 11 <211> 397 <212> PRT <213> Artificial Sequence <220> <223> Fusion Protein SYN000 <400> 11 Met Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met 1 5 10 15 Thr Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln 20 25 30 Ala Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg 35 40 45 Leu Phe Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln 50 55 60 His Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Arg Gln Pro Arg 65 70 75 80 Gly Trp Glu Gln Leu Glu Gln Ser Gly Tyr Pro Val Gln Arg Leu Val 85 90 95 Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp Asn Gln Val Asp Gln Val 100 105 110 Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser Gly Gly Asp Leu Gly Glu 115 120 125 Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg Leu Ala Leu Thr Leu Ala 130 135 140 Ala Ala Glu Ser Glu Arg Phe Val Arg Gln Gly Thr Gly Asn Asp Glu 145 150 155 160 Ala Gly Ala Ala Ser Gly Pro Ala Asp Ser Gly Asp Ala Leu Leu Glu 165 170 175 Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile 180 185 190 Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu 195 200 205 Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr 210 215 220 His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val 225 230 235 240 Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile 245 250 255 Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro 260 265 270 Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg Val Tyr Val 275 280 285 Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg Thr Gly Leu Thr Leu Ala 290 295 300 Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu 305 310 315 320 Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg 325 330 335 Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile 340 345 350 Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp 355 360 365 Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp 370 375 380 Tyr Ala Ser Gln Pro Gly Lys Pro Pro Lys Asp Glu Leu 385 390 395 <210> 12 <211> 396 <212> PRT <213> Artificial sequence <220> <223> Fusion protein SYN001 <400> 12 Met Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met 1 5 10 15 Thr Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln 20 25 30 Ala Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg 35 40 45 Leu Ala Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln 50 55 60 His Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Arg Gln Pro Arg 65 70 75 80 Gly Trp Glu Gln Leu Glu Gln Ser Gly Tyr Pro Val Gln Arg Leu Val 85 90 95 Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp Asn Gln Val Asp Gln Val 100 105 110 Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser Gly Gly Asp Leu Gly Glu 115 120 125 Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg Leu Ala Leu Thr Leu Ala 130 135 140 Ala Ala Glu Ser Glu Arg Phe Val Arg Gln Gly Thr Gly Asn Asp Glu 145 150 155 160 Ala Gly Ala Ala Ser Gly Pro Ala Asp Ser Gly Asp Ala Leu Leu Glu 165 170 175 Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile 180 185 190 Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu 195 200 205 Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr 210 215 220 His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val 225 230 235 240 Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile 245 250 255 Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro 260 265 270 Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg Val Tyr Val 275 280 285 Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg Thr Gly Leu Thr Leu Ala 290 295 300 Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu 305 310 315 320 Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg 325 330 335 Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile 340 345 350 Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp 355 360 365 Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp 370 375 380 Tyr Ala Ser Gln Pro Gly Lys Pro Pro Lys Asp Glu 385 390 395 <210> 13 <211> 421 <212> PRT <213> Artificial Sequence <220> <223> Fusion protein SYN014 <400> 13 Met Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met 1 5 10 15 Thr Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln 20 25 30 Ala Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg 35 40 45 Leu Phe Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln 50 55 60 His Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Gly Gly Gly Ser 65 70 75 80 Leu Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Thr 85 90 95 Phe Thr Arg His Arg Gln Arg Arg Gly Trp Glu Gln Leu Glu Gln Cys 100 105 110 Gly Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu 115 120 125 Ser Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro 130 135 140 Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln 145 150 155 160 Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val 165 170 175 Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Gly Pro Ala 180 185 190 Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu 195 200 205 Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln 210 215 220 Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu 225 230 235 240 Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala 245 250 255 Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp 260 265 270 Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr 275 280 285 Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn 290 295 300 Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe 305 310 315 320 Tyr Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val 325 330 335 Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr 340 345 350 Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro 355 360 365 Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro 370 375 380 Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu 385 390 395 400 Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro 405 410 415 Pro Lys Asp Glu Leu 420 <210> 14 <211> 421 <212> PRT <213> Artificial Sequence <220> <223> Fusion Protein SYN016 <400> 14 Met Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met 1 5 10 15 Thr Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln 20 25 30 Ala Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg 35 40 45 Leu Ala Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln 50 55 60 His Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Gly Gly Gly Ser 65 70 75 80 Leu Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Thr 85 90 95 Phe Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys 100 105 110 Gly Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu 115 120 125 Ser Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro 130 135 140 Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln 145 150 155 160 Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val 165 170 175 Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Gly Pro Ala 180 185 190 Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu 195 200 205 Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln 210 215 220 Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu 225 230 235 240 Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala 245 250 255 Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp 260 265 270 Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr 275 280 285 Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn 290 295 300 Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe 305 310 315 320 Tyr Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val 325 330 335 Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr 340 345 350 Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro 355 360 365 Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro 370 375 380 Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu 385 390 395 400 Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro 405 410 415 Pro Lys Asp Glu Leu 420 <210> 15 <211> 421 <212> PRT <213> Artificial Sequence <220> <223> Fusion Protein SYN017 <400> 15 Met Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met 1 5 10 15 Thr Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln 20 25 30 Ala Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg 35 40 45 Leu Phe Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln 50 55 60 His Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Gly Gly Gly Ser 65 70 75 80 Leu Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Thr 85 90 95 Phe Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys 100 105 110 Gly Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu 115 120 125 Ser Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro 130 135 140 Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln 145 150 155 160 Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val 165 170 175 Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Ser Gly Pro Ala 180 185 190 Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu 195 200 205 Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln 210 215 220 Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu 225 230 235 240 Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala 245 250 255 Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp 260 265 270 Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr 275 280 285 Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn 290 295 300 Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe 305 310 315 320 Tyr Arg Thr Gly Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val 325 330 335 Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr 340 345 350 Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro 355 360 365 Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro 370 375 380 Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu 385 390 395 400 Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro 405 410 415 Pro Lys Asp Glu Leu 420 <210> 16 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> 16 Arg Gln Arg Arg 1 <210> 17 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> 17 Arg Gln Lys Arg 1 <210> 18 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> 18 Arg Ser Lys Arg 1 <210> 19 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> 19 Arg Ser Arg Arg 1 <210> 20 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> 20 Arg Thr Lys Arg 1 <210> twenty one <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> twenty one Arg Thr Arg Arg 1 <210> twenty two <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> twenty two Arg Asn Lys Arg 1 <210> twenty three <211> 4 <212> PRT <213> Artificial sequence <220> <223> Cleavage site <220> <221> Peptides <222> (1)..(4) <223> Cleavage site <400> twenty three Arg Asn Arg Arg 1

Claims

1. A fusion protein consisting of the amino acid sequence of SEQ ID NO:

6.

2. An isolated nucleic acid encoding the fusion protein according to claim 1. A vector comprising the nucleic acid according to claim 2. The vector according to claim 3 , wherein the vector is an expression vector.

5. A recombinant host cell comprising the nucleic acid molecule according to claim 2 or the vector according to any one of claims 3-4. The recombinant host cell according to claim 5 , wherein the host cell is a bacterial cell, a yeast cell, a mammalian cell or an insect cell.

7. The recombinant host cell of claim 5, wherein the host cell is a human cell.

8. A pharmaceutical composition comprising the fusion protein according to claim 1, the nucleic acid according to claim 2, the vector according to any one of claims 3-4 or the recombinant host cell according to any one of claims 5-7 and a pharmaceutically acceptable carrier and / or excipient.

9. A pharmaceutical composition comprising the fusion protein according to claim 1, the nucleic acid according to claim 2, the vector according to any one of claims 3-4 or the recombinant host cell according to any one of claims 5-7, and a pharmaceutically acceptable diluent.

10. The pharmaceutical composition according to claim 8 or 9, further comprising one or more additional pharmaceutical agents.

11. The pharmaceutical composition according to claim 10, wherein the agent is selected from immunosuppressants and antiviral agents.

12. The pharmaceutical composition according to claim 11, wherein the antiviral agent is selected from valganciclovir, ganciclovir, cidofovir, leflunomide, letermovir, maribavir and brincidofovir.

13. Use of the fusion protein according to claim 1, the nucleic acid according to claim 2, the vector according to claim 3, the recombinant host cell according to claim 5 or the pharmaceutical composition according to claim 8 or 9 in the preparation of a medicament for treating CMV infection in an individual.

14. Use according to claim 13, wherein the CMV infection is a latent and / or lytic CMV infection.

15. The use according to claim 13, wherein CMV infection can be detected in: i) one or more tissues selected from the group consisting of retina, cornea, heart, liver, kidney, lung, gastrointestinal tissue, thymus, spleen, skin and muscle, and / or ii) a body fluid selected from one or more of saliva, blood, urine, semen and breast milk.

16. The use according to claim 13, wherein the CMV infection is an infection in an immunocompromised patient selected from HIV patients, neonates and immunosuppressed patients, bone marrow transplant patients, solid organ transplant patients, immunotherapy patients, cancer patients, intensive care patients, trauma patients, stem cell patients, gene therapy patients, cell therapy patients, elderly patients and patients with multiple diseases.

17. The use according to claim 13, wherein the fusion protein, the nucleic acid, the vector, the host cell or the composition is administered intravenously, intratumorally, intraperitoneally, intrathecally and / or intralymphatically.

18. The use according to claim 13, wherein the individual is a human.

19. The use according to claim 18, wherein the human is an immunocompromised patient.

20. The use according to claim 18, wherein the human is a child.

21. The use according to claim 18, wherein the human is an adult.

22. The use according to claim 18, wherein the human is in need of a solid organ transplant and / or a hematopoietic stem cell transplant.

23. A method for producing the fusion protein of claim 1, comprising culturing the host cell of claim 5 under conditions that allow expression of the encoded fusion protein.

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