Vaccine comprising antibody or Fc-containing fusion protein comprising antibody-containing Fc portion

Abstract

Described is a vaccine for use in active immunization of a subject against an infectious or malignant disease, the vaccine comprising: (a) an antibody against an antigen associated with the infectious or malignant disease; or (b) an Fc-containing fusion protein comprising an Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC) selected from the group consisting of Type I: activating Fc [gamma] RI, Fc [gamma] RIIa, Fc [gamma] RIIc, Fc [gamma] RIIIa, Fc [gamma] RIIIb, and inhibitory Fc [gamma] RIIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21. In addition, a vaccine for use in active immunization of a subject against an HSV-related disease is described; and (a) wherein the antibody is an anti-HSV antibody and the subject is suffering from an acute HSV-related disease, preferably an acute HSV infection; or (b) wherein in the Fc-containing fusion protein comprising an Fc portion of an antibody fused to an antigen, the antigen is associated with a HSV-related disease.

Description

[0001] The present invention relates to a vaccine for use in active immunization of a subject against an infectious disease or a malignant disease, the vaccine comprising: (a) an antibody against an antigen associated with the infectious disease or the malignant disease; or (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or the malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21. Furthermore, the present invention relates to a vaccine for use in active immunization of a subject against an HSV-related disease; and (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection; or (b) wherein in the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen, the antigen is associated with an HSV-related disease.

[0002] Vaccines are biological agents used to induce an immune response to protect vaccinated individuals from infection or disease development (which is often induced by pathogenic microbial agents or cancer cells). In order to induce an immune response, the vaccine needs to include a recognition marker corresponding to the target (e.g., a viral protein) against which the immune response is intended to be induced. Active immunization methods are divided into live vaccines and non-live vaccines. Live vaccines are usually attenuated pathogens, while non-live vaccines contain or encode components of the pathogen or the entire pathogen that has been inactivated [1]. These components can also be modified to tune the immune response, for example by expressing them as fusion proteins with effector domains.

[0003] Live vaccines have been in use since the discovery of vaccination and have since been developed to protect against tuberculosis (BCG), typhoid, polio, measles, mumps and rubella (MMR combination vaccine), rotavirus, smallpox, varicella zoster, influenza, Japanese encephalitis and yellow fever virus infection. The principle of attenuating pathogenicity and viral replication fitness is to allow the immune response to be established before the vaccine strain reverts to a pathogenic wild-type strain, however, for example, the case of evolution of wild-type poliovirus after vaccination with the live attenuated poliovirus Sabin vaccine strain is well known and documented.

[0004] The group of non-live vaccines is growing rapidly as newer technologies become available. Approved vaccines using whole-killed pathogens include whole-cell pertussis, poliovirus, influenza, Japanese encephalitis, hepatitis A, and rabies. Subunit vaccines with purified proteins, recombinant native proteins, or artificial fusion constructs, polysaccharides, or peptides derived from pathogens have been developed for pertussis, influenza, hepatitis B, meningococci, pneumococci, typhoid, SARS-CoV-2, and hepatitis A. Protein-polypeptide conjugates have been developed for Haemophilus influenzae type B, pneumococci, meningococci, and typhoid. Virus-like particle vaccines have been developed for Ebola, hepatitis B, malaria, and human papillomavirus, in which pathogen structures are presented on the surface of non-infectious particles derived from the virus. Similarly, polymers coated with epitopes have been shown to elicit neutralizing antibodies, for example for Plasmodium falciparum sporozoites [2].

[0005] Viral vectors have been used to encode and deliver pathogenic information for antigens used for vaccination. Examples of vaccination based on adenoviral vectors are Ebola virus (ChAd3-EBO-Z) and SARS-CoV-2 (e.g., Ad5-nCoV, Ad26.COV2-S, Sputnik V, ChAdOX1-nCoV). In addition, recombinant vesicular stomatitis virus (rVSV), poxvirus (modified Ankara vaccinia), adeno-associated virus (AAV), retroviruses, cytomegalovirus, and Sendai virus have been developed as viral vectors for vaccination. Based on methods for delivering antigens not directly but encoded in nucleic acids for injection in carriers, mRNA and DNA have been used as vaccine platforms [3]. The SARS-CoV-2 pandemic led to the approval of the first mRNA vaccine encoding a modified viral spike protein. The mRNA is delivered to the cytoplasm via lipid particles, and the cellular machinery translates this message into viral proteins. RNA vaccine approaches have also been experimentally tested for other infectious diseases, such as Ebola virus, and in cancer therapy, through the expression of cancer-associated neoantigens. In addition, the first DNA vaccine has been developed for SARS-CoV-2 (ZyCoV-D) and the related coronavirus disease (COVID), and has been approved for emergency use in India [4]. DNA vaccines have several advantages, including easy, rapid, and inexpensive production, stability, and the potential for rapid adaptation, and have been experimentally developed for several pathogens, including Plasmodium and Mycoplasma, as well as influenza virus, hepatitis B virus, human immunodeficiency virus, rabies virus, and lymphocytic choriomeningitis virus.

[0006] The goal of vaccines is to induce long-lasting humoral and cellular immune responses, which are conferred by memory B and T cells that can be easily reactivated. When the B cell receptor of a B cell interacts with Ag, the B cell is activated to secrete antibodies. In humans, the five major antibody isotypes are classified as IgM, IgG, IgA, IgD, and IgE based on their unique Fc fragments. Each class interacts with a specific Fc receptor (FcR). IgG is the most abundant antibody in the blood and is further classified into four subtypes, namely IgG1, IgG2, IgG3, and IgG4; while IgA consists of two subtypes, IgA1 and IgA2. After activation, B cells can undergo class switch recombination, and the expression of antibody isotypes can change from IgM / IgD to IgA, IgE, or IgG, or they can maintain IgM expression. Ag titer indicates the magnitude and composition of the B cell response, whereby high titer enables strong activation and effector differentiation as well as long-lasting B cell memory responses [5]. B cells that have bound to Ag can be activated in the germinal center in a T-cell-independent or T-cell-dependent manner to become plasma cells or memory B cells. Plasma cells migrate to the bone marrow and can reside there for a long time, while memory B cells circulate in the body and can be quickly reactivated when encountering Ag again.

[0007] To induce strong T cell responses, effector differentiation and T memory cells, Ag peptides or proteins alone are generally not very effective as vaccines because these are inefficiently taken up by antigen presenting cells (APCs). The efficiency of Ag peptide or protein uptake by APCs can be greatly increased in the following ways: either by fusing a moiety that targets the APC, such as by expressing the Ag fused to the crystallizable fragment (Fc) of an immunoglobulin (Ig); or when the Ag is coated with antibodies to form an immune complex (IC). For example, when monocyte-derived dendritic cells are pulsed with polyclonal tetanus IgG ICs, stronger activation can be achieved compared to “naked” tetanus toxin Ag [6]. Subsequent proteolytic cleavage via the lysosomal pathway (e.g., in the case of phagocytosed Ag) or the cytoplasmic proteasomal pathway (e.g., pathogen proteins produced in the cytosol) generates Ag-derived peptides that are loaded onto class I or class II major histocompatibility complexes (MHC), with lysosomal-derived peptides typically loaded onto MHC-II and cytoplasmic proteasomal-derived peptides loaded onto MHC-I. However, antigenic peptides generated in lysosomes can also be transferred to the cytoplasm and loaded onto MHC-I complexes, a process known as cross-presentation and most commonly seen in dendritic cells [7]. Peptides presented by MHC-I or MHC-II can stimulate CD8+ or CD4+ T-cell responses, respectively. In addition to peptides presented by MHC, secondary activation signals are required for effective T-cell activation, including expression of T cell CD28 binding molecules (e.g., CD80, CD86) on APCs and secretion of cytokines by APCs [7].

[0008] Fc-fusion proteins are pathogen-derived Ags fused to the Fc of Ig. The goal of this strategy is to direct the antigenic portion to be taken up into the APC via interaction with the Fc receptor (FcR) on the APC. Several Fc-fusion proteins have been experimentally tested for vaccination, and 13 Fc fusion proteins have been approved by the European Union and / or the United States by 2020 [8]. Recombinant vaccines with spike receptor binding domains (RBDs) fused to the Fc from IgG have been developed as vaccines against SARS-CoV-2, and are being tested in Phase I / II human clinical trials [9]. Similarly, oligomeric influenza hemagglutinin HA-Fc fusion proteins induce epitope-specific neutralizing antibody responses in mice

[10] . IgG2a Fc fused to the Epstein-Barr virus (EBV) protein gp350 induces a potent neutralizing immune response in Balb / c mice

[11] . Fusion proteins of Ebola virus glycoproteins with Fc induced robust humoral and cellular responses, including robust CD8+ T cell responses, and protected mice from lethal Ebola virus infection

[12] . Fusion proteins consisting of HSV-2 gD protein fused to an IgG1 Fc fragment induced potent mucosal and systemic B and T cell immunity, including memory responses that were stable for at least 6 months

[13] . All of these experimental approaches demonstrate the suitability of recombinant Fc-fusion proteins for immunization against a variety of viral pathogens.

[0009] Similar experimental approaches have shown the efficacy of Fc-fusion proteins as potential anti-tumor vaccines. For example, the Xcl1-OVA-Fc fusion construct can be efficiently taken up by dendritic cells and stimulates a potent H-2kb / OVA-specific T-cell response that can control tumor progression in mice. This vaccine increases Ag-specific tumor-infiltrating CD8+ T-cells with enhanced IFNg production and degranulation potential

[14] .

[0010] Similarly, approaches have been attempted to use Fc fusion proteins as a vaccination strategy against some bacterial infections. For example, IgG2a Fc-PspA fusion protein polarizes alveolar macrophages (AMs) toward an activated AM1 phenotype, induces Th1 cell polarization, and activates conventional dendritic cells, suggesting potential as a good vaccine candidate against Streptococcus pneumoniae

[15] . Dectin1-Fc (IgG2a), Dectin1-Fc (IgG2b), and wheat germ agglutinin (WGA)-Fc (IgG2a) fusion proteins have been examined for their potential to induce immune responses against several fungal pathogens in order to induce immune responses against common fungal antigens (e.g., chitin and β-glucan), and were able to demonstrate antifungal activity against Histoplasma capsulatum, Cryptococcus neoformans, Candida albicans, and Aspergillus fumigatus

[16] . Finally, Fc fusion proteins have been tested as vaccine strategies against protozoan pathogens (e.g., Plasmodium falciparum, which causes malaria)

[17] .

[0011] Immune complexes (ICs) are generated during the natural humoral immune response when endogenously produced, usually polyclonal, antibodies bind in a multivalent manner to microbial target antigens, which are usually present in multiple copies on the exterior of an invading pathogen, thereby forming large antibody-Ag complexes. Antibodies opsonize microbial factors by binding to exposed epitopes in the target antigen with their Fab regions, which can also result in allosteric or structural Fab-mediated changes in the target antigen and thereby can lead to the exposure of new B-cell epitopes. The potentiating effect of ICs on B-cell responses has been attributed to the function of follicular dendritic cells (FDCs), which can capture and retain intact ICs to act as long-lasting Ag reservoirs for B-cell stimulation [18, 19]. FDCs can be found in primary follicles and germinal centers of secondary and tertiary lymphoid organs (e.g., spleen, lymph nodes, and gut-associated lymphoid tissue), where they can encounter Ag-specific B cells and assist in B cell affinity maturation through somatic hypermutation. These B cells can phagocytose ICs presented by FDCs and can then stimulate CD4+ T-cells through MHC-II presentation of IC-derived Ag peptides, a feedback process that promotes B-cell proliferation and eventual antibody secretion.

[0012] Another key feature of IC is the multivalent binding (opsonization) of many antibody molecules to coat pathogens or tumor cells with Fc domains, which increases effector function through avid binding of Fc to FcRs on APCs. Interaction of IC with FcRs on APCs can lead to uptake of IC via phagocytosis and activation of APCs. The enhancing effects of IC on antigen presentation and T-cell stimulation have long been known

[20] . In addition, under certain circumstances (including antibody excess and antibody polyclonality), IC can also inhibit T-cell activation

[21] . When comparing IC vaccines against Mycobacterium tuberculosis in Ag or antibody excess, animals immunized with IC in Ag excess survived lethal infection, while animals in the antibody excess IC group showed some mortality

[22] .

[0013] The immune complex (IC) vaccination strategy aims to immunize against Ig-opsonized Ag by resembling naturally occurring complexes of target Ag bound to antibodies. Thus, multivalent binding of IgG to its target within the IC is important for inducing a strong and durable humoral immune response. The IC vaccination approach has been experimentally tested against a variety of pathogens. Yeast-derived hepatitis B virus (HBV) surface antigen (HBsAg) was used together with antibodies from hyperimmunized donors to generate an IC vaccine against HBV. In a Phase I clinical study with healthy adults, serum anti-HBV antibodies could be generated and interferon gamma (IFNg) and interleukin 2 (IL-2) levels could be effectively increased without any safety issues

[23] . A subsequent Phase IIa study with HBeAg-positive chronic hepatitis B patients demonstrated a reduction in viral load in half of the treated patients and T-cell cytokine responses in some treated patients, indicating the therapeutic effect of the vaccine. In addition, PBMC-derived dendritic cells from these patients showed activation and maturation upon ex vivo stimulation with the IC vaccine

[24] . Interestingly, the Phase IIb trial did not show statistically different effects between the vaccinated and placebo groups at relevant primary and secondary endpoints

[25] , and the subsequent Phase III aimed to achieve better results by increasing the number of injections of the IC vaccine from six (Phase IIb) to twelve (Phase III). Surprisingly, even worse outcomes were noted compared to Phase IIb, suggesting that simply increasing the dose of the IC vaccine does not necessarily enhance the immune response

[26] . When polyclonal or broadly neutralizing antibody therapy was used, stimulation of de novo neutralizing antibodies and protective CD8+ T cell responses was observed in different SIV / SHIV animal models, and it was proposed that this was the result of the following process: the formation of ICs established by the infused antibodies binding to circulating virions or Ags, and the uptake of these ICs by APCs, followed by MHC-I cross-presentation [27-30]. Vaccines (IC vaccines) have been approved for the protection of poultry from infectious bursal disease virus (IBDV). In addition, IC vaccine strategies have been developed for other veterinary viral diseases induced by equine herpesvirus 1 and porcine parvovirus. Compared to sE protein alone, IC-vaccines using IgG-opsonized sE protein of tick-borne encephalitis virus induced similar antibody responses in mice, but analysis of the fine specificity of the induced antibodies suggested differences, suggesting that sE-IgG IC may reveal new B-cell epitopes through structural changes induced by antibodies

[31] . IC-vaccine strategies have also been explored for other major human pathogens (including human immunodeficiency virus)

[32] and for bacterial infections

[33] . Soluble IC and dendritic cells loaded with IC have also been investigated as anti-tumor vaccines

[34] .

[0014] Interestingly, while the use of ICs as vaccines may provide a reasonable approach for inducing a protective immune response, possible adverse effects have also been described that need to be considered in vaccine design. ICs can dysregulate immune responses in chronic infectious diseases and have been shown to influence inflammatory responses in autoimmune diseases through prolonged FcR engagement. This can lead to aberrant signaling and a sustained inflammatory response, including elevated levels of proinflammatory cytokines, activated lymphocytes, and ultimately immune cell exhaustion. For example, circulating ICs have been proposed to impair antibody-mediated clearance of opsonized target cells in mice infected with chronic lymphocytic choriomeningitis virus (LCMV). When anti-CD20 rituximab was injected into mice chronically infected with LCMV, B cell clearance was substantially reduced due to the presence of ICs, which competitively bind to FcgRs and block their function in rituximab-induced B cell clearance

[35] . In these mice, agonistic anti-CD40 antibodies were also impaired in FcgR-dependent activation of dendritic cells. The authors proposed that in chronic viral infections, FcgR-mediated effector functions are reduced due to circulating ICs. Interestingly, circulating ICs have also been described for other chronic viral infections such as HBV or hepatitis C virus (HCV) infection

[36] , suggesting that the observed effects may also be relevant for other chronic viral infections, possibly also for herpes simplex virus (HSV) infection. Indeed, in serum samples from patients with erythema multiforme, a skin reaction that can be triggered by HSV infection, increased levels of HSV ICs were detected

[37] , suggesting that HSV-ICs can form and may exert adverse effects on FcR function in chronic recurrent conditions.

[0015] The Fc portion of IgG confers the ability to exert effector functions including antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and antibody-dependent phagocytosis (ADCP).

[0016] In ADCC, antibodies that have bound to their targets activate effector cells, which are primarily natural killer (NK) cells, but may also be macrophages, neutrophils, or eosinophils. Activation results in killing of target cells bound to the antibody. In CDC, the complement protein C1q binds to the Fc domain of IgG1, IgG3, or IgM bound to its target Ag, which leads to a chain of events and ultimately to the formation of a membrane attack complex and the lysis of target cells. Different IgG isotypes interact with different FcgRs present on human cells with different binding affinities. IgG1 binds with high affinity to FcgRI and exerts ADCP, binds with intermediate affinity to FcgRIIaH131 and with low affinity to FcgRIIaR131, both of which also mediate ADCP, binds with low affinity to the FcgRIIb variant that is of importance for immunosuppression, and binds with intermediate and low affinity to FcgRIIIaV158 and FcgRIIIaF158, respectively, both of which mediate ADCC

[38] . IgG1 also has the highest affinity for C1q to activate CDC. Antibodies of the IgG2 isotype do not interact with FcgRI, bind to FcgRIIaH131 and FcgRIIaR131 with medium and low affinity, thereby mediating ADCC induced by myeloid cells, do not interact with the FcgRIIb variant, and bind to FcgRIIIaV158 with low affinity, but not to FcgRIIIaF158. Antibodies of the IgG3 isotype bind to FcgRI with high affinity, thereby exerting ADCP, bind to FcgRIIa or FcgRIIb variants with low affinity, and bind to FcgRIIIa variants with medium affinity

[39] . Antibodies of the IgG4 isotype bind to FcgRI with high affinity and can mediate ADCP and cytokine release, interact with FcgRIIa variants with low affinity, thereby inducing receptor clustering, and interact with FcgRIIb variants with low affinity, thereby supporting clearance and immunosuppression of ICs, while no interaction with FcgRIIIa variants has been observed

[38] . To date, only IgG1, IgG2, and IgG4 therapeutic antibodies have been approved clinically. These styles are relatively similar and differ primarily in their hinge and CH2 regions. For example, IgG4 has a unique S228 residue in the hinge region that can induce an interchangeable disulfide bond configuration. This can lead to reorganization in vivo, and lead to monovalent bispecific antibodies through Fab-arm exchange (which can be prevented by modification of the S228P amino acid substitution). IgG2 can adopt different disulfide isoforms (IgG2A, IgG2B, IgG2A / B) in structure, and transitions between different isoforms have been shown to confer FcR-independent superagonist properties attributed to IgG2B

[40] .Although also able to exert Fc effector functions through high affinity to FcgRI and moderate affinity to FcgRIII variants, antibodies of the IgG3 isotype have a long hinge region and complex disulfide bonds that impart greater polymorphism, which is believed to increase their immunogenicity, which has been a reason that has hampered the development of IgG3 for clinical applications to date.

[0017] FcgRI, FcgRIIa and FcgRIIIa act as activating receptors, which have a signal transduction motif, the immunoreceptor tyrosine-based activation motif (ITAM), which is located in the g subunit of FcgRI and FcgRIIIa, while FcgRIIa carries this motif in its cytoplasmic tail. In contrast, cross-linking of the inhibitory receptor FcgRIIb causes phosphorylation of the immunoreceptor tyrosine-based inhibitory motif (ITIM) and inhibits the transduction of signals. FcgRI is a high-affinity receptor for IgG1, IgG3 and IgG4, and is mainly expressed on activated neutrophils and myeloid cells (including dendritic cells and monocytes / macrophages). FcgRI has a high affinity for monomeric IgG1 and IgG3, and it is believed that most FcgRI molecules are occupied by free monomeric IgG1 due to the high serum IgG1 concentration of ~15 mg / ml. In the presence of the proinflammatory cytokines interleukin 3 (IL3), tumor necrosis factor-a (TNFa), or interferon g (IFNg), clustering of FcgRI is induced, resulting in better uptake of ICs and demonstrating that nanoscale reorganization of FcgRI can enhance FcgRI-mediated effector function

[41] . In addition to FcgRI, FcgRIIa is the main mediator for ADCP. Although FcgRIIa has only low and moderate affinities for monomeric IgG1 and IgG3, respectively, it binds strongly to ICs, similar to FcgRI, leading to ADCP. The primary receptor for ADCC-induced cell killing in humans is FcgRIIIa, which is mainly expressed on natural killer (NK) cells and macrophages. In contrast, FcgRIIb is an inhibitory receptor expressed on immune cells (including B cells, dendritic cells, monocytes / macrophages, mast cells and basophils) and on liver sinusoidal endothelial cells (LSECs), where it also plays a key role in the clearance of ICs. Knockout of FcgRIIb in mice severely reduces the clearance of blood-borne ICs

[42] . Indeed, blood-borne HIV-1 ICs have been shown to be cleared by LSECs and transported to lysosomes via FcgRIIb-mediated endocytosis, suggesting that viral ICs can also be cleared by FcgRIIb

[43] . Although FcgRIIb-mediated phagocytosis of ICs in LSECs can promote tolerogenic responses (through, for example, the induction of Tregs), LSECs can also switch towards immunogenic responses, depending on the cytokine environment. To what extent FcgRIIb expressed on other immune cells can also contribute to IC clearance is largely unknown.However, knockout studies suggest that FcgRIIb negatively affects phagocytosis of immune complexes in macrophages

[44] . Mutations in IgG1 at the glycosylation site N297 result in reduced effector function, which is attributed to reduced binding to low-affinity FcgRs. In contrast, in the presence of increased avidity (e.g., by multiple mAb copies within the IC), N297-mutated IgG1 can still bind to FcgRI, but the efficiency depends on the level of free monomeric IgG present that competes for binding to FcgRI. In the presence of high levels of monomeric IgG1, binding of FcgRI to IgG1-N297A ICs is strongly reduced

[45] .

[0018] IgG interacts not only with FcgR but also with nonclassical FcRs, including neonatal FcRn and cytoplasmic tripartite motif protein 21 (TRIM21).

[0019] Neonatal FcRn is a kind of MHC-I-related receptor, which can interact with IgG in the form of a heterodimer composed of heavy a-chain and b2-microglobulin. FcRn involves transport, such as by mediating the endocytic transport, distribution and retention of IgG antibodies. The interaction of FcRn and IgG depends on several histidine residues (such as, His435) on the ring exposed at the CH2-CH3 domain interface, which interact with the acidic residues of FcRn. This interaction gives the characteristic high affinity under acidic pH and the relatively weak affinity under pH 7.3-7.4, which exists for most IgG subclasses. Therefore, FcRn is important for maintaining serum IgG levels, by recycling. Some human IgG3 allotypes have Arg435 and therefore have a relatively high affinity combination with FcRn at pH 6 and at neutral pH, which is owing to the lack of protonation / deprotonation circulation under different pH. In addition, FcRn participates in the transport of ICs for MHC-presentation in hematopoietic cells. ICs are usually taken up by FcgR-induced phagocytosis and are transferred to FcRn in early endosomes once the pH becomes acidic. FcRn then supports the delivery of multimeric IgG-Ag complexes (but not monomeric IgG-Ag complexes) to proteolytic degradation and MHC-I cross-presentation or MHC-II presentation (which stimulates T-cell responses)

[46] . In fact, FcRn knockout mice show reduced CD4+ expansion after IC stimulation, which supports the function of FcRn in MHC-II Ag-presentation for T-cell stimulation

[47] . Supporting this model, FcRn was found in a ternary complex with FcgRIIa and IC, and FcRn blockade with antibodies in a rheumatoid arthritis model reduced IC-induced inflammation, which proves that FcRn is involved in proinflammatory IC-induced immune responses

[48] . FcRn is also important for the uptake of IgG-opsonized pathogens by neutrophils, the most abundant phagocytes of the body. In addition, FcRn overexpression in mice has also been shown to increase T cell responses

[49] . Cross-presentation of Ag-derived peptides on MHC-I following IC engulfment is thought to depend on the APC cell type as well as the nature and uptake pathway of the bound Ag within the IC, and may occur via either vacuolar or cytoplasmic pathways. Not only is the Fc domain of IgG important for FcRn binding, but the variable Fab region can also influence FcRn binding and subsequent effects, suggesting a more complex interplay

[50] .

[0020] The dimeric cytoplasmic non-classical Fc-receptor TRIM21 can bind to the dimeric Fc present in IgG via its C-terminal PRYSPRY domain and has a ubiquitin ligase enzyme domain that allows polyubiquitination and subsequent proteasomal degradation of target proteins bound to IgG. The complex of IgG bound to viral Ag can be taken up into the cytosol by a largely unknown mechanism, and TRIM21 binding and ubiquitination of IgG will induce an antiviral state in the cell, a process that is tightly regulated by autoubiquitination and phosphorylation of TRIM21. Like FcRn, TRIM21 binds to the CH2-CH3 region of the Fc domain. As a result of TRIM21 binding, the IgG-Ag complex is first monoubiquitinated, then polyubiquitinated (via K63-linked ubiquitin chains and later branched K48-linked ubiquitin chains), and directed to the proteasome, where the deubiquitinase Poh1 releases the entire ubiquitin chain and induces NFkB, AP1, IRF3, IRF5, and IRF 7 signaling, which leads to the production of proinflammatory cytokines. There is evidence that TRIM21-mediated proteolytic degradation of adenoviral vectors can affect MHC-I presentation and the induction of T-cell responses

[51] . The induction of proinflammatory cytokines by TRIM21-mediated ubiquitination has been extensively studied in non-enveloped adenoviral vector systems and bacteria, however, non-neutralizing antibodies that bind to the N-protein from LCMV (an enveloped virus) can also be bound by TRIM21 and also alter N-specific T cell responses to confer protection against infection

[52] .

[0021] In addition to the active immunization strategy described above using Fc-fusion proteins or ICs (during which it is sought to induce a host immune response by exposing the host to foreign recognition markers or structures), in passive immunization, antibodies are given as a prevention or therapy for microbial factors, with the purpose of neutralizing their pathogenic activity and providing short-term protection for pathogens and / or related diseases. This type of immunity can protect against measles, mumps, pertussis, poliomyelitis, rabies, rubella, tetanus, varicella and herpes zoster virus infections. The administration of antibodies is also used as a treatment for symptoms associated with snake or spider bites and is also used as an immunosuppressant (Rho-GAM, anti-lymphocyte serum). Passive immunization is often ineffective because the duration of immunization is short-lived and can vary greatly from person to person. Long-term protection after passive antibody therapy has not been described so far.

[0022] The first monoclonal antibody (mAb), muromonab-CD3, is a murine mAb targeting CD3 with the same function as was approved by the FDA in 1986, and the 100th mAb was approved in 2021, approximately one-third of which were developed to treat cancer. Monoclonal antibody therapies have been developed and are clinically approved by the FDA for the treatment of several different indications, including infectious diseases such as SARS-CoV-2 infection / COVID-19 (Bamlanivimab / Etesevimab; Casirivimab / Imdevimab; Sotrovimab, all targeting the spike protein), HIV infection (Ibalizumab, targeting CD4), respiratory syncytial virus (RSV) infection (Palivizumab, targeting the A antigenic site of the RSV F protein), anthrax (Obiltoxaximab, Raxicumab), Clostridium difficile (CDI), and SARS-CoV-2 infection (COVID-19). difficile infection (Bezlotoxumab), various types of cancer, such as bladder cancer (Durvalumab; Atezolizumab, both targeting PD-L1), hairy cell leukemia (Moxetumomab pasudodox, targeting CD22), diffuse large B-cell lymphoma (Polatuzumab vedotin, targeting CD79b), cutaneous squamous cell carcinoma (Cemiplimab, targeting PD-1), acute myeloid leukemia (Gemtuzumab ozogamicin, targeting CD33), Merkel cell carcinoma (Avelumab, targeting PD-L1), acute lymphoblastic leukemia (Inotuzumab ozogamicin, targeting CD33), and leukemia (Herceptin, targeting PD-L1). ozogamicin, targeting CD22; Blinatumomab, bispecific targeting CD19 and CD3), soft tissue sarcoma (olaratumab, targeting PDGFRa), multiple myeloma (daratumumab, targeting CD38; elotuzumab, targeting SLAMF7), neuroblastoma (dinutuximab, targeting GD2), non-small cell lung cancer (necitumumab, targeting EGFR), melanoma (nivolumab);Pembrolizumab, both targeting PD-1), gastric cancer (Ramucirumab, targeting VEGFR2), chronic lymphocytic leukemia (Obinutuzumab; Ofatumumab, both targeting CD20), breast cancer (Pertuzumab; Trastuzumab; Trastuzumab emtansine, both targeting HER2), Hodgkin lymphoma (Brentuximab vedotin, targeting CD30), metastatic melanoma (Ipilimumab, targeting CTLA-4), colorectal cancer (Panitumumab; Cetuximab, both targeting EGFR; Bevacizumab, targeting VEGF-A), non-Hodgkin lymphoma (Ibritumomab tiuxetansine, both targeting HER2), Hodgkin lymphoma (Brentuximab vedotin, targeting CD30), metastatic melanoma (Ipilimumab, targeting CTLA-4), colorectal cancer (Panitumumab; Cetuximab, both targeting EGFR; Bevacizumab, targeting VEGF-A), tiuxetan; Rituximab, both targeting CD20), chronic myeloid leukemia (Alemtuzumab, targeting CD52), and other diseases, including, for example, asthma (Omalizumab, targeting IgE), rheumatoid arthritis (Adalimumab, targeting TNFa), Crohn's disease (Infliximab, targeting TNFa), or transplant rejection (Daclizumab, targeting IL2). In addition to approved mAb therapies, as of December 2019, >570 therapeutic mAbs are under investigation in clinical trials worldwide. ;

[0023] In light of the prior art, there is a need to provide further means for the effective treatment or prevention of infectious diseases, in particular with regard to a lasting or long-term effect.

[0024] The present invention is based in part on the surprising observation that antibodies (having a certain specificity for an antigen) are exclusively capable of inducing antibody-dependent cellular phagocytosis (ADCP) without eliciting antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

[0025] This surprising finding of the present invention leads to the unexpected possibility that antibodies can be used as vaccines to actively immunize subjects against infectious or malignant diseases, particularly associated with the beneficial effect of generating durable and long-term immunity.

[0026] Thus, briefly, based on the observed effects, the present invention in its broadest sense relates to the therapeutic use of antibodies or antibody-derived molecules, such as Fc fusion proteins or combinations thereof, as a strategy for active immunization of individuals against targeted antigens by inducing de novo endogenous T-cell and / or B-cell immune responses.

[0027] According to the present invention, switching Fc-effector functions towards ADCP while avoiding ADCC and CDC induces long effector responses and enhances de novo responses, which are also directed against epitopes revealed by binding of (monoclonal) antibodies.

[0028] The mode of action of the described monoclonal antibodies is binding to a specific target, thereby inactivating or activating said target. This may be solely through masking of the site via mAb-binding, but may also involve the induction of Fc-mediated ADCC or CDC.

[0029] To the best of our knowledge, no therapeutic monoclonal antibodies have been described in the prior art that do not induce CDC or ADCC but exclusively induce ADCP.

[0030] Thus, the present invention is based in part on the proposal that modification of monoclonal antibodies towards activation of ADCP while avoiding CDC and ADCC results in long-lasting active immunization.

[0031] In fact, this is surprising in light of the prior art discussed above and the experimental evidence based on the examples.

[0032] In fact, the present invention surprisingly demonstrates that therapeutic anti-HSV IgG1 has a long-lasting protective effect, including evidence from clinical studies. Without being bound by theory, based on this surprising observation of the present invention, a long-lasting vaccine-like effect can be induced by forming ICs by monoclonal antibody target binding, followed by ADCP, thereby inducing proteolytic processing, which leads to enhanced presentation of the target peptide on MHC-I (via cross-presentation) or MHC-II to induce de novo CD8 and CD4 T-cell responses, respectively. This leads to the basic principle of the present invention.

[0033] Thus, the absence of ADCC and CDC is an important feature to prevent unwanted immunological side effects and imbalances (e.g., tolerance responses caused by killing of target cells). The experimental results of the present invention provide evidence that switching the Fc-function of antibody-opsonized pathogens toward specialized ADCP-inducing functionality and avoiding ADCC and CDC will generate a long-lasting immunological vaccine-like effect.

[0034] ADCC and CDC inevitably lead to the destruction of cells or pathogens by proteolytic digestion, for example, by the serine protease granzyme B secreted by natural killer cells or cytotoxic T cells. In contrast, endocytosed Ag-IC is digested primarily by lysosomal proteases, including the cysteine ​​proteases cathepsin B, K, H, L and S, and the aspartyl protease cathepsin D. We propose that specifically targeting ADCP and avoiding CDC and ADCC will result in more potent and potentially more extensive T-cell peptide presentation on the MHC complex. Changes in T-cell responses following monoclonal therapy have been proposed previously, however, the findings of the present invention reveal a new approach that enhances de novo T-cell and B-cell responses by a combination of the following aspects: specific enhancement of ADCP, induction of specific cytokine responses, and simultaneous eradication of ADCC and CDC functions in therapeutic antibodies, which has never been proposed before.

[0035] In view of the prior art, the technical problem underlying the present invention is to provide further means and methods for the treatment and / or prevention of (recurrent) infectious diseases or malignant diseases, in particular with regard to long-term or lasting effects. The advantages of these further means and methods for the treatment and / or prevention provided by the present invention are in particular that the individual / subject who has to be treated will simultaneously acquire long-term immunity against the infectious disease or malignant disease by means of vaccination.

[0036] The technical problem is solved by providing the embodiments characterized in the claims.

[0037] As mentioned above, it has been surprisingly demonstrated in the appended examples that antibodies (having a certain specificity for an antigen) are exclusively capable of inducing antibody-dependent cellular phagocytosis (ADCP) without triggering antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

[0038] According to the prior art discussed above, this discovery is particularly surprising and unexpected.As mentioned, this surprising discovery of the present invention leads to unexpected possibilities about the following aspects: antibodies can be used as vaccines to actively immunize subjects for infectious diseases or malignant diseases, which is particularly associated with the beneficial effects of generating lasting and long-term immunity.However, the present invention is not limited to the following conception: using antibodies for antigens associated with the infectious diseases or malignant diseases as vaccines for use in actively immunizing subjects for infectious diseases or malignant diseases.Based on this basic principle, it is also envisioned that Fc-containing fusion proteins (which include the Fc part of antibodies fused to antigens associated with the infectious diseases or malignant diseases) can be used as vaccines for use in actively immunizing subjects for infectious diseases or malignant diseases.

[0039] Therefore, in The first aspect The present invention relates to a vaccine for use in active immunization of a subject against an infectious disease or a malignant disease, the vaccine comprising:

[0040] (a) antibodies against antigens associated with said infectious disease or malignant disease; or

[0041] (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of:

[0042] Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and

[0043] Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0044] In addition, in relation to the first aspect above The second aspect , the present invention relates to a vaccine for use in active immunization of a subject against an infectious disease, wherein the infectious disease is an HSV-related disease; and

[0045] (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection; or

[0046] (b) wherein in the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen, the antigen is associated with an HSV-related disease.

[0047] The second aspect is described in more detail further below.

[0048] In the following, we describe in more detail The first aspect .

[0049] Thus, in general, the present invention relates to vaccines.

[0050] Vaccines generally refer to an "immunogenic composition" that contains at least one agent that is similar to a virus or microorganism that causes a disease. Thus, vaccines are generally biological preparations that normally provide active acquired immunity to a specific infectious disease. Vaccines are often prepared from a weakened or killed form of a microorganism, its toxin, or one of its surface proteins. The agent stimulates the body's immune system to recognize the agent as a threat, destroy it, and further recognize and destroy any microorganisms or viruses related to that agent that it may encounter in the future.

[0051] A vaccine or immunogenic portion thereof typically elicits an immunological response (either cellular or antibody-mediated) in a host to the composition.

[0052] However, the term "vaccine" as used herein in accordance with the present invention refers to a composition that does not itself contain at least one agent similar to a virus or microorganism that causes a disease. Instead, the vaccine comprises: (a) an antibody against an antigen associated with the infectious disease or malignant disease; or (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0053] As outlined above and below, the vaccines according to the present invention are proposed to elicit an immunological response (i.e., a cellular or antibody-mediated immune response) in an animal or subject by eliciting antibody-dependent cellular phagocytosis (ADCP), more particularly by mediating Fc-based effector functions via antibody-dependent cellular phagocytosis (ADCP). In other words, the vaccines according to the present invention are proposed to elicit an immunological response by eliciting antibody-dependent cellular phagocytosis (ADCP) via interaction with any of the above-mentioned FcRs, and / or by eliciting antibody-dependent cellular phagocytosis (ADCP) leading to the production of cytokines by antigen presenting cells (APCs). It is also proposed that the vaccine according to the present invention elicits an immunological response in the following manner: by eliciting antibody-dependent cellular phagocytosis (ADCP), thereby leading to the production of de novo HSV-derived peptides and presentation by antigen-presenting cells (APCs); and / or by eliciting antibody-dependent cellular phagocytosis (ADCP), thereby leading to the activation of T-cell (and / or B-cell) immune responses against de novo HSV-derived peptides; and / or by eliciting antibody-dependent cellular phagocytosis (ADCP), thereby leading to the production of HSV-derived peptides and presentation by antigen-presenting cells (APCs).

[0054] In addition, the vaccine may comprise further components typical for pharmaceutical compositions as defined below.

[0055] As used herein, the terms "vaccine" and "vaccine composition" are used interchangeably and specifically refer to a composition that will elicit a protective immune response in a subject exposed to the composition according to the invention, as outlined above and below.

[0056] The vaccine of the present invention is used for active immunization of subjects against infectious diseases or malignant diseases.

[0057] "Active immunization" according to the present invention refers to the induction of immunity following exposure to a vaccine as defined above, ultimately leading to the induction of an immunological response (cellular or antibody-mediated immune response). Typically, active immunization aims to ensure that there is an adequate supply of antibodies or T-cells and B-cells in the body that react to potential infectious agents or toxins, preferably before an infection occurs or a toxin is encountered.

[0058] Active immunization according to the present invention is in contrast to "passive immunization".

[0059] Passive immunity is generally understood as the transfer of active humoral immunity of pre-existing antibodies. Passive immunity can occur naturally, when maternal antibodies are transferred to the fetus across the placenta; and it can also be induced artificially, when high levels of antibodies specific to pathogens or toxins (obtained from humans, horses or other animals) are transferred to unimmunized persons via antibody-containing blood products, such as in immunoglobulin therapy or antiserum therapy.

[0060] The vaccine of the present invention (which comprises (a) an antibody against an antigen associated with the infectious disease or malignant disease; or (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21) is not used in passive immunization of a subject, although it comprises an antibody component and the Fc portion of the antibody component, respectively.

[0061] In contrast, contrary to popular belief, the vaccine of the present invention comprising the above antibody component and the Fc portion of the antibody component, respectively, is used in active immunization of a subject.

[0062] Active immunization can be monitored by regular evaluation by the technician using conventional methods. Without being bound by theory, active immunization can be monitored, for example, by analyzing the antibody level for the antigen used for vaccination in the serum of the vaccinated individual. The serological conversion of immunoglobulin from seronegative to seropositive shows that the vaccine actively induces a humoral immune response. Several methods have been established for testing seroconversion. The best known method is enzyme-linked immunosorbent assay (ELISA). Antigens are coated on a plate and serum is added. If there is seroconversion, the antibodies in the serum will be combined with the antigen on the plate and can be detected by a second antibody that is usually coupled to an enzyme. In contrast to passive immunization, in active immunization, these antibodies are derived from the personnel vaccinated. The addition of substrate causes a color conversion by the enzyme, and this can be measured by using a photometer. Alternative assays include immunoblotting, electroluminescent ELISA, measurement using plasma surface resonance or bio-layer interferometry, flow cytometry or microscopy based on immunofluorescence.

[0063] In addition to this B cell-mediated immune response, active immunization also induces T-cell responses to peptides derived from vaccine antigens. T-cell responses to vaccine antigens can be measured, for example, by incubating primary blood mononuclear cells (PBMCs) with vaccine antigens and measuring interleukin-2 (IL-2) or interferon gamma (IFNγ) secretion by activated T-cells using, for example, ELISpot assays, cytokine ELISAs or flow cytometry assays in which the released cytokines are bound by fluorescent beads.

[0064] Active immunization is also seen through changes in the T-cell receptor (TCR) and B-cell receptor repertoires (BCR). Active immunization will induce clonal expansion of B-cells and T-cells specific for the vaccine antigen, and therefore when the entire repertoire is sequenced, individual clones will become more prominent

[55] .

[0065] The vaccine of the present invention is used for active immunization of subjects against infectious diseases or malignant diseases.

[0066] The term "infectious disease" is generally known in the art, and according to the present invention refers to a condition caused by an organism such as a bacterium, virus, fungus or parasite. Many organisms live in or on our bodies. They are usually harmless, or even helpful. However, under certain conditions, some organisms may cause disease. Infection is generally understood to be the invasion of the body or body tissue of an organism by a disease-causing factor (e.g., a bacterium, virus, fungus or parasite), their multiplication or proliferation, and the response of the host / subject's tissue to the infectious agent and the toxins they produce. Infectious diseases (also referred to as disseminated diseases or infectious diseases) generally refer to illnesses caused by infection. As mentioned, infection can be caused by a wide range of pathogens (most prominently, bacteria and viruses). The host / subject can use its immune cells to fight infection. According to the present invention, the subject / host is preferably a mammalian host / subject, more preferably a human host / subject, which is known to respond to infection in the following manner: an innate response, which often involves inflammation, followed by an adaptive response.

[0067] In preferred embodiments, the infectious disease is caused by an infectious agent (often referred to as a "pathogen", such as a bacterium, virus, fungus or parasite) selected from the group consisting of:

[0068] bacteria (preferably Mycobacterium tuberculosis, Staphylococcus aureus, Escherichia coli, Clostridium botulinum and Salmonella spp.);

[0069] Viruses and / or related agents such as viroids (preferably, HIV, rhinovirus, lyssavirus such as rabies virus, Ebola virus, and severe acute respiratory syndrome coronavirus 2);

[0070] Fungi, which are further subclassified into:

[0071] Ascomycota, which includes yeasts such as Candida (the most common fungal infection), filamentous fungi such as Aspergillus, Pneumocystis species, and dermatophytes, which are a group of organisms that cause infections of the skin and other surface structures in humans,

[0072] Basidiomycota, including the human pathogenic genus Cryptococcus;

[0073] Prions;

[0074] Parasites, which are usually divided into:

[0075] Single-celled organisms (e.g., malaria, Toxoplasma, Babesia),

[0076] Large parasites (worms), nematodes (e.g., roundworms and pinworms), tapeworms, flukes (e.g., Schistosoma,

[0077] Arthropods (e.g., ticks, mites, fleas, and lice),

[0078] They can also cause disease in humans, which is similar in concept to an infection, but the invasion of a person or animal's body by these large parasites is usually referred to as an infestation.

[0079] In a more preferred embodiment, the vaccine defined above is for use in the active immunization of a subject against an infectious disease selected from the group consisting of fungal infections, bacterial infections, protozoan infections and viral infections.

[0080] In a more preferred embodiment, the fungal infection is infection with a Candida strain.

[0081] In another more preferred embodiment, the bacterial infection is infection with a strain of Pseudomonas.

[0082] In another more preferred embodiment, the protozoan infection is infection with a strain of Plasmodium.

[0083] In another more preferred embodiment, the viral infection is an HSV-related disease (eg, an HSV-related disease caused by HSV-1 and / or HSV-2).

[0084] According to the basic principle of the present invention, the present invention is not particularly limited to a certain "infectious disease". The skilled person will understand that the vaccine of the present invention can be adjusted to any "infectious disease", more particularly to the corresponding "antigen associated with the infectious disease".

[0085] The term "malignant disease" is generally known in the art and is understood in its broadest sense as a tendency of a medical condition to gradually become worse.

[0086] Malignancy is the most familiar as a characteristic of cancer. Malignant tumors differ from noncancerous benign tumors in that malignancies are not self-limited in their growth, can invade into adjacent tissues, and may be able to spread to distant tissues. Benign tumors do not have any of those characteristics.

[0087] Malignancy in cancer is characterized by anaplasia, invasiveness and metastasis. Malignant tumors are also characterized by genomic instability, so as assessed by whole genome sequencing, cancer often has 10,000 to 100,000 mutations in its entire genome. Cancer usually shows tumor heterogeneity, including multiple subclones. They also often have the expression of reduced DNA repair enzymes, which is due to the epigenetic methylation of DNA repair genes or the microRNA of the control DNA repair gene expression of the change.

[0088] In a preferred embodiment, the malignant disease is a tumor, more preferably cancer.

[0089] The term "cancer" refers to a group of diseases involving an abnormal increase in the number of cells, which have the potential to invade or spread to other parts of the body. There are more than 100 different known cancers that affect humans.

[0090] Cancers are often described by the part of the body they originate in. However, some parts of the body contain multiple tissue types, so for greater accuracy, cancers are additionally classified by the type of cell from which the tumor cells originate.

[0091] Therefore, the vaccine for use in vaccinating a subject against a malignant disease according to the present invention is such a vaccine, wherein the malignant disease, more particularly cancer, is selected from the group consisting of:

[0092] Carcinoma: Carcinomas are cancers that originate from epithelial cells. This group includes many of the most common cancers that occur in older people. Almost all cancers that develop in the breast, prostate, lung, pancreas, and colon are carcinomas of epithelial origin.

[0093] • Sarcomas: Sarcomas are cancers that arise from connective tissue (ie, for example, bone, cartilage, fat, nerves), each of which develops from cells that originate from mesenchymal cells outside the bone marrow.

[0094] Lymphoma and leukemia: These two categories of cancer arise from immature cells that originate in the bone marrow and are intended to fully differentiate and mature into normal components of the immune system and blood, respectively. A preferred type of leukemia is acute lymphoblastic leukemia. This type of leukemia is the most common type of cancer in children, accounting for approximately 30% of cases. A preferred type of lymphoma is diffuse large B-cell lymphoma, which is the most common form, accounting for approximately 30% of non-Hodgkin lymphomas.

[0095] Germ cell tumors: Germ cell tumors are cancers that arise from pluripotent cells and most often arise in the testicles or ovaries (seminoma and dysgerminoma, respectively).

[0096] Germ cell tumors: Germ cell tumors are cancers that originate from immature "precursor" cells or embryonic tissue. Germ cell tumors are usually more common in children. Preferred germ cell tumors are neuroblastoma, retinoblastoma, Wilms tumor, hepatoblastoma, medulloblastoma.

[0097] In a more preferred embodiment, the vaccine for use in vaccinating a subject against a malignant disease according to the invention is a vaccine wherein the malignant disease, more particularly cancer, is selected from the group consisting of bone and muscular sarcomas, brain cancer and / or cancer of the nervous system, breast cancer, cancer of the endocrine system, cancer of the eye, gastrointestinal cancer, genitourinary or gynecological cancer, head and neck cancer, cancer of the hematopoietic system, skin cancer, cancer of the thoracic and respiratory system, HIV / AIDS-related cancers and unclassified cancers.

[0098] Bone and muscle carcinoma

[0099] In an even more preferred embodiment, the bone and muscle sarcoma is selected from the group consisting of chondrosarcoma, Ewing's sarcoma, malignant fibrous histiocytoma / osteosarcoma of bone, osteosarcoma, rhabdomyosarcoma, leiomyosarcoma, and myxosarcoma.

[0100] Brain cancer and / or nervous system cancer

[0101] In a more preferred embodiment, the brain cancer and / or nervous system cancer is selected from the group consisting of astrocytoma, brain stem glioma, fibrous astrocytoma, ependymoma, primitive neuroectodermal tumor, cerebellar astrocytoma, cerebral astrocytoma, glioblastoma, glioma, medulloblastoma, neuroblastoma, oligodendroglioma, pineal astrocytoma, pituitary adenoma and visual pathway and hypothalamic glioma.

[0102] Breast cancer

[0103] In a more preferred embodiment, the breast cancer (breast cancer) is selected from the group consisting of inflammatory breast cancer, invasive lobular carcinoma, ductal carcinoma, invasive cribriform carcinoma, medullary carcinoma, male breast cancer and phyllodes tumor.

[0104] Endocrine system cancer

[0105] In a more preferred embodiment, the cancer of the endocrine system is selected from the group consisting of: adrenal cortical epithelial carcinoma, islet cell epithelial carcinoma (also known as endocrine pancreas), multiple endocrine neoplasia syndrome, parathyroid carcinoma, pheochromocytoma, thyroid cancer and Merkel cell epithelial carcinoma.

[0106] Eye cancer

[0107] In an even more preferred embodiment, the eye cancer is selected from the group consisting of uveal melanoma, retinoblastoma and optic nerve glioma.

[0108] Gastrointestinal cancer

[0109] In a more preferred embodiment, the gastrointestinal cancer is selected from the group consisting of anal cancer, appendiceal cancer, bile duct carcinoma, carcinoid tumor, gastrointestinal cancer, colon cancer, extrahepatic bile duct cancer, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), hepatocellular carcinoma, pancreatic cancer (islet cell), rectal cancer and small intestine cancer.

[0110] Genitourinary or gynecological cancer

[0111] In a more preferred embodiment, the genitourinary or gynecological cancer is selected from the group consisting of: bladder cancer, cervical cancer, endometrial cancer, extragonadal germ cell tumors, ovarian cancer, ovarian epithelial cancer (surface epithelial-stromal tumors), ovarian germ cell tumors, penile cancer, renal cancer, renal cell epithelial-derived cancer, renal pelvis and ureteral cancer (transitional cell carcinoma), prostate cancer, testicular cancer, gestational trophoblastic tumors, ureteral and renal pelvis cancer (transitional cell carcinoma), urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer and Wilms' tumor (Nephroblastoma).

[0112] Head and Neck Cancer

[0113] In an even more preferred embodiment, the head and neck cancer is selected from the group consisting of esophageal cancer, head and neck cancer, cancer of nasopharyngeal epithelial origin, oral cancer, oropharyngeal cancer, paranasal sinus and nasal cavity cancer, pharyngeal cancer, salivary gland cancer, and hypopharyngeal cancer.

[0114] Hematopoietic cancer

[0115] In a more preferred embodiment, the hematopoietic cancer is selected from the group consisting of acute biphenotypic leukemia, acute eosinophilic leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, acute myeloid dendritic cell leukemia, AIDS-related lymphoma, anaplastic large cell lymphoma, angioimmunoblastic T-cell lymphoma, B-cell prolymphocytic leukemia, Burkitt's lymphoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, cutaneous T-cell lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, hepatosplenic T-cell lymphoma, Hodgkin's lymphoma, intravascular large B-cell lymphoma, Lymphoma, large granular lymphocytic leukemia, lymphoplasmacytic lymphoma, lymphomatoid granulomatosis, mantle cell lymphoma, marginal zone B-cell lymphoma, mast cell leukemia, mediastinal large B-cell lymphoma, multiple myeloma / plasma cell neoplasm, myelodysplastic syndrome, mucosa-associated lymphoid tissue lymphoma, mycosis fungoides, nodular marginal zone B-cell lymphoma, non-Hodgkin lymphoma, precursor B-lymphoblastic leukemia, primary central nervous system lymphoma, primary cutaneous follicular lymphoma, primary cutaneous immunocytoma, primary effusion lymphoma, plasmablastic lymphoma, Sezary syndrome, splenic marginal zone lymphoma, and T-cell prolymphocytic leukemia.

[0116] Skin cancer

[0117] In a more preferred embodiment, the skin cancer is selected from the group consisting of: basal cell carcinoma, squamous cell carcinoma, squamous cell skin carcinoma, skin adnexal tumors (e.g., sebaceous epithelial carcinoma), melanoma, Merkel cell carcinoma, keratoacanthoma, sarcoma of primary skin origin (e.g., dermatofibrosarcoma protuberans), and lymphoma of primary skin origin (e.g., mycosis fungoides).

[0118] Thoracic and respiratory cancers

[0119] In a more preferred embodiment, the thoracic and respiratory cancer is selected from the group consisting of adenocarcinoma of the lung, bronchial adenoma / carcinoid, small cell lung cancer, mesothelioma, non-small cell lung cancer, non-small cell lung epithelial carcinoma, pleuropulmonary blastoma, laryngeal cancer, thymoma and thymic epithelial carcinoma and squamous cell carcinoma of the lung.

[0120] HIV / AIDS-related cancers

[0121] In an even more preferred embodiment, the HIV / AIDS-related cancer is Kaposi's sarcoma.

[0122] Unclassified cancer

[0123] There are also cancers that are known but not classified in any of the above cancer groups. The corresponding cancers are collectively referred to as "unclassified cancers". In a more preferred embodiment, the unclassified cancer is selected from the group consisting of epithelioid hemangioendothelioma (EHE), desmoplastic small round cell tumor and liposarcoma.

[0124] In a more preferred embodiment, the vaccine defined above is for use in the active immunization of a subject against a malignant disease selected from the group consisting of solid tumors and hematological malignancies / hematological oncological diseases.

[0125] Solid tumors are generally tumors that do not contain any fluid or cysts.

[0126] The two main types of solid tumors according to the present invention are sarcomas and carcinomas of epithelial origin.

[0127] Sarcomas are tumors in blood vessels, bones, fatty tissue, ligaments, lymph vessels, muscles, or tendons.

[0128] Preferred sarcomas have been outlined above, and in particular embodiments include Ewing sarcoma and osteosarcoma, which are carcinosarcoma of the bone. Rhabdomyosarcoma, which is a soft tissue sarcoma found in muscle.

[0129] Cancers of epithelial origin are tumors that form in epithelial cells. Epithelial cells are found in the skin, glands, and linings of organs. Those organs include the bladder, ureters, and parts of the kidneys.

[0130] Preferred cancers of epithelial origin have been outlined above, and in one particular embodiment include cancers of adrenal cortex epithelial origin. This is when the tumor develops in one or both adrenal glands (located above each kidney).

[0131] Blood disorders / hemato-oncological disorders are defined above.

[0132] As mentioned above, in accordance with the basic principles of the present invention, the present invention is not particularly limited to a certain "infectious disease" or "malignant disease". The skilled person will understand that the vaccine of the present invention can be adjusted to any of the "infectious diseases" or "malignant diseases" described above, and more particularly to the corresponding "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively.

[0133] With regard to the "antigen associated with the malignant disease" and the "antigen associated with the infectious disease", in a preferred embodiment, the antigen is specific to cancer or is an antigen specific to an infectious disease.

[0134] Those skilled in the art know cell surface structures, epitopes or antigens specific to cells specific for cancer, or antigens specific to infectious diseases. Therefore, the technician can select and choose suitable antigens specific to cancer or antigens specific to infectious diseases.

[0135] Without being bound by theory, the "antigen associated with the malignant disease" can be, for example, a tumor-associated antigen (TAA). Various tumor-associated antigens are known in the art, including but not limited to carbonic anhydrase IX, CCCL19, CCCL21, CSAp, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, IGF-1R, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD45, CD46, CD52, CD54, CD56, CD57, CD58, CD59, CD60, CD61, CD62, CD63, CD64, CD65, CD66, CD67, CD68, CD69, CD70, CD71, CD72, CD73, CD74, CD75, CD76, CD77, CD78, CD79, CD80, CD81, CD82, CD83, CD84, CD85, CD86, CD87, CD88, CD80, CD81, CD82, CD83, CD84 ... 5. CD59, CD64, CD66a-e, CD67, CD70, CD74, CD79a, CD80, CD83, CD95, CD126, CD133, CD138, CD147, CD154, AFP, PSMA, CEACAM5, CEACAM6, B7, ED-B of fibronectin, H factor, FHL-1, Flt-3, folate receptor, GROB, HER2, HMGB-1, hypoxia-inducible factor (HIF), HM1.24, insulin-like growth factor-1 (ILGF-1), IFN-γ, IFN- α, IFN-β, IL-2, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-25, IP-10, mCRP, M CP-1, MIP-1A, MIP-1B, MIF, MUC1, MUC2, MUC3, MUC4, MUC5ac, PAM4 antigen, NCA-95, NCA-90, NY-ESO-1, Ia, HM1.24, EGP-1, EGP-2, HLA -DR, tenascin, Le(y), RANTES, T101, TAC, Tn antigen, Thomson-Friedenreich antigen, tumor necrosis antigen, TNF-α, TRAIL receptor (R1 and R2), VEGFR, EGFR, PlGF, complement factor C3, C3a, C3b, C5a, C5, cancer-testis (CT) antigen SPAG9, CT9, CT10, LAGE, MAGE-B5, MAGE-B6, MAGE-C2, MAGE-C3, MAGE-D, HAGE, SAGE, and oncogene products.

[0136] Without being bound by theory, the "antigen associated with the infectious disease" may, for example, be an Epstein-Barr virus (EBV) (derived) protein or protein domain, preferably EBNA1, EBNA2, EBNA3A, EBNA3B, EBNA3C, EBNA-LP, LMP1, LMP2A / B, gp350, A73, RPMS1, ZEBRA, Rta, major intercalated protein p143, major intercalated protein, intercalated protein, major capsid protein, minor capsid protein, capsid protein p18, p23, p40, gN, gp42, gM, gp60, gp78 / 55, gp150, 53 / 55kd membrane protein, viral IL-10, gB, gH, gL;

[0137] Human papillomavirus (HPV) (derived) proteins or protein domains, preferably E1, E2, E4, E5, E6, E7, L1, L2;

[0138] Human immunodeficiency virus 1 or 2 (derived) proteins or protein domains, preferably Gag, Pol, Env, gp160, gp120, gp41, CA, MA, p2, p6, NC, IN, RTp66, RTp55, RTp51, PR, Rev, Tat, Nef, Vif, Vpr, Vpu, Vpx;

[0139] Herpes simplex virus (HSV) 1 or 2 (derived) proteins or protein domains, preferably gB, gC, gD, gH, gG, gL, gE, gI, gK, gM, VP1-2, ICP32, ICP0, VP11 / 12, UL13, vhs, VP16, US3, VP22, ICP34.5, US11, ICP4, DNA polymerase, major capsid protein, helicase, primase, uracil DNA glycosylase, dUTPase, ribonucleotide reductase, major intercalated protein;

[0140] A human cytomegalovirus (HCMV) (derived) protein or protein domain encoded by an open reading frame (ORF), preferably UL57, UL55, UL54, UL75, UL86, UL85, UL104, UL97, UL98, UL100, UL105, UL102, UL114, UL115, UL72, UL70, UL69, UL44, UL45, UL48;

[0141] Hepatitis A virus (HAV) (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3Cpro, P3-3Dpol;

[0142] Hepatitis B virus (HBV) (derived) proteins or protein domains, preferably M, L, S, Polymerase, TP, Spacer, RT, RNase H, preCore, Core, X, HBeAg;

[0143] Hepatitis C virus (HCV) (derived) proteins or protein domains, preferably E1, E2, core, NS1, NS2, NS3, NS4a, NS4b, NS5a, NS5b, precursor polyprotein;

[0144] Influenza A virus (IAV) (derived) proteins or protein domains, preferably HA, NA, M2, M1, NP, NS1, NS2 / NEP, PA, PB1;

[0145] Measles virus (MV) (derived) protein or protein domain, preferably N, P / C / V, M, F, H, L;

[0146] Respiratory syncytial virus (RSV) (derived) protein or protein domain, preferably NS1, NS2, N, P, M, SH, G, F, M21, M22, L;

[0147] Rotavirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, RdRp, VP5, VP6, VP7, VP8, NSP1, NSP2, NSP3, NSP4, NSP5, NSP6;

[0148] Severe acute respiratory syndrome coronavirus (SARS-Cov) type 1 and 2 (derived) proteins or protein domains, preferably ns1, ns2, PLpro, ns4, 3CL, ns6, ns7, ns8, ns9, ns10, RdRp, Hel, ns14, ns15, ns16, S, S-RBD, 3a, E, M, 6, 7a, 7b, 8N, 9b, 14;

[0149] Varicella zoster virus (VZV) (derived) proteins or protein domains encoded by open reading frames (ORFs), preferably ORF0 to ORF71 (at least 71 proteins in total);

[0150] Human herpesvirus 8 (HHV-8) (derived) proteins or protein domains, preferably K1, K2, K3, K4, K4.1, K5, K6, K7, K8, K8.1, K9, K10, K10.5, K11, K12, K13, K14, K15, gB, gL, gH and proteins encoded in ORFs, preferably ORF2, ORF9, ORF10, ORF16, ORF18, ORF24, ORF30, ORF31, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF42, ORF43, ORF44, ORF45, ORF46, ORF47, ORF48, ORF49, ORF50, ORF51, ORF52, ORF53, ORF54, ORF55, ORF56, ORF57, ORF58, ORF59, ORF60, ORF61, ORF62, ORF63, ORF64, ORF65 F66, ORF21, ORF23, ORF25, ORF26, ORF65, ORF33, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF4 2. ORF45, ORF49, ORF50, ORF52, ORF53, ORF55, ORF57, ORF59, ORF67, ORF69, ORF70, ORF72, ORF73, ORF74, ORF75;

[0151] Human herpesvirus 6 (HHV-6) (derived) protein or protein domain encoded by ORF, preferably DR1, DR6, DR7 / U1, U2, U3, U4, U7, U10, U11, U12, U13, U14, U15, U17, U18, U19, U20, U21, U22, U23, U24, U25, U26, U27, U28, U29, U30, U31, U32, U33, U34, U35, U36, U37, U38, U39, U40 , U41, U42, U43, U44, U45, U46, U47, U48, U49, U50, U51, U52, U53, U54, U55, U56, U57, U58, U59, U61, U62, U63, U64, U65, U66, U69, U70, U71, U72, U73, U74, U75, U76, U77, U79, U81, U82, U83, U85, U86, U88, U90, U91, U94, U95, U100;

[0152] Rabies virus (derived) protein or protein domain, preferably N, P, M, G, L;

[0153] Mumps virus (derived) proteins or protein domains, preferably N, V / P, M, F, HN, L; and

[0154] Rhinovirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3C, P3-3D.

[0155] Therefore, in a preferred embodiment, in the vaccine for use according to the first aspect above, the antigen associated with the malignant disease is selected from the group consisting of the following: tumor-associated antigens known in the art, including but not limited to carbonic anhydrase IX, CCCL19, CCCL21, CSAp, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, IGF-1R, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, C D37, CD38, CD40, CD40L, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD74, CD79a, CD80, CD83, CD95, CD126, CD133, CD138, CD147, CD154, AFP, PSMA, CEACAM5, CEACAM6, B7, ED-B of fibronectin, H factor, FHL-1, Flt-3, folate receptor, GROB, HER2, HMGB-1, hypoxia-inducible factor (HIF), HM1.24, Insulin-like growth factor-1 (ILGF-1), IFN-γ, IFN-α, IFN-β, IL-2, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-6, IL-8, IL-12, IL-15, IL-17, IL -18, IL-25, IP-10, mCRP, MCP-1, MIP-1A, MIP-1B, MIF, MUC1, MUC2, MUC3, MUC4, MUC5ac, PAM4 antigen, NCA-95, NCA-90, NY-ESO-1, Ia, HM1.24, EGP-1, EGP-2, HLA-DR, tenascin, Le(y), RANTES, T101, TAC, Tn antigen, Thomson-Friedenreich antigen, tumor necrosis antigen, TNF-α, TRAIL receptor (R1 and R2), VEGFR, EGFR, PlGF, complement factor C3, C3a, C3b, C5a, C5, cancer-testis (CT) antigen SPAG9, CT9, CT10, LAGE, MAGE-B5, MAGE-B6, MAGE-C2, MAGE-C3, MAGE-D, HAGE, SAGE and oncogene products.

[0156] In addition, regarding the term "antigen associated with the infectious disease", in a preferred embodiment, in the vaccine for use according to the first aspect above, the antigen associated with the infectious disease is selected from the group consisting of:

[0157] Epstein-Barr virus (EBV) (derived) proteins or protein domains, preferably EBNA1, EBNA2, EBNA3A, EBNA3B, EBNA3C, EBNA-LP, LMP1, LMP2A / B, gp350, A73, RPMS1, ZEBRA, Rta, major intercalated protein p143, major intercalated protein, intercalated protein, major capsid protein, minor capsid protein, capsid protein p18, p23, p40, gN, gp42, gM, gp60, gp78 / 55, gp150, 53 / 55kd membrane protein, viral IL-10, gB, gH, gL;

[0158] Human papillomavirus (HPV) (derived) proteins or protein domains, preferably E1, E2, E4, E5, E6, E7, L1, L2;

[0159] Human immunodeficiency virus 1 or 2 (derived) proteins or protein domains, preferably Gag, Pol, Env, gp160, gp120, gp41, CA, MA, p2, p6, NC, IN, RTp66, RTp55, RTp51, PR, Rev, Tat, Nef, Vif, Vpr, Vpu, Vpx;

[0160] Herpes simplex virus (HSV) 1 or 2 (derived) proteins or protein domains, preferably gB, gC, gD, gH, gG, gL, gE, gI, gK, gM, VP1-2, ICP32, ICP0, VP11 / 12, UL13, vhs, VP16, US3, VP22, ICP34.5, US11, ICP4, DNA polymerase, major capsid protein, helicase, primase, uracil DNA glycosylase, dUTPase, ribonucleotide reductase, major intercalated protein;

[0161] A human cytomegalovirus (HCMV) (derived) protein or protein domain encoded by an open reading frame (ORF), preferably UL57, UL55, UL54, UL75, UL86, UL85, UL104, UL97, UL98, UL100, UL105, UL102, UL114, UL115, UL72, UL70, UL69, UL44, UL45, UL48;

[0162] Hepatitis A virus (HAV) (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3Cpro, P3-3Dpol;

[0163] Hepatitis B virus (HBV) (derived) proteins or protein domains, preferably M, L, S, Polymerase, TP, Spacer, RT, RNase H, preCore, Core, X, HBeAg;

[0164] Hepatitis C virus (HCV) (derived) proteins or protein domains, preferably E1, E2, core, NS1, NS2, NS3, NS4a, NS4b, NS5a, NS5b, precursor polyprotein;

[0165] Influenza A virus (IAV) (derived) proteins or protein domains, preferably HA, NA, M2, M1, NP, NS1, NS2 / NEP, PA, PB1;

[0166] Measles virus (MV) (derived) protein or protein domain, preferably N, P / C / V, M, F, H, L;

[0167] Respiratory syncytial virus (RSV) (derived) protein or protein domain, preferably NS1, NS2, N, P, M, SH, G, F, M21, M22, L;

[0168] Rotavirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, RdRp, VP5, VP6, VP7, VP8, NSP1, NSP2, NSP3, NSP4, NSP5, NSP6;

[0169] Severe acute respiratory syndrome coronavirus (SARS-Cov) type 1 and 2 (derived) proteins or protein domains, preferably ns1, ns2, PLpro, ns4, 3CL, ns6, ns7, ns8, ns9, ns10, RdRp, Hel, ns14, ns15, ns16, S, S-RBD, 3a, E, M, 6, 7a, 7b, 8N, 9b, 14;

[0170] Varicella zoster virus (VZV) (derived) proteins or protein domains encoded by open reading frames (ORFs), preferably ORF0 to ORF71 (at least 71 proteins in total);

[0171] Human herpesvirus 8 (HHV-8) (derived) proteins or protein domains, preferably K1, K2, K3, K4, K4.1, K5, K6, K7, K8, K8.1, K9, K10, K10.5, K11, K12, K13, K14, K15, gB, gL, gH and proteins encoded in ORFs, including but not limited to ORF2, ORF9, ORF10, ORF16, ORF18, ORF24, ORF30, ORF31, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF42, ORF43, ORF44, ORF45, ORF46, ORF47, ORF48, ORF49, ORF50, ORF51, ORF52, ORF53, ORF54, ORF55, ORF56, ORF57, ORF58, ORF59, ORF60, ORF61, ORF62, ORF63, ORF64, ORF65, ORF66, ORF67, ORF68, ORF69, ORF70, ORF71, ORF72, ORF73, ORF74, ORF75 RF66, ORF21, ORF23, ORF25, ORF26, ORF65, ORF33, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF 42. ORF45, ORF49, ORF50, ORF52, ORF53, ORF55, ORF57, ORF59, ORF67, ORF69, ORF70, ORF72, ORF73, ORF74, ORF75;

[0172] Human herpesvirus 6 (HHV-6) (derived) protein or protein domain encoded by ORF, preferably DR1, DR6, DR7 / U1, U2, U3, U4, U7, U10, U11, U12, U13, U14, U15, U17, U18, U19, U20, U21, U22, U23, U24, U25, U26, U27, U28, U29, U30, U31, U32, U33, U34, U35, U36, U37, U38, U39, U40 , U41, U42, U43, U44, U45, U46, U47, U48, U49, U50, U51, U52, U53, U54, U55, U56, U57, U58, U59, U61, U62, U63, U64, U65, U66, U69, U70, U71, U72, U73, U74, U75, U76, U77, U79, U81, U82, U83, U85, U86, U88, U90, U91, U94, U95, U100;

[0173] Rabies virus (derived) protein or protein domain, preferably N, P, M, G, L;

[0174] Mumps virus (derived) proteins or protein domains, preferably N, V / P, M, F, HN, L; and

[0175] Rhinovirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3C, P3-3D.

[0176] As mentioned above, in a preferred embodiment, the vaccine used in active immunization of a subject for an infectious disease or a malignant disease requires the presence of an acute infection, i.e., the presence of an antigen. This particularly relates to aspect (a), wherein an antibody for an antigen associated with the infectious disease or malignant disease is used as a component of the vaccine of the present invention. This is because, in order to be able to carry out active immunization of a subject according to the basic principle of the present invention, not only the antibody but also the antigen associated with the infectious disease or malignant disease must be present. With regard to aspect (b), active infection is not necessarily required, because the antigen is fused to Fc, and therefore each antigen is easily present, even in the absence of an acute infection.

[0177] In this regard, "acute" means that the subject has an outbreak of an infectious disease or a malignant disease and thus antigens associated with the infectious disease or the malignant disease are present in the subject's body.

[0178] In a more preferred embodiment, "acute" means that the subject shows symptoms of an infectious disease or a malignant disease. In other words, the subject to be vaccinated actually needs vaccination or treatment, and the term "acute" relates to the situation after the onset of an infectious disease or a malignant disease (for the first time). The term "acute" mentioned in the context of the present invention is relative to prophylactic or preventive (i.e., for disease prevention, for example, measures taken to prevent the onset / outbreak of infection and / or disease). More particularly, prophylactic treatment can be understood in such a way that it prevents free viral particles (from outside the body) from attaching to target cells and thereby preventing viral replication.

[0179] In the context of aspects where an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious or malignant disease is used as a component of the vaccine of the present invention, the presence of an acute infection is not necessarily required for active immunization of a subject against an infectious or malignant disease according to the present invention. This particularly relates to aspects where an antibody against an antigen associated with the infectious or malignant disease is used as a component of the vaccine of the present invention. This is because, in order to be able to actively immunize a subject according to the basic principles of the present invention, not only the antibody but also the antigen associated with the infectious or malignant disease must be present. However, in the context of aspects where an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious or malignant disease is used as a component of the vaccine of the present invention, the antigen associated with the infectious or malignant disease is already there (i.e., part of the Fc-containing fusion protein) and is therefore already contained in the vaccine.

[0180] As already outlined above, the vaccines of the present invention may comprise one of two alternative components (although it is also contemplated that the vaccines comprise both components), namely:

[0181] (a) antibodies against antigens associated with said infectious disease or malignant disease; or

[0182] (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of:

[0183] Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and

[0184] Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0185] These two components are described separately in more detail below.

[0186] Antibodies against antigens associated with the infectious or malignant disease:

[0187] Regarding (a) The term "antibody" is known in the art and is also often referred to as immunoglobulin (Ig), i.e., an antibody directed against an antigen associated with the infectious disease or malignant disease.

[0188] In the context of the present invention, in accordance with the basic principles of the invention, the term "antibody" refers to an antibody that comprises at least one antibody Fc part.

[0189] Therefore, in a preferred embodiment, the antibody of component (a) of the vaccine of the present invention described above (ie, the antibody against an antigen associated with the infectious disease or malignant disease) comprises at least one antibody Fc part.

[0190] The "Fc" portion or "Fc" region of an antibody comprises two heavy chain fragments, which comprise the C H 2 and C H 3 domains. The two heavy chain fragments are connected by two or more disulfide bonds and by C H The 3 domains are held together by hydrophobic interactions.

[0191] In the context of the present invention, the term "antibody" relates to whole immunoglobulin molecules, as well as parts of such immunoglobulin molecules (ie, "antigen-binding fragments thereof").

[0192] Further, as outlined in more detail below, the term relates to modified and / or altered antibody molecules. The term also relates to antibodies generated / synthesized recombinantly or synthetically. The term also relates to complete antibodies and antibody fragments thereof, such as separated light and heavy chains.

[0193] Since the antibodies of the present invention comprise at least one antibody Fc portion, Fab, Fv, Fab', Fab'-SH, F(ab')2 are not contemplated.

[0194] The term "antibody" also includes, but is not limited to, fully human antibodies, chimeric antibodies, humanized antibodies, CDR-grafted antibodies, and antibody constructs, such as single-chain Fv (scFv) or antibody fusion proteins.

[0195] In a preferred embodiment, the antibody according to the present invention is a monoclonal antibody. In a further preferred embodiment, the antibody according to the present invention is a humanized antibody or a fully human antibody. In a further preferred embodiment, the antibody according to the present invention is a murine antibody.

[0196] The term "monoclonal antibody" as used herein refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies constituting the population are identical, except for possible naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific, directed against a single antigenic site. Monoclonal antibodies are advantageous because they can be synthesized by hybridoma cultures, substantially free from contamination by other immunoglobulins. The modifier "monoclonal" indicates that the antibody is characterized by being in a substantially homogeneous antibody population, and should not be construed as requiring the antibody to be produced by any ad hoc method. As mentioned above, the monoclonal antibody to be used according to the present invention can be prepared by the hybridoma method described by Kohler, Nature 256 (1975), 495.

[0197] The term "polyclonal antibody" as used herein refers to an antibody produced in or in the presence of one or more other, non-identical antibodies. Generally, polyclonal antibodies are produced from B-lymphocytes in the presence of several other B-lymphocytes that produce non-identical antibodies. Typically, polyclonal antibodies are obtained directly from immunized animals.

[0198] The term "fully human antibody" as used herein refers to an antibody comprising only human immunoglobulin protein sequences. A fully human antibody may comprise murine carbohydrate chains if it is produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, a "mouse antibody" or "murine antibody" refers to an antibody comprising only mouse / murine immunoglobulin protein sequences. Alternatively, a "fully human antibody" may comprise rat carbohydrate chains if it is produced in a rat, in a rat cell, or in a hybridoma derived from a rat cell. Similarly, the term "rat antibody" refers to an antibody comprising only rat immunoglobulin sequences. Fully human antibodies may also be produced, for example, by phage display (which is a widely used screening technique that enables the production and screening of fully human antibodies). Phage antibodies may also be used in the context of the present invention. Phage display technology is described in, for example, US 5,403,484, US 5,969,108, and US 5,885,793. Another technology that enables the development of fully human antibodies involves improvements in mouse hybridoma technology. Mice are made transgenic to contain human immunoglobulin loci, exchanging their own mouse genes (see, e.g., US 5,877,397).

[0199] The term "chimeric antibody" refers to antibodies comprising the variable regions of the invention fused or chimerized with antibody regions (eg, constant regions) from another human or non-human species (eg, mouse, horse, rabbit, dog, cow, chicken).

[0200] The term "antibody" also relates to recombinant human antibodies, xenoantibodies and heterohybrid antibodies. The term "recombinant human antibody" includes all human sequence antibodies prepared, expressed, created or separated by recombinant means, such as antibodies separated from transgenic animals (e.g., mice) with respect to human immunoglobulin genes; antibodies expressed using recombinant expression vectors transfected into host cells; antibodies separated from recombinant combined human antibody libraries; or antibodies prepared, expressed, created or separated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable regions and constant regions (if present) derived from human germline immunoglobulin sequences. However, such antibodies can undergo in vitro mutagenesis (or, when using transgenic animals with respect to human Ig sequences, in vivo somatic cell mutagenesis), and therefore the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that may not naturally exist in the human antibody germline repertoire in vivo (although derived from and related to human germline VH and VL sequences).

[0201] "Xenogeneic antibodies" are defined in relation to the transgenic non-human organism that produces such antibodies. The term refers to antibodies that have an amino acid sequence or encoding nucleic acid sequence corresponding to that found in an organism not consisting of the transgenic non-human animal, and are typically from a species other than that of the transgenic non-human animal.

[0202] The term "heterohybrid antibody" refers to an antibody having light chains and heavy chains of different biological origin. For example, an antibody having a human heavy chain associated with a murine light chain is a heterohybrid antibody. Examples of heterohybrid antibodies include chimeric antibodies and humanized antibodies.

[0203] The term "antibody" also relates to humanized antibodies. The "humanized" form of non-human (e.g., murine or rabbit) antibodies is a chimeric immunoglobulin, immunoglobulin chain or its fragment comprising a minimal sequence derived from a non-human immunoglobulin. Often, humanized antibodies are such human immunoglobulins (acceptor antibodies), wherein the residues from the complementary determining region (CDR) of the recipient are replaced by residues (which have desired specificity, affinity and ability) from the CDR (donor antibody) of non-human species such as mice, rats or rabbits. In some cases, the Fv framework residues of human immunoglobulins are replaced by corresponding non-human residues. Further, humanized antibodies can include residues that are neither found in the CDR or framework sequences of the recipient antibodies nor in the input. These modifications are carried out to further refine and optimize antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, wherein all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin, and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. A humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see: Jones Nature 321 (1986), 522-525; Reichmann Nature 332 (1998), 323-327; and Presta Curr Op Struct Biol 2 (1992), 593-596.

[0204] Popular methods for humanization of antibodies involve CDR transplantation, in which a functional antigen binding site from a non-human "donor" antibody is transplanted onto a human "acceptor" antibody. In addition, humanization of antibodies involves the replacement of amino acid residues in the framework region or in the transplanted CDRs to improve the adaptation / matching of the transplanted CDRs to the framework scaffold of the acceptor variable region. Such adaptation / matching typically restores the original affinity of the donor antibody, or even improves it; see, e.g., EP 0 451 216. CDR transplantation methods are known in the art and are described, e.g., in US 5,225,539, US 5,693,761, and US 6,407,213. Another related method is to produce humanized antibodies from transgenic animals that are genetically engineered to contain one or more humanized immunoglobulin loci capable of undergoing gene rearrangement and gene conversion (see, e.g., US 7,129,084).

[0205] Thus, in the context of the present invention, the term "antibody" relates to whole immunoglobulin molecules as well as to parts of such immunoglobulin molecules.

[0206] However, as mentioned above, the antibody preferably comprises an Fc region of an antibody.

[0207] Further, as discussed above, the term relates to modified and / or altered antibody molecules. The term also relates to antibodies generated / synthesized recombinantly or synthetically. The term also relates to intact antibodies.

[0208] Since the antibodies of the present invention contain at least one antibody Fc portion, the term does not refer to intact antibodies or antibody fragments thereof, such as separated light and heavy chains, Fab, Fv, Fab', Fab'-SH, F(ab')2.

[0209] The term "antibody" also includes, but is not limited to, fully human antibodies, chimeric antibodies, humanized antibodies, CDR-grafted antibodies, and antibody constructs, such as single-chain Fv (scFv) or antibody fusion proteins.

[0210] In a preferred embodiment, the antibody according to the invention is a full length antibody, ie a complete immunoglobulin molecule, which is often also referred to as a complete antibody.

[0211] In the context of the present invention, a "single-chain Fv" or "scFv" antibody fragment has a V H and V L domains, wherein these domains are present in a single polypeptide chain. Typically, the scFv polypeptide further comprises a V H and V LA polypeptide linker between the domains which enables the scFv to form the desired structure for antigen binding. Techniques for producing single chain antibodies are described, for example, in Plückthun, in The Pharmacology of Monoclonal Antibodies, Rosenburg and Moore, eds. Springer-Verlag, NY (1994), 269-315.

[0212] As used herein, a "Fab fragment" consists of: a light chain; and a C H 1 region and the variable region. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule.

[0213] The "Fc" region comprises two heavy chain fragments, which comprise the C H 2 and C H 3 domains. The two heavy chain fragments are connected by two or more disulfide bonds and by C H The 3 domains are held together by hydrophobic interactions.

[0214] The "Fab' fragment" described in the context of the present invention comprises the following parts: a light chain; and a portion of a heavy chain, which comprises V H Domain and C H 1 domain and also in C H 1 and C H The region between the two Fab' domains allows for the formation of an interchain disulfide bond between the two heavy chains of the two Fab' fragments to form a F(ab')2 molecule.

[0215] The "F(ab')2 fragment" described in the context of the present invention comprises the following parts: two light chains; and two heavy chains, which are contained in C H 1 and C H The F(ab')2 fragment consists of two Fab' fragments held together by the disulfide bond between the two heavy chains.

[0216] The "Fv region" described in the context of the present invention comprises the variable regions from both the heavy and the light chain, but lacks the constant region.

[0217] The antibodies, antibody constructs, antibody fragments, antibody derivatives (all Ig-derived) or their corresponding immunoglobulin chains to be used according to the present invention may be further modified using conventional techniques known in the art, for example by using amino acid deletions, insertions, substitutions, additions, and / or recombination and / or any other modifications known in the art, alone or in combination. Methods for introducing such modifications in the DNA sequence underlying the amino acid sequence of the immunoglobulin chain are well known to those skilled in the art; see, for example, Sambrook (1989) (loc. cit.). The term "Ig-derived domain" particularly relates to a (poly)peptide construct comprising at least one CDR. The described fragments or derivatives of the Ig-derived domain define a (poly)peptide that is part of the above-mentioned antibody molecule and / or modified by chemical / biochemical or molecular biological methods. Corresponding methods are known in the art and are described, inter alia, in laboratory manuals (see Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 2nd edition (1989) and 3rd edition (2001); Gerhardt et al., Methods for General and Molecular Bacteriology ASM Press (1994); Lefkovits, Immunology Methods Manual: The Comprehensive Sourcebook of Techniques, Academic Press (1997); Golemis, Protein-Protein Interactions: A Molecular Cloning Manual, Cold Spring Harbor Laboratory Press (2002)).

[0218] As already outlined above, in accordance with the basic principles of the present invention, the present invention is not particularly limited to a certain "infectious disease" or a certain "malignant disease". As a direct result thereof, the skilled person will understand that the vaccine of the present invention is not particularly limited to "antibodies against specific antigens associated with infectious diseases or malignant diseases".

[0219] Therefore, the term "antigen associated with the infectious disease or malignant disease" according to the present invention is not particularly limited to a specific antigen, but may be any antigen associated with the infectious disease or malignant disease.

[0220] The "antigen associated with the infectious disease" may be derived from the factors causing the infectious disease described above (e.g., the bacteria, virus, fungus or parasite described above), that is, it may be a part or structure or domain of the factor causing the infectious disease that is capable of inducing an immune response.

[0221] The "antigen associated with the malignant disease" may be derived from the malignant disease described above, ie, may be, for example, a part, structure or domain of a malignant cell that is capable of eliciting an immune response.

[0222] Therefore, the skilled person will understand that the vaccine of the present invention can be adjusted to any "infectious disease" or "malignant disease", more particularly to the corresponding "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively.

[0223] Therefore, in a preferred embodiment, the "antibodies against antigens associated with infectious diseases or malignant diseases" according to the present invention are antibodies that specifically bind to "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, or specifically recognize "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, or specifically interact with them.

[0224] The terms "bind to", "specifically bind to", "specifically recognize" or "specifically interact with" as used in the context of the present invention are used interchangeably herein and are defined further below.

[0225] Therefore, the antibody used in the context of the present invention is not particularly limited as long as it is an antibody against "an antigen associated with the infectious disease" and "an antigen associated with the malignant disease", respectively.

[0226] Therefore, the antibody may be any antibody that specifically binds to "antigens associated with the infectious disease" and "antigens associated with the malignant disease", or specifically recognizes or specifically interacts with "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively. Preferably, the antigen, i.e., the domain, is a surface antigen.

[0227] A skilled person is easily able to generate such antibodies against a given domain (i.e., "an antigen associated with the infectious disease" and "an antigen associated with the malignant disease", respectively), and determine whether the respective antibodies are able to detect / bind to the given "an antigen associated with the infectious disease" and "an antigen associated with the malignant disease", respectively, based on the common knowledge of the skilled person and the methods described above.

[0228] The generation of specific antibodies against "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, can be based on, for example, immunization of animals (e.g., mice). However, other animals for producing antibodies / antisera are also contemplated within the present invention. For example, monoclonal and polyclonal antibodies can be produced by rabbits, mice, goats, donkeys, etc. The polynucleotides encoding the correspondingly selected polypeptides of "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, can be subcloned into suitable vectors, wherein the recombinant polypeptides are to be expressed in organisms capable of expression, such as bacteria. Therefore, the expressed recombinant protein can be injected intraperitoneally into mice, and the resulting specific antibodies can be obtained, for example, from mouse serum provided by intracardiac blood puncture. The present invention also contemplates the generation of specific antibodies against natural polypeptides and recombinant polypeptides by using DNA vaccine strategies, as illustrated in the attached examples. DNA vaccine strategies are well known in the art and include liposome-mediated delivery, injection by gene gun or jet injection, and intramuscular or intradermal injection. Therefore, antibodies against polypeptides or proteins or epitopes of "antigens associated with the infectious disease" and "antigens associated with the malignant disease" can be obtained by directly immunizing animals, and the immunization is carried out by directly injecting intramuscularly a vector expressing the desired polypeptide or protein or epitope of "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively. The amount of specific antibodies obtained can be quantified by using ELISA, which is also described below in this article. Further methods for producing antibodies are well known in the art, see, for example, Harlow and Lane, "Antibodies, A Laboratory Manual", CSH Press, Cold Spring Harbor, 1988.

[0229] As used herein, the term "specifically binds" refers to a binding reaction that is decisive for the presence of the desired "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, and antibodies, in the presence of a heterogeneous population of proteins and other biological products.

[0230] Thus, under the specified assay conditions, the specifically specified antibodies and the corresponding "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, bind to each other and do not bind to other components present in the sample in significant amounts. Specific binding to the target analyte under such conditions may require a binding moiety that is selected for its specificity for a particular target analyte. A variety of immunoassay formats can be used to select antibodies that specifically react with a particular antigen. For example, solid phase ELISA immunoassays are routinely used to select monoclonal antibodies that specifically immunoreact with analytes. For a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity, see Shepherd and Dean (2000), Monoclonal Antibodies: A Practical Approach, Oxford University Press; and / or Howard and Bethell (2000) Basic Methods in Antibody Production and Characterization, Crc. Pr. Inc. Typically, a specific or selective reaction will be at least twice the background signal-to-noise ratio, and more typically more than 10 to 100 times the background. Those skilled in the art are able to provide and generate specific binding molecules for new polypeptides. For specific binding assays, it can be easily adopted to avoid undesirable cross-reactivity, for example polyclonal antibodies can be easily purified and selected by known methods (see Shepherd and Dean, loc. cit.).

[0231] According to the present invention, the term "antibodies against antigens associated with the infectious disease or malignant disease" means that the antibody molecule or its antigen-binding fragment is able to specifically recognize at least two amino acids of "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, or specifically interact and / or bind thereto. The term relates to the specificity of the antibody molecule, i.e., its ability to distinguish specific regions of "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively. Therefore, specificity can be experimentally determined by methods known in the art and disclosed and described herein. Such methods include, but are not limited to, Western blotting, ELISA-tests, RIA-tests, ECL-tests, IRMA-tests and peptide scanning. Such methods also include K D-value determination, as especially illustrated in the accompanying examples. Peptide scanning (pepspot assay) is routinely used to map linear epitopes in polypeptide antigens. The primary sequence of the polypeptide is successively synthesized on activated cellulose with overlapping peptides. The antibody to be tested for its ability to detect or recognize a specific antigen / epitope is scored for the recognition of certain peptides by the following formula: conventional color development (secondary antibody with horseradish peroxidase and 4-chloronaphthol and hydrogen peroxide), chemiluminescent reaction, or similar means known in the art. In the case of chemiluminescent reaction, in particular, the reaction can be quantitative. If the antibody reacts with a certain set of overlapping peptides, the minimum amino acid sequence necessary for the reaction can be inferred. The same assay can reveal two long-range clusters of reactive peptides, which indicates the recognition of discontinuous (i.e. conformational) epitopes in the antigen polypeptide (Geysen (1986), Mol. Immunol. 23, 709-715).

[0232] The term "binding to..." used in the context of the present invention defines the binding (interaction) of at least two "antigen interaction sites" to each other. According to the present invention, the term "antigen interaction site" defines a motif of a polypeptide, i.e. a part of an antibody or antigen-binding fragment of the present invention, which exhibits the ability to specifically interact with a specific antigen or a specific group of antigens of "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively. The binding / interaction is also understood to define "specific recognition". According to the present invention, the term "specifically recognizes" means that the antibody is able to specifically interact and / or bind to at least two amino acids of each of the "antigens associated with the infectious disease" and "antigens associated with the malignant disease", respectively, as defined herein. Antibodies can recognize different epitopes, interact with different epitopes and / or bind to different epitopes.

[0233] The term "specific interaction" as used according to the present invention means that the antibody or antigen-binding fragment thereof of the present invention does not or substantially does not cross-react with a (poly)peptide having a similar structure. Thus, the antibody or antigen-binding fragment thereof of the present invention specifically binds / interacts with the structure of "an antigen associated with the infectious disease" and "an antigen associated with the malignant disease", respectively, as defined above. Specific examples of such molecules to which the first and second Ig-derived domains are directed are given herein below.

[0234] The cross-reactivity of the panel of antibodies or antigen-binding fragments thereof under investigation can be tested, for example, by evaluating the binding of the panel of antibodies or antigen-binding fragments thereof to the (poly)peptide of interest and to a number of more or less closely related (poly)peptides (structurally and / or functionally) under conventional conditions (see, for example, Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1988); and Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1999)). Only those constructs (i.e., antibodies, antigen-binding fragments thereof, etc.) that bind to a certain structure of HSV, such as a specific epitope or (poly)peptide / protein of HSV, and not or substantially not to any other epitope or (poly)peptide of the same HSV are considered to be specific for the epitope or (poly)peptide / protein of interest and are selected for further investigation according to the methods provided herein. These methods may include, among others, binding studies, blocking and competition studies with molecules that are closely related structurally and / or functionally. These binding studies also include FACS analysis, surface plasmon resonance (SPR, e.g. ), analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, fluorescence spectroscopy, or radiolabeled ligand binding assays.

[0235] The term "binding to..." refers not only to linear epitopes but also to conformational epitopes, structural epitopes or discontinuous epitopes consisting of two regions of a human target molecule or a portion thereof. In the context of the present invention, a conformational epitope is defined by two or more discrete amino acid sequences separated in the primary sequence, which come together on the surface of the molecule when the polypeptide is folded into a native protein (Sela, Science 166 (1969), 1365; and Laver, Cell 61 (1990), 553-536). In addition, the term "binding to..." is used interchangeably with the term "interacting with..." or "recognizing..." in the context of the present invention.

[0236] Therefore, specificity can be determined experimentally by methods known in the art and described herein. Such methods include, but are not limited to, Western blots, ELISA-tests, RIA-tests, ECL-tests, IRMA-tests and peptide scanning.

[0237] Fc-containing fusion protein comprising an antibody fused to an antigen associated with the infectious disease or malignant disease Fc part:

[0238] Regarding (b) , i.e., an Fc fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0239] The fusion protein according to (b) comprises at least two molecules, namely, "the Fc portion of an antibody" and "an antigen associated with the infectious disease or malignant disease".

[0240] The term "an antigen associated with said infectious disease or malignant disease" has been defined above in the context of (a), ie in the context of antibodies against "an antigen associated with said infectious disease or malignant disease".

[0241] Concerning the preferred embodiments of "antigens associated with said infectious or malignant disease", the same applies mutatis mutandis as already explained above in the context of antibodies against "antigens associated with said infectious or malignant disease" as defined above.

[0242] Therefore, in a preferred embodiment, in the vaccine for use according to the first aspect above, the antigen associated with the malignant disease is selected from the group consisting of the following: tumor-associated antigens known in the art, including but not limited to carbonic anhydrase IX, CCCL19, CCCL21, CSAp, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, IGF-1R, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, C D37, CD38, CD40, CD40L, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD74, CD79a, CD80, CD83, CD95, CD126, CD133, CD138, CD147, CD154, AFP, PSMA, CEACAM5, CEACAM6, B7, ED-B of fibronectin, H factor, FHL-1, Flt-3, folate receptor, GROB, HER2, HMGB-1, hypoxia-inducible factor (HIF), HM1.24, Insulin-like growth factor-1 (ILGF-1), IFN-γ, IFN-α, IFN-β, IL-2, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-6, IL-8, IL-12, IL-15, IL-17, IL -18, IL-25, IP-10, mCRP, MCP-1, MIP-1A, MIP-1B, MIF, MUC1, MUC2, MUC3, MUC4, MUC5ac, PAM4 antigen, NCA-95, NCA-90, NY-ESO-1, Ia, HM1.24, EGP-1, EGP-2, HLA-DR, tenascin, Le(y), RANTES, T101, TAC, Tn antigen, Thomson-Friedenreich antigen, tumor necrosis antigen, TNF-α, TRAIL receptor (R1 and R2), VEGFR, EGFR, PlGF, complement factor C3, C3a, C3b, C5a, C5, cancer-testis (CT) antigen SPAG9, CT9, CT10, LAGE, MAGE-B5, MAGE-B6, MAGE-C2, MAGE-C3, MAGE-D, HAGE, SAGE and oncogene products.

[0243] In addition, regarding the term "antigen associated with the infectious disease", in a preferred embodiment, in the vaccine for use according to the first aspect above, the antigen associated with the infectious disease is selected from the group consisting of:

[0244] Epstein-Barr virus (EBV) (derived) proteins or protein domains, preferably EBNA1, EBNA2, EBNA3A, EBNA3B, EBNA3C, EBNA-LP, LMP1, LMP2A / B, gp350, A73, RPMS1, ZEBRA, Rta, major intercalated protein p143, major intercalated protein, intercalated protein, major capsid protein, minor capsid protein, capsid protein p18, p23, p40, gN, gp42, gM, gp60, gp78 / 55, gp150, 53 / 55kd membrane protein, viral IL-10, gB, gH, gL;

[0245] Human papillomavirus (HPV) (derived) proteins or protein domains, preferably E1, E2, E4, E5, E6, E7, L1, L2;

[0246] Human immunodeficiency virus 1 or 2 (derived) proteins or protein domains, preferably Gag, Pol, Env, gp160, gp120, gp41, CA, MA, p2, p6, NC, IN, RTp66, RTp55, RTp51, PR, Rev, Tat, Nef, Vif, Vpr, Vpu, Vpx;

[0247] Herpes simplex virus (HSV) 1 or 2 (derived) proteins or protein domains, preferably gB, gC, gD, gH, gG, gL, gE, gI, gK, gM, VP1-2, ICP32, ICP0, VP11 / 12, UL13, vhs, VP16, US3, VP22, ICP34.5, US11, ICP4, DNA polymerase, major capsid protein, helicase, primase, uracil DNA glycosylase, dUTPase, ribonucleotide reductase, major intercalated protein;

[0248] A human cytomegalovirus (HCMV) (derived) protein or protein domain encoded by an open reading frame (ORF), preferably UL57, UL55, UL54, UL75, UL86, UL85, UL104, UL97, UL98, UL100, UL105, UL102, UL114, UL115, UL72, UL70, UL69, UL44, UL45, UL48;

[0249] Hepatitis A virus (HAV) (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3Cpro, P3-3Dpol;

[0250] Hepatitis B virus (HBV) (derived) proteins or protein domains, preferably M, L, S, Polymerase, TP, Spacer, RT, RNase H, preCore, Core, X, HBeAg;

[0251] Hepatitis C virus (HCV) (derived) proteins or protein domains, preferably E1, E2, core, NS1, NS2, NS3, NS4a, NS4b, NS5a, NS5b, precursor polyprotein;

[0252] Influenza A virus (IAV) (derived) proteins or protein domains, preferably HA, NA, M2, M1, NP, NS1, NS2 / NEP, PA, PB1;

[0253] Measles virus (MV) (derived) protein or protein domain, preferably N, P / C / V, M, F, H, L;

[0254] Respiratory syncytial virus (RSV) (derived) protein or protein domain, preferably NS1, NS2, N, P, M, SH, G, F, M21, M22, L;

[0255] Rotavirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, RdRp, VP5, VP6, VP7, VP8, NSP1, NSP2, NSP3, NSP4, NSP5, NSP6;

[0256] Severe acute respiratory syndrome coronavirus (SARS-Cov) type 1 and 2 (derived) proteins or protein domains, preferably ns1, ns2, PLpro, ns4, 3CL, ns6, ns7, ns8, ns9, ns10, RdRp, Hel, ns14, ns15, ns16, S, S-RBD, 3a, E, M, 6, 7a, 7b, 8N, 9b, 14;

[0257] Varicella zoster virus (VZV) (derived) proteins or protein domains encoded by open reading frames (ORFs), preferably ORF0 to ORF71 (at least 71 proteins in total);

[0258] Human herpesvirus 8 (HHV-8) (derived) proteins or protein domains, preferably K1, K2, K3, K4, K4.1, K5, K6, K7, K8, K8.1, K9, K10, K10.5, K11, K12, K13, K14, K15, gB, gL, gH and proteins encoded in ORFs, including but not limited to ORF2, ORF9, ORF10, ORF16, ORF18, ORF24, ORF30, ORF31, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF42, ORF43, ORF44, ORF45, ORF46, ORF47, ORF48, ORF49, ORF50, ORF51, ORF52, ORF53, ORF54, ORF55, ORF56, ORF57, ORF58, ORF59, ORF60, ORF61, ORF62, ORF63, ORF64, ORF65, ORF66, ORF67, ORF68, ORF69, ORF70, ORF71, ORF72, ORF73, ORF74, ORF75 RF66, ORF21, ORF23, ORF25, ORF26, ORF65, ORF33, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF 42. ORF45, ORF49, ORF50, ORF52, ORF53, ORF55, ORF57, ORF59, ORF67, ORF69, ORF70, ORF72, ORF73, ORF74, ORF75;

[0259] Human herpesvirus 6 (HHV-6) (derived) protein or protein domain encoded by ORF, preferably DR1, DR6, DR7 / U1, U2, U3, U4, U7, U10, U11, U12, U13, U14, U15, U17, U18, U19, U20, U21, U22, U23, U24, U25, U26, U27, U28, U29, U30, U31, U32, U33, U34, U35, U36, U37, U38, U39, U40 , U41, U42, U43, U44, U45, U46, U47, U48, U49, U50, U51, U52, U53, U54, U55, U56, U57, U58, U59, U61, U62, U63, U64, U65, U66, U69, U70, U71, U72, U73, U74, U75, U76, U77, U79, U81, U82, U83, U85, U86, U88, U90, U91, U94, U95, U100;

[0260] Rabies virus (derived) protein or protein domain, preferably N, P, M, G, L;

[0261] Mumps virus (derived) proteins or protein domains, preferably N, V / P, M, F, HN, L; and

[0262] Rhinovirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3C, P3-3D.

[0263] With respect to the second module, i.e., the Fc portion of an antibody, the term "Fc portion of an antibody" in its broadest sense refers to the portion comprising two C H 2 and C H The two heavy chain fragments are linked by two or more disulfide bonds and by C H The 3 domains are held together by hydrophobic interactions.

[0264] When fused to an antigen associated with the infectious or malignant disease according to the present invention, any Fc region or Fc portion of an antibody can be used, as long as the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0265] Assays for assessing the ability of the Fc portion of an antibody to bind to a receptor present on or in an antigen presenting cell (APC) are known in the art.

[0266] Without being bound by theory, the ability of the Fc portion of an antibody to bind to a receptor present on or in an antigen presenting cell (APC) can be assessed as follows:

[0267] A fusion protein consisting of an FcR extracellular domain coupled to the intracellular signaling domain of the murine T-cell receptor (CD3ζ) is overexpressed in murine BW5147 cells. Interaction of the Fc portion of the antibody within the immune complex with the extracellular FcR domain induces signaling via the CD3ζ domain and IL-2 is secreted into the culture supernatant. IL-2 in the supernatant can be measured using an ELISA, in which anti-IL-2 antibodies are coated on a plate and IL-2 in the supernatant binds to these antibodies and can be detected using a secondary antibody coupled to an enzyme (usually horseradish peroxidase). After addition of a substrate, a photometer can be used to measure the color change. Detailed methods are described in

[56] . The ability to bind to TRIM21 can be measured in vitro using SEC-MALS as described in the art (e.g., in Mallery et al., 2010

[61] ). Cytoplasmic engagement of TRIM21 can be measured by using the TRIMaway assay described in the art (Clift et al., 2017

[62] ) and comparing cells in which TRIM21 expression has been knocked down or knocked out to cells in which TRIM21 is expressed, regarding the fate of the antigen bound by the antibody as described for antibodies interacting with adenovirus and degradation of adenovirus proteins following incubation of cells expressing TRIM21 (but not in cells lacking TRIM21) with adenovirus-antibody complexes; see, e.g., Mallery et al., 2010

[61] .

[0268] In a preferred embodiment, the Fc portion of the antibody is capable of binding to a receptor present on or in an APC, the receptor being selected from the group consisting of: type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0269] In a preferred embodiment, the receptor present on or in the APC may have the following amino acid sequence selected from the group consisting of:

[0270] amino acid sequences of activating FcγRI transcript variants 1 to 9 (SEQ ID NOs: 17 to 25);

[0271] The amino acid sequence of FcγRIIa (SEQ ID NOs: 26 to 29);

[0272] The amino acid sequence of FcγRIIc (SEQ ID NO: 30);

[0273] The amino acid sequence of FcγRIIIa (SEQ ID NOs: 31 to 34);

[0274] The amino acid sequence of FcγRIIIb (SEQ ID NOs: 35 to 38);

[0275] amino acid sequence of inhibitory FcγRIIb (SEQ ID NOs: 39 to 50);

[0276] the amino acid sequence of a neonatal FcR (FcRn) (SEQ ID NO: 51); and

[0277] Amino acid sequence of cytoplasmic TRIM21 (SEQ ID NO: 52).

[0278] The receptor present on or in APC to which the Fc portion of the above-mentioned antibody can bind is not particularly limited to the amino acid sequence of any one of SEQ ID NOs: 17 to 52, but may also be a receptor sequence comprising or consisting of the amino acid sequence of any one of SEQ ID NOs: 17 to 52 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NOs: 17 to 52, as long as the Fc portion of the antibody has the ability to bind to the receptor present on or in the APC described herein above and below.

[0279] The same applies to determining whether the Fc portion of the antibody has the ability to bind to the receptor present on or in the APC, as described above. The corresponding assay for determining this ability has also been described above, which is suitable for determining whether the Fc portion of the antibody has the ability to bind to any of the receptors present on or in the APC described above after appropriate modification.

[0280] In order to determine whether an amino acid sequence has a certain degree of identity with the sequence of any one of SEQ ID NO: 17 to 52, the skilled person can use means and methods well known in the art, such as alignment, either manually or by using computer programs known to those skilled in the art. Such alignment can be performed, for example, using means and methods known to the skilled person, such as by using known computer algorithms, such as the Lipman-Pearson method (Science 227 (1985), 1435) or the CLUSTAL algorithm. Preferably, in such an algorithm, the maximum homology is assigned to the conserved amino acid residues present in the amino acid sequence. In a preferred embodiment, ClustalW2 is used to compare amino acid sequences. In the case of paired comparisons / alignments, the following settings are preferably selected: protein weight matrix: BLOSUM 62; gap opening: 10; gap extension: 0.1. In the case of multiple comparisons / alignments, the following settings are preferably selected: protein weight matrix: BLOSUM 62; gap opening: 10; gap extension: 0.2; gap distance: 5; no end gap.

[0281] According to the present invention, in the context of two or more nucleic acid or amino acid sequences, the term "identical" or "identity percentage" refers to two or more sequences or subsequences that are identical or have a specified percentage of identical amino acid residues or nucleotides (e.g., 60% or 65% identity, preferably 70-95% identity, more preferably at least 95% identity to the nucleic acid sequence or amino acid sequence described above), when compared and aligned for maximum correspondence on a comparison window or on a designated region, as measured by using a sequence comparison algorithm known in the art or by manual alignment and visual inspection. Sequences with, for example, 60% to 95% or greater sequence identity are considered to be substantially identical. Such a definition also applies to the complement of the test sequence. Preferably, the described identity is present in a region of at least about 15 to 25 amino acids or nucleotides in length, more preferably in a region of about 50 to 100 amino acids or nucleotides in length. A person skilled in the art will know how to determine the percent identity between / among sequences by using, for example, algorithms known in the art, such as those based on the CLUSTALW computer program (Thompson Nucl. Acids Res. 2 (1994), 4673-4680) or FASTDB (Brutlag Comp. App. Biosci. 6 (1990), 237-245).

[0282] Although the FASTDB algorithm typically does not take into account internal non-matching deletions or additions, i.e., gaps, in its calculations, this can be manually corrected to avoid overestimation of % identity. However, CLUSTALW takes into account sequence gaps in its identity calculations. BLAST and BLAST 2.0 algorithms are also available to those skilled in the art (Altschul, (1997) Nucl. Acids Res. 25: 3389-3402; Altschul (1993) J. Mol. Evol. 36: 290-300; Altschul (1990) J. Mol. Biol. 215: 403-410). The BLASTN program for nucleic acid sequences uses the following parameters as default values: a word length (W) of 11, an expectation value (E) of 10, M=5, N=4, and a comparison of two chains. For amino acid sequences, the BLASTP program uses the following parameters as default values: a word length (W) of 3, and an expectation value (E) of 10. The BLOSUM62 scoring matrix (Henikoff (1989) PNAS 89:10915) used the following parameters: alignment of 50 (B), expectation of 10 (E), M=5, N=4, and comparison of both strands.

[0283] Preferably, the amino acid substitution is a "conservative substitution", which refers to the substitution of an amino acid in a protein with other amino acids having similar characteristics (e.g., charge, side chain size, hydrophobicity / hydrophilicity, main chain conformation and rigidity, etc.), so that the change can often be made without changing the biological activity of the protein. Those skilled in the art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially change the biological activity (see, e.g., Watson, Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co. 4th edition (1987), 224). In addition, substitutions of amino acids that are structurally or functionally similar are less likely to destroy biological activity. In the context of the present invention, the binding compounds / antibodies of the present invention comprise polypeptide chains having sequences that include up to 0 (no change), 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20 or more conservative amino acid substitutions when compared to specific amino acid sequences disclosed herein, e.g., SEQ ID NO: 9 (which refers to the variable region of the heavy chain of an antibody) and 10 (which refers to the variable region of the light chain of an antibody). As used herein, the phrase "up to X" conservative amino acid substitutions includes 0 substitutions and any number of substitutions up to 10, including 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 substitutions.

[0284] Such exemplary substitutions are preferably made according to those set forth in Table 1 below.

[0285] Table 1 Exemplary conservative amino acid substitutions

[0286]

[0287]

[0288] As outlined above, the Fc portion of the fusion protein is capable of binding to a receptor present on or in an antigen presenting cell (APC), as just defined in the preceding embodiments.

[0289] Without being bound by theory, in a preferred embodiment, the Fc portion of the fusion protein may be an Fc domain known in the art to bind to one or more of the above-mentioned receptors.

[0290] In a further preferred embodiment, the Fc portion of the fusion protein may be an Fc domain having an amino acid sequence selected from the group consisting of:

[0291] HDIT101 Fc (SEQ ID NO: 53);

[0292] HDIT101 Fc G236A (SEQ ID NO: 54) (in SEQ ID NO: 54, the mutation G236A in the Fc region is located at position G16A);

[0293] HDIT101 Fc G236A / S239D / I332E (SEQ ID NO: 55) (in SEQ ID NO: 55, the mutations G236A / S239D / I332E in the Fc region are located at positions G16A / S19D / I112E, respectively).

[0294] Therefore, in a preferred embodiment, the Fc portion of the Fc-containing fusion protein comprises the Fc portion of an antibody, wherein the Fc portion of the antibody preferably is / has the following amino acid sequence selected from the group consisting of: SEQ ID NOs: 53 to 55.

[0295] The Fc portion of the Fc-containing fusion protein is not particularly limited to the amino acid sequence of any one of SEQ ID NOs: 53 to 55, but may also be the Fc portion of an antibody of SEQ ID NOs: 53 to 55 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NOs: 53 to 55, as long as the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in the APC described herein above and below.

[0296] The same applies to determining whether the Fc portion of the fusion protein has the ability to bind to a receptor present on or in an APC, as described above. The corresponding assay for determining this ability has also been described above, which is suitable for determining whether any of the Fc portions of the fusion protein described above has the ability to bind to a receptor present on or in an APC after appropriate modification.

[0297] With regard to the determination of sequence identity, the same applies as already explained above.

[0298] In a further preferred embodiment, the Fc portion of the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO:54, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NO:54, wherein the amino acid residue at position 16 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO:54 is substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 16 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO:54 is substituted with amino acid A.

[0299] In a further preferred embodiment, the Fc portion of the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO:55, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NO:55, wherein the amino acid residues at positions 16, 19 and / or 112 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:55 are substituted with another amino acid residue, and wherein the Fc portion of the antibody has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 16 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:55 is substituted with amino acid A. Preferably, the amino acid residue at position 19 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:55 is substituted with amino acid D. Preferably, the amino acid residue at position 112 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 55 is substituted with amino acid E.

[0300] In a further preferred embodiment, the Fc-containing fusion protein may have an amino acid sequence selected from the group consisting of:

[0301] HSV-1gB extracellular domain fused to HDIT101 Fc domain (SEQ ID NO:56);

[0302] HSV-1 gB extracellular domain fused to HDIT101 Fc domain with G236A (SEQ ID NO: 57) (in SEQ ID NO: 57, the mutation G236A in the Fc region is located at position G724A);

[0303] HSV-1 gB extracellular domain fused to HDIT101 Fc domain with G236A / S239D / I332E (SEQ ID NO:58) (in SEQ ID NO:58, the mutations G236A / S239D / I332E in the Fc region are located at positions G724A / S727D / I820E, respectively);

[0304] HSV-2gB extracellular domain fused to HDIT101 Fc domain (SEQ ID NO:59);

[0305] HSV-2 gB extracellular domain fused to HDIT101 Fc domain with G236A (SEQ ID NO: 60) (in SEQ ID NO: 60, the mutation G236A in the Fc region is located at position G726A); and

[0306] HSV-2 gB extracellular domain fused to HDIT101 Fc domain with G236A / S239D / I332E (SEQ ID NO:61) (in SEQ ID NO:61, the mutations G236A / S239D / I332E in the Fc region are located at G726A / S729D / I822E, respectively).

[0307] Therefore, in a preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence selected from the group consisting of: SEQ ID NOs: 56 to 61.

[0308] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of any one of SEQ ID NOs: 56 to 61, but may also be an Fc-containing fusion protein of SEQ ID NOs: 56 to 61 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NOs: 56 to 61, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described above and below in this document.

[0309] The same applies to determining whether the Fc portion of the fusion protein has the ability to bind to a receptor present on or in an APC, as described above. The corresponding assay for determining this ability has also been described above, which is suitable for determining whether any of the Fc portions of the fusion protein described above has the ability to bind to a receptor present on or in an APC after appropriate modification.

[0310] With regard to the determination of sequence identity, the same applies as already explained above.

[0311] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 57, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of SEQ ID NO: 57, wherein the amino acid residue at position 724 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 57 is substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 724 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 57 is substituted with amino acid A.

[0312] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO:58, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of the sequences in SEQ ID NO:58, wherein the amino acid residues at positions 724, 727 and / or 820 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:58 are substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 724 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:58 is substituted with amino acid A. Preferably, the amino acid residue at position 727 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:58 is substituted with amino acid D. Preferably, the amino acid residue at position 820 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 58 is substituted with amino acid E.

[0313] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO:60, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of SEQ ID NO:60, wherein the amino acid residue at position 726 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO:60 is substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 726 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO:60 is substituted with amino acid A.

[0314] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 61, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of the sequences in SEQ ID NO: 61, wherein the amino acid residues at positions 726, 729 and / or 822 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO: 61 are replaced by another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 726 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO: 61 is replaced by amino acid A. Preferably, the amino acid residue at position 729 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO: 61 is replaced by amino acid D. Preferably, the amino acid residue at position 822 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 61 is substituted with amino acid E.

[0315] The same applies to determining whether the Fc portion of the fusion protein has the ability to bind to a receptor present on or in an APC, as described above. The corresponding assay for determining this ability has also been described above, which is suitable for determining whether any of the Fc portions of the fusion protein described above has the ability to bind to a receptor present on or in an APC after appropriate modification.

[0316] With regard to the determination of sequence identity, the same applies as already explained above.

[0317] The addition of cell penetrating peptides has been shown to induce better MHC-I presentation. Two of the known peptides for inducing this are Tat and PTD-4 peptides from HIV-1 (Del Gaizo et al., 2003

[63] ). Therefore, in a preferred embodiment, it is contemplated to add one or more protein transduction peptides (preferably Tat peptides and / or PTD-4 peptides) to the Ag-Fc fusion protein defined herein above to further enhance MHC-I presentation.

[0318] Therefore, in a preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0319] HSV-1 gB extracellular domain fused to Tat peptide and HDIT101 Fc domain (SEQ ID NO: 62).

[0320] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:62, but may also be an Fc-containing fusion protein of SEQ ID NO:62 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:62, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described herein above and below.

[0321] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0322] HSV-1 gB extracellular domain fused to Tat peptide and HDIT101 Fc domain with G236A (SEQ ID NO: 63) (in SEQ ID NO: 63, the mutation G236A in the Fc region is located at position G733A).

[0323] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 63, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of the sequences in SEQ ID NO: 63, wherein the amino acid residue at position 733 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 63 is substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 733 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 63 is substituted with amino acid A.

[0324] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0325] HSV-1 gB extracellular domain fused to Tat peptide and HDIT101 Fc domain with G236A / S239D / I332E (SEQ ID NO: 64) (in SEQ ID NO: 64, the mutations G236A / S239D / I332E in the Fc region are located at positions G733A / S736D / I829E, respectively).

[0326] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO:64, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NO:64, wherein the amino acid residues at positions 733, 736 and / or 829 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:64 are substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 733 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:64 is substituted with amino acid A. Preferably, the amino acid residue at position 736 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:64 is substituted with amino acid D. Preferably, the amino acid residue at position 829 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 64 is substituted with amino acid E.

[0327] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0328] HSV-2 gB extracellular domain fused to Tat peptide and HDIT101 Fc domain (SEQ ID NO: 65).

[0329] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:65, but may also be an Fc-containing fusion protein of SEQ ID NO:65 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:65, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described herein above and below.

[0330] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0331] HSV-2 gB extracellular domain (SEQ ID NO: 66) fused to Tat peptide and HDIT101 Fc domain with G236A (in SEQ ID NO: 66, the mutation G236A in the Fc region is located at position G735A).

[0332] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 66, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NO: 66, wherein the amino acid residue at position 735 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 66 is substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 735 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 66 is substituted with amino acid A.

[0333] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0334] HSV-2 gB extracellular domain (SEQ ID NO:67) fused to Tat peptide and HDIT101 Fc domain with G236A / S239D / I332E (in SEQ ID NO:67, the mutations G236A / S239D / I332E in the Fc region are located at positions G735A / S738D / I831E, respectively).

[0335] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO:67, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of any one of SEQ ID NO:67, wherein the amino acid residues at positions 735, 738 and / or 831 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:67 are substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 735 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:67 is substituted with amino acid A. Preferably, the amino acid residue at position 738 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO:67 is substituted with amino acid D. Preferably, the amino acid residue at position 831 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 67 is substituted with amino acid E.

[0336] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0337] HSV-1 gB extracellular domain fused to PTD-4 peptide and HDIT101 Fc domain (SEQ ID NO: 68).

[0338] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:68, but may also be an Fc-containing fusion protein of SEQ ID NO:68 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:68, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described above and below in this document.

[0339] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0340] HSV-1 gB extracellular domain fused to PTD-4 peptide and HDIT101 Fc domain with G236A (SEQ ID NO: 69) (in SEQ ID NO: 69, the mutation G236A in the Fc region is located at position G732A).

[0341] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 69, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of the sequences in SEQ ID NO: 69, wherein the amino acid residue at position 732 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 69 is substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in the APC described above and below herein. Preferably, the amino acid residue at position 732 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 69 is substituted with amino acid A.

[0342] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0343] HSV-1 gB extracellular domain fused to PTD-4 peptide and HDIT101 Fc domain with G236A / S239D / I332E (SEQ ID NO: 70) (in SEQ ID NO: 70, the mutations G236A / S239D / I332E in the Fc region are located at positions G732A / S735D / I828E, respectively).

[0344] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 70, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of the sequences in SEQ ID NO: 70, wherein the amino acid residues at positions 732, 735 and / or 828 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO: 70 are substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 732 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 70 is substituted with amino acid A. Preferably, the amino acid residue at position 735 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 70 is substituted with amino acid D. Preferably, the amino acid residue at position 828 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 70 is substituted with amino acid E.

[0345] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0346] HSV-2 gB extracellular domain fused to PTD-4 peptide and HDIT101 Fc domain (SEQ ID NO: 71).

[0347] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:71, but may also be an Fc-containing fusion protein of SEQ ID NO:71 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:71, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described above and below in this document.

[0348] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0349] HSV-2 gB extracellular domain (SEQ ID NO: 72) fused to PTD-4 peptide and HDIT101 Fc domain with G236A (in SEQ ID NO: 72, the mutation G236A in the Fc region is located at position G734A).

[0350] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 72, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of the sequences in SEQ ID NO: 72, wherein the amino acid residue at position 734 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 72 is substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in the APC described above and below herein. Preferably, the amino acid residue at position 734 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 72 is substituted with amino acid A.

[0351] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0352] HSV-2 gB extracellular domain (SEQ ID NO: 73) fused to PTD-4 peptide and HDIT101 Fc domain with G236A / S239D / I332E (in SEQ ID NO: 73, the mutations G236A / S239D / I332E in the Fc region are located at positions G734A / S737D / I830E, respectively).

[0353] In a further preferred embodiment, the Fc-containing fusion protein has an amino acid sequence as shown in SEQ ID NO: 73, or an amino acid sequence having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with any one of the sequences in SEQ ID NO: 73, wherein the amino acid residues at positions 734, 737 and / or 830 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO: 73 are substituted with another amino acid residue, and wherein the Fc portion of the fusion protein has the ability to bind to the Fc receptor present on or in APC as described above and below herein. Preferably, the amino acid residue at position 734 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO: 73 is substituted with amino acid A. Preferably, the amino acid residue at position 737 or at positions corresponding to the positions in the amino acid sequence shown in SEQ ID NO: 73 is substituted with amino acid D. Preferably, the amino acid residue at position 830 or at a position corresponding to the position in the amino acid sequence shown in SEQ ID NO: 73 is substituted with amino acid E.

[0354] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0355] HSV-1F gB extracellular domain fused to IgG1 hinge*-CH2-CH3 (SEQ ID NO: 74);

[0356] HSV-2G gB extracellular domain fused to IgG1 hinge*-CH2-CH3 (SEQ ID NO: 75);

[0357] HSV-1F gB extracellular domain fused to IgG1 hinge*-CH2-CH3-StrepTag (SEQ ID NO: 76);

[0358] HSV-2G gB extracellular domain fused to IgG1 hinge*-CH2-CH3-StrepTag (SEQ ID NO: 77);

[0359] HSV-1F gB extracellular domain fused to StrepTag (SEQ ID NO: 78);

[0360] HSV-2G gB extracellular domain fused to StrepTag (SEQ ID NO: 79).

[0361] The strep-tagged sequence also has a thrombin cleavage site before the StrepTag to remove it. This has beneficial properties, particularly for more efficient production and purification of the Fc-containing fusion protein.

[0362] Therefore, in a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0363] HSV-1 F gB extracellular domain (SEQ ID NO: 74) fused to IgG1 hinge*-CH2-CH3.

[0364] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:74, but may also be an Fc-containing fusion protein of SEQ ID NO:74 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:74, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described herein above and below.

[0365] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0366] HSV-2G gB extracellular domain (SEQ ID NO: 75) fused to IgG1 hinge*-CH2-CH3.

[0367] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:75, but may also be an Fc-containing fusion protein of SEQ ID NO:75 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:75, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described herein above and below.

[0368] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0369] HSV-1 F gB extracellular domain fused to IgG1 hinge*-CH2-CH3-StrepTag (SEQ ID NO: 76).

[0370] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:76, but may also be an Fc-containing fusion protein of SEQ ID NO:76 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:76, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described above and below in this document.

[0371] In a further preferred embodiment, the Fc-containing fusion protein has the following amino acid sequence:

[0372] HSV-2G gB extracellular domain fused to IgG1 hinge*-CH2-CH3-StrepTag (SEQ ID NO: 77).

[0373] The Fc-containing fusion protein is not particularly limited to the amino acid sequence of SEQ ID NO:77, but may also be an Fc-containing fusion protein of SEQ ID NO:77 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:77, as long as the Fc portion of the Fc fusion protein has the ability to bind to the Fc receptor present on or in the APC described herein above and below.

[0374] In a further preferred embodiment, a fusion protein without a part containing Fc is also disclosed. The corresponding protein has the following amino acid sequence:

[0375] HSV-1 F gB extracellular domain fused to StrepTag (SEQ ID NO: 78).

[0376] The protein is not particularly limited to the amino acid sequence of SEQ ID NO:78, but may also be a fusion protein of SEQ ID NO:78 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:78, as long as the protein has the ability to bind to the Fc receptor present on or in the APC described above and below in this document.

[0377] In a further preferred embodiment, a fusion protein without a part containing Fc is also disclosed. The corresponding protein has the following amino acid sequence:

[0378] HSV-2G gB extracellular domain fused to StrepTag (SEQ ID NO: 79).

[0379] The protein is not particularly limited to the amino acid sequence of SEQ ID NO:79, but may also be a protein of SEQ ID NO:79 having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequence of SEQ ID NO:79, as long as the protein has the ability to bind to the Fc receptor present on or in the APC described above and below in this document.

[0380] The same applies to determining whether the Fc portion of the fusion protein has the ability to bind to a receptor present on or in an APC, as described above. The corresponding assay for determining this ability has also been described above, which is suitable for determining whether any of the Fc portions of the fusion protein described above has the ability to bind to a receptor present on or in an APC after appropriate modification.

[0381] With regard to the determination of sequence identity, the same applies as already explained above.

[0382] As mentioned, according to (b), the construct comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease is an "Fc-containing fusion protein".

[0383] Therefore, in a preferred embodiment, the construct according to (b) exists in the form of a fusion protein (i.e., a protein formed by expressing a hybrid gene made by combining at least two gene sequences). Typically, as will be explained in more detail below, this is achieved by cloning the cDNA into an expression vector in a frame-matched manner with an existing gene. Therefore, the construct can be a fusion protein, i.e., a chimeric molecule formed by connecting two or more polypeptides via a peptide bond between the amino terminus of a module and the carboxyl terminus of another molecule. In this way, the Fc part of the above-mentioned antibody and the antigen associated with the infectious disease or malignant disease are connected together in the form of a fusion protein, wherein both parts are proteinaceous. Once cloned in a frame-matched manner, the fusion protein is recombinantly expressed by the corresponding nucleic acid sequence encoding the fusion protein.

[0384] Alternatively, the Fc portion of the antibody and the antigen associated with the infectious disease or malignant disease can also be covalently coupled via a chemical conjugate. Thus, the modules of the construct can be chemically coupled via covalent bonds. In particular, in a preferred embodiment, a multimeric complex having multiple Fc domains and / or multiple antigen molecules is envisioned. A corresponding example of a multimeric complex is shown in the Argentinian AntiCovid Consortium 2022

[64] .

[0385] Therefore, in such an embodiment, both parts of (a), i.e., the Fc part of the antibody and the antigen associated with the infectious disease or malignant disease, can be synthesized independently (either chemically or by recombinant technology), optionally purified, and then chemically coupled with a covalent bond. Therefore, the Fc-containing fusion protein of the present invention can be a construct in which the Fc part of the antibody and the antigen associated with the infectious disease or malignant disease are chemically coupled with a covalent bond.

[0386] The term "chemically coupled with a covalent bond" refers to conjugation techniques well known to the skilled person. Many methods for preparing covalent or non-covalent conjugates with proteins or peptides are known in the art, and any such known methods may be used. Without being bound by theory, constructs according to the invention may be prepared using heterobifunctional cross-linkers such as N-succinyl 3-(2-pyridyldithio) propionate (SPDP) (Yu et al., Int. J. Cancer 56:244 (1994)). The general techniques for such conjugation are well known in the art; see, for example, Wong, Chemistry of Protein Conjugation and Cross-linking (CRC Press 1991); Upeslacis et al., "Modification of Antibodies by Chemical Methods", in Monoclonal Antibodies: Principles and Applications, Birch et al. (eds.), pp. 187-230 (Wiley-Liss, Inc. 1995); Price, "Production and Characterization of Synthetic Peptide-Derived Antibodies", in Monoclonal Antibodies: Production, Engineering and Clinical Application, Ritter et al. (eds.), pp. 60-84 (Cambridge University Press 1995). Therefore, in view of the fact that the method for covalently coupling the Fc portion of the antibody and the antigen associated with the infectious disease or malignant disease to each other (preferably coupled to a protein / peptide) is well known to those skilled in the art, the examples provided here are non-limiting.

[0387] The Fc-containing fusion protein according to the present invention may not only comprise the above two parts, i.e. the Fc part of an antibody and the antigen associated with the infectious disease or malignant disease. On the contrary, it may be desirable to place a linker part between the individual parts (a), which may, for example, facilitate the construction of the fusion protein.

[0388] However, as mentioned above, in a preferred embodiment, the Fc-containing fusion protein of the present invention is a "classical" fusion protein. A variety of methods are known for preparing fusion proteins, including nucleic acid synthesis, hybridization and / or amplification to produce synthetic double-stranded nucleic acids encoding the desired fusion protein. Such double-stranded nucleic acids can be inserted into expression vectors by standard molecular biology techniques for fusion protein production (see, e.g., Sambrook et al., Molecular Cloning, A laboratory manual, 2nd edition, 1989).

[0389] Therefore, nucleic acid molecules can be prepared, which encode the Fc fusion protein containing the Fc part of the antibody defined above and the antigen associated with the infectious disease or malignant disease. The nucleic acid is, for example, DNA, which encodes the Fc fusion protein containing the Fc part of the antibody and the antigen associated with the infectious disease or malignant disease. The above-mentioned nucleic acid molecules of the present invention can be natural nucleic acid molecules and recombinant nucleic acid molecules. Therefore, nucleic acid molecules of the present invention can be natural origin, synthetic or semisynthetic. It can include DNA, RNA and PNA, and it can be a hybrid thereof.

[0390] It is obvious to those skilled in the art that regulatory sequences may be added to the nucleic acid molecules of the present invention, in particular for their (recombinant) expression. For example, promoters, transcription enhancers and / or sequences allowing the induction of expression of the polynucleotides of the present invention may be used. Suitable inducible systems are, for example, gene expression regulated by tetracycline, such as described by Gossen and Bujard, Proc. Natl. Acad. Sci. USA 89 (1992), 5547-5551 and Gossen, Trends Biotech. 12 (1994), 58-62, or dexamethasone inducible gene expression systems, such as described by Crook, EMBO J. 8 (1989), 513-519.

[0391] Further, the nucleic acid molecule may comprise, for example, thioester bonds and / or nucleotide analogs. The modification may be useful for stabilization of the nucleic acid molecule against endonucleases and / or exonucleases in cells. The nucleic acid molecule may be transcribed by a suitable vector comprising a chimeric gene that allows the nucleic acid molecule to be transcribed in the cell. In the context of the present invention, the nucleic acid molecule is labeled. Methods for detecting nucleic acids are well known in the art, such as Southern and Northern blotting, PCR or primer extension.

[0392] The nucleic acid molecule encoding the above-described Fc-containing fusion protein (which comprises the Fc portion of an antibody and an antigen associated with the infectious disease or malignant disease) can be a recombinantly produced chimeric nucleic acid molecule comprising any one of the aforementioned nucleic acid molecules (alone or in combination). Preferably, the nucleic acid molecule of the present invention is part of a vector.

[0393] The vector of the invention may be, for example, a plasmid, a cosmid, a virus, a phage or another vector conventionally used, for example, in genetic engineering, and may contain further genes, such as marker genes, which allow selection of the vector in a suitable host cell and under suitable conditions.

[0394] Further, in addition to the nucleotide sequence of the present invention, the vector of the present invention can also include expression control elements, which allow the correct expression of the coding region in a suitable host. Such control elements are known to the technician and can include promoters, splicing boxes, translation initiation codons, translation and insertion sites for introducing the insert fragment into the vector. Preferably, the nucleic acid molecules of the present invention are operably connected to the expression control sequences, thereby allowing expression in eukaryotic or prokaryotic cells. Therefore, the present invention relates to a vector comprising the nucleic acid of the present invention, wherein the nucleic acid is operably connected to a control sequence recognized by the host cell when the vector is used to transfect eukaryotic and / or prokaryotic (host) cells.

[0395] The control sequences that ensure expression in eukaryotic and prokaryotic (host) cells are well known to those skilled in the art. As mentioned above in this article, they generally include regulatory sequences that ensure transcription initiation, and optionally poly-A signals that ensure transcription termination and stabilization of transcripts. Additional regulatory elements may include transcription and translation enhancers, and / or naturally related or heterologous promoter regions. Possible regulatory elements that allow expression in, for example, mammalian host cells include CMV-HSV thymidine kinase promoters, SV40, RSV-promoter (Rous sarcoma virus), human elongation factor 1α-promoter, glucocorticoid-inducible MMTV-promoter (mouse mammary tumor virus), metallothionein or tetracycline-inducible promoters, or enhancers, such as CMV enhancers or SV40-enhancers. For expression in neural cells, it is envisioned that neurofilament promoters, PGDF promoters, NSE promoters, PrP promoters or thy-1 promoters may be used. Such promoters are known in the art and are described in particular in Charron, J. Biol. Chem. 270 (1995), 25739-25745. For expression in prokaryotes, numerous promoters have been described, including, for example, the tac-lac promoter or the trp promoter. In addition to the elements responsible for transcription initiation, such regulatory elements may also include transcription termination signals downstream of the polynucleotide, such as the SV40-poly-A site or the tk-poly-A site. In this context, suitable expression vectors are known in the art, such as the Okayama-Berg cDNA expression vector pcDV1 (Pharmacia), pRc / CMV, pcDNA1, pcDNA3 (In-vitrogene), pSPORT1 (GIBCO BRL), pX (Pagano, Science 255 (1992), 1144-1147), yeast two-hybrid vectors, such as pEG202 and dpJG4-5 (Gyuris, Cell 75 (1995), 791-803), or prokaryotic expression vectors, such as λgt11 or pGEX (Amersham-Pharmacia). In addition to the nucleic acid molecule of the present invention, the vector may further comprise a nucleic acid sequence encoding a secretion signal. Such sequences are well known to those skilled in the art. Further, depending on the expression system used, a leader sequence capable of directing the peptide of the present invention to a cell compartment may be added to the coding sequence of the nucleic acid molecule of the present invention, and they are well known in the art. The leader sequence is assembled in appropriate stages together with translation, initiation and termination sequences and, preferably, a leader sequence capable of directing the translated protein or secretion of the protein into the periplasmic space or extracellular medium.Optionally, the heterologous sequence may encode a fusion protein comprising a C- or N-terminal identification peptide that confers desired characteristics (e.g., stabilization of the expressed recombinant product or simplified purification). Once the vector has been incorporated into a suitable host, the host is maintained under conditions suitable for high-level expression of the nucleotide sequence, and collection and purification of the antibody molecule of the invention or fragment thereof may then follow, as desired.

[0396] Further, the vector of the present invention can also be an expression vector. The nucleic acid molecules and vectors of the present invention can be designed to be introduced directly or via liposomes, viral vectors (e.g., adenoviral, retroviral), electroporation, projectiles (e.g., gene guns) or other delivery systems into cells. In addition, the baculovirus system can be used as a eukaryotic expression system for the nucleic acid molecules of the present invention.

[0397] For its expression, a host cell comprising the vector of the present invention can be prepared. Therefore, the present invention relates to a host transfected or transformed with the vector of the present invention, or a non-human host carrying the vector of the present invention, i.e., a host cell or host genetically modified with a nucleic acid molecule according to the present invention or with a vector comprising such a nucleic acid molecule. The term "genetically modified" means that, in addition to its natural genome, the host cell or host also comprises a nucleic acid molecule or vector according to the present invention introduced into the cell or host or introduced into one of its ancestors / parents. The nucleic acid molecule or vector can be present in a genetically modified host cell or host as an independent molecule (preferably, as a molecule capable of replication) outside the genome, or it can be stably integrated into the genome of the host cell or host. The transformation of host cells with the vector according to the present invention can be carried out by standard methods, such as, for example, in Sambrook and Russell (2001), Molecular Cloning: A Laboratory Manual, CSH Press, Cold Spring Harbor, NY, USA; Methods in Yeast Genetics, A Laboratory Course Manual, Cold Spring Harbor Laboratory Press, 1990 described. The host cells are cultivated in a nutrient medium adapted to the requirements of the specific host cells used, particularly with regard to pH, temperature, salt concentration, aeration, antibiotics, vitamins, trace elements, etc.

[0398] The corresponding host cell for expressing the Fc fusion protein of the present invention can be any prokaryotic or eukaryotic cell. Suitable prokaryotic cells are those commonly used for cloning such as Escherichia coli or Bacillus subtilis. Further, eukaryotic cells include such as fungi or animal cells. Examples of suitable fungal cells are yeast cells, preferably those of Saccharomyces, and most preferably those of the species of Saccharomyces cerevisiae. Suitable animal cells are such as insect cells, vertebrate cells, preferably mammalian cells, such as HEK293, NSO, CHO, COS-7, MDCK, U2-OSHela, NIH3T3, MOLT-4, Jurkat, PC-12, PC-3, IMR, NT2N, Sk-n-sh, CaSki, C33A. These host cells, such as CHO cells, can provide post-translational (secondary) modifications to the antibody molecules of the invention, including leader peptide removal, folding and assembly of H and C chains, glycosylation of the molecules on the correct side, and secretion of functional molecules. Further suitable cell lines known in the art are available from cell line collections such as the German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ) or the American Type Culture Collection (American Type Culture Collection; ATCC). According to the present invention, it is further envisioned that primary cells / cell cultures can function as host cells. The cells are particularly derived from insects (e.g., insects of the genus Drosophila or Blatta species) or mammals (e.g., humans, pigs, mice or rats). The host cells may also include cells from and / or derived from cell lines such as neuroblastoma cell lines. The above-mentioned primary cells are well known in the art and include in particular primary astrocytes, (mixed) spinal cord cultures or hippocampal cultures.

[0399] The Fc-containing fusion protein comprising the Fc portion of an antibody and an antigen associated with the infectious disease or malignant disease can be produced by methods known in the art. As an example, the above-mentioned host cell with an expression vector can be cultured in a culture medium, and the expression vector encodes the Fc-containing fusion protein comprising the Fc portion of an antibody of the present invention and an antigen associated with the infectious disease or malignant disease. Then, the Fc-containing fusion protein comprising the Fc portion of an antibody and an antigen associated with the infectious disease or malignant disease can be recovered from the host cell or culture medium.

[0400] Host cells, such as CHO cells, can provide post-translational (secondary) modifications on the expressed binding compounds of the invention. These modifications include glycosylation and phosphorylation, among others.

[0401] Methods for recovery and / or subsequent purification of the constructs of the invention are known to those skilled in the art.

[0402] In a preferred embodiment, in the vaccine for said use according to the present invention described herein above, said active immunization induces a lasting or long-term immunity against said infectious disease or malignant disease in said subject, and wherein said subject suffers from an acute infectious disease or malignant disease.

[0403] Preferably, in active immunization, the lasting or long-term immunity has an effect for months to years. In the case of eradication of, for example, tumors or cancer cells, in a preferred embodiment, the term "long-term" relates to a "lifelong" effect. For chronic recurrent infectious diseases, such as, in a preferred embodiment, HSV infection, the term "long-term" is protection from asymptomatic reactivation for months to years.

[0404] In a preferred embodiment of the first aspect of the present invention, in the vaccine for the purposes defined above, the antibody or the Fc-containing fusion protein is capable of inducing antibody-dependent cellular phagocytosis (ADCC) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC). In a more preferred embodiment, in the vaccine for the purposes defined above, the Fc-containing fusion protein is capable of inducing antibody-dependent cellular phagocytosis (ADCP) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC), while at the same time the Fc-containing fusion protein is capable of mediating ADCC and CDC, provided that the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen is capable of bridging to a cell via binding of the antigen to a cell, whereby the Fc is free and can mediate ADCC or CDC.

[0405] In the following, we describe in more detail The second aspect .

[0406] As mentioned above, in a second aspect related to the first aspect above, the present invention relates to a vaccine for use in active immunization of a subject against an infectious disease, wherein the infectious disease is an HSV-related disease; and

[0407] (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection; or

[0408] (b) wherein the Fc-containing fusion protein comprises the Fc portion of an antibody fused to an antigen, wherein the antigen is associated with an HSV-related disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC) selected from the group consisting of:

[0409] Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and

[0410] Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0411] Thus, the vaccine of the second aspect of the invention may comprise one of two alternative components (although it is also contemplated that the vaccine comprises both components), namely:

[0412] (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection; or

[0413] (b) wherein the Fc-containing fusion protein comprises the Fc portion of an antibody fused to an antigen, wherein the antigen is associated with an HSV-related disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC) selected from the group consisting of:

[0414] Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and

[0415] Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0416] These two components are described separately in more detail below.

[0417] anti-HSV antibodies and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection:

[0418] Regarding (a) , i.e., the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection, which also applies mutatis mutandis as already described above in relation to the first aspect of the invention, except that the “antibody against an antigen associated with the infectious disease” is an anti-HSV antibody and the “antigen associated with the infectious disease” is an antigen associated with an acute HSV-related disease, preferably an acute HSV infection.

[0419] As mentioned above, according to the basic principle of the present invention, the antibody comprises at least the Fc part of the antibody.

[0420] The antibody used in the vaccine of the second aspect of the present invention is not particularly limited as long as it is an "anti-HSV antibody". Therefore, the antibody can be any antibody that specifically binds to HSV (i.e., a domain or antigen of HSV), or specifically recognizes HSV (i.e., a domain or antigen of HSV) or specifically interacts with it.

[0421] The term "binding to..." used in the context of the present invention defines the binding (interaction) of at least two "antigen interaction sites" to each other. According to the present invention, the term "antigen interaction site" defines a motif of a polypeptide, i.e. a part of an antibody or antigen-binding fragment of the present invention, which shows the ability to specifically interact with a specific antigen or a specific group of antigens of HSV. The binding / interaction is also understood to define "specific recognition". According to the present invention, the term "specifically recognizes" means that the antibody is able to specifically interact and / or bind to at least two amino acids of each of HSV, as defined herein. The antibody can recognize different epitopes on HSV, interact with different epitopes on HSV and / or bind to different epitopes on HSV. The term relates to the specificity of the antibody molecule, i.e. its ability to distinguish specific regions of HSV.

[0422] The term "specific interaction" as used according to the present invention means that the antibody or antigen-binding fragment thereof of the present invention does not or substantially does not cross-react with a (poly)peptide having a similar structure. Thus, the antibody or antigen-binding fragment thereof of the present invention specifically binds / interacts with the structure of HSV (preferably, HSV-1 or HSV-2). Specific examples of such molecules to which the first and second Ig-derived domains are directed are given herein below.

[0423] The cross-reactivity of the panel of antibodies or antigen-binding fragments thereof under investigation can be tested, for example, by evaluating the binding of the panel of antibodies or antigen-binding fragments thereof to the (poly)peptide of interest and to a number of more or less closely related (poly)peptides (structurally and / or functionally) under conventional conditions (see, for example, Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1988); and Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1999)). Only those constructs (i.e., antibodies, antigen-binding fragments thereof, etc.) that bind to a certain structure of HSV, such as a specific epitope or (poly)peptide / protein of HSV, and not or substantially not to any other epitope or (poly)peptide of the same HSV are considered to be specific for the epitope or (poly)peptide / protein of interest and are selected for further investigation according to the methods provided herein. These methods may include, among others, binding studies, blocking and competition studies with molecules that are closely related structurally and / or functionally. These binding studies also include FACS analysis, surface plasmon resonance (SPR, e.g. ), analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, fluorescence spectroscopy, or radiolabeled ligand binding assays.

[0424] The term "binding to..." refers not only to linear epitopes but also to conformational epitopes, structural epitopes or discontinuous epitopes consisting of two regions of a human target molecule or a portion thereof. In the context of the present invention, a conformational epitope is defined by two or more discrete amino acid sequences separated in the primary sequence, which come together on the surface of the molecule when the polypeptide is folded into a native protein (Sela, Science 166 (1969), 1365; and Laver, Cell 61 (1990), 553-536). In addition, the term "binding to..." is used interchangeably with the term "interacting with..." or "recognizing..." in the context of the present invention.

[0425] Therefore, specificity can be determined experimentally by methods known in the art and described herein. Such methods include, but are not limited to, Western blots, ELISA-tests, RIA-tests, ECL-tests, IRMA-tests and peptide scanning.

[0426] As mentioned above, the vaccine for use in active immunization of a subject against an infectious disease (wherein the infectious disease is an HSV-related disease, and (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection) requires the presence of an acute infection. This is because, in order to be able to actively immunize a subject in accordance with the basic principles of the present invention, not only antibodies but also antigens associated with the HSV infection must be present.

[0427] In this regard, "acute" means that the subject has an outbreak of the disease and, therefore, has HSV present in the body. An outbreak of the disease may follow a primary infection or after a reactivation (outburst) from a latent reservoir of HSV.

[0428] In fact, it is well known that after the first infection, HSV spreads from infected epithelial cells to the axons of sensory neurons of the sites of the first infection of the innervation, and then retrogradely transports to the respective dorsal root ganglia, where HSV sets up a lifelong latent reservoir. The HSV infection of neurons exists in a reversible state, and the event of virus reactivation (outbreak) may occur from time to time. The reactivation of the virus can be triggered by a wide range of stress stimuli (e.g., immunodeficiency, trauma, heating, menstruation, UV light and sexual intercourse) acting on neurons or at peripheral sites innervated by infected ganglia. Intermittent HSV reactivation causes the production of infectious HSV from neurons infected latently. Once reactivated, the virus is transported back to the nerve endings in the epithelium by neurons.

[0429] In a more preferred embodiment, "acute" means that the subject shows symptoms of the disease. In other words, the subject to be vaccinated actually needs vaccination or treatment, and the term "acute" relates to the situation after the (first) onset or reactivation outbreak of the disease. The term "acute" mentioned in the context of the present invention is relative to prophylactic or preventive (i.e. measures taken for disease prevention, for example to prevent infection and / or the onset / outbreak of the disease). More particularly, prophylactic treatment can be understood in such a way that it prevents free viral particles (from outside the body) from attaching to target cells and thereby preventing viral replication. In contrast, in an acute infection (which can be a first or recurrent infection), progeny viruses are already in full swing when HSV replicates.

[0430] As will be outlined in more detail further below, the vaccine for use according to the second aspect of the invention is for use in the active immunization of a subject against an HSV-related disease, wherein the disease is selected from the group consisting of: herpes simplex labialis, herpes simplex genitalis, chronic or disseminated cutaneous herpes simplex infection, traumatic herpes, eczema herpeticum, herpetic keratoconjunctivitis, herpes neonatalis, Alzheimer's disease (AD), HSV pneumonia, Bell's palsy, herpetic esophagitis, herpes encephalitis and herpes meningitis, herpetic whitlow, herpetic gingivostomatitis, the presence of oral relapses, the presence of genital relapses, herpetic eczema, herpes neonatalis, immunodeficiency, immunocompromised patients, resistance to viral inhibitory agents, encephalitis, meningitis, meningoencephalitis, eye infections and / or systemic HSV infections.

[0431] In a more preferred embodiment, the vaccine for use according to the second aspect of the invention is for use in active immunization of a subject against an HSV-related disease, wherein the disease is caused by HSV-1 or HSV-2, more preferably wherein the HSV-related disease is selected from the group consisting of herpes simplex labialis, herpes simplex genitalis, chronic or disseminated cutaneous herpes simplex infection, traumatic herpes and eczema herpeticum, which is to be administered topically.

[0432] In a preferred embodiment, the anti-HSV antibody of the vaccine for use according to the second aspect of the invention is monoclonal. In a further preferred embodiment, the anti-HSV antibody of the vaccine for use according to the second aspect of the invention is a humanized antibody or a fully human antibody. In a further preferred embodiment, the anti-HSV antibody of the vaccine for use according to the second aspect of the invention is a murine antibody.

[0433] As already outlined above in the context of the first aspect of the present invention, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies constituting the population are identical, except for possible naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific, directed against a single antigenic site. Monoclonal antibodies are advantageous because they can be synthesized by hybridoma cultures, substantially free of contamination by other immunoglobulins. The modifier "monoclonal" indicates that the antibody is characterized by being in a substantially homogeneous antibody population, and should not be construed as requiring the antibody to be produced by any ad hoc method. As mentioned above, the monoclonal antibody to be used according to the present invention can be prepared by the hybridoma method described by Kohler, Nature 256 (1975), 495.

[0434] The term "polyclonal antibody" as used herein in the context of the present disclosure refers to an antibody produced in or in the presence of one or more other, non-identical antibodies. Generally, polyclonal antibodies are produced from lymphocytes in the presence of several other B-lymphocytes that produce non-identical antibodies. Typically, polyclonal antibodies are obtained directly from immunized animals.

[0435] The term "fully human antibody" as used herein refers to an antibody comprising only human immunoglobulin protein sequences. A fully human antibody may comprise murine carbohydrate chains if it is produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, a "mouse antibody" or "murine antibody" refers to an antibody comprising only mouse / murine immunoglobulin protein sequences. Alternatively, a "fully human antibody" may comprise rat carbohydrate chains if it is produced in a rat, in a rat cell, or in a hybridoma derived from a rat cell. Similarly, the term "rat antibody" refers to an antibody comprising only rat immunoglobulin sequences. Fully human antibodies may also be produced, for example, by phage display (which is a widely used screening technique that enables the production and screening of fully human antibodies). Phage antibodies may also be used in the context of the present invention. Phage display technology is described in, for example, US 5,403,484, US 5,969,108, and US 5,885,793. Another technology that enables the development of fully human antibodies involves improvements in mouse hybridoma technology. Mice are made transgenic to contain human immunoglobulin loci, exchanging their own mouse genes (see, e.g., US 5,877,397).

[0436] The term "chimeric antibody" refers to antibodies comprising the variable regions of the invention fused or chimerized with antibody regions (eg, constant regions) from another human or non-human species (eg, mouse, horse, rabbit, dog, cow, chicken).

[0437] The term "antibody" also relates to recombinant human antibodies, xenoantibodies and heterohybrid antibodies. The term "recombinant human antibody" includes all human sequence antibodies prepared, expressed, created or separated by recombinant means, such as antibodies separated from transgenic animals (e.g., mice) with respect to human immunoglobulin genes; antibodies expressed using recombinant expression vectors transfected into host cells; antibodies separated from recombinant combined human antibody libraries; or antibodies prepared, expressed, created or separated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable regions and constant regions (if present) derived from human germline immunoglobulin sequences. However, such antibodies can undergo in vitro mutagenesis (or, when using transgenic animals with respect to human Ig sequences, in vivo somatic cell mutagenesis), and therefore the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that may not naturally exist in the human antibody germline repertoire in vivo (although derived from and related to human germline VH and VL sequences).

[0438] "Xenogeneic antibodies" are defined in relation to the transgenic non-human organism that produces such antibodies. The term refers to antibodies that have an amino acid sequence or encoding nucleic acid sequence corresponding to that found in an organism not consisting of the transgenic non-human animal, and are typically from a species other than that of the transgenic non-human animal.

[0439] The term "heterohybrid antibody" refers to an antibody having light chains and heavy chains of different biological origin. For example, an antibody having a human heavy chain associated with a murine light chain is a heterohybrid antibody. Examples of heterohybrid antibodies include chimeric antibodies and humanized antibodies.

[0440] The term "antibody" also relates to humanized antibodies. The "humanized" form of non-human (e.g., murine or rabbit) antibodies is a chimeric immunoglobulin, an immunoglobulin chain comprising a minimal sequence derived from a non-human immunoglobulin. Often, humanized antibodies are such human immunoglobulins (acceptor antibodies), wherein the residues from the complementary determining region (CDR) of the recipient are replaced by residues (which have desired specificity, affinity and ability) from the CDR (donor antibody) of non-human species such as mice, rats or rabbits. In some cases, the Fv framework residues of human immunoglobulins are replaced by corresponding non-human residues. Further, humanized antibodies can include residues that are neither found in the CDR or framework sequences of the recipient antibodies nor in the input. These modifications are carried out to further refine and optimize antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, wherein all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin, and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. A humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see: Jones Nature 321 (1986), 522-525; Reichmann Nature 332 (1998), 323-327; and Presta Curr Op Struct Biol 2 (1992), 593-596.

[0441] As already outlined above in the context of the first aspect of the invention, popular methods for the humanization of antibodies involve CDR transplantation, in which a functional antigen binding site from a non-human "donor" antibody is transplanted onto a human "acceptor" antibody. CDR transplantation methods are known in the art and are described in, for example, US 5,225,539, US 5,693,761, and US 6,407,213. Another related method is to produce humanized antibodies from transgenic animals that are genetically engineered to contain one or more humanized immunoglobulin loci capable of undergoing gene rearrangement and gene conversion (see, for example, US 7,129,084).

[0442] Therefore, in the context of the present invention, the term "antibody" refers to whole immunoglobulin molecules as well as parts of such immunoglobulin molecules (i.e., "antigen-binding fragments thereof"). Further, in general, the term refers to modified and / or altered antibody molecules. The term also refers to antibodies generated / synthesized recombinantly or synthetically. The term also refers to intact antibodies. The term "antibody" also includes, but is not limited to, fully human antibodies, chimeric antibodies, humanized antibodies, CDR-grafted antibodies, and antibody constructs, such as single-chain Fv (scFv) or antibody fusion proteins.

[0443] In a preferred embodiment, the anti-HSV antibody of the vaccine for use according to the second aspect of the invention is a full-length antibody, ie a whole immunoglobulin molecule, which is often also referred to as a complete antibody.

[0444] In the context of the present invention, a "single-chain Fv" or "scFv" antibody fragment (which is not envisaged in the context of the second aspect of the invention) has the V H and V L domains, wherein these domains are present in a single polypeptide chain. Typically, the scFv polypeptide further comprises a V H and V L A polypeptide linker between the domains which enables the scFv to form the desired structure for antigen binding. Techniques for producing single chain antibodies are described, for example, in Plückthun, in The Pharmacology of Monoclonal Antibodies, Rosenburg and Moore, eds. Springer-Verlag, NY (1994), 269-315.

[0445] As used herein, a "Fab fragment" consists of: a light chain; and a C H 1 region and the variable region. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule.

[0446] The "Fc" region comprises two heavy chain fragments, which comprise the C H 2 and C H 3 domains. The two heavy chain fragments are connected by two or more disulfide bonds and by C H The 3 domains are held together by hydrophobic interactions.

[0447] A "Fab' fragment" comprises the following: a light chain; and a portion of a heavy chain comprising V H Domain and C H 1 domain and also in C H 1 and C HThe region between the two Fab' domains allows for the formation of an interchain disulfide bond between the two heavy chains of the two Fab' fragments to form a F(ab')2 molecule.

[0448] The "F(ab')2 fragment" comprises the following parts: two light chains; and two heavy chains, which are contained in C H 1 and C H The F(ab')2 fragment consists of two Fab' fragments held together by the disulfide bond between the two heavy chains.

[0449] The "Fv region" contains the variable regions from both the heavy and light chains, but lacks the constant regions.

[0450] The antibodies, antibody constructs, antibody fragments, antibody derivatives (all Ig-derived) or their corresponding immunoglobulin chains to be used according to the present invention may be further modified using conventional techniques known in the art, for example by using amino acid deletions, insertions, substitutions, additions, and / or recombination and / or any other modifications known in the art, alone or in combination. Methods for introducing such modifications in the DNA sequence underlying the amino acid sequence of the immunoglobulin chain are well known to those skilled in the art; see, for example, Sambrook (1989) (loc. cit.). The term "Ig-derived domain" particularly relates to a (poly)peptide construct comprising at least one CDR. The described fragments or derivatives of the Ig-derived domain define a (poly)peptide that is part of the above-mentioned antibody molecule and / or modified by chemical / biochemical or molecular biological methods. Corresponding methods are known in the art and are described, inter alia, in laboratory manuals (see Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 2nd edition (1989) and 3rd edition (2001); Gerhardt et al., Methods for General and Molecular Bacteriology ASM Press (1994); Lefkovits, Immunology Methods Manual: The Comprehensive Sourcebook of Techniques, Academic Press (1997); Golemis, Protein-Protein Interactions: A Molecular Cloning Manual, Cold Spring Harbor Laboratory Press (2002)).

[0451] The antibody used in the vaccine for the purpose according to the second aspect of the present invention is not particularly limited, as long as it is an "anti-HSV antibody". Therefore, the antibody can be any antibody that specifically binds to HSV (i.e., a domain, an antigen, preferably a surface antigen of HSV), or specifically recognizes HSV (i.e., a domain, an antigen, preferably a surface antigen of HSV) or specifically interacts with it. A technician is easily able to generate such antibodies against a given domain (i.e., an antigen of HSV, preferably a surface antigen), and determine whether the respective antibody is able to detect / bind to a given domain, antigen (preferably a surface antigen) of HSV (preferably, HSV-1 or HSV-2), based on the common knowledge of the technician and the methods described above.

[0452] In a preferred embodiment, the antibodies of the vaccine for the purpose according to the second aspect of the present invention bind / recognize viral antigen glycoprotein D, B, C, H, L, E or I (i.e. gD, gB, gC, gH, gL, gE, gI). Glycoproteins D, B, C, H, L, E and I are surface or envelope proteins of HSV-1 and / or HSV-2. These proteins are not only found on the surface of HSV-1 and / or HSV-2 or in the envelope structure, i.e. on the surface of the released infectious particles (i.e., the envelope of free virus particles), but they can also be present on the surface of infected cells, i.e. on the surface of cells. However, in a more preferred embodiment, the antibodies of the present invention bind / recognize viral surface antigen glycoprotein D, B, C, H, L, E or I (i.e., gD, gB, gC, gH, gL, gE or gI) of the envelope of HSV-1 and / or HSV-2. In a preferred embodiment, the anti-HSV antibody or antigen-binding fragment thereof for use according to the present invention recognizes the surface glycoprotein B (gB) of the HSV-1 and / or HSV-2 envelope, preferably its epitope. The glycoprotein B of HSV-1 and / or HSV-2 is well characterized and is not bound to a specific sequence, and exemplary sequences of various HSV-1 and HSV-2 strains are shown in SEQ ID NOs: 11 to 16, respectively. SEQ ID NO: 11 shows the sequence of glycoprotein B of HSV-1 strain F, SEQ ID NO: 12 shows the sequence of glycoprotein B of HSV-1 strain KOS, SEQ ID NO: 13 shows the sequence of glycoprotein B of HSV-1 strain gC-39-R6, SEQ ID NO: 14 shows the sequence of glycoprotein B of HSV-2 strain HG52, SEQ ID NO: 15 shows the sequence of glycoprotein B of HSV-2 strain 333, and SEQ ID NO: 16 shows the sequence of glycoprotein B of HSV-2 strain MMA. Sequence alignment of these glycoprotein B amino acid sequences showed that the overall amino acid homology (preferably, identity) of gB of HSV-1 and HSV-2 is 85%, while the sequences are least conserved at the N- and C-terminal regions of the protein.

[0453] In a preferred embodiment, the anti-HSV antibody of the vaccine for use according to the second aspect of the present invention is capable of inhibiting the spread of HSV from an infected cell to another adjacent uninfected cell (intercellular spread).

[0454] Cell-to-cell spread is the ability of herpes simplex virus to spread to another uninfected cell in the vicinity without releasing acellular particles. Reducing or eliminating the ability of herpes simplex virus to spread to neighboring cells has the beneficial effect of avoiding damage generation. In order to check whether the antibody can inhibit HSV from spreading to another uninfected cell in the vicinity (cell-to-cell spread), methods well known to those skilled in the art can be used. As an example, the following assay can be used: using 400TCID 50 Vero cells grown to confluence on coverslips in 24-well tissue culture plates were infected with a constant amount of virus per well at 37°C for 4 h. A median tissue infection dose (1 TCID 50 ) is the amount of cytopathic agent (e.g., virus) that will produce a cytopathic effect in 50% of the inoculated cell cultures. The viral inoculum is then removed, the cells are washed twice with PBS, and further incubated for 2 days at 37°C in 1 ml DMEM, 2% FCS, Pen / Strep containing an excess of different anti-HSV antibodies or polyclonal anti-HSV control serum to prevent viral spread via the supernatant. Viral antigens of cells infected with HSV are detected with fluorescently labeled polyclonal goat anti-HSV serum (BETHYL Laboratories, Montgomery, TX USA, catalog number A190-136F, batch number A190-136F-2). Preferably, the antibody inhibits cell-to-cell spread if less than 20% of neighboring cells are infected, preferably wherein less than 15%, less than 10%, less than 5%, more preferably less than 3%, and most preferably less than 1% of neighboring cells are infected in the above assay.

[0455] Cell-to-cell spread can also be tested as follows: the presence of neutralizing antibodies does not necessarily prevent the cell-to-cell spread of Herpesviridae. In order to compare antibodies about the destruction of HSV-1 and HSV-2 cell-to-cell spread, this specific transmission mode can be simulated in vitro using standard test methods. For example, in order to infect individual cells, initially the confluent Vero cell monolayer is incubated with HSV-1 or HSV-2 at low MOI (e.g., 100TCID50), respectively. After adsorption for 4 hours at 37°C, the virus inoculum must be removed. In order to promote direct cell-to-cell spread from individually infected cells but prevent the virus from spreading across the cell culture supernatant through viral particles, the Vero cell monolayer is treated with excessive neutralizing anti-gB antibodies, controls or separate culture media. After 48 hours, viral spread can be detected by immunostaining with a mouse monoclonal antibody specific for a common epitope on glycoprotein D of HSV-1 and HSV-2 (e.g., Acris Antibodies, San Diego, CA, USA) and a secondary antibody conjugated with a fluorescent marker. Immunofluorescence images can be acquired at 100 or 400 times magnification using a fluorescent microscope.

[0456] Furthermore, the anti-HSV antibodies of the vaccine for use according to the second aspect of the invention disclosed only in the context of the present invention may also be able to neutralize HSV. "Neutralization" in this context means that the antibody opsonizes the virus so that it cannot infect any further cells. For testing whether an antibody at a concentration of, for example, up to 20 nM is able to neutralize, for example, 100 TCID 50 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 TCID 100 50 The degree of HSV of the specified amount is more than 80%, preferably more than 90%, such as more than 95%, more preferably more than 96%, such as more than 97%, and most preferably more than 98%, such as more than 99%, or even 100%.

[0457] Furthermore, the anti-HSV antibodies of the vaccine for use according to the second aspect of the invention, disclosed only in the context of the present invention, may also be capable of inhibiting cell-to-cell spread independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

[0458] Since the assays described above for testing whether an antibody is able to inhibit cell-to-cell spread do not include complement and / or cytotoxic effector cells, the same assays can be used to determine whether an antibody is able to inhibit cell-to-cell spread independent of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

[0459] In a preferred embodiment, the anti-HSV antibody for use in the vaccine according to the second aspect of the present invention comprises the following complementarity determining region: V comprising SEQ ID NO: 1 H CDR1, comprising V of SEQ ID NO:2 H CDR2, comprising V of SEQ ID NO:3 H CDR3, comprising V of SEQ ID NO:4 L CDR1, comprising V of SEQ ID NO:5 L CDR2, and V comprising SEQ ID NO:6 L CDR3.

[0460] The term "CDR" as used herein refers to a "complementarity determining region", which is well known in the art. CDR is the portion of an immunoglobulin that determines the specificity of the molecule and contacts a specific ligand. CDR is the most variable portion of the molecule and contributes to the diversity of these molecules. In each V domain there are three CDR regions, namely CDR1, CDR2 and CDR3. CDR-H describes the CDR region of the variable heavy chain, and CDR-L refers to the CDR region of the variable light chain. VH means variable heavy chain, and VL means variable light chain. The CDR regions of the Ig-derived regions can be determined as described in Kabat, "Sequences of Proteins of Immunological Interest", 5th Edition, NIH Publication No. 91-3242, US Department of Health and Human Services (1991); Chothia, J. Mol. Biol. 196 (1987), 901-917; or Chothia, Nature 342 (1989), 877-883.

[0461] Therefore, in the context of the present invention, the antibody molecules described herein above are selected from the group consisting of: whole antibodies (immunoglobulins, such as IgG1, IgG2, IgG2a, IgG2b, IgA1, IgGA2, IgG3, IgG4, IgA, IgM, IgD or IgE), F(ab), Fab'-SH, Fv, Fab', F(ab')2 fragments, chimeric antibodies, CDR-grafted antibodies, fully human antibodies, bivalent antibody constructs, antibody fusion proteins, synthetic antibodies, bivalent single chain antibodies, trivalent single chain antibodies, and multivalent single chain antibodies.

[0462] "Humanization methods" are well known in the art and are particularly described for antibody molecules (e.g., Ig-derived molecules). The term "humanized" refers to a humanized form of a non-human (e.g., murine) antibody or fragment thereof (e.g., Fv, Fab, Fab', F(ab'), scFv, or other antigen-binding portion sequences of an antibody) comprising portions of sequences derived from non-human antibodies. Humanized antibodies include human immunoglobulins in which residues from the complementary determining regions (CDRs) of the human immunoglobulins are replaced by residues from CDRs of non-human species such as mice, rats, or rabbits (which have the desired binding specificity, affinity, and ability). In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, wherein all or substantially all of the CDR regions correspond to those of non-human immunoglobulins, and all or substantially all of the FR regions are those of human immunoglobulin consensus sequences. The humanized antibody will also optimally comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin; see, in particular, Jones et al., Nature 321 (1986), 522-525; Presta, Curr. Op. Struct. Biol. 2 (1992), 593-596. Methods for humanizing non-human antibodies are well known in the art. Typically, a humanized antibody has one or more amino acids introduced into it from a non-human source and still retains the original binding activity of the antibody. Methods for humanizing antibodies / antibody molecules are further described in detail in Jones et al., Nature 321 (1986), 522-525; Reichmann et al., Nature 332 (1998), 323-327; and Verhoeyen et al., Science 239 (1988), 1534-1536. Specific examples of humanized antibodies, such as antibodies against EpCAM, are known in the art, see, for example (LoBuglio, Proceedings of the American Society of Clinical Oncology Abstract (1997), 1562; and Khor, Proceedings of the American Society of Clinical Oncology Abstract (1997), 847).

[0463] Thus, in the context of the present invention, antibody molecules are provided which are humanized and can be successfully used in pharmaceutical compositions.

[0464] In addition, in a preferred embodiment, the antibody of the present invention is an antibody that binds to glycoprotein B (gB) of HSV-1 and / or HSV-2, which comprises or consists of a VH domain (heavy chain variable region) and a VL domain (light chain variable region), i.e., the amino acid sequence of the variable region of the heavy chain of the antibody described in SEQ ID NO:9 and the amino acid sequence of the variable region of the light chain of the antibody described in SEQ ID NO:10.

[0465] However, the antibodies used in the present invention are not particularly limited to such variable heavy and light chain variable regions, but may also be antibodies or antigen-binding fragments thereof that bind to glycoprotein B (gB) of the HSV-1 and / or HSV-2 envelope, which contain or consist of VH domains and VL domains having at least 95%, 90%, 85%, 75%, 70%, 65%, 60%, 55% or 50% sequence homology (preferably, identity) with the sequences of SEQ ID NOs: 9 and 10, respectively, as long as the antibody has the ability to be effective in active immunization of a subject against HSV-related diseases, as described above and below herein. Further, the antibody or antigen-binding fragment thereof is a molecule comprising VH and VL domains having up to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more conservative amino acid substitutions relative to the sequences of SEQ ID NOs: 9 and 10. Furthermore, the antibody or antigen-binding fragment thereof is an antibody fragment selected from the group consisting of Fab, Fab', Fab'-SH, FV, scFV, F(ab')2 and diabody.

[0466] In order to determine whether an amino acid sequence has a certain degree of identity with the sequence of SEQ ID NO: 9 and 10, the skilled person can use means and methods well known in the art, such as alignment, either manually or by using computer programs known to those skilled in the art. Such alignment can be performed, for example, using means and methods known to the skilled person, such as by using known computer algorithms, such as the Lipman-Pearson method (Science 227 (1985), 1435) or the CLUSTAL algorithm. Preferably, in such an algorithm, the maximum homology is assigned to the conserved amino acid residues present in the amino acid sequence. In a preferred embodiment, ClustalW2 is used to compare amino acid sequences. In the case of paired comparisons / alignments, the following settings are preferably selected: protein weight matrix: BLOSUM 62; gap opening: 10; gap extension: 0.1. In the case of multiple comparisons / alignments, the following settings are preferably selected: protein weight matrix: BLOSUM 62; gap opening: 10; gap extension: 0.2; gap distance: 5; no end gap.

[0467] According to the present invention, in the context of two or more nucleic acid or amino acid sequences, the term "identical" or "percent identity" refers to two or more sequences or subsequences that are identical or have a specified percentage of identical amino acid residues or nucleotides (for example, 60% or 65% identity, preferably 70-95% identity, more preferably at least 95% identity to the nucleic acid sequence or amino acid sequence described above; it is capable of binding to gB of HSV-1 or HSV-2, and has the ability to be effective in active immunization of subjects against HSV-related diseases (as described above and below in this article), and / or is capable of inhibiting the spread of HSV from infected cells to another adjacent uninfected cell (intercellular spread), or is capable of inhibiting cell-to-cell spread independent of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC) (as described above and below in this article)), when compared and aligned for maximum correspondence over a comparison window or over a designated area, as measured by using a sequence comparison algorithm known in the art or by manual alignment and visual inspection. Sequences with, for example, 60% to 95% or greater sequence identity are considered to be substantially identical. Such a definition also applies to the complement of the test sequence. Preferably, the described identity is present in a region of at least about 15 to 25 amino acids or nucleotides in length, more preferably in a region of about 50 to 100 amino acids or nucleotides in length. Those skilled in the art will know how to determine the identity percentage between / among sequences by using, for example, algorithms known in the art, such as those based on CLUSTALW computer program (Thompson Nucl.Acids Res.2 (1994), 4673-4680) or FASTDB (Brutlag Comp.App.Biosci.6 (1990), 237-245).

[0468] Although the FASTDB algorithm typically does not take into account internal non-matching deletions or additions, i.e., gaps, in its calculations, this can be manually corrected to avoid overestimation of % identity. However, CLUSTALW takes into account sequence gaps in its identity calculations. BLAST and BLAST 2.0 algorithms are also available to those skilled in the art (Altschul, (1997) Nucl. Acids Res. 25: 3389-3402; Altschul (1993) J. Mol. Evol. 36: 290-300; Altschul (1990) J. Mol. Biol. 215: 403-410). The BLASTN program for nucleic acid sequences uses the following parameters as default values: a word length (W) of 11, an expectation value (E) of 10, M=5, N=4, and a comparison of two chains. For amino acid sequences, the BLASTP program uses the following parameters as default values: a word length (W) of 3, and an expectation value (E) of 10. The BLOSUM62 scoring matrix (Henikoff (1989) PNAS 89:10915) used the following parameters: alignment of 50 (B), expectation of 10 (E), M=5, N=4, and comparison of both strands.

[0469] Preferably, the amino acid substitution is a "conservative substitution", which refers to the substitution of an amino acid in a protein with other amino acids having similar characteristics (e.g., charge, side chain size, hydrophobicity / hydrophilicity, main chain conformation and rigidity, etc.), so that the change can often be made without changing the biological activity of the protein. Those skilled in the art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially change the biological activity (see, e.g., Watson, Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co. 4th edition (1987), 224). In addition, substitutions of amino acids that are structurally or functionally similar are less likely to destroy biological activity. In the context of the present invention, the binding compounds / antibodies of the present invention comprise polypeptide chains having sequences that include up to 0 (no change), 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20 or more conservative amino acid substitutions when compared to specific amino acid sequences disclosed herein, e.g., SEQ ID NO: 9 (which refers to the variable region of the heavy chain of an antibody) and 10 (which refers to the variable region of the light chain of an antibody). As used herein, the phrase "up to X" conservative amino acid substitutions includes 0 substitutions and any number of substitutions up to 10, including 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 substitutions.

[0470] Such exemplary substitutions are preferably made according to those set forth in Table 1 below.

[0471] Table 1 Exemplary conservative amino acid substitutions

[0472]

[0473]

[0474] In addition, in a preferred embodiment, the anti-HSV antibody of the vaccine for the use according to the second aspect of the present invention comprises an amino acid sequence having at least 70% sequence identity with the amino acid residues shown at positions 1 to 30, 38 to 51, 68 to 99 and 112 to 122 of SEQ ID NO:7 and at positions 1 to 23, 41 to 55, 63 to 94 and 104 to 114 of SEQ ID NO:8.

[0475] In a further preferred embodiment, the anti-HSV antibody of the vaccine for use according to the second aspect of the invention comprises an amino acid sequence having at least 75%, at least 80%, more preferably at least 85%, at least 90%, even more preferably at least 95%, and most preferably 98% total sequence identity in the framework regions compared to the amino acid residues shown in positions 1 to 30, 38 to 51, 68 to 99 and 112 to 122 of SEQ ID NO:7 and in positions 1 to 23, 41 to 55, 63 to 94 and 104 to 114 of SEQ ID NO:8. Such antibodies are suitable for vaccination according to the present invention as long as the antibody or antigen-binding fragment binds to gB of HSV-1 or HSV-2 and has the ability to be effective in active immunization of subjects against HSV-related diseases (as described above and below herein), and / or is capable of inhibiting the spread of HSV from an infected cell to another adjacent uninfected cell (cell-to-cell spread), and / or is capable of inhibiting cell-to-cell spread independent of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC) (as described above and below herein).

[0476] Therefore, in a preferred embodiment, the anti-HSV antibody for use in the vaccine according to the second aspect of the present invention comprises a light chain and a heavy chain variable region (i.e., the CDRs defined above, i.e., V comprising SEQ ID NO: 1). H CDR1, comprising V of SEQ ID NO:2 H CDR2, comprising V of SEQ ID NO:3 H CDR3, comprising V of SEQ ID NO:4 LCDR1, comprising V of SEQ ID NO:5 L CDR2, and V comprising SEQ ID NO:6 L CDR3), while at the same time the amino acid sequence has variability in the framework regions, wherein there is at least 75%, at least 80%, more preferably at least 85%, at least 90%, even more preferably at least 95%, and most preferably 98% overall sequence identity in the framework regions compared to the amino acid residues shown in positions 1 to 30, 38 to 51, 68 to 99 and 112 to 122 of SEQ ID NO:7 and in positions 1 to 23, 41 to 55, 63 to 94 and 104 to 114 of SEQ ID NO:8.

[0477] In this context, if SEQ ID NO:7 or SEQ ID NO:8 is aligned with the best matching sequence of the polypeptide of interest and the amino acid identity between those two aligned sequences is at least X% at positions 1 to 30, 38 to 51, 68 to 99 and 112 to 122 of SEQ ID NO:7 and positions 1 to 23, 41 to 55, 63 to 94 and 104 to 114 of SEQ ID NO:8, then the polypeptide has "at least X% sequence identity" with SEQ ID NO:7 or 8 in the framework regions. As mentioned above, such amino acid sequence alignments can be performed by using, for example, publicly available computer homology programs such as the "BLAST" program provided on the homepage of the National Centre for Biotechnology Information (NCBI), using the default settings provided there. Further methods for calculating the percentage of sequence identity of a set of amino acid sequences or nucleic acid sequences are known in the art.

[0478] In addition, in a preferred embodiment, the anti-HSV antibody for the vaccine for use according to the second aspect of the present invention comprises a V of SEQ ID NO: 9. H and V of SEQ ID NO: 10 L .

[0479] The sequence of glycoprotein B of HSV-1 and / or HSV-2 is well characterized and, as defined above, is not bound to a particular sequence, exemplary sequences of various HSV-1 and HSV-2 strains are shown in SEQ ID NOs: 11 to 16, respectively. The epitope recognized by the mAb 2c antibody is highly conserved among various HSV strains and between HSV-1 and HSV-2.

[0480] The antibodies of the vaccine for the use according to the second aspect of the invention are not limited to antibodies detecting the above-mentioned epitopes of glycoprotein B of HSV-1 and HSV-2. In fact, according to the present invention and in accordance with the basic principles of the present invention, other antibodies detecting another epitope of glycoprotein B or even an epitope of another protein or polypeptide of HSV-1 and HSV-2 may also be used in vaccines for active immunization of subjects against HSV-related diseases, as described above and below in this article.

[0481] With the normal skills of those skilled in the art and by conventional methods, those skilled in the art can easily derive relevant epitopes (also referred to as functional fragments) of polypeptides of HSV useful in the generation of antibodies such as polyclonal and monoclonal antibodies from the sequences provided herein. However, those skilled in the art can also easily provide modified antibodies such as CDR-grafted antibodies, or humanized antibodies and fully human antibodies, etc.

[0482] Particularly preferred in the context of the present invention is monoclonal antibody. In order to prepare monoclonal antibody, any technology providing antibodies produced by continuous cell line culture can be used. Examples of such technology include hybridoma technology, trioma technology, human B-cell hybridoma technology and EBV-hybridoma technology to produce human monoclonal antibodies (Shepherd and Dean (2000), Monoclonal Antibodies: A Practical Approach, Oxford University Press; Goding and Goding (1996), Monoclonal Antibodies: Principles and Practice-Production and Application of Monoclonal Antibodies in Cell Biology, Biochemistry and Immunology, Academic Pr Inc, USA).

[0483] Antibody derivatives can also be produced by peptide mimetics. Further, the techniques described for producing single-chain antibodies (see, inter alia, U.S. Pat. No. 4,946,778) can be adapted to produce single-chain antibodies that specifically recognize antigens of HSV. In addition, transgenic animals can be used to express humanized antibodies to polypeptides of HSV.

[0484] Without being bound by theory, the anti-HSV antibodies of the vaccine for the purpose according to the second aspect of the present invention can be produced against any natural polypeptide and recombinant polypeptide of glycoprotein B of HSV-1 and HSV-2 or any other protein or polypeptide. The production is based on, for example, immunization of animals (e.g., mice). However, other animals for producing antibodies / antisera are also contemplated within the present invention. For example, monoclonal and polyclonal antibodies can be produced by rabbits, mice, goats, donkeys, etc. The polynucleotide encoding the correspondingly selected polypeptide of HSV-1 or HSV-2 can be subcloned into a suitable vector, wherein the recombinant polypeptide is to be expressed in an organism capable of expression, such as in bacteria. Therefore, the expressed recombinant protein can be injected intraperitoneally into mice, and the resulting specific antibodies can be obtained, for example, from mouse serum provided by intracardiac blood puncture. The present invention also contemplates the production of specific antibodies against natural polypeptides and recombinant polypeptides by using a DNA vaccine strategy, as illustrated in the attached examples. DNA vaccine strategies are well known in the art, and include liposome-mediated delivery, by gene gun or jet injection, and intramuscular or intradermal injection. Therefore, antibodies to polypeptides or proteins or epitopes of HSV-1 and HSV-2 can be obtained by directly immunizing animals, and the immunization is carried out by directly injecting a carrier expressing the desired polypeptide or protein or epitope (particularly the epitope of gB) of HSV-1 and HSV-2 in an intramuscular manner. The amount of the specific antibody obtained can be quantified by using ELISA, which is also described below in this article. Further methods for producing antibodies are well known in the art, referring to, for example, Harlow and Lane, "Antibodies, A Laboratory Manual", CSH Press, Cold Spring Harbor, 1988.

[0485] As used herein, the term "specifically binds" refers to a binding reaction that is decisive for the presence of the desired polypeptide or protein or epitope of HSV-1 and HSV-2 (particularly the epitope of gB), and antibodies, in the presence of a heterogeneous population of proteins and other biologicals.

[0486] Therefore, under the specified assay conditions, the clearly specified antibodies and the corresponding polypeptides or proteins or epitopes of HSV-1 and HSV-2 (particularly the epitopes of gB) bind to each other and do not bind to other components present in the sample in significant amounts. Specific binding to the target analyte under such conditions may require a binding moiety that is selected for its specificity for a specific target analyte. A variety of immunoassay formats can be used to select antibodies that specifically react with a specific antigen. For example, solid phase ELISA immunoassays are routinely used to select monoclonal antibodies that specifically react with the analyte. For a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity, see Shepherd and Dean (2000), Monoclonal Antibodies: A Practical Approach, Oxford University Press; and / or Howard and Bethell (2000) Basic Methods in Antibody Production and Characterization, Crc. Pr. Inc. Typically, a specific or selective response will be at least twice the background signal to noise ratio, and more typically more than 10 to 100 times the background. Those skilled in the art are able to provide and generate specific binding molecules for new polypeptides. For specific binding assays, it can be easily adopted to avoid undesirable cross-reactivity, for example polyclonal antibodies can be easily purified and selected by known methods (see Shepherd and Dean, loc. cit.).

[0487] According to the present invention, the term "anti-HSV antibody" means that the antibody molecule or its antigen-binding fragment is able to specifically recognize at least two amino acids of the desired polypeptide or protein or epitope (especially the epitope of gB) of HSV-1 and HSV-2, respectively, or specifically interact and / or bind thereto. The term relates to the specificity of the antibody molecule, i.e. its ability to distinguish specific regions of the desired polypeptide or protein or epitope (especially the epitope of gB) of HSV-1 and HSV-2. Therefore, the specificity can be experimentally determined by methods known in the art and disclosed and described herein. Such methods include, but are not limited to, Western blots, ELISA-tests, RIA-tests, ECL-tests, IRMA-tests and peptide scanning. Such methods also include K D-value determination, as especially illustrated in the accompanying examples. Peptide scanning (pepspot assay) is routinely used to map linear epitopes in polypeptide antigens. The primary sequence of the polypeptide is successively synthesized on activated cellulose with overlapping peptides. The antibody to be tested for its ability to detect or recognize a specific antigen / epitope is scored for the recognition of certain peptides by the following formula: conventional color development (secondary antibody with horseradish peroxidase and 4-chloronaphthol and hydrogen peroxide), chemiluminescent reaction, or similar means known in the art. In the case of chemiluminescent reaction, in particular, the reaction can be quantitative. If the antibody reacts with a certain set of overlapping peptides, the minimum amino acid sequence necessary for the reaction can be inferred. The same assay can reveal two long-range clusters of reactive peptides, which indicates the recognition of discontinuous (i.e. conformational) epitopes in the antigen polypeptide (Geysen (1986), Mol. Immunol. 23, 709-715).

[0488] A preferred epitope of the anti-HSV antibodies defined above and below is the same as the epitope recognized by mAb2c.

[0489] In a preferred embodiment, the anti-HSV antibody of the vaccine for use according to the second aspect of the present invention is mAb 2c antibody (or an antigen-binding fragment thereof). This monoclonal antibody MAb 2c has been described elsewhere and has been shown to neutralize the virus by abrogating viral cell-to-cell spread, which is a key mechanism by which HSV-1 / 2 evades humoral immune surveillance (independent of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC)); Eis-Hübinger et al., Intervirology 32:351-360 (1991); Eis-Hübinger et al., Journal of General Virology 74:379-385 (1993); WO2011 / 038933A2; Krawczyk A et al., Journal of virology (2011), 85(4):1793-1803; Krawczyk A et al., Proc Natl Acad Sci USA (2013), 110(17):6760-6765.

[0490] As outlined above, the vaccine of the second aspect of the invention may comprise one of two alternative components (although it is also contemplated that the vaccine comprises both components), namely:

[0491] (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection; or

[0492] (b) wherein the Fc-containing fusion protein comprises the Fc portion of an antibody fused to an antigen, wherein the antigen is associated with an HSV-related disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC) selected from the group consisting of:

[0493] Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and

[0494] Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

[0495] The second component (b) is described in more detail below.

[0496] The Fc-containing fusion protein comprises the Fc portion of an antibody fused to an antigen, wherein the antigen is associated with an HSV-associated disease. Associated with, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC):

[0497] Regarding (b) , i.e., an Fc-containing fusion protein comprising the Fc part of an antibody fused to an antigen associated with an HSV-related disease and corresponding preferred embodiments, which are also applicable after appropriate modifications, as already described above in relation to the first aspect of the present invention, except that the "antigen associated with the infectious disease" is an antigen associated with an acute HSV-related disease, preferably an acute HSV infection.

[0498] With regard to the "antigens associated with acute HSV-related diseases, preferably acute HSV infection" and the corresponding preferred embodiments, the same applies mutatis mutandis as already explained above in relation to alternative (a) of the second aspect of the invention.

[0499] Without being bound by theory and as outlined in Example 2 below, and as in Fig.13 As illustrated in , further, more preferred embodiments of the above-mentioned second aspect of the present invention are envisaged.

[0500] The rationale for inducing de novo T- and B-cell responses to induce long-lasting protection against recurrence of HSV lesions, due to the characteristics of the HDIT101 antibody that does not induce ADCC or CDC and combined with its binding properties to its target protein gB, led to the proposal to combine these effects to generate a vaccine.

[0501] Therefore, in a preferred embodiment, the vaccine of the present invention may comprise whole virus particles complexed with an anti-HSV antibody of the present invention (preferably, HDIT101 antibody).

[0502] Furthermore, in a preferred embodiment, the vaccine of the present invention may comprise a trimeric gB extracellular domain fused to the Fc portion of an anti-HSV antibody (preferably, HDIT101 antibody), which is further complexed with an anti-HSV antibody (preferably, HDIT101 antibody).

[0503] In a further preferred embodiment, ADCP can be even further enhanced by introducing specific amino acid substitutions (eg, G236A) in Fc.

[0504] Therefore, in a preferred embodiment, the antibody of the second aspect of the invention and the Fc-containing fusion protein comprising the Fc part of the antibody respectively comprise the Fc part of such an antibody having an amino acid substitution at position 236, 239 and / or 332 of the Fc part of the antibody.

[0505] The corresponding preferred embodiments and sequences apply mutatis mutandis to the second aspect of the invention as defined above in the context of the first aspect of the invention.

[0506] Without being bound by theory, as an Fc-fusion protein, gB-Fc adopts a trimer structure comprising three Fc domains, two of which dimerize, leaving one free Fc domain that can dimerize with the free Fc domains of other gB-Fc trimers, thereby generating larger complexes of multiple copies of gB-Fc.

[0507] To increase immunogenicity, in a further preferred embodiment, the anti-HSV antibody of the present invention (preferably, HDIT101 antibody) can be contacted with a gB-Fc protein unit to reveal the new epitopes induced by HDIT101 binding and to direct the structure towards ADCP, thereby resulting in a multimeric complex of gB-Fc protein multivalently bound to the anti-HSV antibody (preferably, HDIT101 antibody). These structures combine the characteristics of the gB-Fc Ag target and the properties of the anti-HSV antibody (preferably, HDIT101 antibody) according to the present invention for enhanced ADCP, while importantly avoiding ADCC and CDC induction, which is proposed to lead to a substantially increased T-cell (and B-cell) response and vaccine effect. In a preferred embodiment, the corresponding IC increased by mAb can be used as a carrier for enhanced delivery of irrelevant Ag to the ADCP pathway, resulting in activation of de novo T-cell (and B-cell) responses.

[0508] Therefore, in a preferred embodiment, if it is for example in Fig.13As illustrated in FIG. 1 , in addition to antigens associated with infectious diseases or malignant diseases (preferably HSV-infectious diseases), the Fc-containing fusion protein may also contain unrelated antigens fused thereto.

[0509] The corresponding "irrelevant antigen" may be any antigen described in the context of the first aspect of the present invention. Therefore, with regard to the corresponding preferred embodiments of "irrelevant antigens" containing Fc fusion proteins (which, in addition to antigens associated with infectious diseases or malignant diseases (preferably HSV-infectious diseases), also contain such irrelevant antigens fused thereto), this also applies, mutatis mutandis, as has been described above with regard to the first aspect of the present invention.

[0510] In a preferred embodiment of the second aspect of the present invention, in the vaccine for the purposes defined above, the antibody or the Fc-containing fusion protein is capable of inducing antibody-dependent cellular phagocytosis (ADCC) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC). In a more preferred embodiment, in the vaccine for the purposes defined above, the Fc-containing fusion protein is capable of inducing antibody-dependent cellular phagocytosis (ADCP) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC), while at the same time the Fc-containing fusion protein is capable of mediating ADCC and CDC, provided that the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen is capable of bridging to a cell via binding of the antigen to a cell, whereby the Fc is free and can mediate ADCC or CDC.

[0511] In a further preferred embodiment, the vaccines of the first and second aspects of the invention, more particularly the antibodies and Fc-containing fusion proteins of the invention, respectively, are capable of inducing antibody-dependent cellular phagocytosis (ADCP) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

[0512] In a more preferred embodiment, in the vaccine for said use as defined above, the Fc-containing fusion protein is capable of inducing antibody-dependent cellular phagocytosis (ADCP) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC), while at the same time the Fc-containing fusion protein is capable of mediating ADCC and CDC, provided that the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen is capable of bridging to a cell via binding of the antigen to a cell, such that the Fc is free and can mediate ADCC or CDC.

[0513] "Independent of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC)" means that the antibodies and Fc-containing fusion proteins of the present invention can respectively induce antibody-dependent cellular phagocytosis (ADCP) without inducing antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC). Therefore, in a preferred embodiment, the antibodies and Fc-containing fusion proteins of the present invention respectively do not induce antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

[0514] In a further preferred embodiment, the antibodies and Fc-containing fusion proteins of the present invention are respectively capable of inducing antibody-dependent cellular phagocytosis (ADCP).

[0515] In a further preferred embodiment, the antibodies and Fc-containing fusion proteins of the present invention are respectively capable of mediating Fc-based effector functions by antibody-dependent cellular phagocytosis (ADCP).

[0516] Without being bound by theory, the ability to mediate Fc-based effector functions by antibody-dependent cellular phagocytosis (ADCP) can be tested as follows: fluorescent beads can be coupled to an antigen and incubated with an antibody against the antigen. In the case of an antibody that mediates ADCP, incubation of APCs with the bead-Ag-antibody complex results in enhanced uptake of the fluorescent beads into the APCs, compared to bead-Ag without antibody and compared to bead-Ag mixed with Fab2 fragments, demonstrating Fc-based ADCP. Uptake of fluorescent beads can be measured using a flow cytometry assay.

[0517] In a further preferred embodiment, the antibody and Fc-containing fusion protein of the present invention are respectively capable of inducing antibody-dependent cellular phagocytosis (ADCP) via interaction with any of the above-mentioned Fc receptors.

[0518] Without being bound by theory, the ability to trigger antibody-dependent cellular phagocytosis (ADCP) via interaction with any of the above-mentioned Fc receptors can be tested as follows: In order to demonstrate ADCP via Fc receptors, APCs can be manipulated so that specific Fc receptor expression is reduced, or completely knocked out using gene editing tools such as CRISPR / Cas9. In addition, blocking specific Fc receptors with anti-Fc receptor antibodies can prevent the binding of antibodies or Fc-Ag fusion proteins. Therefore, after incubation with antibodies, for example, the uptake of fluorescent beads coated with antigens will be reduced in such APCs, wherein specific Fc receptors are expressed reduced by using, for example, silencing RNA, microRNA, short hairpin RNA or CRISPR / Cas-mediated knockout, or wherein Fc receptors are blocked by using anti-Fc receptor antibodies. The uptake of fluorescent beads can be measured by flow cytometry.

[0519] In a further preferred embodiment, the antibodies and Fc-containing fusion proteins of the present invention are respectively capable of inducing antibody-dependent cellular phagocytosis (ADCP) leading to the production of cytokines by antigen presenting cells (APCs).

[0520] Without being bound by theory, the ability to induce antibody-dependent cellular phagocytosis (ADCP) leading to cytokine production by antigen presenting cells (APCs) can be tested as follows: APCs (e.g., monocyte-derived dendritic cells or macrophages) can be incubated with immune complexes of antibody-bound antigens, or by incubating with Fc-antigen fusion proteins, or by incubating with a mixture of antibody-crosslinked Fc-Ag immune complexes (e.g., gB-Fc mixed with HDIT101). After one to two days of incubation, the supernatant is analyzed for the presence of cytokines (e.g., IL1β, IP10, TNFα, IL6, IFNγ, etc.) released by the APCs using commercially available assays, such as Luminex-based measurements or ELISA. If the antibody or Fc-Ag fusion protein mediates cytokine release via ADCP, the release is prevented in cells that do not express a specific Fc receptor (i.e., cannot mediate ADCP) or in cells in which Fc receptor binding of the antibody or Fc-Ag fusion protein is inhibited by specific anti-Fc receptor antibodies.

[0521] In a further preferred embodiment, the antibodies and Fc-containing fusion proteins of the present invention are capable of inducing antibody-dependent cellular phagocytosis (ADCP) resulting in the generation of de novo HSV-derived peptides on MHC complexes and presentation by antigen presenting cells (APCs), respectively.

[0522] Without being bound by theory, the ability to induce antibody-dependent cellular phagocytosis (ADCP) leading to the generation of de novo HSV-derived peptides on MHC complexes and presentation by antigen presenting cells (APCs) can be tested as follows: APCs can be incubated with antibodies bound to the antigen (i.e., immune complexes) or with Fc-Ag fusion proteins or mixtures thereof. The cells are then lysed, and the MHC complexes can be immunoprecipitated using specific anti-MHC antibodies, and the MHC immunopeptidome, i.e., the peptides bound and presented by the MHC, can be identified using mass spectrometry. This method will directly reveal the identity of the stimulated peptides. This method can also be combined with stable isotope labeling to separate cellular peptides from antigen-derived peptides loaded on the MHC complex.

[0523] In a further preferred embodiment, the antibodies and Fc-containing fusion proteins of the present invention are respectively capable of inducing antibody-dependent cellular phagocytosis (ADCP) leading to the activation of T-cell (and / or B-cell) immune responses against de novo HSV-derived peptides.

[0524] Without being bound by theory, the ability to elicit antibody-dependent cellular phagocytosis (ADCP) leading to activation of T-cell (and / or B-cell) immune responses against de novo HSV-derived peptides can be tested as follows: The method for showing T-cell stimulation by new peptides presented by MHC complexes after ADCP via the antibodies or Fc-Ag fusion proteins or mixtures thereof described in the present invention is to incubate APCs with immune complexes such as antibodies bound to Ag, or with Fc-Ag fusion proteins or mixtures of Fc-Ag and antibodies, then inject these into animal models of malignant or infectious diseases and after two weeks isolate T cells and amplify T-cell receptor nucleic acid sequences from total DNA or mRNA of circulating T-cells via polymerase chain reaction. The identification of the expansion of specific T cell clones (which were not present before vaccination) in mice vaccinated with immune complexes such as antibodies bound to antigens, or by Fc-Ag fusion proteins or mixtures of Fc-Ag fusion proteins and antibodies indicates presentation of de novo peptides via MHC. Similarly, B-cell receptor nucleic acid sequences can be amplified from circulating B cells, and clonal expansion of B cells, ie, clonal expansion of a specific B-cell receptor sequence that was not present prior to vaccination, indicates a B cell response to a de novo epitope.

[0525] In a further preferred embodiment, the antibodies and Fc-containing fusion proteins of the present invention are capable of inducing antibody-dependent cellular phagocytosis (ADCP) leading to the production of HSV-derived peptides and presentation by antigen presenting cells (APCs), respectively.

[0526] The vaccine for use according to the above first and second aspects of the present invention is used for active immunization of a subject.

[0527] Therefore, the antibodies and Fc-containing fusion proteins for use in vaccines according to the first and second aspects of the invention, respectively, as defined above are particularly useful in a medical context.

[0528] Therefore, in a preferred embodiment, the present invention relates to a vaccine pharmaceutical composition comprising an effective amount of the above-described respective antibodies and Fc-containing fusion proteins, and at least one pharmaceutically acceptable excipient.

[0529] The terms "vaccination," "active immunization," and the like are used herein to generally mean obtaining the desired pharmacological and / or physiological effects as described herein above according to the underlying principles of the present invention.

[0530] The vaccine pharmaceutical composition of the present invention can be administered via a wide range of administration forms known to the skilled person. Administration can be systemic, topical, oral, by aerosol, including but not limited to tablets, needle injection, use of inhalers, creams, foams, gels, lotions and ointments.

[0531] Preferably, the vaccine (or vaccine pharmaceutical composition) for use according to the first aspect of the invention is to be administered intravenously, topically, intradermally, subcutaneously, intradermally, intramuscularly and / or intrathecally.

[0532] Thus, preferably, the antibodies and Fc-containing fusion proteins described in the context of the first aspect of the invention as part of a vaccine (or vaccine pharmaceutical composition) are to be administered intravenously, topically, intradermally, subcutaneously, intradermally, intramuscularly and / or intrathecally, respectively.

[0533] Preferably, the vaccine (or vaccine pharmaceutical composition) for the use according to the second aspect of the invention is to be administered intravenously, topically, intradermally, subcutaneously, intradermally, intramuscularly and / or intrathecally.

[0534] Thus, preferably, the antibodies and Fc-containing fusion proteins described in the context of the second aspect of the invention as part of a vaccine (or vaccine pharmaceutical composition) are to be administered intravenously, topically, intradermally, subcutaneously, intradermally, intramuscularly and / or intrathecally, respectively.

[0535] These routes of administration, ie via the intravenous, topical, intradermal, subcutaneous, intradermal, intramuscular and / or intrathecal routes, are known to the skilled person.

[0536] Excipients or carriers are inactive substances that are formulated with active ingredients (i.e., the antibodies and Fc-containing fusion proteins described above of the present invention) for the purpose of increasing the size of the formulation containing the potent active ingredient. Excipients are often referred to as "fillers," "stuffers," or "diluents." Enlargement allows convenient and accurate distribution of drug substances when producing dosage forms. They can also be used for various therapeutic enhancement purposes, such as promoting drug absorption or dissolution, or other pharmacokinetic considerations. Excipients can also be useful during the preparation process to help the handling of the active substance of interest, such as by promoting powder flowability or non-stick properties, in addition to helping in vitro stability, such as preventing denaturation during the expected shelf life. The selection of suitable excipients also depends on the route of administration and dosage form, as well as the active ingredient and other factors.

[0537] Therefore, the vaccine or vaccine pharmaceutical composition comprising an effective amount of the antibody and Fc fusion protein of the present invention described above can be in solid, liquid or gaseous form, and can be in the form of powder, tablet, solution or aerosol. Preferably, the vaccine pharmaceutical composition is optionally contained in a pharmaceutically acceptable carrier and / or diluent.

[0538] These vaccines or vaccine pharmaceutical compositions can be administered to a subject at a suitable dose. The administration of a suitable composition can be carried out by the different modes outlined above, and can also be carried out, for example, by intravenous, intraperitoneal, subcutaneous, intramuscular, topical, intradermal, intranasal or intrabronchial administration.

[0539] The dosage regimen will be determined by the attending physician and clinical factors. As is well known in the medical field, the dosage for any one patient depends on many factors, including the patient's body size, body surface area, age, specific compound to be administered, sex, time and route of administration, general health, and other drugs being administered simultaneously. Proteinaceous pharmaceutically active substances can be present in an amount of 1 ng to 10 mg / kg body weight per dose; however, doses below or above this exemplary range are envisioned, especially in view of the factors mentioned above. If the regimen is a continuous infusion, it should also be in the range of 1 μg to 10 mg units / kg body weight / minute.

[0540] Examples of suitable pharmaceutical carriers, excipients and / or diluents are well known in the art and include phosphate buffered saline solutions, water, emulsions (e.g., oil / water emulsions), various types of wetting agents, sterile solutions, etc. Compositions comprising such carriers can be formulated by well-known conventional methods. These vaccine pharmaceutical compositions can be administered to subjects at suitable doses, i.e., with an "effective amount," which can be easily determined by a technician by methods known in the art. The dosage regimen will be determined by the attending physician and clinical factors. As is well known in the medical field, the dosage for any one patient depends on many factors, including the patient's or subject's weight, body surface area, age, specific compound to be administered, sex, time and route of administration, general health, and other drugs being administered simultaneously.

[0541] Therefore, preferably, the antibody of the present invention described above and the Fc fusion protein are included in an effective amount, respectively. The term "effective amount" refers to an amount sufficient to induce detectable active immunization in a subject to which the vaccine pharmaceutical composition is to be administered. According to the above, the content of the antibody of the present invention and the Fc fusion protein is not limited in the vaccine pharmaceutical composition, as long as it is useful for the active immunization described above, but preferably comprises 0.0000001-10 weight % / total composition. Further, the antibody described in this article is preferably used in a carrier. Typically, a suitable amount of a pharmaceutically acceptable salt is used in the carrier to make the composition isotonic. Examples of carriers include, but are not limited to, saline, Ringer's solution, and dextrose solution. Preferably, acceptable excipients, carriers or stabilizers are nontoxic at the amounts and concentrations used, and include buffers such as citrate, phosphate and other organic acids; salt-forming counterions such as sodium and potassium; low molecular weight (>10 amino acid residues) polypeptides; proteins such as serum albumin or gelatin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as histidine, glutamine, lysine, asparagine, arginine or glycine; carbohydrates including glucose, mannose or dextrin; monosaccharides; disaccharides; other Sugars, such as sucrose, mannitol, trehalose or sorbitol; chelating agents, such as EDTA; nonionic surfactants, such as Tween, Pluronics or polyethylene glycol; antioxidants, including methionine, ascorbic acid and tocopherol; and / or preservatives (e.g., octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens, such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol). Suitable carriers and their formulations are described in more detail in Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Co.

[0542] Active immunization can be monitored by regular assessments by a skilled artisan using routine methods as outlined above.

[0543] The antibodies and Fc-containing fusion proteins of the vaccines of the present invention or the vaccine pharmaceutical compositions of the present invention can be in sterile aqueous or non-aqueous solutions, suspensions and emulsions, as well as creams and suppositories. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Preservatives and other additives such as antimicrobials, antioxidants, chelating agents and inert gases may also be present. Further, the pharmaceutical compositions of the present invention may include further agents, depending on the intended use of the pharmaceutical compositions. The agents may be, for example, polyoxyethylene sorbitan monolaurate (which is available on the market under the trade name Tween), propylene glycol, EDTA, citrate, sucrose, and other agents well known to those skilled in the art that are suitable for the intended use of the pharmaceutical compositions.

[0544] According to the present invention, the terms "vaccine" and "vaccine pharmaceutical composition" relate to a vaccine composition for administration to a patient, preferably a human patient.

[0545] As shown in the appended examples, the vaccine according to the invention surprisingly leads to a significantly longer time to first relapse and a lower relapse rate in patients with severe infectious diseases (see Example 8).

[0546] Furthermore, pharmacokinetic time-to-event (TTE) modeling analysis revealed that multiple administrations of the vaccine surprisingly resulted in a higher median probability of relapse / longer time free from injury (see Example 9).

[0547] This leads to a further preferred embodiment of the invention wherein, briefly,

[0548] (i) specific subgroups of patients may benefit from vaccination; and / or

[0549] (ii) Provides a specific beneficial prime-boost dosing regimen.

[0550] This is in accordance with the technical problem set out above as the basis of the present invention (according to which further means and methods are provided for the treatment and / or prevention of (recurrent) infectious diseases and malignant diseases, in particular with regard to a long-term or lasting effect, wherein, in particular, the individuals / subjects who have to be treated simultaneously acquire long-term immunity against the infectious disease or malignant disease by means of vaccination).

[0551] These further embodiments of the invention are described below separately for the first and second aspects of the invention.

[0552] first In the following, the present invention is described in more detail. The first aspect (relates to a vaccine for use in active immunization of a subject against an infectious disease or a malignant disease, the vaccine comprising: (a) an antibody against an antigen associated with the infectious disease or the malignant disease; or (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or the malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21) for a specific subgroup of patients and / or a specific prime-boost dosing regimen.

[0553] Therefore, in a preferred embodiment, the present invention relates to a vaccine comprising: (a) antibodies against an antigen associated with said infectious disease or said malignant disease, wherein said vaccine is to be administered to subjects having an acute relapse of said infectious disease or said malignant disease and suffering from a chronic (and / or severe chronic) (infectious) disease or a chronic (and / or severe chronic) (malignant) disease with more than 4 relapses / year.

[0554] As outlined above, in the context of a vaccine for use in active immunization of a subject against an infectious disease or a malignant disease, in a preferred embodiment, it requires the presence of an acute relapse (in terms of acute / persistent infection) or an acute relapse of a malignant disease (in terms of acute / persistent malignant disease), and therefore it requires the presence of an antigen. In particular, this relates to aspect (a), wherein antibodies to an antigen associated with the infectious disease or malignant disease are used as components of the vaccine of the present invention. This is because, in order to be able to actively immunize a subject according to the basic principles of the present invention, not only the antibodies but also the antigen associated with the infectious disease or malignant disease must be present.

[0555] Furthermore, according to this preferred embodiment, the subject preferably suffers from a chronic (preferably, severe chronic) infectious disease or a chronic (preferably, severe chronic) malignant disease with more than 4 relapses / year.

[0556] “Severe infectious disease” / “chronic infectious disease” / “severe chronic infectious disease” or “severe malignant disease” / “chronic malignant disease” / “severe chronic malignant disease” is a disease that presents / shows several recurrences / year.

[0557] The term "chronic" is generally understood in a medical context as a (health) condition or disease that is persistent or long-lasting in its effects (also called chronic disease or chronic illness) or a disease that occurs over time. The term "chronic" is often applied when the course of the disease lasts for more than three months.

[0558] The term "relapse" is generally understood in the art in the context of a medical setting as the return or reappearance of a condition previously characteristic of the disease, and may be, for example, a new or repeated outbreak of the disease and / or symptoms. The skilled person is easily able to determine the onset / appearance of the outbreak or symptoms, i.e. the onset or onset of a relapse according to the present invention.

[0559] The severity of chronic disease depends on the number of relapses / year.

[0560] In preferred embodiments, the subject to be treated has more than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 relapses per year.

[0561] In a more preferred embodiment, the subject to be treated has more than 4 relapses per year.

[0562] In another preferred embodiment, when the severe infectious disease is an HSV-related disease, preferably a severe HSV infection, the recurrence is a recurrence of the lesion.

[0563] As mentioned, in the context of the first aspect of the invention, the invention relates to a specific dosing regimen.

[0564] Therefore, in another preferred embodiment, the present invention relates to a vaccine comprising: (a) antibodies against an antigen associated with said infectious disease or malignant disease, wherein said vaccine is to be administered in a prime-boost regimen by the following administration steps:

[0565] (i) administering a first dose of the vaccine to the subject during the first week of relapse, preferably immediately after the onset of the first relapse (priming administration);

[0566] (ii) administering a further dose of the vaccine to the subject during the first week of the second relapse, preferably immediately after the onset of the second relapse (boosting administration); and

[0567] (iii) optionally, during the first week of each further relapse, preferably immediately after the onset of each further relapse, repeating step (ii) (further booster administrations),

[0568] One initial administration and at least one booster administration are to be performed per year.

[0569] In a further preferred embodiment, in the context of the first aspect of the invention, the specific dosage regimen described above and below is preferably to be administered to subjects suffering from a chronic (and / or severe chronic) (infectious) disease or a chronic (and / or severe chronic) (malignant) disease with more than 4 recurrences / year.

[0570] In the context of the present invention, a "week" refers to a period of 7 days.

[0571] In the context of the present invention, "month" refers to a period of 4 weeks.

[0572] The priming and boosting administrations are each to be administered during the first week of relapse, preferably immediately after the onset of relapse of the disease.

[0573] As explained above, a relapse, return, reappearance or repeated outbreak of the respective disease is easily determined by the skilled person and is preferably a point in time wherein disease-specific symptoms can be detected.

[0574] In a preferred embodiment, when the infectious disease is an HSV-related disease, preferably a severe HSV infection, the recurrence is a recurrence of the lesion.

[0575] The time points of the initial boost administration and the boost administration are not necessarily exactly during the first week of relapse, and are not necessarily exactly immediately after the relapse of the disease, respectively. The appropriate time point can be determined within an appropriate range according to the condition of the subject or patient.

[0576] Thus, in preferred embodiments, the specific priming administration may be 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, or 42 days after the onset of a relapse of the disease.

[0577] In a more preferred embodiment, the specific priming administration is on day 0 (ie, on the same day as the relapse of disease).

[0578] In a more preferred embodiment, the specific priming administration is during the first week, more preferably on day 1, day 2, day 3, day 4, day 5, day 6 or day 7 after the onset of relapse of the disease.

[0579] The same preferred embodiments apply mutatis mutandis to said one or more booster administrations.

[0580] The boosting administrations were repeated for each further relapse as outlined (further boosting administrations).

[0581] Therefore, in a preferred embodiment, the initial boost administration of the vaccine of the present invention is performed at the onset of the disease or after its first relapse (within the limits stated above), and the nth (n is an integer of 1 or greater) boost administration is performed at the onset of the disease or after its nth (n is an integer of 1 or greater) relapse (within the limits stated above).

[0582] As outlined, in the priming and boosting regimen, in a preferred embodiment, one priming administration and at least one boosting administration per year are to be administered.

[0583] In a further preferred embodiment, one priming administration and at least two boosting administrations per year are to be administered.

[0584] In a further preferred embodiment, one priming administration and at least three boosting administrations per year are to be administered.

[0585] In a further preferred embodiment, one priming administration and at least four, five, six, seven, eight, nine, ten, eleven, twelve or more boosting administrations per year are administered.

[0586] In a further preferred embodiment, one priming administration and n-th boosting administration per year (n is an integer of 1 or greater) are to be performed.

[0587] In another preferred embodiment, the present invention relates to a vaccine comprising: (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with said infectious disease or malignant disease, wherein said Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), said receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21, wherein said vaccine is to be administered to subjects suffering from a chronic (and / or severe chronic) (infectious) disease or a chronic (and / or severe chronic) (malignant) disease with more than 4 relapses / year.

[0588] With regard to preferred embodiments, the same applies mutatis mutandis as has been explained above in relation to the above defined vaccines comprising (a) the antibodies of the invention in the context of specific subgroups of patients.

[0589] In another preferred embodiment, when the severe infectious disease is an HSV-related disease, preferably a (severe) chronic HSV infection, the recurrence is a recurrence of the lesion.

[0590] In addition, with regard to a specific dosing regimen (i.e., prime-boost administration) of a vaccine comprising the following components: (b) an Fc-fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and type II: neonatal FcR (FcRn) and cytoplasmic TRIM21, in another preferred embodiment, the present invention relates to the vaccine, wherein the vaccine is to be administered in a prime-boost dosing regimen by the following administration steps:

[0591] (i) administering a first dose of the vaccine to the subject (prime administration);

[0592] (ii) administering further doses of the vaccine to the subject at intervals of 15 to 30 days after the initial boost (boosting administrations); and

[0593] (iii) Optionally, step (ii) is repeated at intervals of 15 to 30 days after the previous booster administration (further booster administration).

[0594] The "interval" (interval between individual administrations) indicates the interval between the administration of the nth dose (n is an integer of 1 or more) and the administration of the (n+1)th dose.

[0595] Suitable intervals between the first (priming) dose and the further (boosting) dose and between the further (boosting) dose and respectively the subsequent further (boosting) dose, respectively, can be determined within a suitable range depending on the condition of the subject or patient. Specific administration intervals between the first (priming) dose and the further (boosting) dose and between the further (boosting) dose and the respective subsequent further (boosting) dose are 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days or 45 days, respectively.

[0596] In a preferred embodiment, the interval between the first dosage and the further (reinforced) dosage and between the further (reinforced) dosage and the corresponding further (reinforced) dosage is 5 to 45 days in stages, respectively. In a further preferred embodiment, the interval is 5 to 45 days. In a further preferred embodiment, the interval is 5 to 40 days. In a further preferred embodiment, the interval is 5 to 35 days. In a further preferred embodiment, the interval is 5 to 30 days. In a further preferred embodiment, the interval is 5 to 25 days. In a further preferred embodiment, the interval is 5 to 20 days. In a further preferred embodiment, the interval is 5 to 15 days. In a further preferred embodiment, the interval is 5 to 10 days.

[0597] In a further preferred embodiment, the interval is from 10 days to 45 days. In a further preferred embodiment, the interval is from 10 days to 40 days. In a further preferred embodiment, the interval is from 10 days to 35 days. In a further preferred embodiment, the interval is from 10 days to 30 days. In a further preferred embodiment, the interval is from 10 days to 25 days. In a further preferred embodiment, the interval is from 10 days to 20 days. In a further preferred embodiment, the interval is from 10 days to 15 days.

[0598] In a further preferred embodiment, the interval is from 15 days to 45 days. In a further preferred embodiment, the interval is from 15 days to 40 days. In a further preferred embodiment, the interval is from 15 days to 35 days. In a further preferred embodiment, the interval is from 15 days to 30 days. In a further preferred embodiment, the interval is from 15 days to 25 days. In a further preferred embodiment, the interval is from 15 days to 20 days.

[0599] In a further preferred embodiment, the interval is 20 to 45 days. In a further preferred embodiment, the interval is 20 to 40 days. In a further preferred embodiment, the interval is 20 to 35 days. In a further preferred embodiment, the interval is 20 to 30 days. In a further preferred embodiment, the interval is 20 to 25 days.

[0600] In a further preferred embodiment, the interval is 25 to 45 days. In a further preferred embodiment, the interval is 25 to 40 days. In a further preferred embodiment, the interval is 25 to 35 days. In a further preferred embodiment, the interval is 25 to 30 days.

[0601] In a further preferred embodiment, the interval is 30 to 45 days. In a further preferred embodiment, the interval is 30 to 40 days. In a further preferred embodiment, the interval is 30 to 35 days.

[0602] In the most preferred embodiments, the interval is 15 or 30 days.

[0603] In other preferred embodiments, the further (boosting) dose and the respective subsequent further (boosting) doses are administered 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days (1 week), 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days (2 weeks), 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days (3 weeks), 22 days, 24 days, 25 days, 26 days, 27 days, 28 days (4 weeks), 29 days, 30 days or more after the first (priming) dose and the further (boosting) dose, respectively.

[0604] In a more preferred embodiment, the interval between the further (boosting) dose and respectively the subsequent further (boosting) dose is 15 days, 20 days, 25 days or 30 days after the first (priming) dose and the further (boosting) dose, respectively.

[0605] Secondly In the following, the present invention is described in more detail. The second aspect (relating to a vaccine for use in active immunization of a subject against an infectious disease, wherein the infectious disease is an HSV-related disease; and (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection; or (b) wherein in the Fc-containing fusion protein comprising the Fc portion of the antibody fused to an antigen, the antigen is associated with an HSV-related disease) to a specific subgroup of patients and / or a specific prime-boost dosing regimen.

[0606] Therefore, in a preferred embodiment, the present invention relates to a vaccine for use in active immunization of a subject against an infectious disease, wherein the infectious disease is an HSV-related disease; and (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection, wherein the vaccine is to be administered to subjects having an acute recurrence of the disease and suffering from a chronic (and / or severe chronic) HSV-related disease, preferably a chronic (and / or severe chronic) HSV infection with more than 4 recurrences / year.

[0607] Concerning the preferred embodiments of this specifically defined group of patients / subjects (i.e. subjects suffering from chronic (and / or severe chronic) HSV-related diseases with more than 4 recurrences / year, preferably chronic (and / or severe chronic) HSV infection), the same applies mutatis mutandis as already explained above in relation to the first aspect of the invention in the context of a specific subgroup of patients.

[0608] As mentioned, in the context of the second aspect of the invention, the invention relates to specific dosing regimens.

[0609] Therefore, in another preferred embodiment, the present invention relates to a vaccine for use in active immunization of a subject against an infectious disease, wherein the infectious disease is an HSV-related disease; and (a) wherein the antibody is an anti-HSV antibody and the subject has an acute relapse of the disease and suffers from a chronic (and / or severe chronic) HSV-related disease, preferably a chronic (and / or severe chronic) HSV infection, wherein the vaccine is to be administered in a prime-boost dosing regimen by the following administration steps:

[0610] (i) administering a first dose of the vaccine to the subject during the first week of relapse, preferably immediately after the onset of the first relapse (priming administration);

[0611] (ii) administering a further dose of the vaccine to the subject during the first week of relapse, preferably immediately after the onset of a second relapse (boosting administration); and

[0612] (iii) optionally, during the first week of each further relapse, preferably immediately after the onset of each further relapse, repeating step (ii) (further booster administrations),

[0613] One initial administration and at least one booster administration are to be performed per year.

[0614] With regard to the preferred embodiments of this specifically defined dosage regimen, the same applies mutatis mutandis as has been explained above in relation to the first aspect of the invention in the context of a specific dosage regimen.

[0615] In another preferred embodiment, the present invention relates to a vaccine for use in active immunization of a subject against an infectious disease, wherein the infectious disease is an HSV-related disease; and wherein the vaccine comprises: (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and type II: neonatal FcR (FcRn) and cytoplasmic TRIM21, wherein in the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen, the antigen is associated with an HSV-related disease, and wherein the vaccine is to be administered to a subject suffering from a (severe) chronic HSV-related disease, preferably a (severe) chronic HSV infection, with more than 4 relapses / year.

[0616] Concerning the preferred embodiments of this specifically defined group of patients / subjects (i.e. subjects suffering from (severe) chronic HSV-related diseases with more than 4 recurrences / year, preferably (severe) chronic HSV infection), the same applies mutatis mutandis as already explained above in relation to the first aspect of the invention in the context of a specific subgroup of patients.

[0617] In addition, regarding a specific dosing regimen (i.e., prime-boost administration) for a vaccine comprising the following components: (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21, wherein in the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen, the antigen is associated with an HSV-related disease, and wherein the vaccine is to be administered in a prime-boost dosing regimen by the following administration steps:

[0618] (i) administering a first dose of the vaccine to the subject (prime administration);

[0619] (ii) administering further doses of the vaccine to the subject at intervals of 15 to 30 days after the initial boost (boosting administrations); and

[0620] (iii) Optionally, step (ii) is repeated at intervals of 15 to 30 days after the previous booster administration (further booster administration).

[0621] With regard to the preferred embodiments of this specifically defined dosage regimen, the same applies mutatis mutandis as has been explained above in relation to the first aspect of the invention in the context of a specific dosage regimen.

[0622] The present invention also relates to a method of actively immunizing a subject against an infectious disease or a malignant disease in a subject as defined herein above in the context of the first aspect of the invention.

[0623] The present invention also relates to a method of actively immunizing a subject against an HSV-related disease in a subject as defined herein above in the context of the second aspect of the invention.

[0624] Concerning preferred embodiments of the method for active immunization, the same applies mutatis mutandis as already explained above in the context of the above defined antibodies and Fc-containing fusion proteins (respectively), vaccines or vaccine pharmaceutical compositions for said use.

[0625] In the present invention, in a preferred embodiment, the subject is a mammal, such as a dog, a cat, a pig, a cow, a sheep, a horse, a rodent (e.g., a rat, a mouse, and a guinea pig) or a primate (e.g., a gorilla, a chimpanzee, and a human). In a most preferred embodiment, the subject is a human.

[0626] In a preferred embodiment, in particular with respect to the second aspect of the invention, the disease is selected from the group consisting of herpes simplex labialis, herpes simplex genitalis, chronic or disseminated cutaneous herpes simplex infection, traumatic herpes, eczema herpeticum, herpetic keratoconjunctivitis, herpes neonatalis, Alzheimer's disease (AD), HSV pneumonia, Bell's palsy, herpetic esophagitis, herpes encephalitis and herpes meningitis, herpetic whitlow, herpetic gingivostomatitis, the presence of oral relapses, the presence of genital relapses, eczema herpeticum, herpes neonatalis, immunodeficiency, immunocompromised patients, resistance to viral inhibitory agents, encephalitis, meningitis, meningoencephalitis, eye infections and / or systemic HSV infections.

[0627] Other aspects and advantages of the present invention will be described in the following examples, which are provided for illustrative purposes and not in a limiting manner. Each publication, patent, patent application or other document cited in this application is hereby incorporated by reference in its entirety.

[0628] Figure 1 :HDIT101 does not induce complement-dependent cytotoxicity (CDC)

[0629] Complement-dependent cytotoxicity (CDC) was measured by measuring the formation of the Cb5-9 terminal complement complex (TCC). 150 plaque-forming units of HSV-2G were incubated with HDIT101 at different concentrations and a complement source (HSV-seronegative serum vs. heat-inactivated HSV-seronegative serum). The HDIT101 concentration was 100 nM (neutralizing concentration), 1000 nM (10× neutralizing concentration, excess antibody), or 10 nM (50% neutralizing concentration). Positive controls (high and medium C5b-9 concentration controls) and negative controls (buffer only) were included. C5b was measured using an enzyme-linked immunosorbent assay. mAb mouse anti-human C5b-9 (clone aE11) was coated on the plate. Samples or controls (which contained complement complexes) were added. Goat anti-human C5 antibody was added, followed by incubation with F(ab)2 (rabbit anti-goat IgG (H+L) conjugated with peroxidase). Substrate was added and the color change was measured using a photometer. Zymosan activated normal human serum calibrated against purified C5b-9 was used as a standard. At the HDIT101 concentrations tested, no substantial induction of C5b-9 was observed compared to the medium and high controls.

[0630] Figure 2 :HDIT101 does not induce antibody-dependent cellular cytotoxicity (ADCC)

[0631] ADCC was measured using a commercially available reporter assay (ADCC Reporter Bioassay, Core Kit, Promega #G7018). Target cells (Vero cells infected with HSV-1 or HSV-2) were incubated with a series of dilutions of HDIT101 or polyclonal serum (as a control) and effector cells (transformed Jurkat cells stably expressing CD16 (FcγRIIIa) and NFAT-luciferase reporter genes). Once ADCC is induced, the NFAT-luciferase reporter gene is activated, and luciferase luminescence can be measured by adding substrate and measuring relative light units using a luminometer. The fold change relative to the sample without antibody or serum was calculated and plotted. At the HDIT101 concentration tested, no substantial induction of FcgRIIIa-induced NFAT signaling was observed in the reporter cells, in contrast to the polyclonal serum.

[0632] Figure 3 :HDIT101 induces antibody-dependent phagocytosis (ADCP) in THP-1 cell line

[0633] THP-1 cells were incubated with antibody-HSV-1F immune complexes for 18 hours. Cells were fixed using paraformaldehyde, permeabilized, and stained with anti-human IgG coupled to FITC. Stained cells were analyzed using flow cytometry (A) or fluorescence microscopy (B). A significant increase in fluorescent cells was observed relative to control antibody, deglycosylated HDIT101N297A, or unstained controls.

[0634] Figure 4 : HSV-infected cells conditioned with HDIT101 bind to cells expressing FcgRI, but not to cells expressing FcgRIIa or FcgRIIIa

[0635] Murine thymoma cells (BW5147) were modified to stably express a chimeric protein consisting of the extracellular domain of human FcgRI, IIa, IIb, or IIIa coupled to a TCR-ζ domain. Activation of the receptor by IgG binding results in IL-2 secretion in this model, which can then be measured in the supernatant via ELISA. Vero cells infected with HSV-1 were incubated with serial dilutions of HDIT101 or a positive control (Cytotect), and IL-2 secretion was measured. Infected cells opsonized with HDIT101 activated IL-2 secretion from cells expressing the FcgRI extracellular domain fusion protein, but not for cells expressing the FcgRIIa or FcgRIIIa fusion proteins.

[0636] Figure 5 HDIT101 induces ADCP in both M1 and M2 monocyte-derived macrophages (MDMs)

[0637] Beads coated with fluorescent streptavidin were decorated with biotinylated gB and then complexed with HDIT101 or mutant HDIT101-N279A. Monocyte-derived M1 or M2 macrophages were incubated with the respective bead complexes for 2 hours and subsequently analyzed via flow cytometry. Mean fluorescence intensity (MFI) is shown as fold change relative to control conditions with untreated macrophages. Data indicate mean ± SD of three different donors. Phagocytosis was increased by HDIT101 to approximately 50-fold, whereas phagocytosis induced by HDIT101-N297A was significantly reduced.

[0638] Figure 6 : HSV-1F-infected cells conditioned with HDIT101 induce IP10 cytokine production in MDM

[0639] 293Tgfp cells were infected with HSV-1F and conditioned with saturating concentrations of HDIT101. Monocyte-derived macrophages (MDMs) of the M2 type were differentiated from CD14-positive cells from a blood donor (#Buco009) using macrophage colony stimulating factor (M-CSF). MDMs were incubated for 2 hours with 293Tgfp cells infected with HSV-1 and conditioned with HDIT101, and ADCP was measured by counting GFP-positive CD11-positive cells using fluorescence microscopy imaging (A). Parallel samples were incubated for 24 hours, and supernatants were harvested and subjected to cytokine measurements using LEGENDplex from Biolegend TM Human M1 / M2 Macrophage Panel (10-plex) (B). Incubation of MDM with HSV-1 infected 293Tgfp cells opsonized with HDIT101 strongly induced cytokines, especially IP-10 (CXCL10), by >1000-fold, whereas unopsonized HSV-1 infected 293Tgfp cells did not.

[0640] Figure 7 HDIT101-opsonized viral particles induce an augmented IP10 cytokine response in the presence of MDM and autologous T cells

[0641] Using macrophage colony stimulating factor (M-CSF), M2-type monocyte-derived macrophages (MDM) were differentiated from CD14-positive cells from a blood donor (#Buco009). HSV-1F immune complexes (ICs) with HDIT101 were generated by incubating the virus for 1 hour in Vero cells at a concentration of HDIT101 that 100% neutralizes viral infectivity. ICs or HDIT101 alone were then transferred to MDM and mixed with autologous T cells (TCs). At the time of monocyte isolation, T cells were pre-activated for 2 days using anti-CD28 / CD3 beads in the presence of IL7 / 15. Activation was terminated after two days by washing off the beads, and the T cells were rested for 7 days in a medium containing IL7 / 15 before being added to MDM. The addition of HSV-1 FIC with HDIT101 strongly induced IFNg (~150-fold), and most strongly induced IP-10 (20,000-fold).

[0642] Figure 8 : IL-2 production by autologous T-cells stimulated with HSV-1:HDIT101 IC is dominantly by M1-type MDMs and moderately by M2-type MDMs after ADCP

[0643] CD14-positive monocytes were isolated from HSV-seropositive blood donors (Buco #0010) and differentiated into M2 type MDMs using M-CSF or into M1 type MDMs using granulocyte-MCSF (GM-CSF). HSV-1F immune complexes (ICs) with HDIT101 were generated by incubating the virus for 1 hour in Vero cells at a concentration of HDIT101 that 100% neutralizes viral infectivity. Autologous T cells were generated and resting, unstimulated T cells were added to macrophages or kept alone (no MDM). HDIT101:HSV-1 IC or HDIT101 antibody was added to MDM T cell co-cultures and Human Th1LEGENDplex from Biolegend was used 24 hours or 48 hours later. TM (5-plex) kit to analyze supernatants for IL-2 concentration. IL-2 levels were substantially increased in the presence of HDIT101:HSV-1 IC, but not in the presence of HDIT101 alone. Notably, M1 MDMs supported much stronger IL-2 induction (>250-fold) compared to M2 MDMs (~40-fold) after 48 hours of stimulation with HDIT101:HSV-1 IC. The presence of MDMs and ICs is necessary for the induction of IL-2 production by T cells.

[0644] Fig. 9 : Proposed mechanism for induction of de novo B- / T-cell responses via HDIT101-induced ADCP

[0645] HSV-1 infected cells or HSV-1 particles bind to HDIT101, and HDIT101 induces ADCP by binding to FcgRI. Notably, HDIT101 does not induce ADCC, or CDC. ADCP of HDIT101-IC results in presentation of viral peptides on MHC I or MHC II complexes, and notably, induction of IP10 (which acts as a chemokine to attract dendritic cells, NK cells, and T cells to sites of infection). Flock dendritic cells (FDCs) take up intact HDIT101-opsonized ICs and transport them to germinal centers (GCs), where de novo B cell responses are induced to epitopes revealed by structural changes induced by Fab binding in gB within the ICs. Existing tissue-resident memory T cell responses (CD8 RM) are enhanced and migrate toward the site of HSV replication, and when circulating naive T cells recognize viral T cell epitopes presented on MHC complexes, de novo T cell responses are induced. The unique characteristics of HDIT101 mediating ADCP and inducing IP10 and IFNg support the generation of de novo T-cell (and B-cell) responses. These characteristics can be used in vaccine design, whereby inactivated viruses or virus-like particles or recombinant gB or gB-Fc proteins are complexed with HDIT101 and act as vaccines.

[0646] Fig.10 : HDIT101 N297A is insufficient to rescue immunocompetent Balb / c mice from lethal HSV-2G infection.

[0647] Eight-week-old female immunocompetent Balb / c OlaHsd mice were infected intravaginally with 5×10e5 plaque forming units of HSV-2G and treated intravenously 14 hours later with 300 μg HDIT101 (n=14), HDIT101 N297A (n=12), murine mAb2c (n=13), or PBS (n=10). Survival was monitored for 30 days post-infection. Treatment with HDIT101 or murine mAb2c increased survival to 50% within 30 days, while all control mice died within 10 days. Mice treated with HDIT101 N297A showed significantly reduced survival compared to mice treated with HDIT101 (p=0.0036, Gehan-Breslow-Wilcoxon statistical test).

[0648] Fig.11 : Intravenous HDIT101 treatment of chronic recurrent genital HSV-2 infection results in a substantial prolongation of the time to first lesion recurrence

[0649] Kaplan Meier curves for the probability of remaining lesion-free over time after treatment are shown for MATCH-2 study participants with confirmed HSV-2 lesions treated with valacyclovir or with intravenous HDIT101. Beginning approximately 30 days after treatment, the probability of remaining lesion-free was substantially increased in patients treated with HDIT101 compared to the valacyclovir treatment arm.

[0650] Fig.12 : Intravenous HDIT101 treatment of chronic recurrent genital HSV-2 infection results in a significant reduction in relapse rates

[0651] The median relapse rate and lower and upper percentiles for patients in the per protocol set are depicted. The mean relapse rate in the per protocol set (PPS) for patients treated with HDIT101 was significantly reduced from 3.789 to 2.696 compared to patients treated with valacyclovir (p=0.008).

[0652] Fig.13 : Strategies for using HDIT101:gB-Fc immune complexes that mediate ADCP as vaccines to induce anti-HSV responses or responses against unrelated Ags

[0653] (A) Generate HSV-1 / 2 gB-Fc fusion proteins that form trimeric complexes with Fc-mediated intertrimer linkages, thereby generating large complexes with multivalent Fc effector domains. In addition, HDIT101 enhances cross-linking and can reveal new B-cell epitopes due to structural changes through Fab binding. Multivalent Fc domains from gB-Fc and HDIT101 IC increase ADCP and anti-HSV responses, i.e., gB peptide presentation on MHC by APCs. Addition of further amino acid substitutions in Fc that enhance ADCP (but not CDC or ADCC) will increase vaccination efficacy.

[0654] (B) As in A), but includes an unrelated Ag fused between gB and Fc for which a potent MHC T-cell response is sought to be induced. The increased Fc effector function of ADCP due to multiple Fc-domains leads to better uptake by APCs (e.g., dendritic cells, including follicular dendritic cells), which carry the ICs to germinal centers where new B-cell responses can develop. De novo T-cell responses are induced due to more efficient uptake via ADCP in APCs and more efficient presentation of ADCP-induced viral peptides on MHC. The gB-Fc:HDIT101 IC with enhanced ADCP and MHC-presentation characteristics can be used to deliver unrelated Ag (uAg) into the same pathway, which will generate enhanced T-cell and B-cell responses against uAg.

[0655] Fig.14 Enhanced phagocytosis and T cell activation by primary human monocyte-derived dendritic cells (MDDCs) in the presence of HDIT101

[0656] MDDCs were generated from PBMCs of independent donors by isolating CD14-positive monocytes and differentiating them for 7 days using interleukin 4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF).

[0657] (A) Phagocytosis assay of MDDC. MDDC from three independent healthy donors were matured overnight using 0.5 μg / ml lipopolysaccharide (LPS). Cells were then incubated overnight with ATTO488 NHS-ester labeled HSV-1 at an MOI of 10 in combination with HDIT101 (150 μg / ml) or acyclovir (50 μg / ml) or left untreated and analyzed the next day by flow cytometry. Average data for three independent donors are shown, with error bars indicating standard deviation. Phagocytosis of labeled virus was significantly increased in the presence of HDIT101 compared to controls (p<0.001). Statistical analysis was performed using a one-way ANOVA test.

[0658] (B) Autologous T cell activation after ADCP by HDIT101. Same as in A), but with four independent HSV-1 seropositive healthy donors and without maturation of MDDCs. Changes in CD69 MFI were normalized to untreated MDDCs. Autologous T cells were added to MDDCs after incubation with HSV-1 in the presence or absence of HDIT101 or acyclovir or without treatment, and analyzed by flow cytometry using fluorochrome-labeled anti-CD69 IgG as a T cell activation marker. In the presence of HDIT101, T cells showed significantly increased levels of the CD69 activation marker, indicating increased activation in the presence of HDIT101. Statistical analysis was performed using a one-way ANOVA test.

[0659] Fig.15 : Enhanced T cell activation by primary human monocyte-derived macrophages (MDM) or dendritic cells (MDDC) in the presence of HDIT101

[0660] MDM and MDDC were generated from PBMC of three independent healthy donors by isolating CD14 positive monocytes and differentiating for 7 days using granulocyte-macrophage colony stimulating factor (GM-CSF) (for MDM) or interleukin 4 (IL-4) + GM-CSF (for MDDC). As above, cells were incubated with HSV-1 in the presence of HDIT101 or acyclovir, or were not treated. T cell activation was determined by measuring CD69 levels on CD4+ and CD8+ cells. The data demonstrated that in MDM and MDDC, CD4+ and CD8+ T cells were activated to a significantly greater extent in the presence of HDIT101. Statistical analysis was performed using a one-way ANOVA test.

[0661] Fig.16: Enhanced T cell activation by primary human monocyte-derived type 1 macrophages (MDM1) or dendritic cells (MDDC) from HSV-seropositive vs. HSV-seronegative donors in the presence of HDIT101

[0662] MDM and MDDC were generated from PBMC of healthy HSV-1-seropositive (A) or HSV-seronegative (B) donors by isolating CD14 positive monocytes and differentiating for 7 days using granulocyte-macrophage colony stimulating factor (GM-CSF) (for MDM) or interleukin 4 (IL-4) + GM-CSF (for MDDC). As above, cells were incubated with HSV-1 in the presence of HDIT101 or acyclovir, or were not treated. T cell activation was determined by measuring CD69 levels on CD3+ cells. Statistical analysis was performed using two-factor ANOVA.

[0663] Fig.17 : Rapid kinetics of ADCP mediated by HDIT101

[0664] Monocytes were isolated from PBMCs by MACS CD14+MicroBead separation (Miltenyi Biotec) according to the manufacturer's instructions. Cells were differentiated for one week by adding M-CSF (50 ng / ml) (MDM2 cells), GM-CSF (80 ng / ml) (MDM1 cells) or GM-CSF (80 ng / ml) + interleukin 4 (IL-4) (20 ng / ml) (MDDC). Differentiated cells were then exposed to HSV-1 labeled with a pH-sensitive dye (IncuCyte pHrodo Orange Cell Labeling Dye, Sartorius) a...

Claims

1. A vaccine for active immunization of a subject against an infectious disease or a malignant disease, the vaccine comprising: (a) antibodies against antigens associated with said infectious disease or malignant disease; or (b) an Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen associated with the infectious disease or malignant disease, wherein the Fc portion is capable of binding to a receptor present on or in an antigen presenting cell (APC), the receptor being selected from the group consisting of: Type I: activating FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, FcγRIIIb, and inhibitory FcγRIIb; and Type II: neonatal FcR (FcRn) and cytoplasmic TRIM21.

2. The vaccine of claim 1(a), wherein the vaccine is to be administered to subjects having acute relapses of the infectious disease and suffering from severe chronic disease with more than 4 relapses / year.

3. The vaccine according to claim 1 or claim 2, wherein the vaccine is to be administered in a prime-boost dosing regimen by the following administration steps: (i) administering a first dose of the vaccine to the subject during the first week of relapse, preferably immediately after the onset of the first relapse (priming administration); (ii) administering a further dose of the vaccine to the subject during the first week of the second relapse, preferably immediately after the onset of the second relapse (boosting administration); and (iii) optionally, during the first week of each further relapse, preferably immediately after the onset of each further relapse, repeating step (ii) (further booster administrations), One initial administration and at least one booster administration are to be performed per year.

4. The vaccine of claim 1(b), wherein the vaccine is to be administered to subjects suffering from a severe chronic disease with more than 4 relapses / year.

5. The vaccine according to claim 1(b) and / or claim 4, wherein the vaccine is to be administered in a prime-boost dosing regimen by the following administration steps: (i) administering a first dose of the vaccine to the subject (prime administration); (ii) administering to the subject a further dose of the vaccine at intervals of 15 to 30 days after the initial boost administration (boosting administration); and (iii) Optionally, repeating step (ii) at intervals of 15 to 30 days after the previous booster administration (further booster administration).

6. The vaccine for use according to any one of claims 1 to 5, wherein the antibody or the Fc-containing fusion protein is capable of inducing antibody-dependent cellular phagocytosis (ADCP) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

7. The vaccine for use according to any one of claims 1 to 6, wherein the infectious disease is selected from the group consisting of fungal infection, bacterial infection, protozoan infection and viral infection, preferably HSV-related disease.

8. The vaccine for use according to claim 1 or 6, wherein the malignant disease is selected from the group consisting of solid tumors and hematological malignancies / hematological oncological diseases.

9. The vaccine for use according to any one of claims 1 to 8, wherein in claim 1 (a) or (b), the antigen associated with the infectious disease is selected from the group consisting of: Epstein-Barr virus (EBV) (derived) proteins or protein domains, preferably EBNA1, EBNA2, EBNA3A, EBNA3B, EBNA3C, EBNA-LP, LMP1, LMP2A / B, gp350, A73, RPMS1, ZEBRA, Rta, major intercalated protein p143, major intercalated protein, intercalated protein, major capsid protein, minor capsid protein, capsid protein p18, p23, p40, gN, gp42, gM, gp60, gp78 / 55, gp150, 53 / 55kd membrane protein, viral IL-10, gB, gH, gL; Human papillomavirus (HPV) (derived) proteins or protein domains, preferably E1, E2, E4, E5, E6, E7, L1, L2; Human immunodeficiency virus 1 or 2 (derived) proteins or protein domains, preferably Gag, Pol, Env, gp160, gp120, gp41, CA, MA, p2, p6, NC, IN, RTp66, RTp55, RTp51, PR, Rev, Tat, Nef, Vif, Vpr, Vpu, Vpx; Herpes simplex virus (HSV) 1 or 2 (derived) proteins or protein domains, preferably gB, gC, gD, gH, gG, gL, gE, gI, gK, gM, VP1-2, ICP32, ICP0, VP11 / 12, UL13, vhs, VP16, US3, VP22, ICP34.5, US11, ICP4, DNA polymerase, major capsid protein, helicase, primase, uracil DNA glycosylase, dUTPase, ribonucleotide reductase, major intercalated protein; A human cytomegalovirus (HCMV) (derived) protein or protein domain encoded by an open reading frame (ORF), preferably UL57, UL55, UL54, UL75, UL86, UL85, UL104, UL97, UL98, UL100, UL105, UL102, UL114, UL115, UL72, UL70, UL69, UL44, UL45, UL48; Hepatitis A virus (HAV) (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3Cpro, P3-3Dpol; Hepatitis B virus (HBV) (derived) proteins or protein domains, preferably M, L, S, Polymerase, TP, Spacer, RT, RNase H, preCore, Core, X, HBeAg; Hepatitis C virus (HCV) (derived) proteins or protein domains, preferably E1, E2, core, NS1, NS2, NS3, NS4a, NS4b, NS5a, NS5b, precursor polyprotein; Influenza A virus (IAV) (derived) proteins or protein domains, preferably HA, NA, M2, M1, NP, NS1, NS2 / NEP, PA, PB1; Measles virus (MV) (derived) protein or protein domain, preferably N, P / C / V, M, F, H, L; Respiratory syncytial virus (RSV) (derived) protein or protein domain, preferably NS1, NS2, N, P, M, SH, G, F, M21, M22, L; Rotavirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, RdRp, VP5, VP6, VP7, VP8, NSP1, NSP2, NSP3, NSP4, NSP5, NSP6; Severe acute respiratory syndrome coronavirus (SARS-Cov) type 1 and 2 (derived) proteins or protein domains, preferably ns1, ns2, PLpro, ns4, 3CL, ns6, ns7, ns8, ns9, ns10, RdRp, Hel, ns14, ns15, ns16, S, S-RBD, 3a, E, M, 6, 7a, 7b, 8N, 9b, 14; Varicella zoster virus (VZV) (derived) proteins or protein domains encoded by open reading frames (ORFs), preferably ORF0 to ORF71 (at least 71 proteins in total); Human herpesvirus 8 (HHV-8) (derived) proteins or protein domains, preferably K1, K2, K3, K4, K4.1, K5, K6, K7, K8, K8.1, K9, K10, K10.5, K11, K12, K13, K14, K15, gB, gL, gH and proteins encoded in ORFs, including but not limited to ORF2, ORF9, ORF10, ORF16, ORF18, ORF24, ORF30, ORF31, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF42, ORF43, ORF44, ORF45, ORF46, ORF47, ORF48, ORF49, ORF50, ORF51, ORF52, ORF53, ORF54, ORF55, ORF56, ORF57, ORF58, ORF59, ORF60, ORF61, ORF62, ORF63, ORF64, ORF65, ORF66, ORF67, ORF68, ORF69, ORF70, ORF71, ORF72, ORF73, ORF74, ORF75 RF66, ORF21, ORF23, ORF25, ORF26, ORF65, ORF33, ORF34, ORF35, ORF36, ORF37, ORF38, ORF39, ORF40, ORF41, ORF 42. ORF45, ORF49, ORF50, ORF52, ORF53, ORF55, ORF57, ORF59, ORF67, ORF69, ORF70, ORF72, ORF73, ORF74, ORF75; Human herpesvirus 6 (HHV-6) (derived) protein or protein domain encoded by ORF, preferably DR1, DR6, DR7 / U1, U2, U3, U4, U7, U10, U11, U12, U13, U14, U15, U17, U18, U19, U20, U21, U22, U23, U24, U25, U26, U27, U28, U29, U30, U31, U32, U33, U34, U35, U36, U37, U38, U39, U40 , U41, U42, U43, U44, U45, U46, U47, U48, U49, U50, U51, U52, U53, U54, U55, U56, U57, U58, U59, U61, U62, U63, U64, U65, U66, U69, U70, U71, U72, U73, U74, U75, U76, U77, U79, U81, U82, U83, U85, U86, U88, U90, U91, U94, U95, U100; Rabies virus (derived) protein or protein domain, preferably N, P, M, G, L; Mumps virus (derived) proteins or protein domains, preferably N, V / P, M, F, HN, L; and Rhinovirus (derived) proteins or protein domains, preferably VP1, VP2, VP3, VP4, P2-2A, P2-2B, P2-2C, P3-3A, P3-VPg, P3-3C, P3-3D; and / or Wherein in claim 1(a) or (b), the antigen associated with the malignant disease is selected from the group consisting of: Tumor-associated antigens, preferably carbonic anhydrase IX, CCCL19, CCCL21, CSAp, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, IGF-1R, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD74, CD79a, CD80, CD83, CD95, CD126, CD133, CD138, CD147, CD154, AFP, PSMA, CEACAM5, CEACAM6, B7, ED-B of fibronectin, H factor, FHL-1, Flt-3, folate receptor, GROB, HER2, HMGB-1, hypoxia-inducible factor (HIF), HM1.24, insulin-like growth factor-1 (ILGF-1), IFN-γ, IFN-α, IFN-β, IL-2, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-25, IP-10, mCRP, MCP-1, M IP-1A, MIP-1B, MIF, MUC1, MUC2, MUC3, MUC4, MUC5ac, PAM4 antigen, NCA-95, NCA-90, NY-ESO-1, Ia, HM1.24, EGP-1, EGP-2, HLA-DR, Tenascin, Le(y), RANTES, T101, TAC, Tn antigen, Thomson-Friedenreich antigen, tumor necrosis antigen, TNF-α, TRAIL receptor (R1 and R2), VEGFR, EGFR, PlGF, complement factor C3, C3a, C3b, C5a, C5, cancer-testis (CT) antigen SPAG9, CT9, CT10, LAGE, MAGE-B5, MAGE-B6, MAGE-C2, MAGE-C3, MAGE-D, HAGE, SAGE and oncogene products.

10. The vaccine for use according to any one of claims 1 to 8, wherein the antibody defined in claim 1(a) is a monoclonal antibody.

11. The vaccine for use according to any one of claims 1 to 8 and 10, wherein the antibody defined in claim 1(a) is a humanized antibody or a fully human antibody.

12. The vaccine for use according to any one of claims 1 to 10, wherein the active immunization induces a lasting or long-term immunity against the infectious disease or malignant disease in the subject, and wherein the subject suffers from an acute infectious disease or malignant disease.

13. The vaccine for use according to any one of claims 1 to 12, wherein the infectious disease is an HSV-related disease; and (a) wherein the antibody is an anti-HSV antibody and the subject suffers from an acute HSV-related disease, preferably an acute HSV infection; or (b) wherein in the Fc-containing fusion protein comprising the Fc portion of an antibody fused to an antigen, the antigen is associated with an HSV-related disease.

14. The vaccine according to claim 13(a), wherein the vaccine is to be administered to subjects with acute relapses of the disease and suffering from severe chronic HSV-related disease with more than 4 relapses / year, preferably severe chronic HSV infection.

15. The vaccine according to claim 13 or claim 14, wherein the vaccine is to be administered in a prime-boost dosing regimen by the following administration steps: (i) administering a first dose of the vaccine to the subject during the first week of relapse, preferably immediately after the onset of the first relapse (priming administration); (ii) administering a further dose of the vaccine to the subject during the first week of relapse, preferably immediately after the onset of a second relapse (boosting administration); and (iii) optionally, during the first week of each further relapse, preferably immediately after the onset of each further relapse, repeating step (ii) (further booster administrations), One initial administration and at least one booster administration are to be performed per year.

16. The vaccine according to claim 13(b), wherein the vaccine is to be administered to subjects suffering from severe chronic HSV-related disease with more than 4 recurrences / year, preferably severe chronic HSV infection.

17. The vaccine according to claim 13(b) and / or claim 16, wherein the vaccine is to be administered in a prime-boost dosing regimen by the following administration steps: (i) administering a first dose of the vaccine to the subject (prime administration); (ii) administering to the subject a further dose of the vaccine at intervals of 15 to 30 days after the initial boost administration (boosting administration); and (iii) Optionally, repeating step (ii) at intervals of 15 to 30 days after the previous booster administration (further booster administration).

18. The vaccine for use according to any one of claims 13 to 17, wherein the antibody or the Fc-containing fusion protein is capable of inducing antibody-dependent cellular phagocytosis (ADCP) independently of antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC).

19. The vaccine for use according to any one of claims 13 to 18, wherein the anti-HSV antibody defined in claim 13(a) is a monoclonal antibody.

20. The vaccine for use according to any one of claims 13 to 19, wherein the anti-HSV antibody defined in claim 13(a) is a humanized antibody or a fully human antibody.

21. The vaccine for use according to any one of claims 13 to 20, wherein the anti-HSV antibody recognizes glycoprotein B (gB) of HSV-1 and / or HSV-2.

22. The vaccine for use according to any one of claims 13 to 21, comprising the following complementarity determining region: V comprising SEQ ID NO: 1 H CDR1, comprising V of SEQ ID NO:2 H CDR2, comprising V of SEQ ID NO: 3 H CDR3, comprising V of SEQ ID NO:4 L CDR1, comprising V of SEQ ID NO:5 L CDR2, and V comprising SEQ ID NO:6 L CDR3.

23. The vaccine for use according to claim 22, wherein the antibody comprises an amino acid sequence having at least 70% sequence identity with the amino acid residues shown at positions 1 to 30, 38 to 51, 68 to 99 and 112 to 122 of SEQ ID NO:7 and at positions 1 to 23, 41 to 55, 63 to 94 and 104 to 114 of SEQ ID NO:

8.

24. The vaccine for use according to claim 22 or 23, wherein the antibody comprises a V of SEQ ID NO:

9. H and V of SEQ ID NO: 10 L .

25. The vaccine for use according to any one of claims 22 to 24, wherein the antibody is a mAb2c antibody.

26. The vaccine for use according to any one of claims 13 to 25, wherein the antibody is to be administered intravenously, topically, intradermally, subcutaneously, intradermally, intramuscularly and / or intrathecally.

27. The vaccine for use according to any one of claims 13 to 26, wherein the disease is selected from the group consisting of: herpes simplex labialis, herpes simplex genitalis, chronic or disseminated cutaneous herpes simplex infection, traumatic herpes, eczema herpeticum, herpetic keratoconjunctivitis, herpes neonatalis, Alzheimer's disease (AD), HSV pneumonia, Bell's palsy, herpetic esophagitis, herpes encephalitis and herpes meningitis, herpetic whitlow, herpetic gingivostomatitis, the presence of oral relapses, the presence of genital relapses, herpetic eczema, herpes neonatalis, immunodeficiency, immunocompromised patients, resistance to viral inhibitory agents, encephalitis, meningitis, meningoencephalitis, eye infections and / or systemic HSV infections.

28. The vaccine for use according to any one of claims 1 to 27, wherein the anti-HSV antibody is a full-length antibody / complete antibody.

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