Poxvirus vaccine

Abstract

The present invention relates to vaccine compositions and uses thereof for the prophylactic or therapeutic treatment of an infection caused by viruses of the Poxviridae family, specifically the Orthopoxvirus genus of viruses in that viral family, which may stimulate a broad and effective adaptive immune response across multiple Poxviridae species and a diverse spectrum of human leukocyte antigen (HLA) alleles.

Description

[0001] POXVIRUS VACCINE

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to vaccine compositions and uses thereof for the prophylactic or therapeutic treatment of an infection caused by viruses of the Poxviridae family, specifically the Orthopoxvirus genus of viruses in that viral family, wherein the above are comprised of one or more epitopes selected for their ability to stimulate a broad and effective adaptive immune response across multiple Poxviridae species and a diverse spectrum of human leukocyte antigen (HLA) alleles.

[0004] BACKGROUND

[0005] The Poxviridae family concerns double-stranded DNA viruses that are hosted by vertebrates and arthropods. The 83 species in the Poxviridae family fall into 22 genera, which are divided into two subfamilies of Chordopoxvirinae and Entomopoxvirinae (Koonin et al., 2019, ICTV Berlin, 2019.003G, EC 51). Of the 22 poxvirus genera, four may infect humans, namely Orthopoxvirus, Parapoxvirus, Yatapoxvirus and Molluscipoxvirus. Notable examples of each virus genus respectively include monkeypox virus, orf virus, tanapox virus and molluscum contagiosum virus. Perhaps the most famous species of the Poxviridae family is the variola virus, the cause of smallpox disease (Breman and Henderson, 2002, The New England Journal of Medicine, 346, 1300-1308). Following vaccination efforts that began with the work of Edward Jenner in 1796 and were intensified by the WHO in 1967, smallpox is the only human disease to be completely eradicated.

[0006] While arguably the most dangerous Poxviridae species has been eradicated, the many remaining family members presently cause widespread infection and disease across the globe. Molluscum contagiosum virus, for example, affects around 200,000 people a year, constituting about 1 % of all diagnosed skin diseases. While cases are not fatal, this contagious viral infection of the skin can often persist for 6-12 months and can prove more problematic for immunocompromised patients (De Clercq et al., 2023, Travel Medicine and Infectious Disease, 52, 102528). There has been no published research into a successful vaccine candidate to date.

[0007] Mpox (formerly known as monkeypox) is an infectious viral disease caused by the monkeypox virus of the Orthopoxvirus genus. While the illness is usually mild, with symptoms including a rash, fever and swollen lymph nodes and those infected recovering within a few weeks without treatment, cases can be severe in children, pregnant women or immunosuppressed individuals (De Clercq et al., 2023, Travel Medicine and Infectious Disease, 52, 102528). Transmission may occur through close contact, including sexual contact, and by means of respiratory droplets. Prior estimates of death with infection with the monkeypox virus were between 0 and 11 % (Bunge et al., 2022, PLOS Neglected Tropical Diseases, 16(2), e0010141).

[0008] The 2022-2023 outbreak of mpox, caused by the monkeypox virus, was declared a public health emergency of international concern (PHEIC) by the WHO on 23 July 2022. Although the WHO declared an end to the global health emergency declared in response to the worldwide outbreak of monkeypox virus in May 2023, the declaration of a PHEIC categorises the recent mpox outbreak alongside diseases such as swine flu, polio, Ebola, Zika virus and COVID-19. As of 5 July 2023, there have been 88,122 confirmed cases and 148 deaths attributed to mpox disease (WHO, 2023). The MVA-BN vaccine, originally developed for smallpox, has been shown to be effective at reducing the risk of mpox illness by inducing a humoral immune response (Greenberg etal., 2016, PLOS ONE, 11 (6) e0157335). However, there is no evidence for its efficacy in other Poxviridae family members beyond the variola and vaccinia viruses.

[0009] Cellular immunity, as an arm of the adaptive immune system that is specialised to resolve infections and prevent reinfection from pathogens, often works in tandem with humoral - antibody-based - immunity upon natural exposure to a foreign body. A cellular immune response involves the interaction of T cells, each providing a variety of immune-related functions to aid in the reduction or elimination of pathogen-infected host cells (Amanna & Slifka 2011 , Virology 411 (2): 206-215). Furthermore, the generation of memory T cells as part of the cellular immune response results in the ability to mount a faster and stronger immune response upon re-exposure to a previously encountered pathogen (Restifo & Tattinoni 2013, Current Opinion in Immunology 25(5): 556-63). As Poxviridae vaccine development has been focused on single virus species and activating a neutralising antibody-based humoral immune response, scope exists to design a vaccine that is more likely to generate a robust cellular immune response in a broad population to multiple Poxviridae species.

[0010] However, when designing vaccines engineered to induce a broad T cell response, there exists a further challenge of human leukocyte antigen (HLA) restriction within an individual and a broader population. The HLA complex is a set of genes encoding the major histocompatibility complex (MHC) proteins in humans, responsible for the regulation of an individual’s immune system, as well as the ability to specifically present at the surface of infected cells, and elicit an immune response against, epitopes generated during a natural infection, or delivered to said individual in the form of a vaccine (Marsh et al. 2010 Tissue Antigens 75(4): 291-455).

[0011] The high polymorphism of HLA alleles and subsequent immune system variability between individuals results in a diverse spectrum of “HLA types” across the population. As an added complication to peptide-based vaccine development, such HLA types can have a significant impact on the efficacy of a potentially prophylactic viral vaccine composition between different individuals. As such, generation of an epitope-based vaccine composition that is compatible with a particular subset of HLA types may prove ineffective with a significant proportion of the global population comprising individuals of different HLA types. Considering this, the generation of T-cell and B-cell epitope vaccines, that target a limited number of HLA types, may only prove advantageous for a narrow, select population.

[0012] The current lack of an approved vaccine composition which is efficacious across multiple Poxviridae species and a wide range of HLA populations, creates significant danger for at-risk populations, including health care workers and patients in acute danger of nosocomial or community-transmitted infections. Furthermore, in the events of lab leaks or intentional release of Orthopoxvirus biothreats such as smallpox, a stockpile of T cell Orthopoxvirus vaccines may be invaluable to preventing widespread disease and death.

[0013] Thus, there exists a need for a safe and effective vaccine for use in the therapeutic or prophylactic treatment of Poxviridae viruses, optimised to incorporate epitopes covering a diverse spectrum of HLA types, with the potential to stimulate a broad adaptive immune response against said viruses across the global human population.

[0014] SUMMARY OF INVENTION

[0015] This invention is based on the surprising discovery that, by using a platform to identify predicted cross-reactive Orthopoxvirus epitopes that bind HLA molecules across a broad spectrum of HLA types, potentially broad spectrum vaccines with predicted high efficacy can be formulated that comprise one or more of said epitopes. Such a vaccine thus has the potential to stimulate a broad adaptive immune response to Orthopoxvirus that is both cellular and potentially humoral in nature, for the therapeutic or prophylactic treatment of infection with Orthopoxvirus in humans across the global population.

[0016] In a first aspect of the invention, there is provided a vaccine composition comprising: i) a polypeptide, or ii) a polynucleotide encoding a polypeptide; wherein the polypeptide comprises one or more epitope sequences, wherein the one or more epitope sequences have the amino acid sequences of any one or more of SEQ ID Nos 1 to 57, or variants thereof, wherein the variant sequences have at least 70% sequence identity to any one or more of SEQ ID Nos 1 to 57.

[0017] The vaccine composition of the invention may comprise any of the polypeptides described herein. It is envisaged that the polypeptide may, for example, comprise at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57, or variants thereof. In the present disclosure, variants of a sequence are envisaged to have at least 70% sequence identity to their respective native sequence.

[0018] It is envisaged that the vaccine composition of the present invention may comprise any number of epitopes as would be suitable for use. In some embodiments, the polypeptide of the composition comprises at least one epitope having the amino acid sequence of SEQ ID No: 1 , or variants having at least 70% sequence identity thereto. In some embodiments, the polypeptide of the composition comprises at least one epitope having the amino acid sequence of SEQ ID No: 56, or variants having at least 70% sequence identity thereto.

[0019] The polypeptide of the vaccine composition may comprise at least 2 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. The at least two epitope sequences may have the amino acid sequence of each of SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto. The at least two epitope sequences may have the amino acid sequence of each of SEQ ID Nos: 2 and 56, or variants having at least 70% sequence identity thereto.

[0020] The polypeptide of the vaccine composition may comprise at least one epitope sequence having the amino acid sequence of SEQ ID No: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16 and / or 17, or a variant having at least 70% sequence identity thereto. Preferably, the polypeptide comprises epitope sequences having the amino acid sequences of each of SEQ Nos: 1 to 17, or variants having at least 70% sequence identity thereto.

[0021] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 1 , or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 1 to 3, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 1 to 4, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 1 to 5, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 1 to 6, or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 1 to 7, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 1 to 8, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 1 to 9, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 1 to 10, or variants having at least 70% sequence identity thereto; xi) SEQ I D Nos: 1 to 11 , or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 1 to 12, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 1 to 13, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 14, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 15, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 16, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 17, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 18, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 19, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 20, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 21 , or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 22, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 23, or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 24, or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 25, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 26, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 27, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 28, or variants having at least 70% sequence identity thereto; xxix) SEQ ID Nos: 1 to 29, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 30, or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 31 , or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 32, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 33, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 34, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 35, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 36, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 37, or variants having at least 70% sequence identity thereto; xxxviii) SEQ ID Nos: 1 to 38, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 39, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 40, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 41 , or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 42, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 43, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 44, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 45, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 46, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 47, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 48, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 49, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 50, or variants having at least 70% sequence identity thereto.

[0022] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 56, or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 2 and 56, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 2, 3 and 56, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 2 to 4 and 56, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 2 to 4, 7 and 56, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 2 to 5, 7 and 56 or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 2 to 7 and 56, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 2 to 7, 56 and 57, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 2 to 7, 9, 56 and 57, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 2 to 7, 9, 11 , 56 and 57, or variants having at least 70% sequence identity thereto; xi) SEQ ID Nos: 2 to 7, 9, 11 , 13, 56 and 57, or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 2 to 9, 11 , 13, 56 and 57, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 2 to 9, 11 to 13, 56 and 57, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 9, 11 to 13, 56 and 57, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 9, 11 to 14, 56 and 57, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 9, 11 to 15, 56 and 57, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 9, 11 to 15, 17, 56 and 57, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 9, 11 to 15, 17, and 55 to 57, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 9, 11 to 15, 17, 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 9, 11 to 15, 17, 19, 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 20, 24, and 55 to 57, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 21 , 24, and 55 to 57, or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 21 , 24, 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 22, 24, 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 25, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 26, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 27, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 54 to 57, or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 53 to 57, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , 32, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35, 36, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 37, and 53 to 57, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: I to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 38, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 39, and 53 to 57, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 40, and 53 to 57, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 9,

[0023] II to 15, 17 to 29, 31 , 32, 35 to 41 , and 53 to 57, or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 42, and 53 to 57, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 43, and 53 to 57, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 44, and 53 to 57, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 45, and 53 to 57, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 46, and 53 to 57, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 47, and 53 to 57, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 48, and 53 to 57, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 49, and 53 to 57, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 50, and 53 to 57, or variants having at least 70% sequence identity thereto.

[0024] It is also envisaged that the polypeptide of the vaccine composition may comprise a concatemer of epitope sequences each having the amino acid sequences of any of SEQ ID Nos 1 to 57, or variants thereof. By “concatemer” we intend that the polypeptide may comprise multiple copies of an indicated epitope sequence linked in series. The number of copies will be dependent on the requirements of the polypeptide but it may be for example, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more copies of the epitope in question. The polypeptide may further comprise one or more spacer regions. It is envisaged that the spacer regions may be positioned between the epitope sequences. Although, as the skilled person will appreciate, depending on the adjacent epitope sequences in question, a spacer may not be required. In the instance of the polypeptide being a concatemer, the polypeptide may optionally include spacer regions, as would be determined by a person of skill in the art. One consideration here is whether undesirable epitopes may be created at the junction of two epitopes when a spacer is not included. Thus, in some embodiments, some epitopes may be separated by a spacer, while others are not. Equally, all epitopes may be separated by spacer regions. Thus, when the polypeptide comprises more than one of the epitope sequences of SEQ ID Nos 1 to 57 or the variants thereof, the one or more spacer regions may be positioned between one or more or each epitope sequence. In an embodiment of the invention, the epitope variant sequences of the polypeptide have at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to any of SEQ ID Nos 1 to 57. In a further embodiment of the invention, the polypeptide is no more than 2500, 2400, 2300, 2200, 2100, 2000, 1900, 1800, 1700, 1600, 1500, 1400, 1300, 1200, 1100, 1000, 900, 800, 700, 600, 500, 400, 300, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10 or 9 amino acids in length.

[0025] The polynucleotide forming part of the vaccine composition may be DNA, RNA or mRNA. Thus, the vaccine composition may be an mRNA vaccine composition. The polynucleotide may include synthetic non-natural nucleotides, as would be understood by a person of skill in the art.

[0026] In a second aspect of the invention, there is provided a vector comprising a polynucleotide as defined according to the first aspect of the invention, wherein the polynucleotide further comprises regulatory elements capable of driving transcription and / or translation of the polynucleotide in a host cell. The vector may be a viral vector or any other suitable vector.

[0027] A vaccine composition according to the first aspect of the invention or a vector according to the second aspect of the invention may be formulated in a nanoparticle or lipid formulation.

[0028] In a third aspect of the invention, there is provided a microorganism comprising: a polypeptide as defined according to the first aspect of the invention, or a polynucleotide as defined according to the first aspect of the invention. The microorganism may be a bacterial microorganism.

[0029] In a fourth aspect of the invention, there is provided a pharmaceutical composition comprising: a vaccine composition according to the first aspect of the invention, a vector according to the second aspect of the invention or a microorganism according to the third aspect of the invention, and a pharmaceutically acceptable carrier, diluent, excipient and / or adjuvant. The pharmaceutical composition may be formulated for parental, oral, sublingual, nasal, naso-oral, or pulmonary administration. Said parental administration may be subcutaneous, intradermal, intramuscular, subdermal, intraperitoneal, or intravenous administration.

[0030] The vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention may be for use in treating or preventing a disease or condition in a subject. Preferably, the disease or condition is an infection with an orthopoxvirus.

[0031] The vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention may be for use in the treatment or prophylaxis of an infection caused by akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, macacapox virus, monkeypox virus, mousepox virus, racoonpox virus, skunkpox virus, taterapox virus, vaccinia virus, variola virus, or volepox virus. Preferably, the the infection is caused by akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, monkeypox virus, variola virus or vaccinia virus.

[0032] The invention also provides the use of the vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention in the manufacture of a medicament for the prevention or treatment of an infection with an orthopoxvirus in a subject in need thereof.

[0033] The invention also provides a method of treating or preventing an infection with an orthopoxvirus may comprise administering an effective amount of the vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention.

[0034] BRIEF DESCRIPTION OF FIGURES

[0035] Figure 1 shows the proteome identifiers used when downloading the amino acid sequences of the proteins for 13 Orthopoxviruses from the UniProt database.

[0036] Figure 2 shows the number of proteins, the number of unique sequences, and protein sequence length distribution for the 13 Orthopoxviruses.

[0037] Figure 3 shows the hotspot detection workflow. Using the AP score map of a given protein (1), where each row represents one of the 156 considered Class I HLA alleles and each column represents an amino acid within the protein sequence, the sum of AP scores across HLAs is calculated for each amino acid position (2), as well as the mean of these summed up AP scores (dashed line). The mean of the AP score sums is used as a baseline and a LOESS smoothing curve is applied on theAP score sums at amino acid level (3). Next, the peak summits along the smoothing curve are identified using a rolling maximum function (circles) and the peak regions identified as windows of consecutive amino acids consistently above the baseline (rectangles). Finally, the hotspot sequences of the peak regions are extracted, as well as their start and end coordinates within the protein sequence (4).

[0038] Figure 4 shows the hotspot extension flow. The aligned hotspots output by Clustal Omega (A) are formatted per cluster (B) and the flanking dashes, internally used during the MSA are replaced by amino acids from the corresponding original protein sequence and the new start and end coordinates of the extended hotspots are extracted (C). Figure 5 shows the conserved region identification flow. The Clustal Omega output of the re-aligned extended hotspots (A) is used to extract the sequence of consensus symbols (rectangle in A). Using run length encoding (RLE), conserved hotspot regions are identified as consecutive amino acids denoted by an “*” or within the Clustal Omega consensus symbol sequence (rectangles in B).

[0039] Figure 6 shows the number of hotspots identified, the number of unique hotspots, the number of hotspots per protein and the hotspot length distribution for the 13 Orthopoxviruses.

[0040] Figure 7 shows the homologous hotspots for different (sub)sets of Orthopoxviruses. Connecting dots in the lower part of the plot denote combinations of Orthopoxviruses and the corresponding bar in the upper part of the plot (together with its associated number) denotes the number of homologous hotspots for that combination of Orthopoxviruses. For instance, second bar from the left tells us that there were 110 clusters of hotspots common to all Orthopoxviruses (i.e., all rows are connected by dots in the corresponding lower part of the plot), while the first bar from the left, tells us that there are 129 hotspots that are specific to only Volepox and Skunkpox. Only combinations of Orthopoxviruses that had five or more homologous hotspots are displayed.

[0041] Figure 8 shows the conserved regions derived from the homologous hotspots for different (sub)sets of Orthopoxviruses. Connecting dots in the lower part of the plot denote combinations of Orthopoxviruses and the corresponding bar in the upper part of the plot (together with its associated number) denotes the number of homologous hotspots for that combination of Orthopoxviruses. For instance, second bar from the left tells us that there were 130 clusters of conserved regions common to all Orthopoxviruses (i.e., all rows are connected by dots in the corresponding lower part of the plot), while the first bar from the left, tells us that there are 133 clusters of conserved regions that are specific to only Volepox and Skunkpox. Only combinations of Orthopoxviruses that had five or more homologous hotspots are displayed.

[0042] Figure 9 shows a table with all 235 hotspot conserved regions identified in the species of interest. These sequences were extracted from the hotspot clusters containing at least all species of interest and they contain species-specific amino acid variations. The first column in the table contains a unique conserved region identifier. The second column contains the amino acid sequence of the conserved region. The third column contains a pipe (“|”) delimited list of species in which a given conserved region was identified.

[0043] Figure 10 shows the results of a Greedy Hill Climbing (GHC) analysis with an allowed budget from 1 to 10 vaccine elements for different geographical regions and worldwide. The NEC / NOI dataset (triangles) contains all 235 hotspot conserved regions. The IEDB dataset (circles) contains the 54 hotspot conserved regions overlapping with linear T cell epitopes hosted in the IEDB database that were identified in Orthopox species of interest and associated with positive outcomes in humans. The X axis shows the allowed number of vaccine elements for each run, while the Y axis shows the estimated population coverage in a given region as well as worldwide.

[0044] Figure 11 shows the results of a Greedy Hill Climbing (GHC) analysis with an allowed budget from 1 to 50 vaccine elements for different geographical regions and worldwide. The NEC / NOI dataset (triangles) contains all 235 hotspot conserved regions. The IEDB dataset (circles) contains the 54 hotspot conserved regions overlapping with linear T cell epitopes hosted in the IEDB database that were identified in Orthopox species of interest and associated with positive outcomes in humans. The X axis shows the allowed number of vaccine elements for each run, while the Y axis shows the estimated population coverage in a given region as well as worldwide.

[0045] Figure 12 shows a table with the 54 hotspot conserved regions identified in the species of interest that overlap with epitopes hosted in the IEDB database annotated as generating positive outcome for the species of interest in humans. The first column in the table contains a unique conserved region identifier. The second column contains the amino acid sequence of the conserved region. The third column contains a pipe (“|”) delimited list of species in which a given conserved region was identified.

[0046] Figure 13 shows a table with results from the GHC analysis with budgets ranging from 1 to 50 vaccine elements for the worldwide population when using the entire set of 235 hotspot conserved regions. The first column contains the allowed budget for a given iteration of the GHC analysis. The second column contains the estimated population coverage for selected vaccine elements within a given budget. The third column contains sequence identifiers of the selected vaccine elements. These identifiers correspond to the ones in Figure 9.

[0047] Figure 14 shows a table with results from the GHC analysis with budgets ranging from 1 to 50 vaccine elements for the African population when using the entire set of 235 hotspot conserved regions. The first column contains the allowed budget for a given iteration of the GHC analysis. The second column contains the estimated population coverage for selected vaccine elements within a given budget. The third column contains sequence identifiers of the selected vaccine elements. These identifiers correspond to the ones in Figure 9.

[0048] Figure 15 shows a table with results from the GHC analysis with budgets ranging from 1 to 50 vaccine elements for the Asian population when using the entire set of 235 hotspot conserved regions. The first column contains the allowed budget for a given iteration of the GHC analysis. The second column contains the estimated population coverage for selected vaccine elements within a given budget. The third column contains sequence identifiers of the selected vaccine elements. These identifiers correspond to the ones in Figure 9.

[0049] Figure 16 shows a table with results from the GHC analysis with budgets ranging from 1 to 50 vaccine elements for the European population when using the entire set of 235 hotspot conserved regions. The first column contains the allowed budget for a given iteration of the GHC analysis. The second column contains the estimated population coverage for selected vaccine elements within a given budget. The third column contains sequence identifiers of the selected vaccine elements. These identifiers correspond to the ones in Figure 9.

[0050] Figure 17 shows a table with results from the GHC analysis with budgets ranging from 1 to 50 vaccine elements for the North American population when using the entire set of 235 hotspot conserved regions. The first column contains the allowed budget for a given iteration of the GHC analysis. The second column contains the estimated population coverage for selected vaccine elements within a given budget. The third column contains sequence identifiers of the selected vaccine elements. These identifiers correspond to the ones in Figure 9.

[0051] Figure 18 shows a table with results from the GHC analysis with budgets ranging from 1 to 50 vaccine elements for the South and Central American population when using the entire set of 235 hotspot conserved regions. The first column contains the allowed budget for a given iteration of the GHC analysis. The second column contains the estimated population coverage for selected vaccine elements within a given budget. The third column contains sequence identifiers of the selected vaccine elements. These identifiers correspond to the ones in Figure 9.

[0052] Figure 19 shows the overlap in vaccine elements identified by the GHC analysis between different regions of the world as well as worldwide when using a budget of 50 vaccine elements. Connecting dots in the lower part of the plot denote combinations of geographical regions and the corresponding bar in the upper part of the plot (together with its associated number) denotes the number of common vaccine elements for that combination of geographical regions. For instance, first bar from the left tells us that there were 41 vaccine elements common to all regions that were selected through GHC when using a maximum budget of 50 vaccine elements (i.e., all rows are connected by dots in the corresponding lower part of the plot).

[0053] Figure 20 shows details of the GHC selected vaccine elements with budgets from 1 to 50 vaccine elements for the worldwide population. The first column in the table contains the SEQ ID NO. corresponding to the vaccine element identifier and its sequence. The second column contains the vaccine element identifier (same as in Figure 9). The third column contains the amino acid sequence of the hotspot conserved region selected as vaccine element. The fourth column contains a pipe (“|”) delimited list of species covered (i.e., presenting this exact amino acid sequence) by a given vaccine element.

[0054] DETAILED DESCRIPTION

[0055] This invention is predicated on the development of a platform that is able to identify Orthopoxvirus epitopes that have a high probability of effectively stimulating a broad adaptive cellular immune response to multiple Orthopoxviruses when used in a vaccine composition. Thus, the incorporation of such epitopes into a vaccine composition may allow for the therapeutic or prophylactic treatment of infection with Orthopoxviruses. Unlike prior orthopoxvirus vaccination approaches, the vaccine compositions of the present invention are designed to stimulate a broad adaptive immune response through the specific activation of CD8+ and CD4+ T cells. It is expected that this will lead to the generation of a more substantial and sustained level of immunity. Furthermore, a surprisingly robust statistical model allows for the identification of those predicted Orthopoxvirus epitopes that are capable of triggering immunogenicity across a wide variety of human leukocyte antigen (HLA) types, hence the vaccine composition may have the potential to elicit protection against Orthopoxvirus across the global human population.

[0056] Orthopoxvirus is a genus of Poxviridae, a family of generally enveloped doublestranded DNA viruses for which vertebrates and arthropods serve as natural hosts. The virion is exceptionally large, with a diameter around 200 nm and a length of around 300 nm. The subfamily Chordopoxvirus has been shown to conserve the central region of the genome and contains around 90 genes (Gubser etal., 2004, Journal of General Virology, 85(1), 105-117). Infection with Poxviridae viruses such as vaccinia virus, molluscum contagiosum virus and Mpox virus is often mild, causing fever and a rash. However, infection can prove fatal as seen in the 2022-2023 Mpox outbreak or indeed the variola virus (causing smallpox disease), which had a 30 % fatality rate before its eradication by vaccination. The Orthopoxvirus genus houses both variola virus and Mpox virus, and as such there exists the possibility of the emergence of a variola virus-like virus could occur during natural evolution of modem zoonotic Orthopoxviruses (Shchelkunov, 2013, PLOS Pathogens 9(12), e1003756).

[0057] Orthopoxviruses replicate by first binding to a glycosaminoglycans or laminin receptor on the host cell surface before entering the cell where the virus uncoats in a two-step process. The outer membrane of the virus is removed as the particle first enters the cell, then the exposed virus particle fuses with the cellular membrane to release the core into the cytoplasm. Orthopoxviruses rely on virus- encoded proteins, enabling them to replicate in the cytoplasm. When the core enters the cytoplasm, early mRNA and protein synthesis occur, followed by DNA replication (Moss, 2013, Cold Spring Harbor Perspectives in Biology, 5(9), a010199). The replicated DNA provides a template for the synthesis of intermediate and late classes of mRNA. The initial infectious form, called a mature virion (MV), has a single external membrane. Some MVs are wrapped in a modified trans-Golgi or endosomal membrane to become triple-membrane particles called wrapped virions (WVs), whereas other MVs remain free or in inclusions within the cytoplasm until liberated by cell lysis. The WVs are transported on microtubules to the periphery of the cell where the outer membrane fuses with the plasma membrane to release an extracellular enveloped virion (EV), consisting essentially of a MV with one additional membrane.

[0058] Thus, in a first aspect of the invention, there is provided a vaccine composition comprising: i) a polypeptide, or ii) a polynucleotide encoding a polypeptide; wherein the polypeptide comprises one or more epitope sequences, wherein the one or more epitope sequences have the amino acid sequences of any one or more of SEQ ID Nos 1 to 57, or variants thereof, wherein the variant sequences have at least 70% sequence identity to any one or more of SEQ ID Nos 1 to 57.

[0059] The term vaccine composition, or vaccine, which from herein may be referred to interchangeably as the “composition”, relates to a biological preparation that induces active acquired immunity to a particular infectious disease, in this case an Orthopoxvirus infection. Typically, the vaccine contains an agent, or “foreign” agent, that resembles the infection-causing pathogen, or part of the infectioncausing pathogen, which within the prior art has often been a weakened or killed form of said pathogen, or recombinant protein or protein fragment from such pathogen, or polynucleotide encoding such a protein or protein fragment (Williamson et al. 1995, FEMS Immunology and Medical Microbiology 12 (3-4): 223-230). Such a foreign agent, protein or protein fragment would be recognised by a vaccine-receiver’s immune system, which in turn would destroy said agent and develop “memory” against the pathogen, inducing a level of lasting protection against future infections from the same or similar pathogenic sub-species. Through the route of vaccination, including those vaccine compositions of the present invention, it is envisaged that once the vaccinated subject again encounters the same pathogen of which said subject was vaccinated against, the individual’s immune system may thereby recognise said pathogen and elicit a more effective defence against infection. A more in-depth description of types of vaccines within the art can be found in US6541003 B1.

[0060] The active acquired immunity that may be induced by the invention is expected to be predominantly cellular. The active acquired immunity that may be induced by the invention may also be humoral. Humoral immunity refers to a response involving B cells which produce antibodies that specifically bind to antigens, or any future antigens, corresponding to those within the administered vaccine composition. B cells, each expressing a unique B cell receptor (BCR), recognise antigens in their native form. Upon this recognition and further interaction with other cells of the immune system, the activated B cell can differentiate into a plasma cell specialised to secrete antibodies against the encountered antigen. The term antibody refers to an immunoglobulin (Ig) that is produced by the immune system to specifically identify and neutralise foreign antigens. A subset of these B-cell derived plasma cells become long-lived antigen-specific memory B cells, as would be well understood by the skilled person.

[0061] Cellular immunity, meanwhile, can be broken into two distinct arms. The first involves helper T cells, or CD4+ T cells, which produce cytokines and orchestrate the activity of other immune cells in the immune response. The second involves killer T cells, also known as cytotoxic T lymphocytes (CTLs), or CD8+ T cells, which are cells capable of recognising antigens / epitopes presented by HLA and eradicate viral or bacterial infected host cells or cancerous or otherwise diseased host cells. In contrast to B cells, T cells only recognise antigens that have been processed into peptides and have been loaded onto histocompatibility complex (MHC) molecule and presented at the cell surface. CD4+ T cells interact with MHC class II molecules (MHC Class II), and are responsible for orchestrating the immune response, recognizing foreign antigens, activating various parts of the immune system and activating B cells and CD8+ T cells. CD8+ T cells interact with MHC Class I receptors and play a role in mounting an immune response against intracellular pathogens. As would be understood by the skilled person, on resolution of the infection, a subset of both CD8+ T cells and CD4+ T cells may remain as memory T cells, contributing to the acquired adaptive immunity, and allowing for a faster and stronger response to any secondary infection from the same foreign body (Bonilla & Oettgen 2010, Journal of Allergy and Clinical Immunology 125: 33-40).

[0062] It is envisaged that the vaccine composition of the present invention may be an epitope-based vaccine, or in other words, is comprised of one or more epitopes. Epitope-based vaccines (EVs) make use of short antigen-derived peptides corresponding to immune epitopes, which are administered to trigger a protective cellular immune response. EVs may also potentially enhance humoral responses. EVs potentially allow for precise control over the immune response activation by focusing on the most relevant — immunogenic and conserved — antigenic regions. Experimental screening of large sets of peptides is time-consuming and costly; therefore, in silico methods that facilitate T-cell epitope mapping of protein antigens are paramount for EV development. The prediction of T-cell epitopes focuses on the presentation of peptides at the infected cell surface by proteins encoded by the major histocompatibility complex (MHC).

[0063] The epitopes of the present invention may interact with MHC Class I and / or MHC Class II molecules to induce a CD8+ T cell and / or CD4+ T cell response, respectively. In a preferred embodiment of the present invention, there may be at least one epitope that interacts with MHC Class I, and at least one epitope that interacts with MHC Class II.

[0064] An “antigen” refers to a molecule capable of being bound by a receptor on a T cell, and may be comprised of one or more epitopes. An “antigen” may also refer to a molecule capable of being bound by an antibody or a B cell. As such, the terms epitope and antigen may be used interchangeably herein. Epitopes may also be referred to by the molecule for which they bind, such as “T cell epitopes”, or more specifically, “MHC Class I epitopes” or “MHC Class II epitopes”.

[0065] Thus in some embodiments of the present invention, the one or more epitopes may have the same length, or same number of amino acids. In other embodiments, the one or more epitopes may differ in length, or the number of amino acids. In some embodiments, the one or more epitopes may overlap with each other at least partly. In other embodiments, the one or more epitopes may overlap across more than one hotspot.

[0066] In other embodiments, one of the epitopes may fully comprise the entirety of another epitope within the same composition. It is envisaged that the one or more epitopes of the present invention are capable of stimulating a broad adaptive immune response across a plurality of human leukocyte antigen (HLA) types. In the context of the present invention, the term “plurality” is used to refer to “at least two”, or “two or more”. The human leukocyte antigen (HLA) complex is a set of genes encoding the MHC proteins in humans. Owing to the highly polymorphic nature of HLA genes, in which the term “polymorphic” refers to a high variability of different alleles, the precise MHC proteins of each human individual coded by varying HLA genes may differ to fine-tune the adaptive immune system. Many thousands of different alleles have been recognised for HLA molecules. As a result, each individual may have a unique “HLA type”, or HLA genotype, that differs across the global population, with a slight variability in the functioning of the immune system. The terms HLA type, HLA allele, or HLA genotype may be used interchangeably herein. HLA types are of particular significance when considering a vaccine comprised of epitopes that interact with MHC class I or class II molecules, as many epitopes are restricted in their capability of binding only particular HLA molecules encoded by particular HLA alleles, or in other words, are restricted to certain HLA types only. It would thus be appreciated by the skilled person that T cell epitopes that are capable of binding to a subject’s MHC Class I or MHC Class II molecules (and be presented at the infect cell surface), compatible with said subject’s HLA type, would thus present as a robust vaccine. A vaccine composition consisting of the same T cell epitopes may not prove effective if given to a subject with a different HLA type, if said HLA type encodes MHC molecules that are not capable of binding or interacting with said T cell epitopes. Such epitopes would not be able to stimulate a broad adaptive immune response across for either MHC Class I and / or MHC Class II immunogenicity in that particular subject. The vaccine composition of the present invention, in contrast, is envisaged to be able to stimulate a broad adaptive immune response across a plurality of HLA types, including alleles such as HLA-A*24:02 and HLA-B*07:01. The HLA alleles as referenced herein are given contemporary HLA nomenclature as standard to the field, wherein HLA-A, for example, refers to the gene loci in chromosome 6, whilst HLA-A*24:02 refers to the protein the allele codes for. An in-depth explanation of the complexities of HLA nomenclature can be found in Marsh et al. 2010, Tissue Antigens 75(4): 291-455. The approach of the present invention analysed 156 of the most frequent HLA molecules in the human population.

[0067] The platform used to identify and predict the one or more epitopes of the present invention was surprisingly robust, as was its integrated statistical analysis. Firstly, epitope mapping of the proteome for Orthopoxviruses for Class I epitopes was carried out using cell-surface antigen presentation and immunogenicity predictors from the “NEC Immune Profiler” suite of tools. Antigen Presentation (AP) was predicted from a machine learning model that integrates in an ensemble machine learning layer information from several HLA binding predictors - trained using empirically measured binding affinity data - and 13 different predictors of antigen processing.

[0068] This approach advantageously uses a statistical model to quantitatively analyse the predicted immunogenic potential of one or more epitopes - in other words the predicted ability of the one or more epitopes to instigate an immunogenic response - within an amino acid sub-sequence, across a set of different HLA types. The candidate regions (or “hotspots”) of the amino acid sequence that are identified by the quantitative statistical analysis may represent regions (or areas) of the one or more source proteins that are most likely to be viable vaccine targets and may be used in vaccine design and creation. These source proteins include those taken from species of the akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, macacapox virus, monkeypox virus, mousepox virus, racoonpox virus, skunkpox virus, taterapox virus, vaccinia virus, variola virus, and volepox virus.

[0069] It is envisaged that each of the hotspots identified herein may comprise one or more epitopes capable of stimulating an adaptive immune response through MHC Class I and / or MHC Class II. A candidate region may comprise a single epitope that is predicted to instigate an immunogenic response across a plurality of the HLA types. Such an epitope may be termed as “overlapping with” a number of HLA types. More typically however, a candidate region comprises a plurality of epitopes that, collectively, overlap with a large proportion of the analysed HLA types. For example, one epitope within a candidate region may overlap with n HLA types and a different epitope within the candidate region may overlap with m HLA types such that the candidate region is predicted to instigate an immunogenic response across the (m+n) HLA types.

[0070] The approach comprised the step of assigning, for each of the set of HLA types, an antigen presentation (AP) score for each amino acid, wherein said score is indicative of the immunogenic potential of an epitope comprising that amino acid, for that HLA type. For a given HLA allele, the score allocated to an amino acid corresponds to the best score obtained by an epitope prediction overlapping with this amino acid. For Class I HLA alleles, 1 represents the best score, wherein the amino acid has a higher likelihood of being naturally presented on the cell surface, whereas a score closer to 0 represents a lower likelihood. For Class II HLA alleles, in contrast, the predictions are of percentile rank binding affinity scores wherein lower scores are best. With a range of possible output scores of 0 to 100 for Class II HLA alleles, a score of 0 represents the best score, with the highest binding affinity.

[0071] The predictions for Class I and Class II HLA types were performed using an antigen presentation and binding affinity prediction algorithm, as well as experimental data. Examples of publicly available databases and tools that may be used for such predictions include the Immune Epitope Database (IEDB) (https: / / www.iedb.org / ), the NetMHC family of prediction tools

[0072] (http: / / www.cbs.dtu.dk / services / NetMHC / ), the TepiTool prediction tool (http: / / tools.iedb.org / tepitool / ), the NetChop prediction tool

[0073] (http: / / www.cbs.dtu.dk / services / NetChop / ) and the MHC-NP prediction tool (http: / / tools.immuneepitope.org / mhcnp / .). Other techniques are disclosed in W02020 / 070307 and WO2017 / 186959. Antigen presentation was predicted from a machine learning model that integrates in an ensemble machine learning layer information from several HLA binding predictors (trained on ic50nam binding affinity data) and a plurality of different predictors of antigen processing (trained on mass spectrometry data).

[0074] Each of the identified epitopes was then preferably allocated a score based on the immunogenic potential predicted using the above techniques. Advantageously, the method not only identified candidate regions comprising epitopes that may bind to an HLA molecule, but also those CD8 epitopes that are naturally processed by a cell’s antigen processing machinery, and presented on the surface of host infected cells.

[0075] The AP scores were assigned by the following protocol. Firstly, a plurality of epitopes were identified across the amino acid sequence, in a “moving window” of amino acids of fixed length. This was performed for each HLA type. For each of the identified first epitopes, a score was generated that is indicative of the immunogenic potential of that epitope, for the respective HLA type. A plurality of further epitopes were subsequently identified across the amino acid sequence, for each HLA type. Again, this was performed using a “moving window approach”. Each of the further epitopes were also assigned a score that was indicative of the immunogenic potential of that epitope, for the respective HLA type. Each amino acid was then assigned, for each HLA type, the score of the epitope that was predicted to have the best immunogenic potential of all the epitopes comprising that amino acid. Hence, for a particular HLA type, if epitope “A” and epitope “B” both comprised a particular amino acid “X”, the amino acid “X” would have been assigned the score of whichever epitope “A” or “B” is predicted to have the best immunogenic potential. In other words, for a given HLA type, the score allocated to an amino acid corresponds to the best score obtained by an epitope overlapping with this amino acid.

[0076] The AP score for each amino acid within a given source protein, or open reading frame, was averaged across HLA types. An AP score is given for each amino acids, wherein it is the average AP score of that amino acid across 156 of the most common HLA-A and HLA-B alleles that correspond to MHC Class I. In total, 156 of the most common human HLA-A and HLA-B across the globe were subjected to analysis. The one or more epitopes of the present invention may, further to being able to interact with the top 156 HLA-A or HLA-B alleles, also be able to stimulate a broad adaptive immune response across a plurality of HLA types including HLA-C, HLA-DP, HLA-DQ and / or HLA-DR alleles.

[0077] The HLA types analysed may further be characterised into HLA types of the same or different human population groups. A population group may be an ethnic population group (e.g. Caucasian, Africa, Asian) or a geographical population group (e.g. Lombardy, Lagos).

[0078] The composition of the present invention may comprise one or more epitopes found within any one or more of the hotspots, including the sequences of any of SEQ ID NOs: 1-57, and wherein said epitopes meet a particular threshold of a mean antigen presentation (AP) cut off value. Said mean AP cut off value is the value, averaged across all amino acids within an epitope, for which said epitope is considered able to stimulate a broad adaptive immune response across a plurality of HLA types, for either MHC Class I and / or MHC Class II immunogenicity.

[0079] For the sake of avoiding confusion, the term “antigen presentation (AP) value” may be used to mean binding affinity or percentile ranking, and the terms shall be used interchangeably. As such, reference to a mean “AP cut off value” in the context of MHC Class II, is to be construed as the mean binding affinity or mean percentile ranking of the relevant epitopes.

[0080] The term “polypeptide” is used to mean a chemical that is made of two or more amino acids, which combine to make protein / polypeptide. As used herein, the term “polypeptide” denotes the polypeptide of the vaccine composition, whereas the term “epitope” is used to denote potentially shorter amino acid sequences making up the polypeptide of the vaccine composition. Though of course, the “epitopes” are also polypeptides as denoted in the art and as would be understood by the skilled person. The term “epitope” as used herein also refers to any part of an antigen that is specifically recognised by receptors on any T cells (T cell receptors), as is its natural meaning. The “epitope” may also refer to any part of an antigen that is specifically recognised by any antibodies or B cells. The term “amino acid” retains its meaning as used in the art and would be understood by the skilled person. The term “variant” is used to refer to amino acid sequences that differ from the indicated SEQ ID No by one or more amnio acid residues. This could be a substitution, addition or deletion of one or more amino acids. An example would be the amino acid sequences ACDEF and ACDEE, whereby the latter is a variant of the former as one amino acid has been substituted. Thus, variants may have amino acids inserted, deleted or substituted from the indicated sequence. It is intended, and would be understood by the skilled person, that the polypeptide of the vaccine composition comprises a chain of epitopes, with their sequences selected from SEQ ID Nos 1 to 57. In an embodiment, each of the one or more epitope variant sequence of the polypeptide have at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to any of SEQ ID Nos 1 to 57. The skilled person would understand that the polypeptide may therefore comprise combinations of epitopes selected from SEQ ID Nos 1 to 57 having any % identity with the respective SEQ ID No. An example of this would be a polypeptide comprising an epitope with 100% identity to SEQ ID No 1 and 70% identity to SEQ ID No 2. This means that the polypeptide may retain its intended qualities and uses thereof if the amino acid sequence changes. Substitutions, deletions and insertions of amino acids may comprise potential manipulations of the polypeptide.

[0081] It is envisaged that the vaccine composition of the present invention may comprise any number of epitopes as would be suitable for use. In some embodiments, the polypeptide of the composition comprises at least one epitope having the amino acid sequence of SEQ ID No: 1 , or variants having at least 70% sequence identity thereto. In some embodiments, the polypeptide of the composition comprises at least one epitope having the amino acid sequence of SEQ ID No: 56, or variants having at least 70% sequence identity thereto.

[0082] In some embodiments, the polypeptide of the vaccine composition may comprise at least 2 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. The at least two epitope sequences may have the amino acid sequence of each of SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto. The at least two epitope sequences may have the amino acid sequence of each of SEQ ID Nos: 2 and 56, or variants having at least 70% sequence identity thereto. In another embodiment, the polypeptide of the vaccine composition may comprise at least 3 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. In another embodiment, the polypeptide of the vaccine composition may comprise at least 4 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. In another embodiment, the polypeptide of the vaccine composition may comprise at least 5 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. In another embodiment, the polypeptide of the vaccine composition may comprise at least 6 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. In another embodiment, the polypeptide of the vaccine composition may comprise at least 7 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. In another embodiment, the polypeptide of the vaccine composition may comprise at least 8 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. In another embodiment, the polypeptide of the vaccine composition may comprise at least 9 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. In another embodiment, the polypeptide of the vaccine composition may comprise at least 10 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57. Preferably, the polypeptide of the vaccine composition may comprise at least 17 epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57.

[0083] The polypeptide may comprise at least one epitope sequence having the amino acid sequence of SEQ ID No: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16 and / or 17, or a variant having at least 70% sequence identity thereto. Preferably, the polypeptide comprises epitope sequences having the amino acid sequences of each of SEQ Nos: 1 to 17, or variants having at least 70% sequence identity thereto.

[0084] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 1 , or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 1 to 3, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 1 to 4, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 1 to 5, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 1 to 6, or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 1 to 7, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 1 to 8, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 1 to 9, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 1 to 10, or variants having at least 70% sequence identity thereto; xi) SEQ I D Nos: 1 to 11 , or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 1 to 12, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 1 to 13, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 14, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 15, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 16, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 17, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 18, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 19, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 20, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 21 , or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 22, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 23, or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 24, or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 25, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 26, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 27, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 28, or variants having at least 70% sequence identity thereto; xxix) SEQ ID Nos: 1 to 29, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 30, or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 31 , or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 32, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 33, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 34, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 35, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 36, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 37, or variants having at least 70% sequence identity thereto; xxxviii) SEQ ID Nos: 1 to 38, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 39, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 40, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 41 , or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 42, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 43, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 44, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 45, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 46, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 47, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 48, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 49, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 50, or variants having at least 70% sequence identity thereto.

[0085] Said one or more epitopes may be advantageous in providing optimised, worldwide protection against Orthopoxvirus. It is estimated that protection with at least one epitope sequence, SEQ ID NO: 1 , may provide protection to at least 70%, 75%, 80%, 85%, 90% of the worldwide population. Preferably, SEQ ID NO: 1 may provide protection to at least 92% of the worldwide population. It is estimated that a combination of said one or more of the epitope sequences may provide protection to at least 70%, 75%, 80%, 85%, 90% or 95% of the worldwide population. Preferably, a combination of SEQ ID Nos: 1 to 17 provide protection to at least 97%, more preferably 97.45% of the worldwide population.

[0086] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 1 , or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 1 to 3, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 1 to 4, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 1-4 and 6, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 1 to 6, or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 1 to 7, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 1 to 7 and 9, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 1 to 7 and 9-10, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 1 to 7 and 9-11 , or variants having at least 70% sequence identity thereto; xi) SEQ ID Nos: 1 to 7, 9-11 and 13, or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 1 to 7 and 9 to 13, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 1 to 13, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 14, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 15, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 16, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 16 and 18, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 18, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 19, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 20, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 21 , or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 22, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 22 and 24, or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 22, 24 and 25, or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 25, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 25 and 38, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 25, 27 and 38, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 27 and 38, or variants having at least 70% sequence identity thereto; xxix) SEQ ID Nos: 1 to 28 and 38, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 29 and 38, or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 29, 31 and 38, or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 29, 31 , 38 and 53, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 31 , 38 and 53, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 32, 38 and 53, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 33, 38 and 53, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 33, 38, 52 and 53, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 33, 38, and 51 to 53, or variants having at least 70% sequence identity thereto; xxxviii) SEQ ID Nos: 1 to 33, 35, 38, and 51 to 53, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 33, 35, 36, 38, and 51 to 53, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 33, 35 to 38, and 51 to 53, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 33, 35 to 39, and 51 to 53, or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 33, 35 to 40 and 51 to 53, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 33, 35 to 41 and 51 to 53, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 33, 35 to 42 and 51 to 53, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 33, 35 to 43 and 51 to 53, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 33, 35 to 44 and 51 to 53, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 33, 35 to 45 and 51 to 53, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 33, 35 to 46 and 51 to 53, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 33, 35 to 47 and 51 to 53, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 33, 35 to 48 and 51 to 53, or variants having at least 70% sequence identity thereto.

[0087] Said one or more epitopes may be advantageous in providing optimised, protection against Orthopoxvirus in an African population. It is estimated that protection with at least one epitope sequence, SEQ ID NO: 1 , may provide protection to at least 70%, 75%, 80%, 85%, 90% of the African population. Preferably, SEQ ID NO: 1 may provide protection to at least 92% of the African population.

[0088] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 1 , or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 1 to 3, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 1 to 4, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 1 to 5, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 1 to 6, or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 1 to 7, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 1 to 8, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 1 to 9, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 1 to 10, or variants having at least 70% sequence identity thereto; xi) SEQ I D Nos: 1 to 11 , or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 1 to 12, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 1 to 13, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 14, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 15, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 15 and 17, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 17, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 18, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 18 and 20, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 20, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 21 , or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 21 and 25, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 21 , 24 and 25, or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 22, 24 and 25, or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 25, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 25 and 27, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 27, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 27 and 38, or variants having at least 70% sequence identity thereto; xxix) SEQ ID Nos: 1 to 28 and 38, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 29 and 38, or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 30 and 38, or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 30, 38 and 53, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 31 , 38 and 53, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 32, 38 and 53, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 32, 38, 53 and 54, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 32, 38, and 52 to

[0089] 54, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 33, 38, and 52 to 54 or variants having at least 70% sequence identity thereto; xxxviii) SEQ ID Nos: 1 to 33, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 34, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 35, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 36, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 39, and 51 to 54, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 40, and 51 to 54, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 41 , and 51 to 54, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 42, and 51 to 54, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 43, and 51 to 54, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 44, and 51 to 54, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 45, and 51 to 54, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 46, and 51 to 54, or variants having at least 70% sequence identity thereto.

[0090] Said one or more epitopes may be advantageous in providing optimised, protection against Orthopoxvirus in an Asian population. It is estimated that protection with at least one epitope sequence, SEQ ID NO: 1 , may provide protection to at least 70%, 75%, 80%, 85%, 90% of the Asian population. Preferably, SEQ ID NO: 1 may provide protection to at least 91% of the Asian population.

[0091] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 1 , or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 1 to 2 and 4, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 1 to 4, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 1 to 5, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 1 to 6, or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 1 to 7, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 1 to 7 and 9, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 1 to 7, 9 and 10, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 1 to 7, and 9 to 11 , or variants having at least 70% sequence identity thereto; xi) SEQ ID Nos: 1 to 11 , or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 1 to 12, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 1 to 13, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 14, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 15, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 16, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 17, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 18, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 19, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 20, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 21 , or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 21 and 24, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 21 , 24 and 25, or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 22, 24 and 25, or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 25, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 26, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 27, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 28, or variants having at least 70% sequence identity thereto; xxix) SEQ ID Nos: 1 to 28 and 38, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 29 and 38, or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 29, 31 and 38, or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 31 and 38, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 31 , 38 and 53, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 32, 38 and 53, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 33, 38 and 53, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 33, 38, 53 and 54, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 33, 38, and 52 to 54, or variants having at least 70% sequence identity thereto; xxxviii) SEQ ID Nos: 1 to 33, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 34, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 35, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 36, 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 38, and 51 to 54, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 39, and 51 to 54, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 40, and 51 to 54, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 41 , and 51 to 54, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 42, and 51 to 54, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 43, and 51 to 54, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 44, and 51 to 54, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 45, and 51 to 54, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 46, and 51 to 54, or variants having at least 70% sequence identity thereto.

[0092] Said one or more epitopes may be advantageous in providing optimised, protection against Orthopoxvirus in a European population. It is estimated that protection with at least one epitope sequence, SEQ ID NO: 1 , may provide protection to at least 70%, 75%, 80%, 85%, 90% of the European population. Preferably, SEQ ID NO: 1 may provide protection to at least 91 % of the European population.

[0093] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 56, or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 2 and 56, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 2, 3 and 56, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 2 to 4 and 56, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 2 to 4, 7 and 56, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 2 to 5, 7 and 56 or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 2 to 7 and 56, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 2 to 7, 56 and 57, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 2 to 7, 9, 56 and 57, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 2 to 7, 9, 11 , 56 and 57, or variants having at least 70% sequence identity thereto; xi) SEQ ID Nos: 2 to 7, 9, 11 , 13, 56 and 57, or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 2 to 9, 11 , 13, 56 and 57, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 2 to 9, 11 to 13, 56 and 57, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 9, 11 to 13, 56 and 57, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 9, 11 to 14, 56 and 57, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 9, 11 to 15, 56 and 57, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 9, 11 to 15, 17, 56 and 57, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 9, 11 to 15, 17, and 55 to 57, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 9, 11 to 15, 17, 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 9, 11 to 15, 17, 19, 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 20, 24, and 55 to 57, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 21 , 24, and 55 to 57, or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 21 , 24, 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 22, 24, 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 25, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 26, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 27, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 54 to 57, or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 53 to 57, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , 32, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35, 36, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 37, and 53 to 57, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos:

[0094] I to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 38, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 39, and 53 to 57, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 40, and 53 to 57, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 9,

[0095] II to 15, 17 to 29, 31 , 32, 35 to 41 , and 53 to 57, or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 42, and 53 to 57, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 43, and 53 to 57, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 44, and 53 to 57, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 45, and 53 to 57, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 46, and 53 to 57, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 47, and 53 to 57, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 48, and 53 to 57, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 49, and 53 to 57, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 50, and 53 to 57, or variants having at least 70% sequence identity thereto.

[0096] Said one or more epitopes may be advantageous in providing optimised, protection against Orthopoxvirus in a North American population. It is estimated that protection with at least one epitope sequence, SEQ ID NO: 56, may provide protection to at least 70%, 75%, 80%, 85%, 90% of the North American population. Preferably, SEQ ID NO: 56 may provide protection to at least 92% of the North American population.

[0097] The polypeptide of the vaccine composition may comprise one or more epitopes having the amino acid sequence of each of: i) SEQ ID No: 1 , or variants having at least 70% sequence identity thereto; ii) SEQ ID Nos: 1 and 2, or variants having at least 70% sequence identity thereto; iii) SEQ ID Nos: 1 to 3, or variants having at least 70% sequence identity thereto; iv) SEQ ID Nos: 1 to 4, or variants having at least 70% sequence identity thereto; v) SEQ ID Nos: 1 to 5, or variants having at least 70% sequence identity thereto; vi) SEQ ID Nos: 1 to 6, or variants having at least 70% sequence identity thereto; vii) SEQ ID Nos: 1 to 7, or variants having at least 70% sequence identity thereto; viii) SEQ ID Nos: 1 to 7 and 10, or variants having at least 70% sequence identity thereto; ix) SEQ ID Nos: 1 to 7, 9 and 10, or variants having at least 70% sequence identity thereto; x) SEQ ID Nos: 1 to 7, and 9 to 11 , or variants having at least 70% sequence identity thereto; xi) SEQ ID Nos: 1 to 7, 9 to 11 , and 13, or variants having at least 70% sequence identity thereto; xii) SEQ ID Nos: 1 to 7, and 9 to 13, or variants having at least 70% sequence identity thereto; xiii) SEQ ID Nos: 1 to 13, or variants having at least 70% sequence identity thereto; xiv) SEQ ID Nos: 1 to 14, or variants having at least 70% sequence identity thereto; xv) SEQ ID Nos: 1 to 15, or variants having at least 70% sequence identity thereto; xvi) SEQ ID Nos: 1 to 16, or variants having at least 70% sequence identity thereto; xvii) SEQ ID Nos: 1 to 16 and 19, or variants having at least 70% sequence identity thereto; xviii) SEQ ID Nos: 1 to 17 and 19, or variants having at least 70% sequence identity thereto; xix) SEQ ID Nos: 1 to 19, or variants having at least 70% sequence identity thereto; xx) SEQ ID Nos: 1 to 20, or variants having at least 70% sequence identity thereto; xxi) SEQ ID Nos: 1 to 21 , or variants having at least 70% sequence identity thereto; xxii) SEQ ID Nos: 1 to 21 and 24, or variants having at least 70% sequence identity thereto; xxiii) SEQ ID Nos: 1 to 22 and 24 or variants having at least 70% sequence identity thereto; xxiv) SEQ ID Nos: 1 to 22, 24 and 25 or variants having at least 70% sequence identity thereto; xxv) SEQ ID Nos: 1 to 25, or variants having at least 70% sequence identity thereto; xxvi) SEQ ID Nos: 1 to 25 and 27, or variants having at least 70% sequence identity thereto; xxvii) SEQ ID Nos: 1 to 27, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 28, or variants having at least 70% sequence identity thereto; xxix) SEQ ID Nos: 1 to 29, or variants having at least 70% sequence identity thereto; xxx) SEQ ID Nos: 1 to 29 and 31 , or variants having at least 70% sequence identity thereto; xxxi) SEQ ID Nos: 1 to 29, 31 , and 53, or variants having at least 70% sequence identity thereto; xxxii) SEQ ID Nos: 1 to 29, 31 , 32, and 53, or variants having at least 70% sequence identity thereto; xxxiii) SEQ ID Nos: 1 to 29, 31 , 32, 52 and 53, or variants having at least 70% sequence identity thereto; xxxiv) SEQ ID Nos: 1 to 29, 31 to 33, 52 and 53, or variants having at least 70% sequence identity thereto; xxxv) SEQ ID Nos: 1 to 29, 31 to 33, 38, 52 and 53, or variants having at least 70% sequence identity thereto; xxxvi) SEQ ID Nos: 1 to 29, 31 to 34, 38, 52 and 53, or variants having at least 70% sequence identity thereto; xxxvii) SEQ ID Nos: 1 to 29, 31 to 35, 38, 52 and 53, or variants having at least 70% sequence identity thereto; xxviii) SEQ ID Nos: 1 to 29, 31 to 36, 38, 52 and 53, or variants having at least 70% sequence identity thereto; xxxix) SEQ ID Nos: 1 to 29, 31 to 38, 52 and 53, or variants having at least 70% sequence identity thereto; xl) SEQ ID Nos: 1 to 29, 31 to 39, 52 and 53, or variants having at least 70% sequence identity thereto; xli) SEQ ID Nos: 1 to 29, 31 to 40, 52 and 53, or variants having at least 70% sequence identity thereto; xlii) SEQ ID Nos: 1 to 29, 31 to 41 , 52 and 53, or variants having at least 70% sequence identity thereto; xliii) SEQ ID Nos: 1 to 29, 31 to 42, 52 and 53, or variants having at least 70% sequence identity thereto; xliv) SEQ ID Nos: 1 to 29, 31 to 43, 52 and 53, or variants having at least 70% sequence identity thereto; xlv) SEQ ID Nos: 1 to 29, 31 to 44, 52 and 53, or variants having at least 70% sequence identity thereto; xlvi) SEQ ID Nos: 1 to 29, 31 to 45, 52 and 53, or variants having at least 70% sequence identity thereto; xlvii) SEQ ID Nos: 1 to 29, 31 to 46, 52 and 53, or variants having at least 70% sequence identity thereto; xlviii) SEQ ID Nos: 1 to 29, 31 to 47, 52 and 53, or variants having at least 70% sequence identity thereto; xlix) SEQ ID Nos: 1 to 29, 31 to 48, 52 and 53, or variants having at least 70% sequence identity thereto; or I) SEQ ID Nos: 1 to 29, 31 to 49, 52 and 53, or variants having at least 70% sequence identity thereto.

[0098] Said one or more epitopes may be advantageous in providing optimised, protection against Orthopoxvirus in a South and Central American population. It is estimated that protection with at least one epitope sequence, SEQ ID NO: 1 , may provide protection to at least 70%, 75%, 80%, 85%, 90% of the South and Central American population. Preferably, SEQ ID NO: 1 may provide protection to at least 92% of the South and Central American population.

[0099] In an embodiment of the invention, the polypeptide of the vaccine composition may comprise at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57 or variants thereof. For the avoidance of doubt, the polypeptide of the vaccine composition, according to any embodiment, may comprise at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57, or any variant thereof having at least 70% sequence identity thereto.

[0100] In a further embodiment of the invention, the polypeptide of the vaccine composition may comprise a concatemer of one or more epitope sequences having the amino acid sequences of any of SEQ ID Nos 1 to 57 or variants thereof. The term “concatemer” is used to mean a linear arrangement of repeated sequences that are connected in a continuous chain, with or without spacers as required. The skilled person would understand that the polypeptide of the vaccine composition may alternatively comprise combinations of epitopes selected from SEQ ID No 1 to 57 wherein certain epitopes are repeated. For example, the polypeptide may comprise the SEQ ID Nos 5, 5, 5, 8 and 9 in order. Thus, the polypeptide of the vaccine composition may comprise a combination of concatenated and non-concatenated sequences. Concatenated polypeptide sequences may comprise 2, 3, 4, 5, 6, 7, 8, 9, 10 or more repeats of any of the epitopes, as would be appreciated by the person of skill in the art. This applies to all embodiments of the invention.

[0101] The order of the sequences of the polypeptide may also be important, and can be tailored to optimise the immunogenicity of the polypeptide, whilst minimising the creation of off-target or autoimmune epitopes.

[0102] The term “spacer regions” is used to describe additional stretches of amino acids between the epitope sequences. These may be important to avoid additional off- target or autoimmune epitopes being created at the junction between the intended epitopes. The polypeptide may further comprise one or more spacer regions; preferably wherein when the polypeptide comprises more than one of the epitope sequences of SEQ ID Nos 1 to 57 or the variants thereof, the one or more spacer regions are positioned between one or more or each epitope sequence. Alternatively, spacer regions may be positioned between only selected epitope sequences as would be determined according to the specific polypeptide design, as explained above. Therefore, the polypeptide may comprise spacer regions between some epitope sequences and no spacer region between other epitope sequences. The spacer regions of the polypeptide may be present in instances of either one or multiple of the epitope sequences of SEQ ID Nos 1 to 57 comprising the polypeptide. A spacer region may be at least 5, 10, 15, 20 or 25 amino acids in length. A spacer region may be less than or equal to 25, 20, 15, 10 or 5 amino acids in length. Preferably, the spacer region is 5 amino acids in length. The inclusion of spacer regions in the polypeptide is applicable to all embodiments of the invention.

[0103] In an embodiment of the invention, the epitope variant sequences of the polypeptide of the vaccine composition have at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to any of SEQ ID Nos 1 to 57.

[0104] In a further embodiment of the invention, the polypeptide of the vaccine composition is no more than 2500, 2400, 2300, 2200, 2100, 2000, 1900, 1800, 1700, 1600, 1500, 1400, 1300, 1200, 1100, 1000, 900, 800, 700, 600, 500, 400, 300, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10 or 9 amino acids in length. The varying lengths of the polypeptide are intended to encapsulate the respective beneficial characteristics of having a polypeptide that is shorter, usually termed an “oligopeptide” or one that is longer, termed a “polypeptide”.

[0105] It is envisaged that the present invention may further comprise tertiary protein structures, or domains thereof, of species including the akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, macacapox virus, monkeypox virus, mousepox virus, racoonpox virus, skunkpox virus, taterapox virus, vaccinia virus, variola virus, or volepox virus.

[0106] The vaccine composition may of course comprise a polynucleotide encoding the polypeptide. The polynucleotide may be DNA, RNA or mRNA. Preferably, the vaccine composition is a mRNA vaccine composition. The term “polynucleotide” refers to a linear polymer whose molecule is composed of many nucleotide units, constituting a section of a nucleic acid molecule. The terms “DNA”, “RNA” and “mRNA” retain their meaning as used in the art and would be familiar to the skilled person.

[0107] In a second aspect of the invention, there is provided a vector comprising a polynucleotide as defined according to the first aspect of the invention, in that the polynucleotide encodes a polypeptide, wherein the polypeptide comprises one or more epitope sequences, wherein the one or more epitope sequences have the amino acid sequences of any one or more of SEQ ID Nos 1 to 57, or variants thereof, wherein the variant sequences have at least 70% sequence identity to any one or more of SEQ ID Nos 1 to 57. The polynucleotide of the vector may further comprise regulatory elements capable of driving transcription and / or translation of the polynucleotide in a host cell. The vector may also be a viral vector. By “vector” we intend any vehicle that is capable of supporting (i.e. carrying, encapsulating, incorporating and / or protecting) the polynucleotide and facilitating stable or transient transfection of a target host cell with the polynucleotide. The polynucleotide may also include the indicated regulatory elements to drive expression of a polypeptide encoded by the polynucleotide in the host cell. Suitable vectors may be, for example, plasmids, viral particles (e.g. lentiviruses, adenoviruses, AAVs and any number of other viruses, as would be understood by a person of skill in the art), nanoparticles, lipid-nanoparticles etc. An example of a vector suitable for the invention is a lentivirus, capable of inserting DNA into host cell genomes. Examples may also include one or more adenovirus vectors, vesicular stomatis virus vectors, influenza virus vectors or measles virus vectors. When viral particles are used as vectors, they are typically modified from their wild-type form to remove viral antigen encoding nucleic acid and / or to prevent viral replication in the host cell. The vector may be a naked DNA such as a plasmid or a virus, among other vectors used in the art and that would be familiar to the skilled person. The term “regulatory elements” refers to nucleotide sequences of genes that are involved in regulation of transcription.

[0108] A vaccine composition as defined according to the first aspect of the invention or a vector according to the second aspect of the invention may be formulated in a nanoparticle or lipid formulation. The term “nanoparticle” refers to a nanoscale particle between 1 and 100 nanometres in diameter, often with a very high surface area to volume ratio. Formulation in a nanoparticle may improve delivery of the vaccine composition, polynucleotide or vector of the invention to a subject and / or the immune system of a subject. The nanoparticle may be a lipid nanoparticle, designed to facilitate encapsulation and delivering of the vaccine composition or vector of the invention.

[0109] In a third aspect of the invention, there is provided a microorganism comprising a polypeptide or a polynucleotide as defined according to the first aspect of the invention. The microorganism may be a bacterium. The term “microorganism” refers to a microscopic organism, such as a bacterium, virus or fungus. Microorganisms may be used to further propagate the polynucleotide or polypeptide or to be administered as part of therapeutic or prophylactic use in order to deliver the polynucleotide or polypeptide to a specific tissue or cell type such as an antigen presenting cell. A microorganism comprising a polypeptide may be suited to industrial production of the polypeptide, and may include: Escherichia coli, Bacillus subtilis, Streptomyces spp., Corynebacterium glutamicum, Pseudomonas putida, Clostridium spp. and Lactobacillus. The microorganism of the invention may also be genetically modified. The polypeptide may also be produced by synthetic methods, of which the skilled person would be aware. The term “prophylactic treatment”, as used herein, refers to a medical procedure whose purpose is to prevent or reduce the morbidity or duration of (rather than treat or cure) a disease, such as an infection. In contrast, the term “therapeutic treatment” refers to a medical procedure with the purpose of treating or curing a viral infection or the associated symptoms thereof, as would be appreciated within the art.

[0110] In a fourth aspect of the invention, there is provided a pharmaceutical composition comprising: a vaccine composition according to the first aspect of the invention, a vector according to the second aspect of the invention or a microorganism according to the third aspect of the invention, and a pharmaceutically acceptable carrier, diluent, excipient and / or adjuvant, preferably further comprising an adjuvant. The vaccine composition, vector, microorganism or pharmaceutical composition may be formulated for parental, oral, sublingual, nasal, naso-oral, or pulmonary administration. Said parental administration may be subcutaneous, intradermal, intramuscular, subdermal, intraperitoneal, or intravenous administration.

[0111] In an embodiment of the invention, the pharmaceutical composition is an mRNA vaccine.

[0112] The term “mRNA vaccine” describes a vaccine that uses a copy of an mRNA molecule to produce an immune response. The vaccine delivers molecules of antigen-encoding mRNA into host immune cells, which use the designed mRNA as a blueprint to build foreign protein, or fragment of a protein, that would normally be produced by a pathogen such as a virus. These protein molecules stimulate an adaptive immune response that teaches the body to identify and destroy the corresponding pathogen. In an embodiment of the invention, the foreign protein produced is of an Orthopoxvirus. The mRNA may be delivered by a co-formulation of the mRNA encapsulated in lipid nanoparticles. The mRNA may also be formulated for nasal or intratracheal delivery via a nanoparticle-delivery-based system. This system may comprise a biodegradable poly(amine-co-ester) polymer that forms polyplexes with mRNA so that the vaccine is inhalable.

[0113] The phrase "pharmaceutically acceptable" refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a human, as appropriate. The preparation of a pharmaceutical composition that contains the vaccine composition of the present invention will be known to those of skill in the art in light of the present disclosure. Moreover, for human administration, it will be understood that preparations should meet sterility, pyrogenicity, general safety and purity standards. A specific example of a pharmacologically acceptable carrier as described herein is borate buffer or sterile saline solution (0.9% NaCI).

[0114] As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives {e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavouring agents, dyes, such like materials and combinations thereof, as would be known to one of ordinary skill in the art (see, for example, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329).

[0115] Examples of adjuvants which may be effective include but are not limited to: unmethylated cytosine-guanine dinucleotide (CpG) motifs, granulocytemacrophage colony-stimulating factor (GM-CSF), aluminium hydroxide, N-acetyl- muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-nor-muramyl-L-alanyl-D- isoglutamine (CGP 11637, referred to as nor-MDP), N-acetylmuramyl-L-alanyl-D- isoglutaminyl-L-alanine-2-(1'-2'-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)- ethylamine (CGP I9835A, referred to as MTP-PE), and RIBI, which contains three components extracted from bacteria, monophosphoryl lipid A, trehalose dimycolate and cell wall skeleton (MPL+TDM+CWS) in a 2% squalene / Tween 80 emulsion. Further examples of adjuvants and other agents include aluminium hydroxide, aluminium phosphate, aluminium potassium sulfate (alum), beryllium sulfate, silica, kaolin, carbon, water-in-oil emulsions, oil-in-water emulsions, muramyl dipeptide, bacterial endotoxin, lipid X, Corynebacterium parvum (Propionobacterium acnes), Bordetella pertussis, polyribonucleotides, sodium alginate, lanolin, lysolecithin, vitamin A, saponin, liposomes, levamisole, DEAB- dextran, blocked copolymers or other synthetic adjuvants. Such adjuvants are available commercially from various sources, for example, Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.) or Freund's Incomplete Adjuvant and Complete Adjuvant (Difco Laboratories, Detroit, Mich.).

[0116] In some embodiments of the present invention, the vaccine composition may further comprise a pharmaceutically acceptable carrier, diluent, excipient and / or adjuvant, as well as minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, and / or adjuvants which enhance the effectiveness of the vaccine.

[0117] The vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention may be for use in treating or preventing a disease or condition in a subject. Preferably, the disease or condition is an infection with an orthopoxvirus.

[0118] The vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention may be for use in the therapeutic or prophylactic treatment of any Orthopoxvirus infection. The terms “treatment” and “prophylaxis” are to be used interchangeably with “therapeutic treatment” and “prophylactic treatment” respectively. It is envisaged that the vaccine composition, vector, microorganism or pharmaceutical composition may be used against any Orthopoxvirus infection.

[0119] It is envisaged that the polypeptide, polynucleotide, vector, microorganism, composition or vaccine composition of the present invention may aid in the therapeutic or prophylactic treatment of an Orthopoxvirus infection in a human subject. The virus may be selected from: akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, macacapox virus, monkeypox virus, mousepox virus, racoonpox virus, skunkpox virus, taterapox virus, vaccinia virus, variola virus, or volepox virus. Said composition may comprise one or more epitopes as defined according to the first aspect of the invention that are capable of stimulating a broad adaptive immune response across a variety of HLA types. The term “subject” refers to a person subjected to treatment, observation or experiment and would be understood by the skilled person as such.

[0120] The vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention may be for use in the treatment or prophylaxis of an infection caused by akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, macacapox virus, monkeypox virus, mousepox virus, racoonpox virus, skunkpox virus, taterapox virus, vaccinia virus, variola virus, or volepox virus. Preferably, the infection is caused by akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, monkeypox virus, variola virus or vaccinia virus.

[0121] As Orthopoxvirus subgenera that are known to pathogenically infect humans, akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, monkeypox virus, variola virus or vaccinia virus provide preferential viruses to be targeted by the invention.

[0122] The invention also provides for the use of the vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention in the manufacture of a medicament for the prevention or treatment of an infection with an orthopoxvirus in a subject in need thereof.

[0123] The invention also provides a method of treating or preventing an infection with an orthopoxvirus comprising administering to a subject in need thereof an effective amount of the vaccine composition according to the first aspect of the invention, the vector according to the second aspect of the invention, the microorganism according to the third aspect of the invention or the pharmaceutical composition according to the fourth aspect of the invention.

[0124] It is envisaged that administration of the vaccine composition, vector, microorganism or pharmaceutical composition according to the present invention would be carried out following an appropriate immunisation regimen. The term “appropriate immunisation regimen” is to be construed as a schedule or timescale of one or more administrations of the compositions of the present invention, which may resultantly yield the most effective results in consideration of immunisation efficacy and safety of the subject to which the composition is being administered. For example, for the therapeutic or prophylactic treatment of an Orthopoxvirus infection, an immunisation regimen should be chosen that yields as effective immunisation against the Orthopoxvirus as possible, whilst still maintaining suitable safety for the subject.

[0125] In some embodiments of the present invention, the immunisation regimen may comprise a single administration. In other embodiments, the immunisation regimen may comprise multiple administrations, either concomitantly or over an appropriate period of time.

[0126] It is envisaged that the appropriate dosage regimen may be repeated for a subject at a suitable time.

[0127] There exists further the possibility to further administer boost immunisations after a more extended period of time. This may be selected as an appropriate measure if a subject’s T-cell response falls below determined protective levels. The boost immunisations may be administered if a subject’s immunoglobulin G (IgG) antibody levels fall below determine protective levels. Thus in some embodiments, an appropriate dosage regimen may be given as a “boost immunisation” after 6 months.

[0128] In some embodiments of the present invention, the vaccine composition, vector, microorganism or pharmaceutical composition may be administered for the treatment or prevention of infections caused by a virus in combination with one or more other antiviral therapies or other appropriate therapies such as stem cell therapies. Such antiviral therapies may include administration of Tecovirimat (Tpoxx ®), Brincidofovir (Tembexa ®), Cidofovir (Vistide ®), VIGIV (Vaccinia Immune Globulin Intravenous), orTrifluridine (Viroptic ®). Such antiviral therapies may be administered simultaneously, separately or sequentially with the composition of the present invention. In a further embodiment, the antiviral therapy is administered via the same or different route of administration as the vaccine composition, vector, microorganism or pharmaceutical composition of the present invention, for example via intradermal injection.

[0129] The invention will now be explained with reference to the following examples.

[0130] Example 1

[0131] The sets of protein sequences for each of the 13 Orthopoxvirus proteomes are shown in Figures 1 and 2. As shown in Figure 3, the first part of the data processing to identify potential epitopes involved the generation of epitope scores for each amino acid position in all the proteins in the 13 Orthopoxvirus proteomes, for 156 H LA types.

[0132] The amino acid sequences of the proteins were split into minimal epitopes (9- and 10-mers) by means of a sliding window. The important determinants of antigen presentation (AP) were assessed for each peptide of each protein and each virus in the Orthopoxvirus genus for their potential to be efficiently presented. These determinants consisted of: (1) the predicted binding affinity between the candidate peptide and 156 of the most frequent HLA molecules in the human population, (2) the predicted potential of the candidate peptide to be efficiently processed by the antigen processing machinery of the host infected cell, and (3) the predicted probability of the candidate peptide to be presented on the host infected cell surface, which, among other factors, takes binding and processing into account. The Al prediction platform used was the NEC Immune Profiler (NIP), which provided the Al predictions for these key determinants, such as the AP scores. The AP score is in the range between 0 and 1 , with 1 being the maximum of the likelihood that a specific candidate peptide was presented on the host infected cell surface.

[0133] The AP scores were calculated based on a set of 156 most frequent Class I HLA alleles (A and B) for all the possible combinations between an HLA allele and each epitope, for each Orthopoxvirus. As such, each amino acid within a protein sequence was assigned an AP score.

[0134] Example 2

[0135] To ascertain whether specific regions in a given viral protein were enriched with higher immunogenic scores as compared to the rest of the protein, a framework was implemented based on a peak calling algorithm able to identify sub-regions of a protein which present higher immunogenicity than the rest of the protein, as shown in Figure 3. These regions were labelled as “hotspots”.

[0136] The entire map of AP score predictions of a given protein (see Figure 3, (1)) was used as input data. First, the AP scores for each amino acid within the protein sequence were summed across the 156 considered HLA alleles and the mean of these sums was calculated (see Figure 3, (2)). This mean represents the baseline or “waterline” for the peak detection algorithm. A local polynomial regression (LOESS) smoothing curve with a span of 10% was applied on the individual AP score sums (i.e., at amino acid level) thus generating a presentation potential "landscape" of the entire protein sequence (see Figure 3, (3)).

[0137] A rolling function was applied on the “landscape” to identify all local maxima within the protein. The default window size for the rolling function was set to nine amino acids (i.e., 9-mers), in agreement with the framework used to predict the presentation potential of each peptide within the protein. The identified peak maxima represent the peak summits across the “landscape”. Only peak summits that are above the “waterline” (i.e., the baseline) were considered.

[0138] The peak summits were then input into a run length encoding (RLE) algorithm to identify the peak regions. A peak region was defined as the entire region flanking a peak summit on both sides that is consistently above the “waterline”. The minimum required size of a peak region was set to 15 amino acids, to conjointly account for Class II HLAs, and the maximum allowed length was set to 100 amino acids, to meet the limitations of peptide synthesis. Peak regions that were shorter than the minimum required length were flanked on both sides with the remaining number of amino acids required to meet the minimum length. Then, the best window of 15 amino acids, with respect to the sum of AP scores across its constituting amino acids was kept as the corresponding hotspot. For peak regions exceeding 100 amino acids, but up to 150 amino acids in length, the same approach was used: the best window of 100 amino acids was kept. The peak regions longer than 150 amino acids were recursively split into shorter regions by re-calculating the baseline within the region (i.e., raising the “waterline”) and subsequent de novo detection of the peak summits and peak regions.

[0139] Finally, the amino acid sequences corresponding to the peak regions identified across the proteins are extracted, together forming the entire set of hotspots for a given viral species (see Figure 3(4)).

[0140] Example 3

[0141] To ascertain whether there were highly immunogenic regions that were shared by multiple Orthopoxviruses, a framework was developed to identify and cluster hotspots that were common to more than one viral species.

[0142] For that, all possible pairwise combinations of identified hotspots were generated across all proteins and all Orthopoxviruses.

[0143] {{x. y } I y EO / \X + y } with x and y being two non-identical hotspots and O the entire set of Orthopoxviruses. Next, the hotspot sequences were converted into ordered vectors of amino acid frequencies. For instance, the frequency vector of hotspot x would be denoted as:

[0144] Fx = { (fai, .. ,fan) I a, EA / i1 < / < |A| } where A is the complete, ordered set of amino acids compiled based on the entire protein sequence input data and fa, is the frequency of amino acid / within the hotspot sequence. The similarity between the two amino acid frequency vectors within each pair was assessed by means of Pearson correlation coefficient (p):

[0145] P(X,y)= corr Fx, Fy)

[0146] This allowed for the comparison of the amino acid proportions between two hotspot sequences. Only hotspot pairs with a p > 0.95 were kept for further analysis:

[0147] C = { (x,y) | p(x,y)> 0.95 }

[0148] The chosen cutoff value is a stringent threshold in itself. However, it still allowed for point mutations and / or differences in the flanking regions (i.e., shifts in the start and / or end coordinates of the hotspot sequence) of the two hotspot sequences being compared.

[0149] Potential false positives can arise as (1) hotspot sequences with the same amino acid composition, but in a different order (e.g., mirrored or shuffled) or (2) very short hotspot sequences (e.g. , 15 amino acids) for which the amino acid frequency profile is quite flat (i.e., a frequency vector with most values being 0 and / or 1). To discard false positives, the pairwise hotspot sequence similarity was calculated for the entire set of pairs with a p > 0.95:

[0150] S(x,y)= { 1 - dist(x, y) / max(dist(x, y)) | (x, y) E C }

[0151] The distance dist between sequence x and y is calculated as the restricted Damerau-Levenshtein distance. The sequence similarity is a value between 0 and 1 , with 0 denoting two completely different character sequences and 1 denoting identical character sequences.

[0152] Next, the difference between the Pearson correlation coefficient and sequence similarity was calculated for all considered hotspot pairs: (X,y)~ { P(*,y) " (*,y) I (x> Y) C }

[0153] Based on the distribution of D^yj, the 99thpercentile was used as a cutoff to discard hotspot pairs presenting extreme differences (i.e. , equal to or above the 99thpercentile). In other words, pairs of hotspots with a high p but low S(X,y> were considered false positives and discarded. As such, the final set of hotspots was defined as:

[0154] H = { (x, y) | D(x,y)< P99 (D) x\ (x, y) E C }

[0155] These hotspots were then placed under a directed graph representation, where each hotspot was represented by a vertex and an edge between two vertices was defined for each (x, y) EH. Finally, clusters of homologous hotspots were identified by extracting all connected vertices within the graph. Due to the nature of this implementation, the minimum size of a hotspot cluster was two. In other words, a hotspot was considered for further analysis if it is common to at least two Orthopoxvirus species.

[0156] Example 4

[0157] The clusters of homologous hotspots were converted to the FASTA format and the hotspot sequences were aligned using Clustal Omega, a third party tool for multiple sequence alignment (MSA), using default parameters.

[0158] The output of the MSA tool (see Figure 4, (A)) was then used to extend individual hotspot sequences (i.e., per viral species) where possible, such that the “real estate” of the homologous hotspots is maximized. This was done by replacing the flanking dashes (“-”) output by Clustal Omega (i.e., placeholders used during the alignment of the amino acid sequences) surrounding a hotspot sequence, with the actual amino acids at those positions taken from the original, corresponding protein sequence (see Figure 4, (C)).

[0159] The rationale behind this step was that minor differences in the baseline used in peak detection (i.e., “waterline”) would cause a hotspot region within a given protein and a given viral species to be trimmed left and / or right with respect to the same hotspot region in an analogous protein in a different viral species. However, such small differences in the “waterline” level should not have a notable impact on the overall hotspot region, with respect to its presentation potential.

[0160] The clusters of extended hotspots were then re-aligned using the same MSA third party tool, using the default parameter settings. Based on the sequence of consensus symbols that Clustal Omega outputs, each cluster of homologous hotspots was processed to identify which hotspot sub-regions are the most conserved across the viral species accounted for within the cluster. An amino acid position within a hotspot was considered conserved if designated with an or a in the Clustal Omega consensus symbol sequence.

[0161] Next, the consensus symbol sequence was transformed into a binary vector, with all conserved amino acid positions converted to a 1 and all other amino acid positions converted to a 0. Run length encoding (RLE) was then used to identify all the conserved regions within a cluster of homologous hotspots (see Figure 5). In other words, the regions of consecutive 1s were delineated. Only conserved regions that were at least 15 amino acids in length were kept. Note that one hotspot cluster can yield one, several, or no highly conserved regions.

[0162] A conservation score was assigned to each highly conserved hotspot region, calculated between the representative sequence within a hotspot cluster and the hotspot sequence of a given Orthopoxvirus species within the same cluster. The representative sequence was defined as the most prevalent hotspot sequence within a given cluster. The conservation score was calculated as a value between 0 and 1 , with 1 being perfect conservation. The mean AP score across the entire set of considered HLA alleles and across the constituting amino acids of a given hotspot sequence was also calculated for all conserved hotspot sequences. This score is considered as an indicator of immunogenicity.

[0163] Example 5

[0164] The sets of protein sequences for each of the thirteen Orthopoxviruses were assessed fortheir potential to be presented on the host infected cell surface. This allowed for each amino acid within a protein sequence to be assigned an antigen presentation (AP) score. Based on these AP scores, regions within the viral proteins that show high predicted presentation potential were identified. These regions were labelled as “hotspots”.

[0165] A total of 17,563 across all 13 Orthopoxviruses were identified, with more than 1 ,000 hotspots for each viral species based on their entire set of protein sequences. The minimum length of a hotspot region was set to 15 amino acids, to account for Class II HLA alleles and the maximum length was set to 100 to meet the limitations of peptide manufacturing. The number of hotspots per protein ranged from 1 to 27, depending on protein length and distribution of AP scores within the protein sequence. There were no notable outliers present in the data, with respect to the number of hotspots and their length distribution. Figure 6 contains more granular descriptive statistics regarding the identified hotspots.

[0166] The entire set of hotspots was used to check if there were any homologous protein regions, with a high presentation potential across multiple Orthopoxviruses, which could lead to the identification of cross-reactive epitopes. Clusters of highly similar hotspots across different species were therefore identified. There were 2,219 hotspot clusters identified in total, the minimum size of a cluster being 2. In other words, only hotspots that were homologous in at least two Orthopoxvirus species were kept.

[0167] Out of those, 110 hotspot clusters accounted for all thirteen Orthopoxvirus species (see Figure 7). When looking at a subset of viruses of interest, for instance Monkeypox, Cowpox, Vaccinia, and Variola, 429 hotspots that are homologs in at least these four Orthopoxvirus species were found. Interestingly enough, Volepox, Skunkpox, and Racoonpox seem to share more hotspots among themselves than the rest of the Orthopoxviruses. 86 hotspots homologous to all these three Orthopoxviruses were identified, and another 129 hotspots were shared between Volepox and Skunkpox.

[0168] The highly conserved hotspot regions within each hotspot cluster were then identified. There were 2,216 conserved regions of at least 15 amino acids in length coming from 2,049 (out of 2,219) hotspot clusters. Note that one hotspot cluster can yield one, several, or none highly conserved regions. Out of those, 130 were homologous to all thirteen Orthopoxviruses (see Figure 8). For the same example subset of viruses of interest as described above, 445 highly conserved regions were shared between the four Orthopoxviruses. Across all 130 highly conserved hotspot regions that were shared among all Orthopoxes, the median sequence conservation score was 1 (i.e., identical sequences) with a mean of 0.995.

[0169] Taken together, these results show that there is a substantial set of protein subsequences, homologous to all Orthopoxviruses, that show high sequence conservation, thus representing potential candidates for a broadly protective T-cell vaccine against all Orthopoxviruses.

[0170] Example 6

[0171] To assess which is the optimal (sub)set of conserved hotspot regions (i.e., candidate vaccine elements) such that the expected population coverage is maximized, the inventors employed a Greedy Hill Climbing (GHC) algorithm. Figures 7 and 8 illustrate the results of GHC for a number of vaccine elements, ranging from 1 to 10 and from 1 to 50 respectively out of the 235 possible candidate vaccine elements identified in the species of interest.

[0172] This analysis was performed on simulated individual haplotypes (Digital Twins) sampled from a Worldwide population, as well as on different regions of the world: Africa, Asia, Europe, North America, and South and Central America to assess response likelihood variation at geographical subregion level. For this purpose, the HLA haplotype data from The Allele Frequency Net Database was leveraged (https: / / academic.oup.com / nar / article / 48 / D1 / D783 / 5624967).

[0173] The Digital Twin framework allowed for the assessment of hotspot performance by estimating the response likelihood of individuals to these peptides based on their HLA haplotype. This is achieved by (i) simulating populations for a given geographical region of interest based on the data derived from the AFND and (ii) selecting candidate vaccine elements from the set of sequences that maximize the likelihood of response given the HLA haplotype makeup of the individuals in the underlying population, at the same time accounting for rare HLA alleles. More specifically, the population for each geographical region of interest was generated by sampling ten times 10,000 individuals from the total number of individuals available for that region in the data derived from AFND. This means that for each geographical region, the hotspot performance was assessed across 100,000 individuals in total. For the Worldwide population, the individuals were sampled from all regions available in AFND with the sample size for each of these regions being proportional to the current global distribution.

[0174] Two datasets were individually evaluated with GHC. A first dataset, denoted NEC / NOI in Figures 10 and 11 , that contains the entire set of 235 conserved hotspot regions (as in Figure 9) of potential vaccine element candidates identified in the species of interest. A second dataset which contains only the conserved hotspot regions that overlap with linear T cell epitopes from species of interest hosted in the IEDB database (www.iedb.org ; https: / / pubmed.ncbi.nlm.nih.gov / 30357391 / ) that were found to generate positive outcomes in humans (as listed in Figure 12). The same 1 to 50 vaccine element budget range was used for both datasets. The larger budget (e.g. , up to 50 vaccine elements) was used to assess convergence of the GHC optimization. These results show that in almost all geographical regions (except Europe) the vaccine elements from the entire NEC / NOI dataset will achieve maximal population coverage with fewer vaccine elements as compared to restricting the set of vaccine elements to the ones overlapping known and reported epitopes (i.e. , IEDB dataset).

[0175] Upon comparison of the results of the two sets, the inventors observed that for most of the assessed regions, maximal estimated population coverage was achieved with fewer vaccine elements when using the entire set of conserved regions from SOI “NEC / NOI” than when using only the conserved regions that overlap with IEDB epitopes. Detailed results of the GHC optimization analysis with a budget ranging from 1 to 50 vaccine elements are shown in Figure 13, where the first column contains the allowed number of vaccine elements for a given run of GHC on a worldwide population. The second contains the estimated population coverage in percentages for the selected vaccine elements for a given budget (i.e. , allowed number of vaccine elements). The last column contains the sequence identifiers for the selected vaccine elements for a given budget. These sequence identifiers correspond to the identifiers in Figure 9.

[0176] The data presented in Figure 13 demonstrate that with only one selected vaccine element, the inventors observed an expected population coverage of -92% on a worldwide population. Importantly, the optimal combination was found to be using 17 vaccine elements with an expected population coverage of -97.45%. Adding more vaccine elements beyond that does not increase the expected coverage in a significant manner. However, it is possible that the rest of the vaccine element sequences would account for more rare, specific HLA haplotypes.

[0177] Similar to Figure 13, Figures 14-18 contain the results of the GHC analysis and the corresponding selected vaccine element sequences for the African, Asian, European, North American, and South and Central American populations.

[0178] The inventors assessed how similar the sets of selected vaccine elements are between the different geographical regions studied. Figure 19 shows the overlap between selected vaccine elements. Out of maximum 50 possible vaccine elements for a given region, 41 were found to be common between the African, Asian, European, North American, South and Central American, and Worldwide populations. In total, 57 out of 235 unique vaccine elements were selected by GHC across all geographical regions. This demonstrates the potential of a broadly protective vaccine by selecting a subset of vaccine elements from the results of the GHC analysis.

[0179] The table in Figure 20 shows the mapping between the identifiers of the vaccine elements selected by the GHC optimization analysis, their amino acid sequence and corresponding SEQ ID NO. In brief, given a set of conserved hotspot sequences as candidate vaccine elements and a fixed budget of elements to be selected, GHC will aim at selecting the optimal subset of vaccine elements that fits the number of allowed vaccine elements and maximizes the expected population coverage in a given geographical region.

[0180] The sequences forming part of the description are the following:

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

Claims

CLAIMSA vaccine composition comprising: i) a polypeptide, or ii) a polynucleotide encoding a polypeptide; wherein the polypeptide comprises one or more epitope sequences, wherein the one or more epitope sequences have the amino acid sequences of any one or more of SEQ ID Nos 1 to 57, or variants thereof, wherein the variant sequences have at least 70% sequence identity to any one or more of SEQ ID Nos 1 to 57; wherein SEQ ID Nos 1 to 57 have the following sequences:DMSSSQDLSFIYRKPETNYYIHPILMALFGIKLPALENAYVHGDTYSLIQQLYEFRRVKSYNYMLLVNR L (SEQ ID NO: 1),VNGHFFVSNKVFDKSLLYKYENDIITVPFRLSYE PFVWGVNFRK (SEQ ID NO: 2),QNNRYRFAFLYLLVKWYKKYHIPIMKLYPTPEEIPDFAFYLKIGTLLVSSSVKHIPLMTDLSKKGYILYD NVVTLPL (SEQ ID NO: 3), d) RRVIGQINQPTATVTEVHAATSLSVYTKPYYGNT (SEQ ID NO: 4), e) DITKLDITKTELEHISKYMKYYTTYIDHIVNIVLQNNYIDILASIID (SEQ ID NO: 5),YGSSIRLELVNLIQAKTKNFTIDFKLKYFLGSGAQ SKSSLLHAIN (SEQ ID NO: 6), g) SGIKTKYYKFDYIQETIRSDTFVSSVREVFYFGKFNIIDWQFAIHYSFHPRHYATVMNNLSELTASGG (SEQ ID NO: 7),ERPQHMLMRVAVGIHQWDIES (SEQ ID NO: 8),AMRHLSLAGLLSDHKSNVEGINFIIKSSYVFKRYLAIYGFGVTFKDLRPNSTFTN KLEAI N VEKI ELI KD AYAKYLKDV (SEQ ID NO: 9),SNINQPWIKTISKRMRVDIINHSIVTRGKSSILQTI EIIFTNRTC (SEQ ID NO: 10), k) YKASTVI KGPLLKLLLETKTI LVRSETKQKFPYEGGKVF (SEQ ID NO: 11),I) YTYRIIKSSFPVPTI (SEQ ID NO: 12), m) IPTILPHQLATLDYLVRTIIDE (SEQ ID NO: 13), n) DSIRTLVSWFFSNGKLKDNFSLSSIRFHIKELENEYYFRNEVFHC (SEQ ID NO: 14), o) QNNRYRFAFLYLLVKWYRKYHVPIMKLYPTPEEIPDFAFYLKIGTLLVSSSVKHIPLMTDLSKKGYILYDNVVTLPL (SEQ ID NO: 15), p) IITPIVFYRSGTETKITFALKKLIIDREWANVIGLSGD (SEQ ID NO: 16), q) VDTEFINKFLEFSNKVYEALYYVHSLLYS (SEQ ID NO: 17), r) NLDISSVQLSNNKYVLFVKKMLPKIILFQN (SEQ ID NO: 18), s) DSKKIIEIVSHLRASTTENAAYQVLQQN (SEQ ID NO: 19), t) RNLEELTTIFIKYVNGWVKKGGHVTLFIDRGSI (SEQ ID NO: 20), u) INDDFYHISTGGYGIVFKIDNYVVKFVFEATKLYSPMETTAEFTVPKFLYNN (SEQ ID NO: 21), v) DADVYVKIDNVPDNMAVYLHTNLLMFGTRKNSFIYNISKKFSAITGTYSEATKRTIFSQISH (SEQ ID NO: 22), w) GFMRFFQLLRLMADKPHETAIKEVVMAY (SEQ ID NO: 23), x) MNDYLGIFKNNDVRTLIGLILFVLALYSPPLISILMIFISSFLLPLTSLVIT (SEQ ID NO: 24), y) RLVETLPENMDFRSDHLTTFECFEEIITLAKKYIYIA (SEQ ID NO: 25), z) PKSFKSLKKVYTFFKFLADKKMTLFKSILFNL (SEQ ID NO: 26), aa) EIKTGISSFRILRRMATTFTFDSF (SEQ ID NO: 27), bb) VNGHFFVTNKVFDKSLLYKYENDIITVPFRLSYEPFVWGVNFRK (SEQ ID NO: 28),cc) FYDLPPFTQHLLNIRLTDTEYRARFIGG (SEQ ID NO: 29), dd) YIDYALRRTTNIPVEMMGTDVVRLKDYQHFVARVFLGLDSMHSLLLFHETGVGKTMTTVYILKHLKDIYTNWAIILLVKKALIEDPWMNTILRYAPEITKD (SEQ ID NO: 30), ee) SVNNLQMDKTSSLRL (SEQ ID NO: 31), ff) TFQQKISKYFNSRLFGHDIESFINR (SEQ ID NO: 32), gg) YIRNTLTEKIFVNAFTVDKVKQLLASNQVKFYFNKRLNQLTRI RQGKFI KN KI H LLPGDWVEVAVQEYTSIIFGRQPSLHR (SEQ ID NO: 33), hh) AFKSLIKIDSIPGLKTYNMKDITYEKSND (SEQ ID NO: 34), ii) ISTIQESFIRFTVTNKEGIKIRTKIPLSKVHGLDVKNVQLVDAID (SEQ ID NO: 35), jj) ISTIQESFIRFTVTNKEGVKIRTKIPLSKVHGLDVKNVQLVDAID (SEQ ID NO: 36), kk) ITNKLIEGTNIWYSNS (SEQ ID NO: 37),KIDKLRQIVAYFSEF (SEQ ID NO: 38), mm) KIHILIQEIVHEVMIVKKKE (SEQ ID NO: 39), nn) KKDYKLVFEIINQVKDE (SEQ ID NO: 40), oo) KLAI KLGFKSLVQYI KFI FLQMALLYI K (SEQ ID NO: 41),PP) KLSTQIEPQRNLTVQ (SEQ ID NO: 42), qq) KNNSNQLVWENFLAHM (SEQ ID NO: 43), rr) KQVPFM RTDM LQNM FAAN RDN VASRL (SEQ ID NO: 44), ss) AIYDSMQYTYKIIANSVYGLMGFRNSALYS (SEQ ID NO: 45),«) KQVPFM RTDM LQNM FAAN RDN VTSRL (SEQ ID NO: 46), uu) KQVPFMRTDVLQNMFAANRDNVASRL (SEQ ID NO: 47), vv) KYLVPNVIGGVFINK (SEQ ID NO: 48), ww) LALRFFLQRLYFLDHREIHYLFR (SEQ ID NO: 49),XX) LCIRISMVISLLSMITMSAFLIVRLNQ (SEQ ID NO: 50), yy) DDTPIDVSPEIMLNGIMYR (SEQ ID NO: 51), zz) AVLIMAIKVFTNSQIFNIISYIIL (SEQ ID NO: 52), aaa) AMGLNYKLTFLHTLYKRVLHMLLL (SEQ ID NO: 53), bbb) SFIISTLNKILSDENYLLKIIAVFDSKLISEKETLNEYKQLYTISSESLI (SEQ ID NO: 54), ccc) I ITPVVFYRSGTETKITFALKKLM I DREVVAN VIGLSGD (SEQ ID NO: 55), ddd) YIRNTLTEKIFVNAFTVNKVKQLLASNQVKFYFNKRLNQLTRIRQGKFIKNKIHLLPGDWVEVAVQEYT (SEQ ID NO: 56), SIIFGRQPSLHR and eee) SNINQPWIKTISKRMRVNIINHSIVTRGKSSILQTIEIIFTNRTC (SEQ ID NO: 57).

2. The vaccine composition according to claim 1 , wherein the polypeptide comprises at least one epitope sequence having the amino acid sequence of SEQ ID No: 1 or a variant having least 70% sequence identity thereto.

3. The vaccine composition according to claim 1 , wherein the polypeptide comprises at least one epitope sequence having the amino acid sequence of SEQID No: 56 or a variant having least 70% sequence identity thereto.

4. The vaccine composition according to claim 1 , wherein the polypeptide comprises at least one epitope sequence having the amino acid sequence of SEQ ID No: 1 and at least one epitope sequence having the amino acid sequence of SEQ ID No: 2; or a variant having at least 70% sequence identity thereto.

5. The vaccine composition according to claim 1 , wherein the polypeptide comprises at least one epitope sequence having the amino acid sequence of SEQ ID No: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16 and / or 17, or a variant having at least 70% sequence identity thereto; preferably wherein the polypeptide comprises epitope sequences having the amino acid sequences of each of SEQNos: 1 to 17, or variants having at least 70% sequence identity thereto.

6. The vaccine composition according to claim 1 , wherein the polypeptide comprises one or more epitopes having the amino acid sequence of each of: i. SEQ ID Nos: 1 to 3, or variants having at least 70% sequence identity thereto; ii. SEQ ID Nos: 1 to 4, or variants having at least 70% sequence identity thereto; iii. SEQ ID Nos: 1 to 5, or variants having at least 70% sequence identity thereto; iv. SEQ ID Nos: 1 to 6, or variants having at least 70% sequence identity thereto; v. SEQ ID Nos: 1 to 7, or variants having at least 70% sequence identity thereto; vi. SEQ ID Nos: 1 to 8, or variants having at least 70% sequence identity thereto; vii. SEQ ID Nos: 1 to 9, or variants having at least 70% sequence identity thereto; viii. SEQ ID Nos: 1 to 10, or variants having at least 70% sequence identity thereto; ix. SEQ ID Nos: 1 to 11 , or variants having at least 70% sequence identity thereto; x. SEQ ID Nos: 1 to 12, or variants having at least 70% sequence identity thereto; xi. SEQ ID Nos: 1 to 13, or variants having at least 70% sequence identity thereto;xii. SEQ ID Nos: 1 to 14, or variants having at least 70% sequence identity thereto; xiii. SEQ ID Nos: 1 to 15, or variants having at least 70% sequence identity thereto; xiv. SEQ ID Nos: 1 to 16, or variants having at least 70% sequence identity thereto; xv. SEQ ID Nos: 1 to 17, or variants having at least 70% sequence identity thereto; xvi. SEQ ID Nos: 1 to 18, or variants having at least 70% sequence identity thereto; xvii. SEQ ID Nos: 1 to 19, or variants having at least 70% sequence identity thereto; xviii. SEQ ID Nos: 1 to 20, or variants having at least 70% sequence identity thereto; xix. SEQ ID Nos: 1 to 21 , or variants having at least 70% sequence identity thereto; xx. SEQ ID Nos: 1 to 22, or variants having at least 70% sequence identity thereto; xxi. SEQ ID Nos: 1 to 23, or variants having at least 70% sequence identity thereto; xxii. SEQ ID Nos: 1 to 24, or variants having at least 70% sequence identity thereto; xxiii. SEQ ID Nos: 1 to 25, or variants having at least 70% sequence identity thereto;xxiv. SEQ ID Nos: 1 to 26, or variants having at least 70% sequence identity thereto; xxv. SEQ ID Nos: 1 to 27, or variants having at least 70% sequence identity thereto; xxvi. SEQ ID Nos: 1 to 28, or variants having at least 70% sequence identity thereto; xxvii. SEQ ID Nos: 1 to 29, or variants having at least 70% sequence identity thereto; xxviii. SEQ ID Nos: 1 to 30, or variants having at least 70% sequence identity thereto; xxix. SEQ ID Nos: 1 to 31 , or variants having at least 70% sequence identity thereto; xxx. SEQ ID Nos: 1 to 32, or variants having at least 70% sequence identity thereto; xxxi. SEQ ID Nos: 1 to 33, or variants having at least 70% sequence identity thereto; xxxii. SEQ ID Nos: 1 to 34, or variants having at least 70% sequence identity thereto; xxxiii. SEQ ID Nos: 1 to 35, or variants having at least 70% sequence identity thereto; xxxiv. SEQ ID Nos: 1 to 36, or variants having at least 70% sequence identity thereto; xxxv. SEQ ID Nos: 1 to 37, or variants having at least 70% sequence identity thereto;xxxvi. SEQ ID Nos: 1 to 38, or variants having at least 70% sequence identity thereto; xxxvii. SEQ ID Nos: 1 to 39, or variants having at least 70% sequence identity thereto; xxxviii. SEQ ID Nos: 1 to 40, or variants having at least 70% sequence identity thereto; xxxix. SEQ ID Nos: 1 to 41 , or variants having at least 70% sequence identity thereto; xl. SEQ ID Nos: 1 to 42, or variants having at least 70% sequence identity thereto; xli. SEQ ID Nos: 1 to 43, or variants having at least 70% sequence identity thereto; xlii. SEQ ID Nos: 1 to 44, or variants having at least 70% sequence identity thereto; xliii. SEQ ID Nos: 1 to 45, or variants having at least 70% sequence identity thereto; xliv. SEQ ID Nos: 1 to 46, or variants having at least 70% sequence identity thereto; xlv. SEQ ID Nos: 1 to 47, or variants having at least 70% sequence identity thereto; xlvi. SEQ ID Nos: 1 to 48, or variants having at least 70% sequence identity thereto; xlvii. SEQ ID Nos: 1 to 49, or variants having at least 70% sequence identity thereto; orxlviii. SEQ ID Nos: 1 to 50, or variants having at least 70% sequence identity thereto.

7. The vaccine composition according to claim 1 , wherein the polypeptide comprises one or more epitopes having the amino acid sequence of each of: i. SEQ ID Nos: 2 and 56, or variants having at least 70% sequence identity thereto; ii. SEQ ID Nos: 2, 3 and 56, or variants having at least 70% sequence identity thereto; iii. SEQ ID Nos: 2 to 4 and 56, or variants having at least 70% sequence identity thereto; iv. SEQ ID Nos: 2 to 4, 7 and 56, or variants having at least 70% sequence identity thereto; v. SEQ ID Nos: 2 to 5, 7 and 56 or variants having at least 70% sequence identity thereto; vi. SEQ ID Nos: 2 to 7 and 56, or variants having at least 70% sequence identity thereto; vii. SEQ ID Nos: 2 to 7, 56 and 57, or variants having at least 70% sequence identity thereto; viii. SEQ ID Nos: 2 to 7, 9, 56 and 57, or variants having at least 70% sequence identity thereto; ix. SEQ ID Nos: 2 to 7, 9, 11 , 56 and 57, or variants having at least 70% sequence identity thereto; x. SEQ ID Nos: 2 to 7, 9, 11 , 13, 56 and 57, or variants having at least 70% sequence identity thereto;xi. SEQ ID Nos: 2 to 9, 11 , 13, 56 and 57, or variants having at least 70% sequence identity thereto; xii. SEQ ID Nos: 2 to 9, 11 to 13, 56 and 57, or variants having at least 70% sequence identity thereto; xiii. SEQ ID Nos: 1 to 9, 11 to 13, 56 and 57, or variants having at least 70% sequence identity thereto; xiv. SEQ ID Nos: 1 to 9, 11 to 14, 56 and 57, or variants having at least 70% sequence identity thereto; xv. SEQ ID Nos: 1 to 9, 11 to 15, 56 and 57, or variants having at least 70% sequence identity thereto; xvi. SEQ ID Nos: 1 to 9, 11 to 15, 17, 56 and 57, or variants having at least 70% sequence identity thereto; xvii. SEQ ID Nos: 1 to 9, 11 to 15, 17, and 55 to 57, or variants having at least 70% sequence identity thereto; xviii. SEQ ID Nos: 1 to 9, 11 to 15, 17, 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xix. SEQ ID Nos: 1 to 9, 11 to 15, 17, 19, 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xx. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 20, and 55 to 57, or variants having at least 70% sequence identity thereto; xxi. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 20, 24, and 55 to 57, or variants having at least 70% sequence identity thereto; xxii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 21 , 24, and 55 to 57, or variants having at least 70% sequence identity thereto;80 xxiii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 21 , 24, 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxiv. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 22, 24, 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxv. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 25, and 55 to 57, or variants having at least 70% sequence identity thereto; xxvi. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 25, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxvii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 26, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxviii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 27, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxix. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 55 to 57, or variants having at least 70% sequence identity thereto; xxx. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 54 to 57, or variants having at least 70% sequence identity thereto; xxxi. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , and 53 to 57, or variants having at least 70% sequence identity thereto; xxxii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 28, 31 , 32, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxiii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxiv. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35, and 53 to 57, or variants having at least 70% sequence identity thereto;81 xxxv. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35, 36, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxvi. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 37, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxvii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 38, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxviii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 39, and 53 to 57, or variants having at least 70% sequence identity thereto; xxxix. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 40, and 53 to 57, or variants having at least 70% sequence identity thereto; xl. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 41 , and 53 to 57, or variants having at least 70% sequence identity thereto; xli. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 42, and 53 to 57, or variants having at least 70% sequence identity thereto; xlii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 43, and 53 to 57, or variants having at least 70% sequence identity thereto; xliii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 44, and 53 to 57, or variants having at least 70% sequence identity thereto; xliv. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 45, and 53 to 57, or variants having at least 70% sequence identity thereto; xlv. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 46, 53 to 57, or variants having at least 70% sequence identity thereto; xlvi. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 47, and 53 to 57, or variants having at least 70% sequence identity thereto;82 xlvii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 48, and 53 to 57, or variants having at least 70% sequence identity thereto; xlviii. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 49, and 53 to 57, or variants having at least 70% sequence identity thereto; or xlix. SEQ ID Nos: 1 to 9, 11 to 15, 17 to 29, 31 , 32, 35 to 50, and 53 to 57, or variants having at least 70% sequence identity thereto.

8. The vaccine composition according to any preceding claim, wherein the polypeptide comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of the epitope sequences, or the variants thereof.

9. The vaccine composition according to any preceding claim, wherein the polypeptide comprises a concatemer of one or more of the epitope sequences, or the variants thereof.

10. The vaccine composition according to any preceding claim, wherein the polypeptide comprises one or more spacer regions; preferably wherein when the polypeptide comprises more than one of the epitope sequences or the variants thereof, one or more spacer regions are positioned between one or more or each epitope sequence.11 . The vaccine composition according to any preceding claim, wherein the one or more epitope variant sequences have at least 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity to their respective sequence selected from SEQ ID Nos: 1 to 57.

12. The vaccine composition according to any preceding claim, wherein the polypeptide is no more than 2500, 2400, 2300, 2200, 2100, 2000, 1900, 1800, 1700, 1600, 1500, 1400, 1300, 1200, 1100, 1000, 900, 800, 700, 600, 500, 400, 300, 200, 150, 140, 130, 120, 110, 100, 90, 80, 70, 60, 50, 40, 30, 20, 15, 10 or 9 amino acids in length.

13. The vaccine composition according to any preceding claim, wherein the polynucleotide is DNA or RNA, preferably wherein the vaccine composition is a mRNA vaccine composition.8314. A vector comprising a polynucleotide as defined in any preceding claim, preferably wherein the polynucleotide further comprises regulatory elements capable of driving transcription and / or translation of the polynucleotide in a host cell.

15. The vector according to claim 14, wherein the vector is a viral vector.

16. The vaccine composition according to any of claims 1 to 13 or the vector according to claim 14 or 15, formulated in a nanoparticle or lipid formulation.

17. A microorganism comprising a polypeptide as defined in any of claims 1 to 12, or a polynucleotide as defined in any of claims 1 to 13, preferably wherein said microorganism is a bacterial microorganism.

18. A pharmaceutical composition comprising the vaccine composition according to any of claims 1 -13, the vector according to any of claims 14-16 or the microorganism according to claim 17, and a pharmaceutically acceptable carrier, diluent, excipient and / or adjuvant.

19. The pharmaceutical composition according to claim 18, wherein said pharmaceutical composition is formulated for parental, oral, sublingual, nasal, naso-oral, or pulmonary administration.

20. The pharmaceutical composition according to claim 19, wherein said parental administration is subcutaneous, intradermal, intramuscular, subdermal, intraperitoneal, or intravenous administration.

21. The vaccine composition according to any of claims 1-13, vector according to any of claims 14-16, microorganism according to claim 17 or pharmaceutical composition according to any of claims 18-20, for use in treating or preventing a disease or condition in a subject, preferably wherein the disease or condition is an infection with an orthopoxvirus.

22. The vaccine composition, vector, microorganism or pharmaceutical composition for use according to claim 21 , wherein the vaccine composition, vector, microorganism or pharmaceutical composition is for use in the treatment84 or prophylaxis of an infection caused by akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, macacapox virus, monkeypox virus, mousepox virus, racoonpox virus, skunkpox virus, taterapox virus, vaccinia virus, variola virus, or volepox virus.

23. The vaccine composition, vector, microorganism or pharmaceutical composition for use according to claim 22, wherein the infection is caused by akhmeta virus, alaskapox virus, camelpox virus, cowpox virus, monkeypox virus, variola virus or vaccinia virus.

24. Use of a vaccine composition according to any of claims 1-13, vector according to any of claims 14-16, microorganism according to claim 17 or pharmaceutical composition according to any of claims 18-20 in the manufacture of a medicament for the prevention or treatment of an infection with an orthopoxvirus in a subject in need thereof.

25. A method of treating or preventing an infection with an orthopoxvirus, the method comprising administering an effective amount of a vaccine composition according to any of claims 1-13, vector according to any of claims 14-16, microorganism according to claim 17 or pharmaceutical composition according to any of claims 18-20 to a subject in need thereof.

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