MX2008005193A - Use of vaccines for the treatment/ prevention of the transmission of influenza pathogens between species - Google Patents

Abstract

The present invention relates to a method for prevention of the transmission of a pathogen of an animal of a first species to an animal of a second species characterized in that antigen of the pathogen of the animal of the first species is used for the immunization of the animal of the second species against the pathogen of the animal of the first species, wherein the administration of said antigen results in the reduction or the absence of the reproduction of pathogen of the animal of the first species in the animal of the second species.

Description

USE OF VACCINES FOR THE TREATMENT / PREVENTION OF THE TRANSMISSION OF PATHOGENS FIELD OF THE INVENTION The present invention relates to the field of medicine, preferably to the field of infectious diseases. In particular, the present invention relates to the vaccination of animals of a first species in order to prevent intra-species transmission ("horizontal transmission") as well as inter-species transmissions ("vertical transmission") of pathogens. More particularly, the present invention relates to vaccines against influenza and its use for the treatment and prevention of influenza infections, additionally for the prevention of intra- and inter-species transmission of influenza. BACKGROUND OF THE INVENTION Influenza infections continue to be an important infection in animals and in humans. Influenza is caused by viruses that go through changes / modifications continuous antigens and that have an animal reservoir. Therefore, in the future new epidemics and pandemics may appear, and the eradication of the disease will be difficult to achieve. Influenza viruses are well known in the art and are described in more detail by P. Palese, Nature Medicine, vol. REF..190868 10, no. 12, pp. S 82 to S 86 of December 2004, with other references. Briefly, the genome of influenza A virus consists of eight segments of a single strand, and the viral particles have two major glycoproteins on their surface: hemagglutinin (H) and neuraminidase (N). With at least 15 different hemagglutinin subtypes (Hl to H15) and 9 different neuraminidase subtypes (Ni to N9), there is considerable antigenic variation among influenza viruses. It has been demonstrated that the influenza virus of the type of Avian Plague H5N1 virus infects both pigs and man. Viruses can also be transmitted directly from avian species to humans (Claas et al., Lancet 1998, 351: 472; Suarez et al. , J. Virol. 1998, 72: 6678; Subbarao et al. , Science 1998, 279: 393; Shortridge, Vaccine 1999, 1 7 (Suppl 1): S26-S29). Mortality in known human clinical cases approaches approximately 50%.

During the last century, pigs have been an important vector for influenza pandemics. Pigs, camels and seals, preferably pigs, can serve as a "mixing chamber" for avian influenza viruses, and, therefore, represent a potential risk factor for overcoming the difficulties of the species. poultry, the natural reservoir of influenza viruses, to mammals. This normally occurs through double infections of susceptible animals, eg. , pig, with both, a mammalian flu virus (porcine), as well as an established avian flu virus. This double infection can create new recombinant viruses that can be the cause of human or swine pandemics. However, recent evidence would indicate that a recombination of current avian H5 strains with influenza viruses in mammals would not result in highly virulent recombinants. On the other hand, the avian flu virus can affect pigs and by spontaneous mutations can adapt to pigs. The critical obstacle would be overcome as soon as the virus can cause horizontal infections within a porcine (or other mammal) population. However, a majority of the pigs in Southeast Asia have been infected with strains of the bird flu virus (H5) that originate from neighboring poultry hatchlings. As these infections to date have been subclinical, they can only be diagnosed by laboratory methods and, therefore, are often overlooked. There is a high risk that these subclinically infected pigs serve as an opportunity for the virus to adapt to the mammalian system, spread within the swine population, and also infect humans.

As the obstacle in species between pigs and humans is expected to be low, the risk of horizontal infections of the "swine" influenza viruses in humans is dramatically increased. The currently available vaccines against influenza A infection are preparations of killed virus vaccines, which contain variants of the influenza Hl, H2 and H3 subtype. The use of these vaccines is limited to the vaccination of human beings in order to prevent transmission from man to man. Therefore, those vaccines, including the current vaccination strategy to prevent human-to-human transmission, are not preventive with respect to the transmission and adaptation of non-mammalian influenza viruses to mammals. Those vaccines, including the current vaccination strategy, do not sufficiently consider the fact that non-mammalian influenza viruses, for example, avian flu viruses are capable of infecting non-human mammals, such as pigs, camels, seals, etc. ., and of recombining with mammalian influenza virus in those non-human mammals. Therefore, there is a need to increase the availability of new higher vaccines and new vaccination methods to provide better methods to control influenza infections and to have a positive impact on the burden of the disease.

BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows the idea of the invention set forth herein. Immunization of animals of a second species (eg, in swine) against a pathogen (eg, influenza H5N1) of an animal of a first species (poultry) prevents, or at least reduces, transmission of the pathogen of the first species to another animal of the same species (eg, from pig to pig) or to a third species (eg, from pig to human). Figure 2 shows the amino acid sequences of two H5 polypeptides.

DETAILED DESCRIPTION OF THE INVENTION It has surprisingly been discovered that pigs can be effectively vaccinated against avian influenza virus (See Example 2, RESULTS). By vaccinating the pigs with a suitable vaccine comprising an antigen of the avian influenza virus, preferably Hl, H3, H5, H7 and / or H9, more preferably H5 and / or H7, the risk of generating strains of influenza variants adapted to pigs can be reduced or completely eliminated. As a result, the adaptation of an avian influenza virus in humans can be reduced or completely eliminated due to the elimination of the virus within a natural mammalian reservoir of influenza virus. Accordingly, one aspect of the present invention pertains to the generation of a vaccine, preferably a recombinant vaccine, based on hemagglutinin from avian influenza of subtypes 1, 3, 5, 7 and / or 9 (ie, H1, H3). , H5, H7 and / or H9) for the vaccination of pigs in order to prevent or reduce the reproduction of avian influenza viruses in pigs, when they are infected with an avian influenza virus comprising the same antigen or an antigen , which shows cross-reactivity with the antigen (s) used for vaccination. The term "reproduction", as used in the present invention, includes, but is not limited to, the replication of the viral genome and / or the assembly of particles. The term "reduce" or "reduction of reproduction" as used in this invention means that the rate of replication in vaccinated animals is statistically significantly lower than in unvaccinated animals. In other words, the titres of the avian flu virus in pigs challenged with an infectious influenza virus are in the arithmetic average lower than in pigs that have not been vaccinated before the challenge. By the term "in the lower arithmetic average" is meant a reduction of more than 20%, preferably of more than 40%, still preferably more than 50%, even more preferably more than 80%, even more preferably more than 100%. With respect to the latter, the term "deleted" means that no replication of the virus is detected in vaccinated pigs challenged with the infectious influenza virus after day 10, preferably after day 9, more preferably after day 8, even more preferably after day 7, still more preferably after day 6, still more preferably after day 5, even more preferably after day 4, even more preferably after day 6, even more preferably after day 3, even more preferably after day 2, most preferably after day 1 of the challenge. According to a more general aspect, the present invention relates to the use of an antigenic composition comprising an antigen of a pathogen of an animal of a first species for the preparation of a pharmaceutical composition for the immunization of an animal of a second species. , wherein said pharmaceutical composition, when administered to the animal of the second species, results in a reduction or absence of the reproduction of an infectious pathogen to an animal of the first species, in an animal of the second species, when said animal of the second species is infected with said infectious pathogen of the animal of the first species, preferably provided that the antigen of the antigenic composition of the pharmaceutical composition is present in or within the infectious pathogen. The first species and the second species can be of the same species or of different species. Preferably, the first and second species are different, even more preferably the first species is a poultry, for example, a bird, chicken, duck, turkey etc., and the second species is a mammal, preferably a pig, cattle , horse, seals, camels, dog, cat, hamster, mouse, human, more preferably a pig. Therefore, according to another aspect, the present invention relates to the use of an antigenic composition comprising an antigen of a poultry pathogen for the preparation of a pharmaceutical composition for immunization of a mammal, wherein said pharmaceutical composition , when administered to the mammal results in a reduction or absence of reproduction of the infectious pathogen for poultry in a mammal, when said mammal is infected with said infectious poultry pathogen, as long as the antigen of the composition antigenic of the pharmaceutical composition is present in the infectious pathogen. The term "pharmaceutical composition" as described in this invention, includes, but is not limited to, vaccines for the reduction or prevention of an infection or a composition of matter for the treatment and decrease of an infection. The term "pathogen", as used includes but is not limited to a microorganism or any pathogenic part thereof, which normally causes diseases or conditions to its host, or at least disrupts the normal physiology of its host. For example, a poultry pathogen is the bird flu virus. Accordingly, according to a further aspect of the present invention, the pathogen according to the invention being infectious for poultry is an influenza virus, preferably the avian influenza virus. According to a further aspect of the present invention, the pathogen being infectious for poultry is an avian influenza virus of subtype Hl, H3, H5, H7 or H9, more preferably an avian influenza virus of subtype H5. or H7, most preferably an avian influenza virus of subtype H5N1. The term "antigenic composition" as used in the present invention means a composition of matter comprising one or more antigens. The term "antigen" as used in the present invention means but not limited to peptides, polypeptides, glycopeptides, or polysaccharides which are capable of interacting specifically with an antigen recognition molecule of the immune system, such as an immunoglobulin (antibody) or T cell antigen receptor in order to produce, activate or stimulate an immune response directed to said antigen in a host to which said antigen is administered. The term "antigen" also refers to nucleic acid molecules, preferably DNA or RNA molecules, each of which codes for and expresses a peptide, polypeptide or glycopeptide that is capable of specifically interacting with an antigen recognition molecule of the immune system, such as an immunoglobulin (antibody) or T cell antigen receptor in order to produce, activate or stimulate an immune response against the antigen that is encoded by the nucleic acid molecule. The antigen used for the preparation of the pharmaceutical composition which is used according to the invention is a microorganism or an antigenic part and / or preparation of said microorganism. With respect to the latter, the term "immunization", as used in this invention, means but is not limited to any cause or increase in an immune response. The term "immune response" as used means, but is not limited to an immune response mediated by antibodies and / or cellular to a composition or vaccine of interest. Generally, an "immune response" includes, but is not limited to, one or more of the following effects: the production or activation of antibodies, B cells, helper T cells, suppressor T cells and / or cytotoxic T cells and / or T cells and d, specifically directed to an antigen or antigens included in the composition or vaccine of interest. Preferably, the host will exhibit either a therapeutic or protective immune response so that resistance to a new infection will be increased and / or the clinical severity of the disease will be reduced. That type of protection will be demonstrated by a reduction or lack of symptoms associated with host infections as described above. The term "antigenic part and / or its preparation" as used in the present invention means that at least one molecule of said part and / or preparation is antigenic or possesses antigenic properties. The antigenic part and / or preparation of a microorganism includes, but is not limited to, peptides, polypeptides, glycopeptides, and / or polysaccharides that include any fragment thereof, which possesses antigenic property. A molecule is "antigenic" or possesses "antigenic properties" when it is capable of interacting specifically with an antigen recognition molecule of the immune system, such as an immunoglobulin (antibody) or T cell antigen receptor. A polypeptide antigen for example it contains an epitope of at least about five, and preferably at least about 10 amino acids. An antigenic portion of a polypeptide, also referred to herein as an "epitope", can be that portion of the polypeptide that is immunodominant for the antibody or recognition of the T cell receptor, or it can be a portion of a polypeptide used to generate an antibody against the molecule by conjugating the antigenic portion to a transporter polypeptide for immunization. A molecule that is antigenic does not necessarily have to be immunogenic by itself, that is, capable of producing an immune response without a transporter. In other words, an antigenic portion also includes but is not limited to a hapten which must be conjugated to a carrier in order to become immunogenic. As mentioned above, the antigen can be a microorganism or an antigenic part and / or its preparation. Preferably, said microorganism is a virus, or an antigenic part and / or its preparation, and most preferably a flu virus., or an antigenic part and / or its preparation. According to a further aspect of the invention, the antigen used for the preparation of the pharmaceutical composition is an avian influenza virus, or an antigenic part and / or its preparation. According to a further aspect of the invention, the antigen used for the preparation of the pharmaceutical composition is haemagglutinin (H) and / or neuraminidase (N) of the influenza virus. Preferably, said antigen is H1, H3, H5, H7, and / or H9 of the avian influenza virus. In this context, the term "and / or" means, that a single antigen or any combination of the antigens mentioned above, can be used for the preparation of the pharmaceutical composition. In contrast to Hl or H3 strains, currently present in the porcine population, H5 and probably H7 seem to be more conserved. Therefore, the probability of cross protection seems to be more favorable. As a result, H5 and / or H7, most preferably H5 of the avian influenza virus, are used as an antigen for the preparation of the pharmaceutical composition. The term "hemagglutinin 5 (H5)" or "H5 of the avian influenza virus" as used in the present invention means but is not limited to any naturally occurring H5 and any modified form of H5, including any deletion mutant, substitution and / or insertion of H5. Additionally it means any antigenic part of H5, which means any peptide fragment which shows antigenic properties in a standard hemagglutinin inhibition assay. Typically, said antigenic part thereof comprises 35, 30, 25, 20, 18, 15, 13, 10, 9, or more preferably 8 contiguous amino acids of the amino acid sequence encoding H5, modified or unmodified, which shows antigenic properties in a standard hemagglutinin inhibition assay. A standard hemagglutinin inhibition assay for example is described in Stephenson et al., Virus Research vol. 103, pp. 91-95 (2004) with other references or is described in the Examples. In the case of questionable results, the Hl assay as described in Example 2 will be understood as the relevant reference assay in connection with all aspects of the invention as described in this invention. Preferred H5 antigens that can be used according to the invention include any modified H5 antigen that shows superior antigenic properties compared to an unmodified H5, where the antigenic property is measured in a standard hemagglutinin inhibition assay, for example as described in Example 2. Briefly, the Hl assay was carried out to detect the presence of specific HA antibodies. A heterologous H5N1 virus, A / chicken / Mexico / 232/94, was used at a concentration of four hemagglutination units "hemagglutinating units" [4 HA units] in the Hl assay. In U-bottom microtiter plates, double serial serum dilutions in PBS were subsequently mixed with equal volumes (25 μl) containing 4 HA units of virus, and incubated at 37 eC for one hour. Chicken red blood cells, at a concentration of 0.5% in PBS, were added to the wells containing serum virus and incubated for 40 min. at room temperature. The Hl titers were determined as reciprocal of the highest serum dilutions in which the inhibition of haemagglutination was observed. Of note, Haesebrouck and Pensaert (1986) discovered "that there may be a correlation between the Hl titles against the challenge virus and the protection of the challenge". Haesebrouck and Pensaert (1986) also determined that pigs with Hl titers of >40 were "completely resistant to the challenge and there was no replication of the virus in the respiratory tract in challenge". Accordingly, the development of Hl > 40 in the vaccinated pig correlates with protection. (F. Haesebrouck and MB Pensaert, 1986. Effect of intratracheal challenge of fattening pigs previously immunized with an inactivated influenza H1N1 vaccine (Veterinary My crobiology, 11 (1986) 239- -249 239) 9. Titles should be assumed of H5 Hl equivalents or at least almost equivalent would also result in complete immune protection of pigs against the avian influenza virus Lower titers, at least result in a seroconversion of the vaccinated animals and result in partial immune protection of those animals, which can also drastically reduce the risk of a pandemic.Most preferably, the H5 antigens used according to the invention include modified H5 antigens, which show an increase in antigenic property compared to the antigen of H5 encoded by the sequence of SEQ ID NO: 2, where the antigenic property is measured in a hemagglutinin inhibition assay is standard, for example as described in Lüschow et al. , Vaccine 19 (2001), pp. 4249-4259 with other references. The term "higher or increased antigenic property" as used in this invention refers to an inhibition of haemagglutinin which is at least 20%, preferably at least 50%, more preferably at least 75%, even more preferably at least 100% higher to that of the reference H5 antigen, ie, the unmodified H5 antigen or the H5 antigen having the sequence of SEQ ID NO: 2. Additionally, preferred H5 antigens that can be used according to the invention include H5 antigens. which comprise a peptide comprising i. the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3; SEC ID NO:; SEQ ID NO: 5 or SEQ ID NO: 6 O; OR. any peptide having at least 85% sequence homology, more preferably at least about 90% sequence homology, even more preferably at least about 95% sequence homology, even more preferably at least about 97% sequence homology , even more preferably at least about 98% sequence homology, and even more preferably at least about 99% sequence homology with the polypeptide of i) comprising the inhibition of hemagglutinin in a standard hemagglutinin inhibition as described above; or iii. any antigenic part of the polypeptides of i) or ii) comprising at least 35, 30, 25, 20, 18, 15, 13, 10, 9, or most preferably 8 contiguous amino acids of any of the peptides of i) or ii). iv. any of the peptides of i), ii) or iii) having the amino acids 223N, 36T / 223N, 36K / 223N, 83A / 223N, 83T / 223N, 83D / 223N, 86A / 223N, 86V / 223N, 120N / 223N , 120S / 223N, 155N / 223N, 155S / 223N, 156A / 223N, 156T / 223N, 189R / 223N, 189K / 223N, 212K / 223N, 212R / 223N, 212E / 223N, 223N / 263A, 223N / 263T, or 120N / 155N / 223N. v. any peptide of i), ii), iii) or iv) having the amino acid 223N and one or more of the following groupings of amino acids selected from the group consisting of: a. aa 93-95: GNF b. aa 123-125: SDH c. aa 128 - 130: SSG d. aa 138 - 140: GSS e. aa 226 - 228: MDF f. aa 270 - 272: EVE g. aa 309-311: NKL; or vi. any peptide of i), ii) iii) or iv) having the amino acid 223N and one or more of the following groupings of amino acids selected from the group consisting of: a. aa 93-95: GNF b. aa 128 - 130: SSG c. aa 138 - 140: GSS.

Other preferred H5 antigens that can be used according to the invention include H5 antigens which comprise the peptides provided in Table 1, or any immunogenic part thereof: TABLE 1 H5 antigens # the amino acid positions that appear in TABLE 1 refers to the positions as defined by way of example in SEQ ID NO: 1. In other words, amino acid 223 of TABLE 1 refers to amino acid 223 of the sequence SEQ. ID NO: l. - means that the amino acids in these positions are variable compared to the reference sequence. Other preferred H5 antigens that can be used according to the invention include H5 antigens which comprise i. a peptide having the sequences Accession Number to NCBI AAT65209, CAJ32556, ABC47656, CAF21874, CAF21870, AAC58998, AAC58997, AAC58996, AAC58994, AAC58993, AAC58992, AAC58991, AAC58990, AAC58995, AAS45134, AAN17270, AAN17269, AAN17268, AAN17267, AAN17266, AAN17265, AAN17264, AAN17263, AAN17262, AAN17261, AAN17260, AAN17259, AAN17257, AAN17256, AAN17255, AAN17254, AAA43083, AAA43082, AAB19079, BAE48696, BAE48693, BAE48696, BAE48695, BAE48694, BAE48692, BAE48691, BAE48690, BAE48689, BAE48688, BAE48687, BAE48686, BAE48685, BAE48684, BAE48683, AAC58999, ABC72082, AAV91149, AAP71993, AAP71992, AAP71991, AAP71990, AAP71989, AAP72011, AAP72010, AAP72009, AAP72008, AAP72007, AAP72006, AAP72005, AAP72004, AAP72003, AAP72002, AAP72001, AAP72000, AAP71999, AAP71998, AAP71997, AAP71996, AAP71995, AAP71994, AAF99718, ABF58847, AAG38534, AAC32102, AAC32099, AAL75847, AAC32101, AAC32098, AAC32088, AAC32078, AAR99628, AAC32100, AAM49555, AAL75843, AAL75839, AAD13573, AAD13568, AAF04720, AAF04 719, AAC34263, AAR16155, AAD13574, AAD13570, AAD13575, AAD13572, AAD13569, AAD13567, AAD13566, AAK57506, AAG01225, AAG01215, AAG01205, AAG01195, or ABD83813, or any peptide having at least 85% sequence homology, more preferably at less about 90% sequence homology, even more preferably at least about 95% sequence homology, even more preferably at least about 97% sequence homology, still more preferably at least about 98% sequence homology, and even more preferably at least about 99% sequence homology to the polypeptide of i) and showing inhibition of hemagglutinin in a standard hemagglutinin inhibition as described above; iii. any of the peptides of i) or ii) having amino acids 223N, 36T / 223N, 36K / 223N, 83A / 223N, 83T / 223N, 83D / 223N, 86A / 223N, 86V / 223N, 120N / 223N, 120S / 223N, 155N / 223N, 155S / 223N, 156A / 223N, 156T / 223N, 189R / 223N, 189K / 223N, 212K / 223N, 212R / 223N, 212E / 223N, 223N / 263A, 223N / 263T, or 120N / 155N / 223N; or iv. any antigenic part of the peptides of i), ii) comprising at least 35, 30, 25, 20, 18, 15, 13, 10, 9, or most preferably 8 contiguous amino acids of any such peptides of i), ii) or iii); or v. any such antigenic parts of iv), wherein those antigenic portions comprise amino acid 223 of H5; or vi. any such antigenic parts of v) wherein those antigenic portions comprise amino acid 223N; or vii. any such peptides of i), ii), iii), iv), v) or vi) having the amino acid 223N and one or more of the following amino acid groupings selected from the group consisting of: h. aa 93-95: GNF i. aa 123 - 125SDH j. aa 128 - 130: SSG k. aa 138 - 140: GSS 1. aa 226 - 228: MDF m. aa 270 - 272: EVE n. aa 309 - 311: NKL; or vüi. any peptide of i), ii) iii) or iv) having the amino acid 223N and one or more of the following amino acid groupings selected from the group consisting of: d. aa 93-95: GNF e. aa 128 - 130: SSG f. aa 138-140: GSS "Sequence Homology", as used in the present invention, refers to a method for determining the ratio of two sequences. To determine the sequence homology, two or more sequences are optimally aligned, and spaces are introduced if necessary. In contrast to sequence identity, conservative amino acid substitutions are counted as a match when sequence homology is determined. In other words, to obtain a polypeptide or polynucleotide having 95% sequence homology with a reference sequence, 85%, preferably 90%, even more preferably 95% of the amino acid or nucleotide residues in the reference sequence must match or comprising a conservative substitution with another amino acid or nucleotide, or an amount of amino acids or nucleotides up to 15%, preferably up to 10%, even more preferably up to 5% of the amino acid residues or total nucleotides, not including conservative substitutions, in the sequence of reference can be inserted in the reference sequence. Preferably, the homologous sequence comprises at least one stretch of 50, even more preferably 100, even more preferably 250, even more preferably 500 nucleotides. After such alignment, the sequence homology is determined on the basis of position by position, for example, the sequences are "homologous" at a particular position if at that position, the nucleotides or amino acid residues are identical. The total number of such position identities is then divided by the total number of nucleotides or amino acid residues in the reference sequence to give the% sequence homology. Sequence homology can be readily calculated by known methods, including, but not limited to, those described in Computational Molecular Biology, Lesk, A. N., ed. , Oxford University Press, New York (1988), Biocomputing: Informatics and Genome Projects, Smith, D.W. , ed. , Academic Press, New York (1993); Computer Analysis of Sequence Data, Part I, Griffin, A.M., and Griffin, H.G., eds., Humana Press, New Jersey (1994); Sequence Analysis in Molecular Biology, von Heinge, G., Academic Press (1987); Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York (1991); and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48: 1073 (1988), the disclosures of which are incorporated herein by reference. Preferred methods for determining sequence homology are designed to give the greatest match between the sequences tested. The methods for determining the sequence homology are encoded in computer programs available to the public which determine the identity of sequences between determined sequences. Examples of such programs include, but are not limited to, the GCG software package (Devereux, J., et al., Nucleic Acids Research, 12 (1): 387 (1984)), BLASTP, BLASTN and FASTA (Altschul , SF et al., J. Molec. Biol., 215: 403-410 (1990) The BLASTX program is available to the public of the NCBI firm and other sources (BLAST Manual, Altschul, S. et al., NCVI NLM NIH Bethesda, MD 20894, Altschul, SF et al., J. Molec. Biol., 215: 403-410 (1990), the disclosures of which are incorporated herein by reference.) These programs optimally align sequences using space weights. of error (default gap weights) in order to produce the highest level of sequence homology between the given sequence and the reference sequence Other preferred H5 antigens that can be used according to the invention are i. mentioned above with the amino acid 223N and the modification 328K +; ii. any of those mentioned above with amino acid 94N / 223N and modification 328K +; iii. any H5 antigen of avian origin having the amino acid 223N, and the 328K + modification, where avian origin means that the H5 sequence was derived from a viral isolate that was originally isolated from a poultry bird infected with the avian influenza virus type 5; or iv. any H5 antigen of avian origin that has the amino acids 94N / 223N and the 328K + modification, where avian origin means that the H5 sequence was derived from a viral isolate that was originally isolated from a poultry bird infected with the bird flu virus kind ; or. V. any H5 antigen of avian origin having the amino acids 155N / 223N and the 328K + modification, where avian origin means that the H5 sequence was derived from a viral isolate that was originally isolated from a poultry infected with the virus. bird flu type 5; or vi. any H5 antigen of avian origin having the amino acid 120N / 155N / 223N and the 328K + modification, where avian origin means that the H5 sequence was derived from a viral isolate that was originally isolated from a poultry infected with the H5N1 virus. bird flu type 5; or vii. any H5 antigen that has the 94N / 223N modifications and the 328K + modification; or viii. any H5 antigen having the modifications 94N / 155N / 223N and the modification 328K +; or; X. any H5 antigen having the modifications 94N / 120N / 155N / 223N and the modification 328K +; or X. any H5 having modifications 223N / modification 328K +, and one or more of the following groupings of amino acids selected from the group consisting of: a. aa 93-95: GNF b. aa 123-125: SDH c. aa 128 - 130: SSG d. aa 138 - 140: GSS e. aa 226 - 228: MDF f. aa 270 - 272: EVE g. aa 309-311: NKL; or xi. any H5 antigen having the amino acid 223N, and the 328K + modification, and one or more of the following groupings of amino acids selected from the group consisting of: a. aa 93-95: GNF b. aa 128 - 130: SSG c. aa 138-140: GSS; or xii. any H5 antigen having the amino acid sequence of SEQ ID NO: 4. Other preferred H5 antigens that can be used according to the invention include H5 antigens as described by Hoffmann et al, PNAS, vol. 106, no. 36, pp. 12915-12920 of September 6, 2005. The description of this reference will be included in its entirety as a reference. The numbering of the amino acid positions of the H5 protein as used in this invention refers to the position of amino acids as provided by way of example in SEQ ID NO: 1. SEQ ID NO: 1 represents the amino sequence of the hemagglutinin of duck strain / China / E319-2 / 03 although it lacks signal peptide. In other words, if reference is made to the amino acid at position 223 (amino acid 223), it is meant the amino acid residue corresponding to amino acid 223 of SEQ ID NO: 1. In the present case, amino acid 223 would be Serine (S). The terms "223N", or "155N" mean by way of example, that at the amino acid positions 223 and 155, respectively - the numbering according to the amino acid positions of SEQ ID NO: 1 -, will encode the amino acid Asparagine ( N) In other words, if reference is made to "H5 antigen having amino acid 223N", an amino acid molecule of H5 that normally encodes Serine at amino acid position 223 - numbering according to the amino acid positions of SEQ ID NO. : 1 - that amino acid will be replaced by an Asparagine (N). The term "328K +" or "328K + modification" means that at the amino acid position 328 of the H5 antigen - numbering according to the amino acid positions of SEQ ID NO: 1-, a second Lysine (K +) is inserted. In cases where the amino acid sequences at positions 328 and 329 naturally encode Lysine-Lysine, no other Lysine (K) will be inserted. Nevertheless, most of the known H5 sequences encode amino acid positions 328 and 329 for Lysine-Arginine. In either of those cases, the term 328K + modification means, that a second Lysine (K) will be inserted between Lysine at position 328 and Arginine at position 329. The modified sequence would then be read Lysine-Lysine-Arginine (KKR). It is also within the meaning of the present invention, that any other antigen, particularly any antigen H and N, can be used in a modified or unmodified version. As mentioned with respect to the H5 antigen, the most preferred is the modified antigens, which show a high antigenic property in a standard hemagglutinin inhibition assay compared to the unmodified antigens. The methods of how to introduce any of the aforementioned modifications within the nucleotide sequence of an influenza virus are well known in the art. The genomic sequence of the whole influenza virus can be modified according to the invention, for example, according to the methods described in U.S. Patent No. 6,951,754, with other references. Additionally, molecular biology, microbiology, and conventional recombinant DNA techniques can be employed within the skill in the art to modify a nucleic acid sequence encoding an antigen as described in this invention. Such techniques are explained in their entirety in the technical literature. ease, e.g. , Sambrook et al. , Molecular Cloning: A Laboratory Manual, Second Edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; DNA Cloning: A Practical Approach, Volumes I and II (D. N. Glover ed., 1985); Oligonucleotide Synthesis (M. J. Gai t ed, 1984); Nucleic Acid Hybridization [B. D. Hames & S. J. Higgins eds. (1985)]; Transcription And Translation [B. D. Hames & S. J. Higgins, eds. (1984)]; Animal Cell Culture [R. I. Freshney, ed. (1986)]; Immobilized Cells And Enzymes [IRL Press, (1986)]; B. Perbal, A Practical Guide to Molecular Cloning (1984); F.

M. Ausubel et al. (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, Inc. 1994). The immune response against a hemagglutinin antigen, eg. , H5 in a vaccinated animal can be measured by conventional methods well known in the art, for example as described in Example 2. Briefly, the serum titers of antibodies, mainly those determined by the inhibition of hemagglutinin (Hl) and / or assays of viral neutralization, are accepted substitute measures of immune protection. Conventional Hl assays and viral neutralization can be carried out in Madin Darby canine kidney cells as described in Palmer et al., 1975, Advanced Laboratory Techniques for Influenza Diagnosis, U.S. Department of Health, Education and Welfare (Washington, DC, and / or Kida et al., 1982, Virology, vol. 122, pp. 38-47. The reference assay used with respect to the current invention is described in Lüschow et al., Vaccine 19 (2001), pp. 4249-4259 with other references. As mentioned above, in the case of questionable results, the Hl assay as described in Example 2 will be understood as the relevant reference test in connection with all aspects of the invention as described in this invention. According to a further general aspect, the present invention relates to the use of an antigenic composition comprising an antigen of a pathogen of an animal of a first species for the preparation of a pharmaceutical composition for the immunization of an animal of a second species. , characterized in that the administration of a pharmaceutical composition, comprising an antigen of a pathogen of an animal of a first species, to an animal of the second species results in a reduction or absence of the replication of a pathogen, which is infectious for an animal of the first species, in an animal of the second species, when said animal of the second species is infected with the infectious pathogen of the animal of the first species, provided that the antigen of the antigenic composition of the pharmaceutical composition is present in or within the infectious pathogen. The term "replication" as used in this invention means inter alia, the duplication / multiplication of the pathogen's genome. The meaning of the term reduction of replication is described above. According to a further general aspect, the present invention relates to the use of an antigenic composition comprising an antigen of a pathogen of an animal of a first species for the preparation of a pharmaceutical composition for the immunization of an animal of a second species. , characterized in that the administration of the pharmaceutical composition, comprising an antigen of a pathogen of an animal of a first species, to the animal of the second species results in the prevention of the adaptation of the infectious pathogen of the first animal to the second animal, when the animal of the second species is infected with said infectious pathogen of the animal of the first species. The term "adaptation" as used in this invention means, among other things, that the pathogen is adapted to replication in the new species, which means that the pathogen has overcome the obstacles of the species. It is considered that the obstacle of species is overcome, if the pathogen is spread vertically, which means from animal to animal of the same species (also called: inter-species transmission or vertical transmission). According to a further aspect, the invention relates to the use of hemagglutinin Hl, H3, H5, H7 and / or H9 of the avian influenza virus for the preparation of a pharmaceutical composition for administration to a pig., wherein said pharmaceutical composition, when administered to a pig, results in a reduction or absence of the reproduction of an infectious avian influenza virus of subtype Hl, H3, H5, H7 or H9 in a pig, when it is infected with said infectious avian influenza virus of subtype Hl, H3, H5, H7 or H9. More preferred is the use of the H5 and / or H7 antigen. According to a further aspect, the invention relates to the use of hemagglutinin Hl, H3, H5, H7 and / or H9 of the avian influenza virus for the preparation of a pharmaceutical composition, when administered to a pig, given as This resulted in the prevention of the adaptation of an infectious avian influenza virus of subtype 5 from the swine to the swine, when it is infected with this infectious avian influenza virus of subtype Hl, H3, H5, H7 or H9. More preferred is the use of the H5 and / or H7 antigen. According to a further aspect, the use of hemagglutinin H5 and / or H7 of the avian influenza virus for the preparation of said pharmaceutical composition is even more preferred. According to a further aspect, the present invention relates to the use of a pharmaceutical composition comprising an antigen of the avian influenza virus of subtypes Hl, H3, H5, H7 and / or H9 for the preparation of a vaccine for the treatment and / or prevention of an animal against bird flu. Preferably, the antigen to be used is Hl, H3, H5, H7, and / or H9 of the avian influenza virus. The most preferred antigen to be used is H5, and / or H7 of the avian influenza virus. According to a further aspect of this embodiment, the animal to be treated is poultry, for example, birds, ducks or geese. According to another aspect of the invention, the animals to be treated are mammals, preferably pigs, cattle, horses, seals, camels, dogs, cats, hamsters, mice or humans. According to a further aspect, the present invention relates to a method for the prevention of the transmission of a pathogen of an animal of a first species to an animal of a second species, characterized in that antigen of a pathogen of an animal is used. of the first species for the immunization of an animal of the second species against said pathogen of the animal of the first species, where the administration of said antigen results in the reduction or absence of the reproduction of said pathogen of the animal of the first species in an animal of the second species. According to a further embodiment of the method described above, the first species is poultry. According to a further aspect of said method, the animal of the second species is a mammal, preferably a pig, cattle, horse, seal, camel, dog, cat, hamster, mouse or human being, more preferred is the pig. The antigens according to this method are those described supra. Preferably, said antigen is a microorganism or an antigenic part and / or preparation of said microorganism.

More preferably, said microorganism is of viral origin, even more preferably said microorganism is the avian influenza virus. The pathogens of the animal of the first species are preferably those mentioned above. Briefly, such a pathogen is the avian flu virus. According to a further aspect, said pathogen is the avian influenza virus of subtype Hl, H3, H5, H7 or H9. More preferably, said pathogen is the avian influenza virus of subtype H5N1. According to a further aspect, the present invention relates to a method for the prevention of the transmission of a pathogen of an animal of a first species to an animal of a second species, characterized in that the antigen of a pathogen of an animal of a first species is used for the immunization of an animal of a second species against the pathogen of the animal of the first species, where the administration of said antigen results in the reduction or absence of pathogen reproduction of the animal of the first species in an animal of the second species, where the antigen is hemagglutinin (H) and / or neuraminidase (N) of the influenza virus, preferably of avian origin. According to a further aspect of said method, the antigen is Hl, H3, H5, H7, and / or H9 of the avian influenza virus, while H5 and / or H7 are more preferred. A more detailed description of those antigens is found above. According to a further important aspect, the present invention relates to a method for the prevention of the transmission of a pathogen of an animal of a first species to a human, characterized in that an antigen of a pathogen of an animal of a first species it is used for the immunization of an animal that is not a human of a second species, which is not against the pathogen of the animal of the first species, where the administration of the antigen results in the reduction or absence of reproduction of the pathogen of the animal of the first species in said animal of the second species. Preferably, the first species is poultry, eg, bird, chicken, duck, etc., and the animal of the second species is a mammal that is not a human being, preferably a pig, cattle, horse, seal, camel, dog, cat, hamster, mouse, most preferably a pig. The pathogens of the animal of the first species are preferably those mentioned above. Briefly, such a pathogen is the avian flu virus. According to a further aspect, said pathogen is the avian influenza virus of subtype Hl, H3, H5, H7 or H9. More preferably, said pathogen is the avian influenza virus of subtype H5N1.

According to a further aspect, the present invention relates to a method for preventing the transmission of a pathogen from an animal of a first species to a human, characterized in that antigen from a pathogen of an animal of a first species is used. for the immunization of an animal of the second species against the pathogen of the animal of the first species, where the administration of said antigen results in the reduction or absence of reproduction of the pathogen of the animal of the first species in an animal of the same species. second species, where the antigen is hemagglutinin (H) and / or neuraminidase (N) of the influenza virus, preferably of avian origin. According to a further aspect of said method, the antigen is Hl, H3, H5, H7, and / or H9 of the avian influenza virus, while H5 and / or H7 are most preferred. A more detailed description of those antigens is found above. A further aspect of the present invention relates to a method for the prevention or reduction of recombination between pathogens of an animal of a first species and an animal of a second species in an animal, wherein a pharmaceutical composition comprising antigen of the pathogen of an animal of the first species is used for the immunization of animals of the second species against the pathogen of the animal of the first species. The recombination event is often responsible for overcoming the obstacles of species. Reducing or preventing these recombination events would reduce the risk of overcoming species barriers. The obstacle of species is considered overcome, if the pathogen spreads vertically, which means from animal to animal of the same species (also called: inter-species transmission or vertical transmission). Preferably, the pathogens are of the family or genus. More preferably, those pathogens are viruses, preferably influenza viruses. According to a further embodiment of this method, the first pathogen is the avian influenza virus and the second pathogen is a mammalian influenza virus. Additionally, if the first pathogen is the avian influenza virus and the second pathogen is a mammalian influenza virus, a preferred pharmaceutical composition comprises the avian influenza virus antigen H1, H3, H5, H7, and / or H9. , even more preferably H5 and / or H7. According to a further aspect, the invention relates to a method for the reduction or prevention of recombination between a pathogen of an animal of a first species and a pathogen of an animal of a second species in an animal, wherein a pharmaceutical composition which comprises antigen of the pathogen of an animal of the first species is used for the immunization of an animal of the second species against the pathogen of the animal of the first species, and where the administration of said antigen results in the reduction or absence of the reproduction of the pathogen of the animal of the first species in an animal of the second species. Preferably, those pathogens are of the family or gender. More preferably, those pathogens are viruses, preferably influenza viruses. Also, if the first pathogen is the avian influenza virus and the second pathogen is a mammalian influenza virus, a preferred pharmaceutical composition comprises avian influenza virus antigen Hl, H3, H5, H7, and / or H9, even more preferably H5 and / or H7. The antigen, composition or antigenic compositions as used in accordance with the invention, can be prepared in recombinant form, for example, by the use of the baculovirus expression system in insect cells. Other examples of well-established recombinant expression systems are bacterial expression systems such as E. coli or B. subtilis, yeast-based expression systems such as S. cerevisiae or S. pombe, or mammalian cell expression systems such as BHK-, CHO- and / or NSO-based expression systems. Such systems are well known in the art and are generally available, for example, commercially through Clontech Laboratories, Inc. 4030 Fabian Way, Palo Alto, California 94303-4607, USA. Other expression strategies are, for example, described in Lüschow et al. , Vaccine no. 19 (2001), pp. 4249-4259, or Vei t et al. , PNAS vol. 103 (2006), pp. 8197-8202. Additionally, recombinant adeno-associated virus systems are well established and for example are described in U.S. Patent No. 5,436,146 or WO200203872 with other references. Likewise, vaccinia virus expression systems (pustular rash), for example as described in U.S. Patent No. 6,265,183 together with other references, are also well established and are suitable for producing antigen or antigens. recombinants, composition or antigenic compositions as used according to the invention. Additional suitable expression systems make use of recombinant popova virus, such as SV40, fowlpox virus, pseudorabies virus and retroviruses. According to a further preferred aspect of the present invention, the antigen (s) or at least one antigen is / are produced in insect cells using the baculovirus expression system. More preferably, the recombinant baculovirus encodes the antigen and produces the antigen. Therefore, according to another aspect of the invention, the Hl, H3, H5, H7 and / or H9 antigens of the avian influenza virus are encoded and produced by the recombinant baculovirus.

However, the relevant antigen (s), composition or antigenic compositions can also be inactivated viruses comprising antigen, an apatogenic version of a live virus comprising an antigen, preparation and / or fragments of a virus, wherein said preparation and / or fragment comprises antigen or antigens. Another important aspect of the present invention is the preparation of pharmaceutical compositions such as vaccines, which comprises at least one antigen of a pathogen as described above. The skilled person knows the additional components which may be comprised in said compositions / vaccines together with antigen (see, for example, Remington's Pharmaceuti cal Sci ences, (1990), 18th ed Mack Publ., Easton). The skilled person can use sterile, physiologically acceptable, known injectable solutions. To prepare a ready-to-use solution, aqueous isotonic solutions, such as, e.g. , saline solution or corresponding plasma protein solutions are readily available. The pharmaceutical composition / vaccine can be present as lyophilizates or dry preparations, which can be reconstituted with a known injectable solution directly before use under sterile conditions, e.g. as a kit of parts. Additionally, the pharmaceutical compositions / vaccines of the present invention may include one or more carriers acceptable in veterinary. As used in this invention, "an acceptable veterinary carrier" includes, but is not limited to, any and all solvents, dispersion media, coatings, adjuvants, stabilizing agents, diluents, preservatives, antibacterial and antifungal agents, isotonic agents, adsorption retardation agents and the like. The diluents may include water, saline, dextrose, ethanol, glycerol and the like. Isotonic agents may include sodium chloride, dextrose, mannitol, sorbitol, and lactose, among others. The stabilizers include albumin and alkaline salts of ethylenediaminetetraacetic acid, among others. "Adjuvants" as used in this invention may include aluminum hydroxide and aluminum phosphate, saponins, e.g. , Quil A, QS-21 (Cambridge Biotech Inc., Cambridge MA), GPI-0100 (Galenica Pharmaceuticals, Inc., Birmingham, AL), water-in-oil emulsion, oil-in-water emulsion, water-in-oil-in-water emulsion. The emulsion may be based in particular on light liquid paraffin oil (European Pharmacopoeia type); isoprenoid oil such as squalane or squalene; oil resulting from the theoligomerization of alkenes, in particular of isobutene or decene; esters of acids or alcohols containing a linear alkyl group, more particularly vegetable oils, ethyl oleate, di- (caprylate / caprate) of propylene glycol, tri- (caprylate / caprate) glyceryl or propylene glycol dioleate; esters of branched fatty acids or alcohols, in particular esostearic acid esters. The oil is used in combination with emulsifiers to form the emulsion. The emulsifiers are preferably nonionic surfactants, in particular sorbitan, mannide (eg, anhydromanitol oleate), glycol, polyglycerol, propylene glycol and oleic, isostearic, ricinoleic or hydroxystearic esters, which are optionally ethoxylated , and polyoxypropylene-polyoxyethylene copolymer blocks, in particular the Pluronic products, especially L121. See Hunter et al., The Theory and Practical Application of Adjuvants (Ed. Stewart-Tull, D. E.S.). JohnWiley and Sons, NY, pp51-94 (1995) t Todd et al., Vaccine 15: 564-570 (1997). Examples of suitable oil-in-water emulsions are emulsigen-based adjuvants, such as EMULSIGEN®, EMULSIGEN-D®, EMULSIGEN-P®, EMULSIGEN-75® (MVP Laboratories, Inc. Omaha, NE, USA). It has been surprisingly discovered that the pharmaceutical compositions / vaccines comprising the H5 antigen, preferably the recombinant H5 antigen, were effectively applied in an adjuvant of oil-in-water emulsions, preferably emulsigen-based adjuvants, more preferably EMULSIGEN® and EMULSIGEN-P ®.

Additionally, it is possible to use the SPT emulsion described on page 147 of "Vaccine Design, The Subunit and Adjuvant Approach" edited by M. Powell and M. Newman, Plenum Press, 1995, and the MF59 emulsion described on page 183 of this same book. A further instance of an adjuvant is a compound selected from the polymers of acrylic or methacrylic acid and the copolymers of maleic anhydride and alkenyl derivative. Advantageous adjuvant compounds are polymers of acrylic or methacrylic acid which are crosslinked, especially with polyalkenyl ethers of sugars or polyalcohols. These compounds are known by the term carbomer (Phameuropa Vol. 8, No. 2, June 1996). Those skilled in the art may also refer to U.S. Patent No. 2,909,462 which discloses that type of acrylic polymers crosslinked with a polyhydroxy compound having at least 3 hydroxyl groups, preferably not more than 8, carbon atoms. Hydrogen of at least three hydroxyls are replaced with unsaturated aliphatic radicals having at least 2 carbon atoms. Preferred radicals are those containing between 2 and 4 carbon atoms, eg. , vinyl, allyl and other ethylenically unsaturated groups. The unsaturated radicals themselves may contain other substituents, such as methyl. The products sold under the name Carbopol; (BF Goodrich, Ohio, E.U.A.) are particularly appropriate. They are crosslinked with an allyl sucrose or with allyl pentaerythritol. Among these, the Carbopol 974P, 934P and 971P can be mentioned. More preferred is the use of Cabopol 971P. Among the copolymers of maleic anhydride and alkenyl derivative, EMA copolymers (Monsanto) which are copolymers of maleic anhydride and ethylene. The dissolution of these polymers in water leads to an acid solution which will be neutralized, preferably at physiological pH, in order to give the adjuvant solution in which the immunogenic, immunological or vaccine composition itself will be incorporated. Other suitable adjuvants include, but are not limited to, the adjuvant system RIBI (Ribi Inc.), block copolymer (CytRx, Atlanta GA), SAF-M (Chiron, Emeryville CA), monophosphoryl lipid A, lipid-amine adjuvant of Avridine, heat-labile enterotoxin of E. coli (recombinant or otherwise), cholera toxin, or muramyl dipeptide, among many others. Preferably, the adjuvant is added in an amount of about 100 μg to about 10 mg per dose. Even more preferably, the adjuvant is added in an amount of about 500 μg to about 5 mg per dose. Even more preferably, the adjuvant is added in an amount of about 750 μg to about 2.5 mg per dose. Most preferably, the adjuvant is added in an amount of about 1 mg per dose. The pharmaceutical compositions / vaccines may also include one or more other immunomodulatory agents such as, for example, interleukins, interferons or other cytokines. The pharmaceutical compositions / vaccines may also include Gentamicin and Mertiolate. While the amounts and concentrations of adjuvants and additives useful in the context of the present invention can be readily determined by the skilled artisan, the present invention contemplates compositions comprising between about 50 ug and about 2000 ug of adjuvant and preferably about 250 ug / 1 ml. of dose of the vaccine composition. In another preferred embodiment, the present invention contemplates vaccine compositions comprising between about lug / ml and about 60 ug / ml of antibiotics, and more preferably less than about 30 ug / ml of antibiotics. Administration strategies for influenza vaccines are well known in the art. Mucosal vaccination strategies are considered for inactivated and attenuated virus vaccines. Although the mucosa can be reached through the local administration of a vaccine, several strategies have been used to administer immunogenic proteins to the mucosa.

In a specific embodiment, the vaccine can be administered in a mixture with, or as a conjugate or chimeric fusion protein with, cholera toxin, such as cholera toxin B or a cholera toxin A / B chimera (Hajishengallis, J Immunol., 154: 4322-32, 1995; Jobling and Holmes, Infect Immun., 60: 4915-24, 1992). Mucosal vaccines have been described based on the use of the cholera toxin B subunit (Lebens and Holmgren, Dev Biol Stand 82: 215-27, 1994). In another embodiment, a mixture with heat-labile enterotoxin (LT) can be prepared for mucosal vaccination. Other mucosal immunization strategies include encapsulating the virus in microcapsules (U.S. Patent No. 5,075,109, U.S. Patent No. 5,820,883, and U.S. Patent No. 5,853,763) and use of a membranous immunopotentiation carrier (WO 98/0558). The immunogenicity of orally administered immunogens can be increased using red blood cells (rbc) or red blood cell ghosts rbc (U.S. Patent No. 5,643,577), or by the use of blue tongue antigen (Patent of the United States No. 5,690,938).

EXAMPLES The following examples show preferred materials and processes according to the present invention. However, it should be understood that these examples are provided by way of illustration only, and nothing of them should be considered a limitation to the general scope of the invention.

EXAMPLE 1 Construction of a recombinant baculovirus encoding and expressing HA H5 antigens The recombinant baculovirus containing the H5 HA antigen was generated in the following manner: the coding sequences of H5 HA (SEQ ID NO: 2) were chemically synthesized and subcloned in the PVL1392 transfer vector (BD Biosciences Pharmingen, San Diego, CA). H5 HA MutK + (SEQ ID NO: 4) was generated using oligonucleotide primers and the Mutagenesis Kit Directed to the QuikChange® Site (Stratagene, La Jolla, CA) and subcloned into the transfer vector pVLl392 (BD Biosciences Pharmingen, San Diego , CA). Plasmids pVLl392 containing the genes encoding H5 HA antigen (SEQ ID NO: 2) and H5 HA MutK + (SEQ ID NO: 4) were then co-transfected with DiamondBac® baculovirus DNA (Sigma) into Sf9 insect cells (BD Biosciences Pharmingen) to generate the recombinant baculovirus containing the H5 HA genes encoding SEQ ID NO: 2 and H5 HA mutK + coding SEQ ID NO: 4. Recombinant baculoviruses containing the genes encoding H5 HA (SEQ ID NO. : 2) and H5 HA MutK + (SEQ ID NO: 4) were purified on plates and the Virus Seeds (Master Seed Viruses) (MSVs) were propagated in the SF9 cell line, aliquoted and stored at -70 ° C. . Insect cells infected with baculovirus H5 HA as described above to generate MSV or working virus viruses (Working Seed Viruses) express antigen H5 HA (SEQ ID NO: 2) and antigen H5 HA MutK + (SEQ ID NO: 4) ) as detected by polyclonal serum or monoclonal antibodies in an indirect fluorescent antibody or Western blot assay. After sowing with the appropriate amounts of recombinant baculoviruses (H5 HA and H5 HA MutK +, respectively), then stirring flasks containing SF + cells (Protein Sciences, Inc., Meriden, CT) were incubated at 27 ± 2 ° C for 7 days. days and with agitation lOOrpm during that time. The bottles used vented lids to allow air flow. The crude whole cell culture containing SF + cells infected with baculovirus and the cell culture supernatants of each culture were harvested.

EXAMPLE 2 Preparation of pharmaceutical compositions (vaccines) comprising HA H5 antigens H5 HA protein from crude whole cells and H5 HA Mutk + protein expressed in insect cells were harvested by the baculovirus-based expression system. The baculoviruses were inactivated in the presence of 5 mM cycled binary ethylenenin (BEI) (final concentration) between approximately 32 and 39 ° C for 72 to 96 hours. After the inactivation was completed, a sodium thiosulfate solution was added., 3 M at a final concentration of 5 mM to neutralize any residual EIB. After neutralization, various adjuvants were added and the following pharmaceutical / vaccine compositions were generated.G.

VACCINES EXAMPLE 3 Vaccination of pigs (swine) against avian influenza 1. INTRODUCTION The purpose of this study was to determine the capacity of experimental vaccines containing a crude extract of recombinant H5 hemagglutinin (HA) antigen to induce titers of inhibition of the hemagglutination (Hl) in pigs. Several adjuvants were evaluated with H5 HA antigens. The HA H5 prototypes evaluated in this study contained either conventional H5 HA antigen or H5 HA MutK +. H5 HA conventional was derived from A / duck / China / E319-2 / 03, while H5 HA MutK + consisted of conventional H5 HA which was constructed to contain three specific amino acid changes in S120N, D150N, S223N and 328mutK +. It also contains the amino acid 94N. Changes of particular amino acids in H5 HA Mut K + result in an H5 HA that more closely resembles the HA of A / HK / 213/03. The amino acid composition of H5 HA of A / HK / 213/03 is currently thought to aid in the recognition of antibodies to H5 HA.

STUDY DESIGN: Table 1. Review of the Study, The small pigs had 3 weeks ± 5 days at the beginning of the study. The small pigs were clinically healthy at the beginning of the study. Blood samples were obtained in Study Days 0, 21, and 35. All animals in the study were observed daily in Study Days 1 through 35 with respect to general health status. For seven days after each vaccination, injection sites were investigated daily and visible reactions were recorded. At the conclusion of the animal phase of the study on Study Day 35, all animals were humanely euthanized. 3. VACCINES Vaccines 501 to 514 as described in EXAMPLE 2 were used for the vaccination study of pigs. 4. HEMAGLUTININ INHIBITION TEST The pigs were vaccinated with the prototypes containing H5 HA on Days 0 and 21. Sera were collected from the pigs for evaluation by hemagglutination inhibition (Hl) assay on Days 0, 21, 35. The Hl assay was performed to detect the presence of HA-speci fi c antibodies. A heterologous H5N1 virus, A / chicken / Mexico / 232/94, was used at a concentration of four haemagglutination units [4 HA units] in the Hl assay. In U-bottom microtiter plates, serial duplicate serum dilutions in PBS were subsequently mixed with equal volumes (25 μl) containing 4 HA units of virus, and incubated at 37 ° C for one hour. Red chicken blood cells, at a concentration of 0.5% in PBS, were added to the wells containing virus in serum and incubated for 40 min. at room temperature. The Hl titers were determined as reciprocal of the highest serum dilutions in which haemagglutination inhibition was observed.

. RESULTS The Hl assay used the official H5N1 antigen of the Mexican government (A / chicken / Mexico / 232/94) [4 HA units] Vaccination regimen of 1 x 1 mL on Days 0 and 21.

BIV H5 (derived from Influenza A virus (A / duck / China / E319-2 / 03 (H5N1)) BIV H5 K + (BIV H5 mutated to include S120N, D155N, S223N, and 328K + added) The results show that the Most vaccine compositions activate an immune response in vaccinated pigs In particular, most vaccine compositions result in seroconversion, which means that most vaccinated pigs developed specific antibodies against the influenza virus The test results clearly prove that the idea of the claimed invention works very well: the risk of pandemic infection of pigs (animal of a second species), with the influenza virus avian influenza (pathogen of a first species) can be significantly reduced by vaccination of pigs with a relevant antigen of the avian influenza virus, this has been clearly demonstrated., through this concept of vaccination, the transmission and adaptation of the avian flu virus to mammals, including humans, is significantly reduced. Pigs are one of the most important reservoirs for avian pathogens, including the avian flu virus. If viral replication in pigs and, therefore, the risk of adaptation of avian influenza to pigs is significantly reduced and controlled, the risk of any adaptation of the avian influenza virus to humans is also significantly reduced. In the case, in which the administration of the antigen results in a lower Hl titer, which means a titer of less than 30, additional boosts with antigen will be necessary to further improve the Hl titer and to increase the immune protection in the vaccinated pigs Therefore, low titer does not mean that no protection can be achieved, it just teaches that additional reinforcements seem to be necessary to improve the immune response. The fact that an immune response could be measured in vaccinated pigs demonstrates that the inventive idea on which the present invention is based works very well. In other words, the experiments provided in this invention clearly and undoubtedly provide evidence that the inventive idea of the present invention works. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (76)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. The use of an antigenic composition comprising antigen of a pathogen of an animal of a first species for the preparation of a pharmaceutical composition for the immunization of an animal of a second species, wherein said pharmaceutical composition, when administered to the animal of the second species. species results in a reduction or absence of reproduction of the infectious pathogen to an animal of the first species, in said animal of the second species, when said animal of the second species is infected with the infectious pathogen of the animal of the first species .

2. The use according to claim 1, wherein the antigen of the antigenic composition of the pharmaceutical composition is present in or within the infectious pathogen.

3. The use according to claim 1 or 2, wherein the first species and the second species are different.

4. The use according to claim 3, wherein the first species is a poultry and the second species is a mammal.

5. The use according to claim 4, wherein the mammal is a pig, cattle, horse, seal, camel, dog, cat, hamster, mouse, human.

6. The use according to claim 5, wherein the mammal is a pig.

The use according to any of claims 1 to 6, wherein the antigen is a microorganism or an antigenic part of said microorganism.

8. The use according to claim 7, wherein the microorganism is a virus.

9. The use according to claim 8, wherein the virus is the influenza virus.

10. The use according to claim 9, wherein the influenza virus is the avian influenza virus.

The use according to any of claims 1 to 10, wherein the infectious pathogen for the animal of the first species is the avian influenza virus.

12. The use according to claim 11, wherein the infectious pathogen for the animal of the first species is the avian influenza virus of subtype Hl, H3, H5, H7 or H9.

13. The use according to claim 11, wherein the infectious pathogen for the animal of the first species is the avian influenza virus of subtype H5N1.

14. The use according to claim 11, wherein the antigen is hemagglutinin (H) and / or neuraminidase (N) of influenza virus.

15. The use according to claim 14, wherein the antigen is Hl, H3, H5, H7, and / or H9 of the avian influenza virus.

16. The use according to claim 14, wherein the antigen is H5, and / or H7, of the avian influenza virus.

17. The use according to any of claims 1 to 16, wherein the antigenic composition comprising the antigen of a pathogen of an animal of a first species comprises a recombinant baculovirus encoding an antigen.

18. The use according to claim 17, wherein the recombinant baculovirus produces the antigen.

19. The use according to claim 17, wherein the recombinant baculovirus expresses the antigen H1, H3, H5, H7 and / or H9.

The use according to any of claims 1 to 17, wherein the administration of a pharmaceutical composition, comprising antigen from a pathogen of an animal of a first species to an animal of the second species results in a reduction or absence of the replication of an infectious pathogen to an animal of the first species, in an animal of the second species, when said animal of the second species is infected with the infectious pathogen of the animal of the first species.

The use according to claim 20, wherein the antigen of the antigenic composition of the pharmaceutical composition is present within or in the infectious pathogen.

22. The use according to any of claims 1 to 18, wherein the administration of the pharmaceutical composition, comprising the antigen of a pathogen of an animal of a first species, to the animal of the second species results in the prevention of the adaptation of the infectious pathogen of the first animal to the second animal, when the animal of the second species is infected with said infectious pathogen of the animal of the first species.

23. The use of hemagglutinin H5 and / or H7 of the avian influenza virus for the preparation of a pharmaceutical composition for administration to a pig, wherein said pharmaceutical composition, when administered to a pig, results in a reduction or absence of the reproduction of an infectious avian influenza virus of subtype Hl, H3, H5, H7 or H9 in a pig, when it is infected with said infectious avian influenza virus of subtype Hl, H3, H5, H7 or H9.

24. The use according to claim 22, wherein the administration of the pharmaceutical composition, comprising hemagglutinin Hl, H3, H5, H7 and / or H9 of the avian influenza virus to a pig results in the prevention of the adaptation of an infectious avian influenza virus of subtype H5 from the bird to the pig, when said pig will be infected with said infectious avian influenza virus of subtype Hl, H3, H5, H7 or H9.

25. The use according to claim 22, wherein the administration of the pharmaceutical composition, comprising the hemagglutinin H5, H7 and / or H9 of the avian influenza virus, to a pig results in the prevention of the adaptation of an infectious avian influenza virus of subtype H5 from the bird to the pig, when said pig will be infected with said infectious avian influenza virus of subtype H5.

26. The use of an antigenic composition comprising an antigen of the avian influenza virus of subtype Hl, H3, H5, H7 and / or H9 for the preparation of a vaccine for the treatment and / or prevention of an animal against influenza avian.

27. The use according to claim 26, wherein the antigenic composition comprises antigen of the avian influenza virus of subtype H5 and / or H7.

28. The use according to claim 27, wherein the antigen is Hl, H3, H5, H7, and / or H9 of the avian influenza virus.

29. The use according to claim 28, wherein the antigen is H5, and / or H7 of the avian influenza virus.

30. The use according to any of claims 26 to 29, wherein the antigenic composition comprises an antigen of the avian influenza virus, comprises a recombinant baculovirus encoding the antigen.

31. The use according to claim 30, wherein the recombinant baculovirus produces the antigen.

32. The use according to claim 31, wherein the recombinant baculovirus expresses the antigen.

33. Use according to any of claims 26 to 32, wherein said animal is a poultry.

34. The use according to any of claims 26 to 32, wherein the animal is a mammal.

35. The use according to claim 34, wherein the mammal is a pig, cattle, horse, seal, camel, dog, cat, hamster, mouse or human being.

36. A method for the prevention of the transmission of a pathogen of an animal of a first species to an animal of a second species, characterized in that the antigen of a pathogen of an animal of a first species is used for the immunization of an animal of a second species against the pathogen of the animal of the first species, where the administration of said antigen results in the reduction or absence of reproduction of the pathogen of the animal of the first species in an animal of the second species.

37. The method according to claim 36, characterized in that the first species is a poultry.

38. The method according to claim 36 or 37, characterized in that the second species is a mammal.

39. The method according to claim 36, characterized in that the mammal is a pig, cattle, horse, seals, camels, dog, cat, hamster, mouse or human being.

40. The method according to claim 39, characterized in that the mammal is a pig.

41. The method according to any of claims 36 to 40, characterized in that the antigen is a microorganism or an antigenic part of said microorganism.

42. The method according to claim 41 characterized in that the microorganism is a virus.

43. The method according to claim 42, characterized in that the virus is the influenza virus.

44. The method according to claim 43, characterized in that the influenza virus is the avian influenza virus.

45. The method according to any of claims 36 to 44, characterized in that the pathogen is the avian influenza virus.

46. The method according to claim 45, characterized in that the pathogen is the avian influenza virus of subtype Hl, H3, H5, H7 or H9.

47. The method according to claim 46, characterized in that the pathogen is the avian influenza virus of subtype H5N1.

48. The method according to claim 43, characterized in that the antigen is hemagglutinin (H) and / or neuraminidase (N) of influenza virus.

49. The method according to claim 48, characterized in that the antigen is Hl, H3, H5, H7, and / or H9 of the avian influenza virus.

50. The method according to claim 49, characterized in that the antigen is H5, and / or H7, of the avian influenza virus.

51. The method according to any of claims 36 to 50, characterized in that the antigen is a recombinant antigen.

52. The method according to claim 51, characterized in that the recombinant antigen is produced by a recombinant baculovirus.

53. The method according to claim 52, characterized in that the recombinant baculovirus expresses antigen Hl, H3, H5, H7 and / or H9.

54. A method for the prevention of the transmission of a pathogen of an animal of a first species to a human, characterized in that antigen of a pathogen of an animal of a first species is used for the immunization of an animal of a second species that it is not a human being, where the administration of the antigen results in the reduction or absence of reproduction of the pathogen of the animal of the first species in said animal of the second species, and reduces / prevents further transmission of the pathogen to a human.

55. The method according to claim 54, characterized in that the first species is a poultry.

56. The method according to claim 54 or 55, characterized in that the second species is a mammal that is not a human being.

57. The method according to claim 56, characterized in that the mammal is a pig, cattle, horse, seal, camel, dog, cat, hamster or mouse.

58. The method according to claim 57, characterized in that the mammal is a pig.

59. The method according to any of claims 54 to 58, characterized in that the antigen is a microorganism or an antigenic part of said microorganism.

60. The method according to claim 59, characterized in that the microorganism is a virus.

61. The method according to claim 60, characterized in that the virus is the influenza virus.

62. The method according to claim 61, characterized in that the influenza virus is the avian influenza virus.

63. The method according to any of claims 54 to 62, characterized in that the pathogen is the avian influenza virus.

64. The method according to claim 63, characterized in that the pathogen is the avian influenza virus of subtype Hl, H3, H5, H7 or H9.

65. The method according to claim 63, characterized in that the pathogen is the avian influenza virus of subtype H5N1.

66. The method according to claim 63, characterized in that the antigen is hemagglutinin (H) and / or neuraminidase (N) of influenza virus.

67. The method according to claim 66, characterized in that the antigen is Hl, H3, H5, H7, and / or H9 of the avian influenza virus.

68. The method according to claim 67, characterized in that the antigen is H5, and / or H7, of the avian influenza virus.

69. The method according to claim 49, characterized in that the antigen is H5, and / or H7 of the avian influenza virus.

70. The method according to any of claims 54 to 69, characterized in that the antigen is a recombinant antigen.

71. The method according to claim 70, characterized in that the recombinant antigen is produced by a recombinant baculovirus.

72. The method according to claim 71, characterized in that the recombinant baculovirus expresses antigen Hl, H3, H5, H7 and / or H9

73. A method characterized in that it is for the prevention or reduction of recombination between a pathogen of an animal of a t species and a pathogen of an animal of a second species in an animal, wherein a pharmaceutical composition comprising antigen of the pathogen of a animal of the t species is used for the immunization of an animal of the second species against the pathogen of the animal of the t species.

74. The method according to claim 73, characterized in that the administration of said antigen results in the reduction or absence of reproduction of the pathogen of the animal of the t species in an animal of the second species

75. The method of compliance with claim 73 or 74, characterized in that the t pathogen is the avian influenza virus and the second pathogen is a mammalian influenza virus.

76. The method according to claim 75, characterized in that the pharmaceutical composition comprises antigen of the avian influenza virus Hl, H3, H5, H7, and / or H9.