OUTER MEMBRANE VESICLE (OMV) VACCINE COMPRISING N. MENINGITIDIS SEROGROUP B OUTER MEMBRANE PROTEINS

Abstract: A composition comprising (a) <i>Neisseria meningitidis </i> serogroup B outer membrane vesicles (OMVs), and (b) an immunogenic component selected from other <i>Neisseria</i> proteins, or immunogenic fragments thereof. Component (b) preferably includes a protein from a different NmB strain from that from which the OMV of component (a) is derived. The OMVs are preferably obtained by deoxycholate extraction. Optionally, the composition may also comprise a protective antigen against other pathogens.

Description: OUTER MEMBRANE VESICLE (OMV) VACCINE COMPRISING N. MENINGITIDIS SEROGROUP B OUTER MEMBRANE PROTEINS All documents cited herein are hereby incorporated by reference in their entirety. TECHNICAL FIELD This invention relates to vaccines against Neisseria meningitidis, serogroup B (NmB). BACKGROUND ART Neisseria meningitidis is a non-motile, Gram-negative diplococcus human pathogen. It colonises the pharynx, causing meningitis and, occasionally, septicaemia in the absence of meningitis. In the United States the attack rate is 0.6-1 per 100,000 persons per year, and it can be much greater during outbreaks (see Lieberman et al. (1996) JAMA 275(19): 1499-1503; Schuchat et al (1997) N Engl J Med 337(14):970-976). In developing countries, endemic disease rates are much higher and during epidemics incidence rates can reach 500 cases per 100,000 persons per year. Mortality is extremely high, at 10-20% in the United States, and much higher in developing countries. Following the introduction of the conjugate vaccine against Haemophilus influenzae, N. meningitidis is the major cause of bacterial meningitis at all ages in the United States (Schuchat et al (1997) supra). Based on the organism's capsular polysaccharide, 12 serogroups of N. meningitidis have been identified. The meningococcal vaccine currently in use is a tetravalent polysaccharide vaccine composed of serogroups A, C, Y and W135. Following the success of the vaccination against H.inβuenzae, however, conjugate vaccines against serogroups A and C have been developed Serogroup B remains a problem, however, and it is currently responsible for approximately 50% of total meningitis in the United States, Europe, and South America. The polysaccharide approach cannot be used because the menB capsular polysaccharide is a polymer of α(2-8)- linked N-acetyl neuraminic acid that is also present in mammalian tissue. This results in tolerance to the antigen; indeed, if a response were elicited, it would be anti-self, and therefore undesirable. In order to avoid induction of autoimmunity and to induce a protective immune response, the capsular polysaccharide has, for instance, been chemically modified substituting the N-acetyl groups with N-propionyl groups, leaving the specific antigenicity unaltered (Romero & Outschoorn (1994) Clin Microbiol Rev 7(4):559-575). An efficacious outer-membrane vesicle (OMV) vaccine against serogroup B has been produced by the Norwegian National Institute of Public Health [e.g. Bjune et al. (1991) Lancet 338(8775): 1093-96]. Whilst this vaccine is safe and prevents NmB disease, its efficacy is limited to the strain used to make the vaccine. Other vaccines based around outer-membrane preparations have also been reported. It is an object of the present invention to broaden the efficacy of these vaccines to other strains. DISCLOSURE OF THE INVENTION Surprisingly, it has been found that the addition of further defined components to OMV vaccines significantly broadens their efficacy. Thus the present invention provides a composition comprising (a) a NmB outer membrane preparation, and (b) an immunogenic component selected from one or more of the following: • a protein disclosed in WO99/57280, or an immunogenic fragment thereof; • a protein disclosed in WO99/36544, or an immunogenic fragment thereof; • a protein disclosed in WO99/24578, or an immunogenic fragment thereof; • a protein disclosed in WO00/66791, or an immunogenic fragment thereof; • a protein disclosed in Tettelin et al. [Science (2000) 287:1809-1815], or an immunogenic fragment thereof; • a protein disclosed in Parkhill et al. [Nature (2000) 404:502-506], or an immunogenic fragment thereof; • a protein disclosed in WO97/28273, or an immunogenic fragment thereof; • a protein disclosed in WO96/29412, or an immunogenic fragment thereof; • a protein disclosed in WO95/03413, or an immunogenic fragment thereof; • a protein disclosed in WO99/31132, or an immunogenic fragment thereof; • a protein disclosed in WO99/58683, or an immunogenic fragment thereof; • a protein disclosed in WO99/55873, or an immunogenic fragment thereof; and/or • Neisseria meningitidis protein PorA, TbpA, TbpB, PilC, OpA, or Omp85. If the composition comprises a protein disclosed in WO99/24578, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160 162,164,166, 168,170,172,174 176,178, 180,182, 184,186 196, 198, 200, 202 204,206,208, 210,212,214,216 218,220, 222,224, 226,228 238, 240, 242, 244 246,248,250, 252,254,256,258 260,262, 264,266, 268,270 280, 282, 284, 286 288,290,292, 294,296,298,300 302,304, 306,308, 310,312 322, 324, 326, 328 330,332,334, 336,338,340,342 344,346, 348,350, 352,354 364, 366, 368, 370 372,374,376, 378,380,382,384 386,388, 390,392, 394,396 406, 408, 410, 412 414,416,418, 420,422,424,426 428,430, 432,434, 436,438 448, 450, 452, 454 456,458,460, 462,464,466,468 470,472, 474,476, 478,480 490, 492, 494, 496 498,500,502, 504,506,508,510 512,514, 516,518, 520,522 532, 534, 536, 538 540,542,544, 546,548,550,552 554,556, 558,560, 562,564 574, 576, 578, 580 582,584,586, 588,590,592,594 596,598, 600,602, 604,606 616, 618, 620, 622 624,626,628, 630,632,634,636 638,640, 642,644, 646,648 658, 660, 662, 664 666,668,670, 672,674,676,678 680,682, 684,686, 688,690 700, 702, 704, 706 708,710,712, 714,716,718,720 722,724, 726,728, 730,732 742, 744, 746, 748 750,752,754, 756,758,760,762 764,766, 768,770, 772,774 784, 786, 788, 790 792,794,796, 798,800,802,804 806,808, 810,812, 814,816 826, 828, 830, 832 834,836,838, 840,842,844,846 848, 850, 852, 854, 856, 858 868, 870, 872, 874 876, 878, 880, 882, 884, 886, 888, 890, & 892, as disclosed in WO99/24578 (or a protein comprising an immunogenic fragment of one or more of these SEQ IDs, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to one of these SEQ IDs). If the composition comprises a protein disclosed in WO99/36544, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, & 90, as disclosed in WO99/36544 (or a protein comprising an immunogenic fragment of one or more of these SEQ IDs, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to one of these SEQ IDs). If the composition comprises a protein disclosed in Tettelin et al. {i.e. a protein encoded by one of the genes disclosed therein), said protein preferably comprises an amino acid sequence selected from the group consisting of NMB0001 to NMB2160 (or a protein comprising an immunogenic fragment of one or more of these 2160 genes, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to one of these 2160 genes). If the composition comprises a protein disclosed in Parkhill et al., said protein preferably comprises an amino acid sequence selected from the group consisting of the 2121 coding sequences disclosed therein (or a protein comprising an immunogenic fragment of one or more of these 2121 sequences, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to one of these 2121 sequences). If the composition comprises a protein disclosed in WO99/57280, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ IDs 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, , 66, 68,70,72,74 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 10 116, 120,122 124 126 128, 130 132 134, 136 138 140, 142, 144 158, 160 162,164 166 168 170, 172174 176, 178 180 182, 184, 186 200, 202 204,206 208 210 212, 214216 218, 220222 224, 226, 228 242, 244 246,248 250 252 254, 256258 260, 262264 266, 268, 270 284, 286 288,290 292 294 296, 298300 302, 304306 308, 310, 31 326, 328 330,332 334 336 338, 340342 344, 346348 350, 352, 354 368, 370 372,374 376 378 380, 382384 386, 388390 392, 394, 39 410, 412 414,416 418 420 422, 424426 428, 430432 434, 436, 438 452, 454 456,458 460 462 464, 466468 470, 472474 476, 478, 48 494, 496 498,500 502 504 506, 508510 512, 514516 518, 520, 52 536, 538 540,542 544 546 548, 550552 554, 556558 560, 562, 56 578, 580 582,584 586 588 590, 592594 596, 598600 602, 604, 60 620, 622 624,626 628 630 632, 634636 638, 640642 644, 646, 648 662, 664 666,668 670 672 674, 676678 680, 682684 686, 688, 690 704, 706 708,710 712 714 716, 718720 722, 724726 728, 730, 732 746, 748 750,752 754 756 758, 760762 764, 766768 770, 772, 774 788, 790 792,794 796 798 800, 802804 806, 808810 812, 814, 816 830, 832 834,836 838 840 842, 844846 848, 850852 854, 856, 858 872, 874 876,878 880 882 884, 886888 890, 892894 896, 898, 90 914, 916 918,920 922 924 926, 928930 932, 934936 938, 940, 94 956, 2120, 2154, 2188, 2222, 2256, 2290, 2324, 2358, 2392, 2426, 2460, 2494, 2528, 2562, 2596, 2630, 2664, 2698, 2732, 2766, 2800, 2834, 2868, 2902, 2936, 2970, 3004, comprising an immunogenic fragment of one or more of these SEQ IDs, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to one of these SEQ IDs). If the composition comprises a protein disclosed in WO99/28273, said protein is preferably the protein disclosed in Figure 4 or Figure 13 of WO97/28273. If the composition comprises a protein disclosed in WO96/29412, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ IDs 1-8 disclosed in WO96/29412 (or a protein comprising an immunogenic fragment of one or more of these SEQ IDs, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to one of these SEQ IDs). If the composition comprises a protein disclosed in WO95/03413, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ IDs 1-23 disclosed in WO95/03413 (or a protein comprising an immunogenic fragment of one or more of these SEQ IDs, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to one of these SEQ IDs). If the composition comprises a protein disclosed in WO99/31132, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ ID 2 disclosed in WO99/31132 (or a protein comprising an immunogenic fragment of SEQ ID 2, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to SEQ ID 2). If the composition comprises a protein disclosed in WO99/58683, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ ID 2 or SEQ ID 4 disclosed in WO99/58683 (or a protein comprising an immunogenic fragment of SEQ ID 2 or SEQ ED 4, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to SEQ ID 2 or SEQ ID 4). If the composition comprises a protein disclosed in WO99/55873, said protein preferably comprises an amino acid sequence selected from the group consisting of SEQ ID 2 or SEQ ID 4 disclosed in WO99/55873 (or a protein comprising an immunogenic fragment of SEQ ID 2 or SEQ D 4, or a protein comprising a sequence having sequence identity (preferably greater than 50% eg. 60%, 70%, 80%, 90%, 95%, 99% or more) to SEQ ID 2 or SEQ ID 4). Details of Opa and PorA can be found in Wiertz et al. [Infect. Immun. (1996) 61:298-304]. PilC is disclosed in Nassif et al. [PNAS USA (1994) 91:3769-73]. Omp85 is disclosed in Manning et al. [Microb. Pathog. (1998) 25:11-21]. TbpA and TbpB are disclosed in Ala'Aldeen & Borriello [Vaccine (1996) 14:49-53] and also in Legrain et al. [Prorein Expr Purif (1995) 6:570-578]. Preferred proteins for component (b) are: • protein '919', typified by SEQ IDs 3069-3074 and 3207-3241 of WO99/57280 (see also Figure 23 and Example 15 therein). • protein '235', typified by SEQ IDs 869-874 and 3149-3178 of WO99/57280 (see also Figure 20 and Example 12 therein). • protein '519', typified by SEQ IDs 3045-3056 and 3185-3206 of WO99/57280 (see also Figure 22 and Example 14 therein). • protein '225', typified by SEQ IDs 793-804 and 3115-3148 of WO99/57280 (see also Figure 19 and Example 11 therein). • protein 'ORF40', typified by example 1 (SEQ IDs 1-6) of WO99/36544 (see also Figure 1 of WO00/66741; see also WO99/31132 and WO99/58683). • protein '287', typified by example 9 of WO99/57280 (see SEQ IDs 1199-1204, 3103- 3108 and 3179-3184 therein). • protein 'ORF1', typified by example 77 (SEQ IDs 647-654) of WO99/24578 (see also WO99/55873 and accession number AJ242535). • protein 'ORF4', typified by example 26 (SEQ IDs 215-226) of WO99/24578 (see also Figure 2 of WO00/66741). • protein 'ORF46', typified by example 55 (SEQ IDs 457-466) of WO99/24578 (see also Figure 12 of WO00/66741). Component (b) of the composition is preferably a NmB protein. It is preferred that component (b) includes a protein from a different NmB strain from that from which the OMV of component (a) is derived i.e. the OMV in component (a) is preferably supplemented by immunogenic component (b) from a different NmB strain. One or more of the components (or all of them) may be adsorbed on Al(OH) 3 . The outer membrane preparation component The compositions of the invention include a NmB outer membrane preparation as component (a). This is preferably in the form of outer membrane vesicles (OMVs). The preparation of OMVs from NmB is well-known in the art. Methods for obtaining suitable preparations are disclosed in, for instance: Claassen et al. [Vaccine (1996) 14:1001-1008]; Cartwright et al. [Vaccine (1999) 17:2612-2619]; Peeters et al. [Vaccine (1996) 14: 1009- 1015]; Fu et al. [Biotechnology NY (1995) 12:170-74]; Davies et al. [J.Immunol.Meth. (1990) 134:215-225]; Saunders et al. [Infect. Immun. (1999) 67: 113-119]; Draabick et al. [Vaccine (2000) 18:160-172]; Moreno et al. [Infect. Immun. (1985) 47:527-533]; Milagres et al. [Infect. Immun. (1994) 62:4419-4424]; Naess et al. [Infect. Immun. (1998) 66:959-965]; Rosenqvist et al. [Dev.BiolStand. (1998) 92:323-333]; Haneberg et al. [Infect. Immunn. (1998) 66: 1334-41]; Andersen et al. [Vaccine (1997) 15:1225-34]; Bjune et al. [Lancet (1991) 338:1093-96] etc. OMVs are preferably a deoxycholate extract from NmB (i.e. obtained from NmB by deoxycholate extraction). The preferred extraction protocol is that described by Fredriksen et al. [Production, characterization and control of MenB -vaccine "Folkehelsa": an outer membrane vesicle vaccine against group B meningococcal disease (1991) NIPH Ann. 14(2):67-80]. A preferred strain from which to extract OMVs is the 44/76 strain (B:15:P1.7,16:P5.5:L3,7,9) of N. meningitidis. Further details of the OMV component can be found in, for instance, Bjune et al. [Lancet (1991) 338(8775): 1093-96], or Fredriksen et al. [Characterization of high molecular weight component in MenB-vaccine 'Folkehelsa', an outer membrane vesicle vaccine against group B meningococcal disease. Pages 818-824 of Pathobiology and immunobiology of Neisseriaceae (eds. Conde-Glez et al.) ISBN 968-6502-13-0]. The OMV component may be adsorbed to aluminium hydroxide adjuvant. A preferred protein:adjuvant ratio is 1:67 (wt/wt). A typical dose of vaccine for a human contains 25μg protein, 2μg LPS and 1.67mg Al(OH) 3 , and can be injected in 0.5ml volumes into the deltoid muscle. The OMV component (e.g. as obtained by deoxycholate extraction) may be treated to remove certain components. For instance, pyrogens or toxic components may be removed (e.g. LPS). It is preferred that the OMV component should retain the 80kDa antigenic component described by Fredriksen et al. [pages 818-824 of Pathobiology and immunobiology of Neisseriaceae], More preferably, the OMV component should retain a protein comprising one or more of the following amino acid sequences: SEQ ID 3, SEQ ID 5, SEQ ID 7, SEQ ID 9, SEQ ID 11, SEQ ID 13 [or (i) a protein having sequence identity to SEQ ID 3, SEQ ID 5, SEQ ID 7, SEQ ID 9, SEQ ID 11, or SEQ ID 13 - depending on the particular SEQ ID, the degree of sequence identity is preferably greater than 50% (eg. 60%, 70%, 80%, 90%, 95%, 99% or more), which includes mutants and allelic variants, or (ii) a protein comprising an immunogenic fragment of SEQ ID 1, SEQ ID 3, SEQ ID 5, SEQ ID 7, SEQ ID 9, SEQ ID 11, or SEQ ID 13 - the fragment should comprise at least n consecutive amino acids from the sequence and, depending on the particular sequence, n is 7 or more (eg. 8, 10, 12, 14, 16, 18, 20 or more).] Combining components (a) and (b) Components (a) and (b) can be combined by simply mixing component (a) with an outer- membrane preparation (e.g. by mixing ORF4 with Norwegian OMVs). As an alternative, they can be combined by manipulating a bacterium such that it produces (preferably hyperproduces) component (a) in its outer membrane - an outer-membrane preparation from such a recombinant bacterium will comprise both component (a) and component (b). Suitable bacteria for manipulation in this way include Neisseria meningitidis (any serogroup or strain), Neisseria lactamica, Neisseria cinerea or any other non-typable Neisseria. Other Gram-negative bacteria can also be used, such as E.coli, Salmonella, Shigella, Bordetella, Yersinia, Helicobacter, etc. Transformation methods are well known in the art. Multivalent vaccines Optionally, the composition of the invention may also comprise one or more of the following components: • a protective antigen against Neisseria meningitidis serogroup A; • a protective antigen against Neisseria meningitidis serogroup C; • a protective antigen against Neisseria meningitidis serogroup Y; • a protective antigen against Neisseria meningitidis serogroup W; • a protective antigen against Haemophilus inβuenzae; • a protective antigen against pneumococcus • a protective antigen against diphtheria; • a protective antigen against tetanus; • a protective antigen against whooping cough; • a protective antigen against Helicobacter pylori; • a protective antigen against polio; and/or • a protective antigen against hepatitis B virus. Preferred examples of these optional components are: • a polysaccharide antigen against Neisseria meningitidis serogroup A; • a polysaccharide antigen against Neisseria meningitidis serogroup C, such as that described in Costantino et al. (1992) Vaccine 10:691-698; • a polysaccharide antigen against Neisseria meningitidis serogroup Y; • a polysaccharide antigen against Neisseria meningitidis serogroup W; • a polysaccharide antigen against Haemophilus influenzae; • a polysaccharide antigen against pneumococcus; • a protective antigen against diphtheria, consisting of a diphtheria toxoid, such as the CRM 197 mutant [eg. Del Guidice et al. (1998) Molecular Aspects of Medicine 19: 1 -70]. • a protective antigen against tetanus, consisting of a tetanus toxoid [eg. Wassilak & Orenstein, Chapter 4 of Vaccines (eds. Plotkin & Mortimer), 1988] • a protective antigen against whooping cough, comprising pertussis holotoxin (PT) and filamentous haemagglutinin (FHA); optionally further comprising pertactin and/or agglutinogens 2 and 3 [eg. Gustafsson et al. (1996) N. Engl. J. Med. 334:349-355; Rappuoli et /. (1991) TIBTECH 9:232-238]. • a protective antigen against H.pylori, comprising one or more of CagA (eg. WO93/18150), VacA (eg. WO93/18150), NAP (eg. WO99/53310), HopX (eg. WO98/04702), HopY (eg. WO98/04702), urease. • a protective antigen against hepatitis B virus, consisting of a HBV surface antigen and/or a HBV core antigen. Where the composition comprises an antigen against diphtheria, it preferably also comprises antigens against tetanus and polio. Where the composition comprises an antigen against tetanus, it preferably also comprises antigens against diphtheria and polio. Where the composition comprises an antigen against polio, it preferably also comprises antigens against diphtheria and tetanus. Pertussis toxin is a toxic protein and, when present in the composition, it is preferably detoxified. Detoxification may be by chemical and/or genetic means. A preferred detoxified mutant is the 9K/129G double mutant [eg. Rappuoli (1997) Nature Medicine 3:374-376]. Where the composition includes a protein that exists in different nascent and mature forms, the mature form of the protein is preferably used. For example, where NspA is included, (WO96/29412; see also Martin et al. (1997) J. Exp. Med 185 1173-1183) the mature form of the protein lacking the signal peptide is preferably used. Where the composition includes a polysaccharide antigen, the polysaccharide is preferably conjugated to a carrier protein. Therapy, prophylaxis, diagnosis The composition of the invention is preferably a vaccine. Vaccines according to the invention may either be prophylactic (ie. to prevent infection) or therapeutic (ie. to treat disease after infection). The invention also provides the compositions of the invention for use as medicaments (preferably as vaccines) or as diagnostic reagents. It also provides the use of a composition according to the invention in the manufacture of: (i) a medicament for treating or preventing infection due to Neisserial bacteria; (ii) a diagnostic reagent for detecting the presence of Neisserial bacteria or of antibodies raised against Neisserial bacteria; and/or (iii) a reagent which can raise antibodies against Neisserial bacteria. Said Neisserial bacteria may be any species or strain (such as N.gonorrhoeae) but are preferably N. meningitidis, especially serogroup B (NmB). The invention also provides a method of treating a patient, comprising administering to the patient a therapeutically effective amount of a composition according to the invention. The method is preferably immunisation. Processes According to further aspects, the invention provides various processes. A process for producing a composition of the invention is provided, comprising the step of extraction (e.g. deoxycholate extraction) of OMVs from N. meningitidis. SEQUENCE LISTING The sequences in the sequence listing are: MODES FOR CARRYING OUT THE INVENTION A summary of standard techniques and procedures which may be employed in order to perform the invention (eg. to utilise the disclosed sequences for vaccination or diagnostic purposes) follows. This summary is not a limitation on the invention but, rather, gives examples that may be used, but are not required. General The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology, microbiology, recombinant DNA, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature eg. Sambrook Molecular Cloning; A Laboratory Manual, Second Edition (1989); DNA Cloning, Volumes I and ii (D.N Glover ed. 1985); Oligonucleotide Synthesis (M J. Gait ed, 1984); Nucleic Acid Hybridization (B .D. Hames & S.J. Higgins eds. 1984); 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); the Methods in Enzymology series (Academic Press, Inc.), especially volumes 154 & 155; Gene Transfer Vectors for Mammalian Cells (J.H. M iller and M .P. Calos eds. 1987, Cold Spring Harbor Laboratory); M ayer and W alker, eds. (1987), Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); Scopes, (1987) Protein Purification: Principles and Practice, Second Edition (Springer-Verlag, N .Y .), and Handbook of Experimental Immunology, Volumes 1-IV (D .M . W eir and C. C. B lackwell eds 1986). Standard abbreviations for nucleotides and amino acids are used in this specification. Proteins used with the invention can be prepared by various means {eg. recombinant expression, purification from cell culture, chemical synthesis etc.) and in various forms (eg. native, fusions etc.). They are preferably prepared in substantially pure form (ie. substantially free from other Neisseria or host cell proteins). Nucleic acid used with the invention can be prepared in many ways (eg. by chemical synthesis, from genomic or cDNA libraries, from the organism itself etc.) and can take various forms (eg. single stranded, double stranded, vectors, probes etc.). The term "nucleic acid" includes DNA and RNA , and also their analogues, such as those containing modified backbones, and also peptide nucleic acids (PNA ) etc. Definitions A composition containing X is "substantially free of Y when at least 85% by weight of the total X+Y in the composition is X . Preferably, X comprises at least about 90% by weight of the total of X+Y in the composition, more preferably at least about 95% or even 99% by weight. The term "comprising" means "including" as well as "consisting" eg. a composition "comprising" X may consist exclusively of X or may include something additional to X, such as X+Y . The term "heterologous" refers to two biological components that are not found together in nature. The components may be host cells, genes, or regulatory regions, such as promoters. Although the heterologous components are not found together in nature, they can function together, as when a promoter heterologous to a gene is operably linked to the gene. Another example is where a Neisserial sequence is heterologous to a mouse host cell. A further examples would be two epitopes from the same or different proteins which have been assembled in a single protein in an arrangement not found in nature. An "origin of replication" is a polynucleotide sequence that initiates and regulates replication of polynucleotides, such as an expression vector. The origin of replication behaves as an autonomous unit of polynucleotide replication w ithin a cell, capable of replication under its own control. A n origin of replication may be needed for a vector to replicate in a particular host cell. W ith certain origins of replication, an expression vector can be reproduced at a high copy number in the presence of the appropriate proteins within the cell. Examples of origins are the autonomously replicating sequences, which are effective in yeast; and the viral T-antigen, effective in COS-7 cells. Identity between proteins is preferably determined by the Smith-W aterman homology search algorithm as implemented in the M PSRCH program (Oxford M olecular), using an affine gap search with parameters gap open penalty= 12 and gap extension penalty= l . Typically, 50% identity or more between two proteins is considered to be an indication of functional equivalence. As used herein, an "allelic variant" of a nucleic acid molecule, or region, for which nucleic acid sequence is provided herein is a nucleic acid molecule, or region, that occurs essentially at the same locus in the genome of another or second isolate, and that, due to natural variation caused by, for example, mutation or recombination, has a similar but not identical nucleic acid sequence. A coding region allelic variant typically encodes a protein having similar activity to that of the protein encoded by the gene to which it is being compared. An allelic variant can also comprise an alteration in the 5' or 3' untranslated regions of the gene, such as in regulatory control regions (eg. see US patent 5,753,235). Expression systems The Neisserial nucleotide sequences can be expressed in a variety of different expression systems; for example those used with mammalian cells, baculoviruses, plants, bacteria, and yeast. i. M ammalian Systems M ammalian expression systems are known in the art. A mammalian promoter is any DNA sequence capable of binding mammalian RNA polymerase and initiating the downstream (3') transcription of a coding sequence (eg. structural gene) into mRNA . A promoter will have a transcription initiating region, which is usually placed proximal to the 5' end of the coding sequence, and a TA TA box, usually located 25-30 base pairs (bp) upstream of the transcription initiation site. The TATA box is thought to direct RNA polymerase II to begin RNA synthesis at the correct site. A mammalian promoter will also contain an upstream promoter element, usually located within 100 to 200 bp upstream of the TATA box. An upstream promoter element determines the rate at which transcription is initiated and can act in either orientation [Sambrook et al. (1989) "Expression of Cloned Genes in M ammalian Cells." In Molecular Cloning: A Laboratory Manual, 2nd ed.]. M ammalian viral genes are often highly expressed and have a broad host range; therefore sequences encoding mammalian viral genes provide particularly useful promoter sequences. Examples include the SV40 early promoter, mouse mammary tumor virus LTR promoter, adenovirus major late promoter (Ad M LP), and herpes simplex virus promoter. In addition, sequences derived from non-viral genes, such as the murine metallotheionein gene, also provide useful promoter sequences. Expression may be either constitutive or regulated (inducible), depending on the promoter can be induced with glucocorticoid in hormone-responsive cells. The presence of an enhancer element (enhancer), combined with the promoter elements described above, will usually increase expression levels. An enhancer is a regulatory DNA sequence that can stimulate transcription up to 1000-fold when linked to homologous or heterologous promoters, with synthesis beginning at the normal RNA start site. Enhancers are also active when they are placed upstream or downstream from the transcription initiation site, in either normal or flipped orientation, or at a distance of more than 1000 nucleotides from the promoter [M aniatis et al. ( 1987) Science 236: 1237 ; Alberts et al. (1989) Molecular Biology of the Cell, 2nd ed.]. Enhancer elements derived from viruses may be particularly useful, because they usually have a broader host range. Examples include the SV40 early gene enhancer [Dijkema et al (1985) EMBO J. 4:161 ) and the enhancer/promoters derived from the long terminal repeat (LTR) of the Rous Sarcoma V irus [Gorman et al. (1982b) Proc. Natl. Acad. Sci. 79:6111] and from human cytomegalovirus [Boshart et al. (1985) Cell 47 :521 ]. Additionally, some enhancers are regulatable and become active only in the presence of an inducer, such as a hormone or metal ion [Sassone-Corsi and Borelli (1986) Trends Genet. 2:215 ; M aniatis et al. (1987) Science 236: 1237]. A DNA molecule may be expressed intracellularly in mammalian cells. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus of the recombinant protein will always be a methionine, which is encoded by the ATG start codon. If desired, the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide. Alternatively, foreign proteins can also be secreted from the cell into the growth media by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provides for secretion of the foreign protein in mammalian cells. Preferably, there are processing sites encoded between the leader fragment and the foreign gene that can be cleaved either in vivo or in vitro. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the secretion of the protein from the cell. The adenovirus triparite leader is an example of a leader sequence that provides for secretion of a foreign protein in mammalian cells. Usually, transcription termination and polyadenylation sequences recognized by mammalian cells are regulatory regions located 3' to the translation stop codon and thus, together with the promoter elements, flank the coding sequence. The 3' terminus of the mature mRNA is formed by site-specific post- transcriptional cleavage and polyadenylation [B irnstiel et al. (1985) Cell 41 -349; Proudfoot and W hitelaw (1988) "Termination and 3' end processing of eukaryotic RNA . In Transcription and splicing (ed. B .D . Hames and D .M . Glover); Proudfoot (1989) Trends Biochem. Sci. 74:105]. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA . Examples of transcription terminater/polyadenylation signals include those derived from SV40 [Sambrook et al (1989) "Expression of cloned genes in cultured mammalian cells." In Molecular Cloning: A Laboratory Manual]. Usually, the above described components, comprising a promoter, polyadenylation signal, and transcription termination sequence are put together into expression constructs. Enhancers, introns with functional splice donor and acceptor sites, and leader sequences may also be included in an expression construct, if desired. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as mammalian cells or bacteria. M ammalian replication systems include those derived from animal viruses, which require trans-acting factors to replicate. For example, plasmids containing the replication systems of papovaviruses, such as SV40 [Gluzman (1981 ) Cell 23: 175] or polyomavirus, replicate to extremely high copy number in the presence of the appropriate viral T antigen. Additional examples of mammalian replicons include those derived from bovine papillomavirus and Epstein-B arr virus. Additionally, the replicon may have two replicaton systems, thus allowing it to be maintained, for example, in mammalian cells for expression and in a prokaryotic host for cloning and amplification. Examples of such mammalian-bacteria shuttle vectors include pM T2 [Kaufman et al. (1989) Mol. Cell. Biol. 9:946] and pHEB O [Shimizu et al. (1986) Mol. Cell. Biol. 6: 1074]. The transformation procedure used depends upon the host to be transformed. M ethods for introduction of heterologous polynucleotides into mammalian cells are known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, and direct microinjection of the DNA into nuclei. M ammalian cell lines available as hosts for expression are known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC), including but not limited to, Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (B H K) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (eg. Hep G2), and a number of other cell lines. ii. Baculovirus Systems The polynucleotide encoding the protein can also be inserted into a suitable insect expression vector, and is operably linked to the control elements w ithin that vector. Vector construction employs techniques which are known in the art. Generally, the components of the expression system include a transfer vector, usually a bacterial plasm id, which contains both a fragment of the baculovirus genome, and a convenient restriction site for insertion of the heterologous gene or genes to be expressed; a wild type baculovirus with a sequence homologous to the baculovirus-specific fragment in the transfer vector (this allows for the homologous recombination of the heterologous gene in to the baculovirus genome); and appropriate insect host cells and growth media. After inserting the DNA sequence encoding the protein into the transfer vector, the vector and the wild type viral genome are transfected into an insect host cell where the vector and viral genome are allowed to recombine. The packaged recombinant virus is expressed and recombinant plaques are identified and purified. M aterials and methods for baculovirus/insect cell expression systems are commercially available in kit form from , inter alia, Invitrogen, San Diego CA ("M axB ac" kit). These techniques are generally known to those skilled in the art and fully described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No. 1555 ( 1987) (hereinafter "Summers and Smith"). Prior to inserting the DNA sequence encoding the protein into the baculovirus genome, the above described components, comprising a promoter, leader (if desired), coding sequence of interest, and transcription termination sequence, are usually assembled into an intermediate transplacement construct (transfer vector). This construct may contain a single gene and operably linked regulatory elements; multiple genes, each with its owned set of operably linked regulatory elements; or multiple genes, regulated by the same set of regulatory elements. Intermediate transplacement constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as a bacterium . The replicon will have a replication system, thus allowing it to be maintained in a suitable host for cloning and amplification. Currently, the most commonly used transfer vector for introducing foreign genes into AcNPV is pAc373. M any other vectors, known to those of skill in the art, have also been designed. These include, for example, pVL985 (which alters the polyhedrin start codon from ATG to A TT, and which introduces a B amHI cloning site 32 basepairs downstream from the ATT; see Luckow and Summers, Virology (1989) 77:31. The plasmid usually also contains the polyhedrin polyadenylation signal (M iller et al. (1988) Ann. Rev. Microbiol, 42:111) and a prokaryotic ampicillin-resistance (amp) gene and origin of replication for selection and propagation in E. coli. B aculovirus transfer vectors usually contain a baculovirus promoter. A baculovirus promoter is any DNA sequence capable of binding a baculovirus RNA polymerase and initiating the downstream (5' to 3') transcription of a coding sequence (eg. structural gene) into mRNA . A promoter will have a transcription initiation region which is usually placed proximal to the 5' end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site and a transcription initiation site. A baculovirus transfer vector may also have a second domain called an enhancer, which, if present, is usually distal to the structural gene. Expression may be either regulated or constitutive. Structural genes, abundantly transcribed at late times in a viral infection cycle, provide particularly useful promoter sequences. Examples include sequences derived from the gene encoding the viral polyhedron protein, Friesen et al., (1986) "The Regulation of B aculovirus Gene Expression," in: The Molecular Biology of Baculoviruses (ed. W alter Doerfler); EPO Publ. Nos. 127 839 and 155 476; and the gene encoding the plO protein, V lak et al., (1988), J. Gen. Virol. 69:765. DNA encoding suitable signal sequences can be derived from genes for secreted insect or baculovirus proteins, such as the baculovirus polyhedrin gene (Carbonell et al. (1988) Gene, 73:409). Alternatively, since the signals for mammalian cell posttranslational modifications (such as signal peptide cleavage, proteolytic cleavage, and phosphorylation) appear to be recognized by insect cells, and the signals required for secretion and nuclear accumulation also appear to be conserved between the invertebrate cells and vertebrate cells, leaders of non-insect origin, such as those derived from genes encoding human -interferon, M aeda et al., (1985), Nature 575:592; human gastrin-releasing peptide, Lebacq-Verheyden et al., (1988), Molec. Cell. Biol. 8:3129; human IL-2, Smith et al, (1985) Proc. Nat'l Acad. Sci. USA, 82:8404; mouse IL-3, (M iyajima et al., (1987) Gene 58:273 ; and human glucocerebrosidase, M artin et al. (1988) DNA, 7:99, can also be used to provide for secretion in insects. A recombinant polypeptide or polyprotein may be expressed intracellularly or, if it is expressed with the proper regulatory sequences, it can be secreted. Good intracellular expression of nonfused foreign proteins usually requires heterologous genes that ideally have a short leader sequence containing suitable translation initiation signals preceding an ATG start signal. If desired, methionine at the N-terminus may be cleaved from the mature protein by in vitro incubation with cyanogen bromide. Alternatively, recombinant polyproteins or proteins which are not naturally secreted can be secreted from the insect cell by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provides for secretion of the foreign protein in insects. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the translocation of the protein into the endoplasmic reticulum . A fter insertion of the DNA sequence and/or the gene encoding the expression product precursor of the protein, an insect cell host is co-transformed with the heterologous DNA of the transfer vector and the genomic DNA of wild type baculovirus - usually by co-transfection. The promoter and transcription termination sequence of the construct will usually comprise a 2-5kb section of the baculovirus genome. M ethods for introducing heterologous DNA into the desired site in the baculovirus virus are known in the art. (See Summers and Smith supra; Ju et al. (1987); Smith et al., Mol. Cell. Biol. (1983) 5:2156; and Luckow and Summers (1989)). For example, the insertion can be into a gene such as the polyhedrin gene, by homologous double crossover recombination; insertion can also be into a restriction enzyme site engineered into the desired baculovirus gene. M iller et al, (1989), Bioessays 4:91. The DNA sequence, when cloned in place of the polyhedrin gene in the expression vector, is flanked both 5' and 3' by polyhedrin-specific sequences and is positioned downstream of the polyhedrin promoter. The newly formed baculovirus expression vector is subsequently packaged into an infectious recombinant baculovirus. Homologous recombination occurs at low frequency (between about 1 % and about 5% ); thus, the majority of the virus produced after cotransfection is still wild-type virus. Therefore, a method is necessary to identify recombinant viruses. A n advantage of the expression system is a visual screen allowing recombinant viruses to be distinguished. The polyhedrin protein, which is produced by the native virus, is produced at very high levels in the nuclei of infected cells at late times after viral infection. Accumulated polyhedrin protein forms occlusion bodies that also contain embedded particles. These occlusion bodies, up to 15 μm in size, are highly refractile, giving them a bright shiny appearance that is readily visualized under the light microscope. Cells infected with recombinant viruses lack occlusion bodies. To distinguish recombinant virus from wild-type virus, the transfection supernatant is plaqued onto a monolayer of insect cells by techniques known to those skilled in the art. Namely, the plaques are screened under the light microscope for the presence (indicative of wild-type virus) or absence (indicative of recombinant virus) of occlusion bodies. "Current Protocols in M icrobiology" Vol. 2 (Ausubel et al. eds) at 16.8 (Supp. 10, 1990); Summers and Sm ith, supra; M iller et al. (1989). Recombinant baculovirus expression vectors have been developed for infection into several insect cells. For example, recombinant baculoviruses have been developed for, inter alia: Aedes aegypti , Autographa californica, Bombyx mori, Drosophila melanogaster, Spodoptera frugiperda, and Trichoplusia ni (W O 89/046699; Carbonell et al, (1985) J. Virol. 56: 153 ; W right (1986) Nature 527 :718; Smith et al, (1983) Mol. Cell. Biol. 5:2156; and see generally, Fraser, et al. (1989) In Vitro Cell. Dev. Biol. 25:225). Cells and cell culture media are commercially available for both direct and fusion expression of heterologous polypeptides in a baculovirus/expression system; cell culture technology is generally known to those skilled in the art. See, eg. Summers and Smith supra. The modified insect cells may then be grown in an appropriate nutrient medium, which allows for stable maintenance of the plasmid(s) present in the modified insect host. Where the expression product gene is under inducible control, the host may be grown to high density, and expression induced. Alternatively, where expression is constitutive, the product will be continuously expressed into the medium and the nutrient medium must be continuously circulated, while removing the product of interest and augmenting depleted nutrients. The product may be purified by such techniques as chromatography, eg. HPLC , affinity chromatography, ion exchange chromatography, etc.; electrophoresis; density gradient centrifugation; solvent extraction, or the like. A s appropriate, the product may be further purified, as required, so as to remove substantially any insect proteins which are also secreted in the medium or result from lysis of insect cells, so as to provide a product which is at least substantially free of host debris, eg. proteins, lipids and polysaccharides. In order to obtain protein expression, recombinant host cells derived from the transformants are incubated under conditions which allow expression of the recombinant protein encoding sequence. These conditions will vary, dependent upon the host cell selected. However, the conditions are readily ascertainable to those of ordinary skill in the art, based upon what is known in the art. iii. Plant Systems There are many plant cell culture and whole plant genetic expression systems known in the art. Exemplary plant cellular genetic expression systems include those described in patents, such as: US 5,693,506; US 5,659,122; and US 5 ,608,143. Additional examples of genetic expression in plant cell culture has been described by Zenk, Phytochemistry 30:3861 -3863 (1991 ). Descriptions of plant protein signal peptides may be found in addition to the references described above in Vaulcombe et al, Mol. Gen. Genet. 209:33-40 (1987); Chandler et al. Plant Molecular Biology 3 :407-418 (1984); Rogers, J. Biol. Chem. 260:3731-3738 (1985); Rothstein et al. Gene 55 :353-356 (1987); W hittier et al. Nucleic Acids Research 15 :2515-2535 (1987); W irsel et al. Molecular Microbiology 3 :3-14 (1989); Yu et al. Gene 122:247-253 (1992). A description of the regulation of plant gene expression by the phytohormone, gibberellic acid and secreted enzymes induced by gibberellic acid can be found in R.L. Jones and J. M acM illin, Gibberellins: in: Advanced Plant Physiology,. M alcolm B . W ilkins, ed, 1984 Pitman Publishing Limited, London, pp. 21 -52. References that describe other metabolically-regulated genes: Sheen, Plant Cell, 2:1027-1038(1990); M aas et al, EMBO J. 9:3447-3452 (1990); B enkel and Hickey, Proc. Natl. Acad. Sci. 84:1337-1339 (1987) Typically, using techniques known in the art, a desired polynucleotide sequence is inserted into an expression cassette comprising genetic regulatory elements designed for operation in plants. The expression cassette is inserted into a desired expression vector with companion sequences upstream and downstream from the expression cassette suitable for expression in a plant host. The companion sequences will be of plasmid or viral origin and provide necessary characteristics to the vector to permit the vectors to move DNA from an original cloning host, such as bacteria, to the desired plant host. The basic bacterial/plant vector construct will preferably provide a broad host range prokaryote replication origin; a prokaryote selectable marker; and, for Agrobacterium transformations, T DNA sequences for Agrobacterium-mediated transfer to plant chromosomes. W here the heterologous gene is not readily amenable to detection, the construct will preferably also have a selectable marker gene suitable for determining if a plant cell has been transformed. A general review of suitable markers, for example for the members of the grass family, is found in W ilmink and Dons, 1993, Plant Mol. Biol. Reptr, 1 1 (2): 165- 185. Sequences suitable for permitting integration of the heterologous sequence into the plant genome are also recommended. These might include transposon sequences and the like for homologous recombination as well as Ti sequences which permit random insertion of a heterologous expression cassette into a plant genome. Suitable prokaryote selectable m arkers include resistance toward antibiotics such as ampicillin or tetracycline. Other DNA sequences encoding additional functions may also be present in the vector, as is known in the art. The nucleic acid molecules of the subject invention may be included into an expression cassette for expression of the protein(s) of interest. Usually, there will be only one expression cassette, although two or more are feasible. The recombinant expression cassette will contain in addition to the heterologous protein encoding sequence the following elements, a promoter region, plant 5' untranslated sequences, initiation codon depending upon whether or not the structural gene comes equipped with one, and a transcription and translation termination sequence. Unique restriction enzyme sites at the 5' and 3' ends of the cassette allow for easy insertion into a pre-existing vector. A heterologous coding sequence may be for any protein relating to the present invention. The sequence encoding the protein of interest will encode a signal peptide which allows processing and translocation of the protein, as appropriate, and will usually lack any sequence which m ight result in the binding of the desired protein of the invention to a membrane. Since, for the most part, the transcriptional initiation region will be for a gene which is expressed and translocated during germination, by employing the signal peptide which provides for translocation, one may also provide for translocation of the protein of interest. In this way, the protein(s) of interest w ill be translocated from the cells in which they are expressed and may be efficiently harvested. Typically secretion in seeds are across the aleurone or scutellar epithelium layer into the endosperm of the seed. W hile it is not required that the protein be secreted from the cells in which the protein is produced, this facilitates the isolation and purification of the recombinant protein. Since the ultimate expression of the desired gene product will be in a eucaryotic cell it is desirable to determine whether any portion of the cloned gene contains sequences which will be processed out as introns by the host's splicosome machinery. If so, site-directed mutagenesis of the "intron" region may be conducted to prevent losing a portion of the genetic message as a false intron code, Reed and M aniatis, Oe// 41 :95-105, 1985. The vector can be microinjected directly into plant cells by use of micropipettes to mechanically transfer the recombinant DNA . Crossway, Mol. Gen. Genet, 202:179- 185, 1985. The genetic material may also be transferred into the plant cell by using polyethylene glycol, Krens, et al. Nature, 296, 72-74, 1982. Another method of introduction of nucleic acid segments is high velocity ballistic penetration by small particles with the nucleic acid either within the matrix of small beads or particles, or on the surface, Klein, et al. Nature, 327, 70-73, 1987 and Knudsen and M uller, 1991 , Planta, 185 :330-336 teaching particle bombardment of barley endosperm to create transgenic barley. Yet another method of introduction would be fusion of protoplasts with other entities, either minicells, cells, lysosomes or other fusible lipid-surfaced bodies, Fraley, et al, Proc. Natl. Acad. Sci. USA, 79, 1859-1863, 1982. The vector may also be introduced into the plant cells by electroporation. (Fromm et al, Proc. Natl Acad. Sci. USA 82:5824, 1985). In this technique, plant protoplasts are electroporated in the presence of plasmids containing the gene construct. Electrical impulses of high field strength reversibly permeabilize biomembranes allowing the introduction of the plasmids. Electroporated plant protoplasts reform the cell wall, divide, and form plant callus. All plants from which protoplasts can be isolated and cultured to give whole regenerated plants can be transformed by the present invention so that whole plants are recovered which contain the transferred gene. It is known that practically all plants can be regenerated from cultured cells or tissues, including but not limited to all major species of sugarcane, sugar beet, cotton, fruit and other trees, legumes and vegetables. Some suitable plants include, for example, species from the genera Fragaria, Lotus, Medicago, Onobrychis, Trifolium, Trigonella, Vigna, Citrus, Linum, Geranium, Manihot, Daucus, Arabidopsis, Brassica, Raphanus, Sinapis, Atropa, Capsicum, Datura, Hyoscyamus, Lycopersion, Nicotiana, Solanum , Petunia, Digitalis, Majorana, Cichorium, Helianthus, Lactuca, Bromus, Asparagus, Antirrhinum, Hererocallis, Nemesia, Pelargonium, Panicum, Pennisetum, Ranunculus, Senecio, Salpiglossis, Cucumis, Browaalia, Glycine, Lolium , lea, Triticum, Sorghum, and Datura. M eans for regeneration vary from species to species of plants, but generally a suspension of transformed protoplasts containing copies of the heterologous gene is first provided. Callus tissue is formed and shoots may be induced from callus and subsequently rooted. A lternatively, embryo formation can be induced from the protoplast suspension. These embryos germinate as natural embryos to form plants. The culture media will generally contain various amino acids and hormones, such as auxin and cytokinins. It is also advantageous to add glutamic acid and proline to the medium, especially for such species as corn and alfalfa. Shoots and roots normally develop simultaneously. Efficient regeneration will depend on the medium, on the genotype, and on the history of the culture. If these three variables are controlled, then regeneration is fully reproducible and repeatable. In some plant cell culture systems, the desired protein of the invention may be excreted or alternatively, the protein may be extracted from the whole plant. Where the desired protein of the invention is secreted into the medium , it may be collected. Alternatively, the embryos and embryoless-half seeds or other plant tissue may be mechanically disrupted to release any secreted protein between cells and tissues. The mixture may be suspended in a buffer solution to retrieve soluble proteins. Conventional protein isolation and purification methods will be then used to purify the recombinant protein. Parameters of time, temperature pH , oxygen, and volumes will be adjusted through routine methods to optimize expression and recovery of heterologous protein. iv. B acterial Systems B acterial expression techniques are known in the art. A bacterial promoter is any DNA sequence capable of binding bacterial RNA polymerase and initiating the downstream (3' ) transcription of a coding sequence (eg. structural gene) into mRNA . A promoter will have a transcription initiation region which is usually placed proximal to the 5 ' end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site and a transcription initiation site. A bacterial promoter may also have a second domain called an operator, that may overlap an adjacent RNA polymerase binding site at which RNA synthesis begins. The operator permits negative regulated (inducible) transcription, as a gene repressor protein may bind the operator and thereby inhibit transcription of a specific gene. Constitutive expression may occur in the absence of negative regulatory elements, such as the operator. In addition, positive regulation may be achieved by a gene activator protein binding sequence, which, if present is usually proximal (5') to the RNA polymerase binding sequence. An example of a gene activator protein is the catabolite activator protein (CAP), which helps initiate transcription of the lac operon in Escherichia coli (E. coli) [Raibaud et al. (1984) Annu. Rev. Genet. 78:173]. Regulated expression may therefore be either positive or negative, thereby either enhancing or reducing transcription. Sequences encoding metabolic pathway enzymes provide particularly useful promoter sequences. Examples include promoter sequences derived from sugar metabolizing enzymes, such as galactose, lactose (lac) [Chang et al. (1977) Nature 798: 1056], and maltose. Additional examples include promoter sequences derived from biosynthetic enzymes such as tryptophan (trp) [Goeddel et al. ( 1980) Nuc. Acids Res. 8:4057 ; Yelverton et al. (1981 ) Nucl. Acids Res. 9:731 ; US patent 4,738,921 ; EP-A-0036776 and EP-A -0121775]. The g-laotamase (bla) promoter system [W eissmann (1981 ) "The cloning of interferon and other mistakes." In Interferon 5 (ed. I. Gresser)], bacteriophage lambda PL [Shimatake et al. (1981) Nature 292: 128] and T5 [US patent 4,689,406] promoter systems also provide useful promoter sequences. In addition, synthetic promoters which do not occur in nature also function as bacterial promoters. For example, transcription activation sequences of one bacterial or bacteriophage promoter may be joined with the operon sequences of another bacterial or bacteriophage promoter, creating a synthetic hybrid promoter [US patent 4,551 ,433]. For example, the tac promoter is a hybrid trp-lac promoter comprised of both trp promoter and lac operon sequences that is regulated by the lac repressor [Amann et al. (1983) Gene 25:167; de Boer et al. (1983) Proc. Natl. Acad. Sci. 80:21 ]. Furthermore, a bacterial promoter can include naturally occurring promoters of non-bacterial origin that have the ability to bind bacterial RNA polymerase and initiate transcription. A naturally occurring promoter of non-bacterial origin can also be coupled with a compatible RNA polymerase to produce high levels of expression of some genes in prokaryotes. The bacteriophage T7 RNA polymerase/promoter system is an example of a coupled promoter system [Studier et al. (1986) J. Mol. Biol. 789: 1 13 ; Tabor et al. (1985) Proc Natl. Acad. Sci. 82:1074]. In addition, a hybrid promoter can also be comprised of a bacteriophage promoter and an E. col; operator region (EPO-A-O 267 851 ). In addition to a functioning promoter sequence, an efficient ribosome binding site is also useful for the expression of foreign genes in prokaryotes. In E. coli, the ribosome binding site is called the Shine- Dalgarno (SD) sequence and includes an initiation codon (ATG) and a sequence 3-9 nucleotides in length located 3-1 1 nucleotides upstream of the initiation codon [Shine et al. (1975) Nature 254:34]. The SD sequence is thought to promote binding of mRNA to the ribosome by the pairing of bases between the SD sequence and the 3' and of E. coli 16S rRNA [Steitz et al. (1979) "Genetic signals and nucleotide sequences in messenger RNA ." In Biological Regulation and Development: Gene Expression (ed. R.F. Goldberger)]. To express eukaryotic genes and prokaryotic genes with weak riboso e-binding site [Sambrook et al. (1989) "Expression of cloned genes in Escherichia coli." In Molecular Cloning: A Laboratory Manual]. A DNA molecule may be expressed intracellularly. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus will always be a methionine, which is encoded by the A TG start codon. If desired, methionine at the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide or by either in vivo on in vitro incubation with a bacterial methionine N-terminal peptidase (EPO-A-O 219 237). Fusion proteins provide an alternative to direct expression. Usually, a DNA sequence encoding the N- terminal portion of an endogenous bacterial protein, or other stable protein, is fused to the 5' end of heterologous coding sequences. Upon expression, this construct will provide a fusion of the two amino acid sequences. For example, the bacteriophage lambda cell gene can be linked at the 5' terminus of a foreign gene and expressed in bacteria. The resulting fusion protein preferably retains a site for a processing enzyme (factor Xa) to cleave the bacteriophage protein from the foreign gene [Nagai et al. (1984) Nature 309:810]. Fusion proteins can also be made with sequences from the lad [Jia et al. (1987) Gene 60:197], trpE [Allen et al. (1987) J. B iotechnol. 5 :93 ; M akoff et al. (1989) J. Gen. Microbiol. 755:1 1 ], and Chey [EP-A-O 324 647] genes. The DNA sequence at the junction of the two amino acid sequences may or may not encode a cleavable site. Another example is a ubiquitin fusion protein. Such a fusion protein is made with the ubiquitin region that preferably retains a site for a processing enzyme (eg. ubiquitin specific processing-protease) to cleave the ubiquitin from the foreign protein. Through this method, native foreign protein can be isolated [M iller et al. (1989) Bio/Technology 7:698]. Alternatively, foreign proteins can also be secreted from the cell by creating chimeric DNA molecules that encode a fusion protein comprised of a signal peptide sequence fragment that provides for secretion of the foreign protein in bacteria [US patent 4,336,336]. The signal sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the secretion of the protein from the cell. The protein is either secreted into the growth media (gram-positive bacteria) or into the periplasmic space, located between the inner and outer membrane of the cell (gram-negative bacteria). Preferably there are processing sites, which can be cleaved either in vivo or in vitro encoded between the signal peptide fragment and the foreign gene. DNA encoding suitable signal sequences can be derived from genes for secreted bacterial proteins, such as the E. coli outer membrane protein gene (ompA) [M asui et al. (1983), in: Experimental Manipulation of Gene Expression; Ghrayeb et al. (1984) EMBO J. 3:2437] and the E. coli alkaline phosphatase signal sequence (phoA) [Oka et al. (1985) Proc. Natl. Acad. Sci. 82:7212]. A s an additional example, the signal sequence of the alpha-am ylase gene from various B acillus strains can be used to secrete heterologous proteins from B. subtilis [Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 244 042]. Usually, transcription termination sequences recognized by bacteria are regulatory regions located 3' to the translation stop codon, and thus together with the promoter flank the coding sequence. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA . Transcription termination sequences frequently include DNA sequences of about 50 nucleotides capable of forming stem loop structures that aid in terminating transcription. Examples include transcription termination sequences derived from genes with strong promoters, such as the trp gene in E. coli as well as other biosynthetic genes. Usually, the above described components, comprising a promoter, signal sequence (if desired), coding sequence of interest, and transcription termination sequence, are put together into expression constructs. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as bacteria. The replicon will have a replication system, thus allowing it to be maintained in a prokaryotic host either for expression or for cloning and amplification. In addition, a replicon may be either a high or low copy number plasmid. A high copy number plasmid will generally have a copy number ranging from about 5 to about 200, and usually about 10 to about 150. A host containing a high copy number plasmid will preferably contain at least about 10, and more preferably at least about 20 plasmids. Either a high or low copy number vector may be selected, depending upon the effect of the vector and the foreign protein on the host. Alternatively, the expression constructs can be integrated into the bacterial genome with an integrating vector. Integrating vectors usually contain at least one sequence homologous to the bacterial chromosome that allows the vector to integrate. Integrations appear to result from recombinations between homologous DNA in the vector and the bacterial chromosome. For example, integrating vectors constructed with DNA from various B acillus strains integrate into the B acillus chromosome (EP-A- 0 127 328). Integrating vectors may also be comprised of bacteriophage or transposon sequences. Usually, extrachromosomal and integrating expression constructs may contain selectable markers to allow for the selection of bacterial strains that have been transformed. Selectable markers can be expressed in the bacterial host and may include genes which render bacteria resistant to drugs such as ampicillin, chloramphenicol, erythromycin, kanamycin (neomycin), and tetracycline [Davies et al. (1978) Annu. Rev. Microbiol. 52:469]. Selectable markers may also include biosynthetic genes, such as those in the histidine, tryptophan, and leucine biosynthetic pathways. A lternatively, some of the above described components can be put together in transformation vectors. Transformation vectors are usually comprised of a selectable market that is either maintained in a replicon or developed into an integrating vector, as described above. Expression and transformation vectors, either extra-chromosomal replicons or integrating vectors, have been developed for transformation into many bacteria. For example, expression vectors have been developed for, inter alia, the following bacteria: B acillus subtilis [Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 036 259 and EP-A -0 063 953 ; W O 84/04541 ], Escherichia coli [Shimatake et al. (1981) Nature 292: 128; Amann et al. (1985) Gene 40:183 ; Studier et al. (1986) J. Mol. Biol. 789:1 13 ; EP-A-0 036 776,EP-A-0 136 829 and EP-A-0 136 907], Streptococcus cremoris [Powell et al. (1988) Appl. Environ. Microbiol. 54:655]; Streptococcus lividans [Powell et al. (1988) Appl. Environ. Microbiol. 54:655], Streptom yces lividans [US patent 4,745,056]. M ethods of introducing exogenous DNA into bacterial hosts are well-known in the art, and usually include either the transformation of bacteria treated with CaCh or other agents, such as divalent cations and DM SO. DNA can also be introduced into bacterial cells by electroporation. Transformation procedures usually vary with the bacterial species to be transformed. See eg. [M asson et al. (1989) FEMS Microbiol. Lett. 60:273 ; Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 036 259 and EP- A-0 063 953; W O 84/04541 , Bacillus], [M iller et al. (1988) Proc. Natl. Acad. Sci. 85:856; W ang et al. (1990) J. Bacterial. 772:949, Campylobacter], [Cohen et al. (1973) Proc. Natl. Acad. Sci. 69:21 10; Dower et al. (1988) Nucleic Acids Res. 76:6127; Kushner (1978) "An improved method for transformation of Escherichia coli with ColE l -derived plasmids. In Genetic Engineering: Proceedings of the International Symposium on Genetic Engineering (eds. H .W . B oyer and S . Nicosia); M andel et al. (1970) J. Mol. Biol. 55:159; Taketo (1988) Biochim. Biophys. Acta 949:318 ; Escherichia], [Chassy et al. (1987) FEMS Microbiol. Lett. 44:173 Lactobacillus]; [Fiedler et al. (1988) Anal. Biochem 770:38, Pseudomonas]; [Augustin et al. (1990) FEMS Microbiol. Lett. 66:203, Staphylococcus], [Barany et al. (1980) J. Bacteriol. 744:698; Harlander (1987) "Transformation of Streptococcus lactis by electroporation, in: Streptococcal Genetics (ed. J. Ferretti and R. Curtiss III); Perry et al. (1981 ) Infect. Immun. 52:1295 ; Powell et al. (1988) Appl. Environ. Microbiol. 54:655; Somkuti et al. (1987) Proc. 4th Evr. Cong. Biotechnology 7 :412, Streptococcus]. v. Yeast Expression Yeast expression systems are also known to one of ordinary skill in the art. A yeast promoter is any DNA sequence capable of binding yeast RNA polymerase and initiating the downstream (3') transcription of a coding sequence (eg. structural gene) into mRNA . A promoter will have a transcription initiation region which is usually placed proximal to the 5' end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site (the "TA TA B ox") and a transcription initiation site. A yeast promoter may also have a second domain called an upstream activator sequence (UAS), which, if present, is usually distal to the structural gene. The UA S permits regulated (inducible) expression. Constitutive expression occurs in the absence of a UAS. Regulated expression may be either positive or negative, thereby either enhancing or reducing transcription. Yeast is a fermenting organism with an active metabolic pathway, therefore sequences encoding enzymes in the metabolic pathway provide particularly useful promoter sequences. Examples include alcohol dehydrogenase (AD H) (EP-A -0 284 044), enolase, glucokinase, glucose-6-phosphate isomerase, glyceraldehyde-3-phosphate-dehydrogenase (GAP or GA PDH), hexokinase, phosphofructokinase, 3- phosphoglycerate mutase, and pyruvate kinase (PyK) (EPO-A-O 329 203). The yeast PH05 gene, encoding acid phosphatase, also provides useful promoter sequences [M yanohara et al. (1983) Proc. Natl. Acad. Sci. USA 80:1 ]. In addition, synthetic promoters which do not occur in nature also function as yeast promoters. For example, UA S sequences of one yeast promoter may be joined with the transcription activation region of another yeast promoter, creating a synthetic hybrid promoter. Examples of such hybrid promoters include the ADH regulatory sequence linked to the GAP transcription activation region (US Patent Nos. 4,876,197 and 4,880,734). Other examples of hybrid promoters include promoters which consist of the regulatory sequences of either the ADH2, GAL4, GAL10, OR PH05 genes, combined with the transcriptional activation region of a glycolytic enzyme gene such as GAP or PyK (EP-A-0 164 556). Furthermore, a yeast promoter can include naturally occurring promoters of non-yeast origin that have the ability to bind yeast RNA polymerase and initiate transcription. Examples of such promoters include, inter alia, [Cohen et al. (1980) Proc. Natl. Acad. Sci. USA 77:1078; Henikoff et al. (1981) Nature 285:835 ; Hollenberg et al. (1981 ) Curr. Topics Microbiol. Immunol. 96: 1 19; Hollenberg et al. ( 1979) "The Expression of B acterial Antibiotic Resistance Genes in the Yeast Saccharomyces cerevisiae," in: Plasmids of Medical, Environmental and Commercial Importance (eds. K.N . Timmis and A. Puhler); M ercerau-Puigalon et al. (1980) Gene 77 :163 ; Panthier et α/. (1980) Curr. Genet. 2: 109;]. A DNA molecule may be expressed intracellularly in yeast. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus of the recombinant protein will always be a methionine, which is encoded by the ATG start codon. If desired, methionine at the N- terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide. Fusion proteins provide an alternative for yeast expression systems, as well as in mammalian, baculovirus, and bacterial expression systems. Usually, a DNA sequence encoding the N-terminal portion of an endogenous yeast protein, or other stable protein, is fused to the 5' end of heterologous coding sequences. Upon expression, this construct will provide a fusion of the two amino acid sequences. For example, the yeast or human superoxide dismutase (SOD) gene, can be linked at the 5' terminus of a foreign gene and expressed in yeast. The DNA sequence at the junction of the two amino acid sequences may or may not encode a cleavable site. See eg. EP-A -0 196 056. Another example is a ubiquitin fusion protein. Such a fusion protein is made with the ubiquitin region that preferably retains a site for a processing enzyme (eg. ubiquitin-specific processing protease) to cleave the ubiquitin from the foreign protein. Through this method, therefore, native foreign protein can be isolated (eg. W O88/024066). Alternatively, foreign proteins can also be secreted from the cell into the growth media by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provide for secretion in yeast of the foreign protein. Preferably, there are processing sites encoded between the leader fragment and the foreign gene that can be cleaved either in vivo or in vitro. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic am ino acids which direct the secretion of the protein from the cell. DNA encoding suitable signal sequences can be derived from genes for secreted yeast proteins, such as the yeast invertase gene (EP-A -0 012 873; JPO . 62,096,086) and the A -factor gene (U S patent 4,588,684). Alternatively, leaders of non-yeast origin, such as an interferon leader, exist that also provide for secretion in yeast (EP-A -0 060 057). A preferred class of secretion leaders are those that employ a fragment of the yeast alpha-factor gene, which contains both a "pre" signal sequence, and a "pro" region. The types of alpha-factor fragments that can be employed include the full-length pre-pro alpha factor leader (about 83 amino acid residues) as well as truncated alpha-factor leaders (usually about 25 to about 50 amino acid residues) (US Patents 4,546,083 and 4,870,008 ; EP-A-0 324 274). Additional leaders employing an alpha-factor leader fragment that provides for secretion include hybrid alpha-factor leaders made with a presequence of a first yeast, but a pro-region from a second yeast alphafactor. (eg. see W O 89/02463.) U sually, transcription termination sequences recognized by yeast are regulatory regions located 3' to the translation stop codon, and thus together with the promoter flank the coding sequence. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA . Examples of transcription terminator sequence and other yeast-recognized termination sequences, such as those coding for glycoly tic enzymes. Usually, the above described components, comprising a promoter, leader (if desired), coding sequence of interest, and transcription termination sequence, are put together into expression constructs. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as yeast or bacteria. The replicon may have two replication systems, thus allowing it to be maintained, for example, in yeast for expression and in a prokaryotic host for cloning and amplification. Examples of such yeast-bacteria shuttle vectors include YEp24 [Botstein et al. (1979) Gene 8:17-24], pC l/1 [Brake et al. (1984) Proc. Natl. Acad. Sci USA 87 :4642-4646], and YRp l 7 [Stinchcomb et al. (1982) J. Mol. Biol. 758: 157]. In addition, a replicon may be either a high or low copy number plasmid. A high copy number plasmid w ill generally have a copy number ranging from about 5 to about 200, and usually about 10 to about 150. A host containing a high copy number plasmid will preferably have at least about 10, and more preferably at least about 20. Enter a high or low copy number vector may be selected, depending upon the effect of the vector and the foreign protein on the host. See eg. Brake et al, supra. Alternatively, the expression constructs can be integrated into the yeast genome with an integrating vector. Integrating vectors usually contain at least one sequence homologous to a yeast chromosome that allows the vector to integrate, and preferably contain two homologous sequences flanking the expression construct. Integrations appear to result from recombinations between homologous DNA in the vector and the yeast chromosome [Orr-W eaver et al. (1983) Methods in Enzymol. 707 :228-245]. An integrating vector may be directed to a specific locus in yeast by selecting the appropriate homologous sequence for inclusion in the vector. See Orr-W eaver et al., supra. One or more expression construct may integrate, possibly affecting levels of recombinant protein produced [Rine et al. ( 1983) Proc. Natl. Acad. Sci. USA 80:6750]. The chromosomal sequences included in the vector can occur either as a single segment in the vector, which results in the integration of the entire vector, or two segments homologous to adjacent segments in the chromosome and flanking the expression construct in the vector, which can result in the stable integration of only the expression construct. Usually, extrachromosomal and integrating expression constructs may contain selectable markers to allow for the selection of yeast strains that have been transformed. Selectable markers may include biosynthetic genes that can be expressed in the yeast host, such as ADE2, HIS4, LEU2, TRP1 , and ALG 7, and the G418 resistance gene, which confer resistance in yeast cells to tunicamycin and G418, respectively. In addition, a suitable selectable marker may also provide yeast with the ability to grow in the presence of toxic compounds, such as metal. For example, the presence of CUP1 allows yeast to grow in the presence of copper ions [B utt et al. (1987) Microbiol, Rev. 57 :351 ]. Alternatively, some of the above described components can be put together into transformation vectors. Transformation vectors are usually comprised of a selectable marker that is either maintained in a replicon or developed into an integrating vector, as described above. Expression and transformation vectors, either extrachromosomal replicons or integrating vectors, have been developed for transformation into many yeasts. For example, expression vectors have been developed for, inter alia, the following yeasts:Candida albicans [Kurtz, et al. (1986) Mol. Cell. Biol. 6:142], Candida maltosa [Kunze, et al. (1985) J. Basic Microbiol. 25:141 ], Hansenula polymorpha [Gleeson, et al. (1986) J. Gen. Microbiol. 752:3459; Roggenkamp et al. (1986) Mol. Gen. Genet. 202:302], Kluyveromyces fragilis [Das, et al. (1984) J. Bacteriol. 758:1 165], Kluyverom yces lactis [De Louvencourt et al. (1983) J. Bacteriol. 754:737 ; Van den Berg et al. (1990) Bio/Technology 8:135], Pichia guillerimondii [Kunze et al. (1985) /. Basic Microbiol. 25: 141 ], Pichia pastoris [Cregg, et al. (1985) Mol. Cell. Biol. 5:3376; US Patent Nos. 4,837 ,148 and 4,929,555], Saccharomyces cerevisiae [Hinnen et al. (1978) Proc. Natl. Acad. Sci. USA 75: 1929; Ito et al. (1983) J. Bacteriol. 755: 163], Schizosaccharomyces pombe [Beach and Nurse (1981) Nature 500:706], and Yarrowia lipolytica [Davidow , et al. (1985) Curr. Genet. 70:380471 Gaillardin, et al. (1985) Curr. Genet. 70:49]. M ethods of introducing exogenous DNA into yeast hosts are well-know n in the art, and usually include either the transformation of spheroplasts or of intact yeast cells treated with alkali cations. Transformation procedures usually vary with the yeast species to be transformed. See eg. [Kurtz et al. (1986) Mol. Cell. Biol. 6: 142; Kunze et al. (1985) /. Basic Microbiol. 25: 141 ; Candida]; [Gleeson et al. (1986) J. Gen. Microbiol. 752:3459; Roggenkamp et al. (1986) Mol. Gen. Genet. 202:302; Hansenula]; [Das et α/. (1984) 7. Bacteriol. 758:1 165 ; De Louvencourt et al. (1983) 7. Bacteriol. 754:1 165 ; Van den Berg et al. (1990) Bio/Technology 8:135; Kluyveromyces]; [Cregg et al. (1985) Mol. Cell. Biol. 5:3376; Kunze et al. (1985) J. Basic Microbiol. 25:141 ; US Patent Nos. 4,837,148 and 4,929,555 ; Pichia]; [H innen et al. (1978) Proc. Natl. Acad. Sci. USA 75;1929; Ito et al. (1983) /. Bacteriol. 755:163 Saccharomyces]; [B each and Nurse (1981 ) Nature 500:706; Schizosaccharomyces]; [D avidow et al. ( 1985) Curr. Genet. 70:39; Gaillardin et al. (1985) Curr. Genet. 70:49; Yarrowia], Antibodies A s used herein, the term "antibody" refers to a polypeptide or group of polypeptides composed of at least one antibody combining site. An "antibody combining site" is the three-dimensional binding space with an internal surface shape and charge distribution complementary to the features of an epitope of an antigen, which allows a binding of the antibody with the antigen. "Antibody" includes, for example, vertebrate antibodies, hybrid antibodies, chimeric antibodies, humanised antibodies, altered antibodies, univalent antibodies, Fab proteins, and single domain antibodies. Antibodies against the proteins of the invention are useful for affinity chromatography, immunoassays, and distinguishing/identifying Neisserial proteins. Antibodies to the proteins of the invention, both polyclonal and monoclonal, may be prepared by conventional methods. In general, the protein is first used to imm unize a suitable animal, preferably a mouse, rat, rabbit or goat. Rabbits and goats are preferred for the preparation of polyclonal sera due to the volume of serum obtainable, and the availability of labeled anti-rabbit and anti-goat antibodies. Immunization is generally performed by mixing or emulsifying the protein in saline, preferably in an adjuvant such as Freund' s complete adjuvant, and injecting the mixture or emulsion parenterally (generally subcutaneously or intramuscularly). A dose of 50-200 μg/injection is typically sufficient. Immunization is generally boosted 2-6 weeks later with one or more injections of the protein in saline, preferably using Freund's incomplete adjuvant. One may alternatively generate antibodies by in vitro immunization using methods known in the art, which for the purposes of this invention is considered equivalent to in vivo immunization. Polyclonal antisera is obtained by bleeding the immunized animal into a glass or plastic container, incubating the blood at 25 °C for one hour, followed by incubating at 4°C for 2-18 hours. The serum is recovered by centrifugation (eg. l ,000g for 10 minutes). About 20-50 ml per bleed may be obtained from rabbits. M onoclonal antibodies are prepared using the standard method of Kohler & M ilstein [Nature (1975) 256:495-96], or a modification thereof. Typically, a mouse or rat is immunized as described above. However, rather than bleeding the animal to extract serum, the spleen (and optionally several large lymph nodes) is removed and dissociated into single cells. If desired, the spleen cells may be screened (after removal of nonspecifically adherent cells) by applying a cell suspension to a plate or well coated with the protein antigen. B -cells expressing membrane-bound immunoglobulin specific for the antigen bind to the plate, and are not rinsed away with the rest of the suspension. Resulting B -cells, or all dissociated spleen cells, are then induced to fuse w ith myeloma cells to form hybridomas, and are cultured in a selective medium (eg. hypoxanthine, aminopterin, thymidine medium, "HAT"). The resulting hybridomas are plated by limiting dilution, and are assayed for the production of antibodies which bind specifically to the immunizing antigen (and which do not bind to unrelated antigens). The selected M Ab-secreting hybridomas are then cultured either in vitro (eg. in tissue culture bottles or hollow fiber reactors), or in vivo (as ascites in mice). If desired, the antibodies (whether polyclonal or monoclonal) may be labeled using conventional techniques. Suitable labels include fluorophores, chromophores, radioactive atoms (particularly P and l 25 I), electron-dense reagents, enzymes, and ligands having specific binding partners. Enzymes are typically detected by their activity. For example, horseradish peroxidase is usually detected by its ability to convert 3,3',5 ,5'-tetramethylbenzidine (TM B) to a blue pigment, quantifiable with a spectrophotometer. "Specific binding partner" refers to a protein capable of binding a ligand molecule with high specificity, as for example in the case of an antigen and a monoclonal antibody specific therefor. Other specific binding partners include biotin and avidin or streptavidin, IgG and protein A , and the numerous receptor-ligand couples known in the art. It should be understood that the above description is not meant to categorize the various labels into distinct classes, as the same label may serve in several different modes. For example, 125 I may serve as a radioactive label or as an electron-dense reagent. HRP may serve as enzyme or as antigen for a M Ab. Further, one may combine various labels for desired effect. For example, M Abs and avidin also require labels in the practice of this invention: thus, one might label a M Ab with biotin, and detect its presence with avidin labeled with l25 I, or with an anti-biotin M Ab labeled w ith HRP. Other permutations and possibilities will be readily apparent to those of ordinary skill in the art, and are considered as equivalents w ithin the scope of the instant invention. Pharmaceutical Compositions Pharmaceutical compositions can comprise either polypeptides, antibodies, or nucleic acid of the invention. The pharmaceutical compositions will comprise a therapeutically effective amount of either polypeptides, antibodies, or polynucleotides of the claimed invention. The term "therapeutically effective amount" as used herein refers to an amount of a therapeutic agent to treat, ameliorate, or prevent a desired disease or condition, or to exhibit a detectable therapeutic or preventative effect. The effect can be detected by, for example, chem ical markers or antigen levels. Therapeutic effects also include reduction in physical symptoms, such as decreased body temperature. The precise effective amount for a subject will depend upon the subject's size and health, the nature and extent of the condition, and the therapeutics or combination of therapeutics selected for administration. Thus, it is not useful to specify an exact effective amount in advance. However, the effective amount for a given situation can be determined by routine experimentation and is within the judgement of the clinician. For purposes of the present invention, an effective dose w ill be from about 0.01 mg/ kg to 50 mg/kg or 0.05 mg/kg to about 10 mg/kg of the DNA constructs in the individual to which it is administered. A pharmaceutical composition can also contain a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to a carrier for administration of a therapeutic agent, such as antibodies or a polypeptide, genes, and other therapeutic agents. The term refers to any pharmaceutical carrier that does not itself induce the production of antibodies harmful to the individual receiving the composition, and which may be administered without undue toxicity. Suitable carriers may be large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, and inactive virus particles. Such carriers are well known to those of ordinary skill in the art. Pharmaceutically acceptable salts can be used therein, for example, mineral acid salts such as hydrochlorides, hydrobromides, phosphates, sulfates, and the like; and the salts of organic acids such as acetates, propionates, malonates, benzoates, and the like. A thorough discussion of pharmaceutically acceptable excipients is available in Remington's Pharmaceutical Sciences (M ack Pub. Co, N.J. 1991 ). Pharmaceutically acceptable carriers in therapeutic compositions may contain liquids such as water, saline, glycerol and ethanol. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, and the like, may be present in such vehicles. Typically, the therapeutic compositions are prepared as injectables, either as liquid solutions or suspensions; solid form s suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. Liposomes are included within the definition of a pharmaceutically acceptable carrier. Delivery Methods Once formulated, the compositions of the invention can be administered directly to the subject. The subjects to be treated can be animals; in particular, human subjects can be treated. Direct delivery of the compositions will generally be accomplished by injection, either subcutaneously, intraperitoneally, intravenously or intramuscularly or delivered to the interstitial space of a tissue. The compositions can also be administered into a lesion. Other modes of administration include oral and pulmonary administration, suppositories, and transdermal or transcutaneous applications (eg. see W O98/20734), needles, and gene guns or hyposprays. Dosage treatment may be a single dose schedule or a multiple dose schedule. Vaccines Vaccines comprise immunising antigen(s), immunogen(s), polypeptide(s), protein(s) or nucleic acid, usually in combination with "pharmaceutically acceptable carriers," which include any carrier that does not itself induce the production of antibodies harmful to the individual receiving the composition. Suitable carriers are typically large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, am ino acid copolymers, lipid aggregates (such as oil droplets or liposomes), and inactive virus particles. Such carriers are well known to those of ordinary skill in the art. Additionally, these carriers may function as immunostimulating agents ("adjuvants"). Furthermore, the antigen or immunogen may be conjugated to a bacterial toxoid, such as a toxoid from diphtheria, tetanus, cholera, 77. pylori, etc. pathogens. Preferred adjuvants to enhance effectiveness of the composition include, but are not limited to: (1) aluminum salts (alum), such as aluminum hydroxide, aluminum phosphate, aluminum sulfate, etc; (2) oil-in-water emulsion formulations (with or without other specific immunostimulating agents such as muramyl peptides (see below) or bacterial cell wall components), such as for example (a) M F59™ (W O 90/14837; Chapter 10 in Vaccine design: the subunit and adjuvant approach, eds. Powell & Newman, Plenum Press 1995), containing 5% Squalene, 0.5% Tween 80, and 0.5 % Span 85 (optionally containing various amounts of M TP-PE (see below), although not required) formulated into submicron particles using a microfluidizer such as M odel H OY microfluidizer (M icrofluidics, Newton, M A), (b) SAF, containing 10% Squalane, 0.4% Tween 80, 5% pluronic-blocked polymer L 121 , and thr-M DP (see below) either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion, and (c) Ribi™ adjuvant system (RA S), (Ribi Immunochem, Hamilton, M T) containing 2% Squalene, 0.2% Tween 80, and one or more bacterial cell wall components from the group consisting of monophosphorylipid A (M PL), trehalose dimycolate (TDM ), and cell wall skeleton (CW S), preferably T \Λ Υ Λ M PL + CW S (Detox ); (3) saponin adjuvants, such as Stimulon (Cambridge B ioscience, W orcester, M A) may be used or particles generated therefrom such as ISCOM s (immunostimulating complexes); (4) Complete Freund' s Adjuvant (CFA) and Incomplete Freund' s Adjuvant (IFA); (5) cytokines, such as interleukins (eg. IL-1 , IL-2, IL-4, IL-5, IL-6, IL-7, IL- 12, etc.), interferons (eg. gamma interferon), macrophage colony stim ulating factor (M -CSF), tumor necrosis factor (TNF), etc; and (6) other substances that act as immunostimulating agents to enhance the effectiveness of the composition. A lum and M F59™ are preferred. As mentioned above, muramyl peptides include, but are not limited to, N-acetyl-muramyl-L-threonyl-D- isoglutamine (thr-M DP), N-acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-M DP), N-acetylmuramyl- L-alanyl-D-isoglutaminyl-L-alanine-2-(l '-2'-dipalmitoyl-in-glycero-3-hydroxyphosphoryloxy)- ethylamine (M TP-PE), etc. The immunogenic compositions (eg. the immunising antigen/immunogen/polypeptide/protein/ nucleic acid, pharmaceutically acceptable carrier, and adjuvant) typically will contain diluents, such as water, saline, glycerol, ethanol, etc. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, and the like, may be present in such vehicles. Typically, the immunogenic compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. The preparation also may be emulsified or encapsulated in liposomes for enhanced adjuvant effect, as discussed above under pharmaceutically acceptable carriers. Immunogenic compositions used as vaccines comprise an immunologically effective amount of the antigenic or immunogenic polypeptides, as well as any other of the above-mentioned components, as needed. B y "immunologically effective amount", it is meant that the administration of that amount to an individual, either in a single dose or as part of a series, is effective for treatment or prevention. This amount varies depending upon the health and physical condition of the individual to be treated, the taxonomic group of individual to be treated (eg. nonhuman primate, primate, etc.), the capacity of the individual's immune system to synthesize antibodies, the degree of protection desired, the formulation of the vaccine, the treating doctor's assessment of the medical situation, and other relevant factors. It is expected that the amount will fall in a relatively broad range that can be determined through routine trials. The immunogenic compositions are conventionally administered parenterally, eg. by injection, either subcutaneously, intramuscularly, or transdermally/transcutaneously (eg. W 098/20734). Additional formulations suitable for other modes of administration include oral and pulmonary formulations, suppositories, and transdermal applications. Dosage treatment may be a single dose schedule or a m ultiple dose schedule. The vaccine may be administered in conjunction with other immunoregulatory agents. A s an alternative to protein-based vaccines, DNA vaccination may be employed [eg. Robinson & Torres (1997) Seminars in Immunology 9:271 -283 ; Donnelly et al. (1997) Annu Rev Immunol 15 :617-648; see later herein]. Gene Delivery Vehicles Gene therapy vehicles for delivery of constructs including a coding sequence of a therapeutic of the invention, to be delivered to the mammal for expression in the mammal, can be administered either locally or systemically. These constructs can utilize viral or non-viral vector approaches in in vivo or ex vivo modality. Expression of such coding sequence can be induced using endogenous mammalian or heterologous promoters. Expression of the coding sequence in vivo can be either constitutive or regulated. The invention includes gene delivery vehicles capable of expressing the contemplated nucleic acid sequences. The gene delivery vehicle is preferably a viral vector and, more preferably, a retroviral, adenoviral, adeno-associated viral (AAV), herpes viral, or alphavirus vector. The viral vector can also be an astrovirus, coronavirus, orthomyxovirus, papovavirus, paramyxovirus, parvovirus, picornavirus, poxvirus, or togavirus viral vector. See generally, Jolly (1994) Cancer Gene Therapy 1 :51-64; Kimura (1994) Human Gene Therapy 5:845-852; Connelly (1995) Human Gene Therapy 6:185-193 ; and Kaplitt (1994) Nature Genetics 6:148-153. Retroviral vectors are well known in the art and we contemplate that any retroviral gene therapy vector is employable in the invention, including B , C and D type retroviruses, xenotropic retroviruses (for example, NZB-X 1 , NZB -X2 and NZB 9-1 (see O'Neill (1985) J. Virol. 53 : 160) polytropic retroviruses eg. M CF and M CF-M LV (see Kelly (1983) J. Virol. 45 :291 ), spumaviruses and lentiviruses. See RNA Tumor Viruses, Second Edition, Cold Spring H arbor Laboratory, 1985. Portions of the retroviral gene therapy vector may be derived from different retroviruses. For example, retrovector LTRs may be derived from a M urine Sarcoma Virus, a tRNA binding site from a Rous Sarcoma Virus, a packaging signal from a M urine Leukemia Virus, and an origin of second strand synthesis from an Avian Leukosis Virus. These recombinant retroviral vectors may be used to generate transduction competent retroviral vector particles by introducing them into appropriate packaging cell lines (see US patent 5,591 ,624). Retrovirus vectors can be constructed for site-specific integration into host cell DNA by incorporation of a chimeric integrase enzyme into the retroviral particle (see W 096/37626). It is preferable that the recombinant viral vector is a replication defective recombinant virus. Packaging cell lines suitable for use with the above-described retrovirus vectors are well known in the art, are readily prepared (see W O95/30763 and W O92/05266), and can be used to create producer cell lines (also termed vector cell lines or "VCLs") for the production of recombinant vector particles. Preferably, the packaging cell lines are made from human parent cells (eg. H T 1080 cells) or mink parent cell lines, which eliminates inactivation in human serum . Preferred retroviruses for the construction of retroviral gene therapy vectors include Avian Leukosis V irus, Bovine Leukemia, V irus, M urine Leukemia Virus, M ink-Cell Focus-Inducing Virus, M urine Sarcoma Virus, Reticuloendotheliosis Virus and Rous Sarcoma V irus. Particularly preferred M urine Leukemia Viruses include 4070A and 1504A (H artley and Rowe ( 1976) J Virol 19:19-25), Abelson (ATCC No. VR-999), Friend (ATCC No. VR-245), Graffi, Gross (ATCC Nol VR-590), Kirsten, Harvey Sarcoma Virus and Rauscher (ATCC No. VR-998) and M oloney M urine Leukemia Virus (ATCC No. VR-190). Such retroviruses may be obtained from depositories or collections such as the American Type Culture Collection ("ATCC") in Rockville, M aryland or isolated from known sources using commonly available techniques. Exemplary known retroviral gene therapy vectors employable in this invention include those described in patent applications GB 2200651 , EP0415731 , EP0345242, EP0334301 , W O89/02468; W O89/05349, W O89/09271 , W O90/02806, W O90/07936, W O94/03622, W 093/25698, W 093/25234, W O93/1 1230, W O93/10218, W O91/02805, W O91/02825, W O95/07994, US 5,219,740, U S 4,405,712, US 4,861 ,719, US 4,980,289, US 4,777,127, US 5,591 ,624. See also Vile (1993) Cancer Res 53 :3860-3864; Vile (1993) Cancer Res 53 :962-967 ; Ram (1993) Cancer Res 53 (1993) 83-88; Takamiya (1992) J Neurosci Res 33:493-503; Baba (1993) J Neurosurg 79:729-735; M ann (1983) Cell 33:153; Cane (1984) Proc Natl Acad Sci 81 :6349; and M iller (1990) Human Gene Therapy 1. Human adenoviral gene therapy vectors are also known in the art and employable in this invention. See, for example, Berkner (1988) Biotechniques 6:616 and Rosenfeld (1991) Science 252:431 , and W O93/07283, W O93/06223, and W O93/07282. Exemplary known adenoviral gene therapy vectors employable in this invention include those described in the above referenced documents and in W 094/12649, W O93/03769, W 093/19191 , W 094/28938, W 095/1 1984, W O95/00655, W O95/27071 , W 095/29993, W 095/34671 , W O96/05320, W O94/08026, W O94/1 1506, W O93/06223, W 094/24299, W O95/14102, W 095/24297, W O95/02697, W 094/28152, W 094/24299, W O95/09241 , W O95/25807, W O95/05835, W 094/18922 and W O95/09654. A lternatively, administration of DNA linked to killed adenovirus as described in Curiel (1992) Hum. Gene Ther. 3: 147-154 may be employed. The gene delivery vehicles of the invention also include adenovirus associated virus (AA V ) vectors. Leading and preferred examples of such vectors for use in this invention are the AAV-2 based vectors disclosed in Srivastava, W O93/09239. M ost preferred AAV vectors comprise the two AAV inverted terminal repeats in which the native D-sequences are modified by substitution of nucleotides, such that at least 5 native nucleotides and up to 18 native nucleotides, preferably at least 10 native nucleotides up to 18 native nucleotides, most preferably 10 native nucleotides are retained and the remaining nucleotides of the D-sequence are deleted or replaced with non-native nucleotides. The native D-sequences of the AAV inverted terminal repeats are sequences of 20 consecutive nucleotides in each AAV inverted term inal repeat (ie. there is one sequence at each end) which are not involved in HP formation. The non-native replacement nucleotide may be any nucleotide other than the nucleotide found in the native D-sequence in the same position. Other employable exemplary AAV vectors are pW P-19, pW N-1 , both of which are disclosed in Nahreini (1993) Gene 124:257-262. Another example of such an AAV vector is psub201 (see Samulski (1987) J. Virol. 61 :3096). Another exemplary AAV vector is the Double-D ITR vector. Construction of the Double-D ITR vector is disclosed in US Patent 5 ,478,745. Still other vectors are those disclosed in Carter US Patent 4,797,368 and M uzyczka US Patent 5,139,941 , Chartejee US Patent 5,474,935, and Kotin W 094/288157. Yet a further example of an AAV vector employable in this invention is SSV9AFAB TKneo, which contains the AFP enhancer and albumin promoter and directs expression predominantly in the liver. Its structure and construction are disclosed in Su (1996) Human Gene Therapy 7:463-470. Additional AAV gene therapy vectors are described in U S 5,354,678, US 5,173,414, US 5,139,941 , and US 5,252,479. The gene therapy vectors of the invention also include herpes vectors. Leading and preferred examples are herpes simplex virus vectors containing a sequence encoding a thymidine kinase polypeptide such as those disclosed in US 5,288,641 and EP0176170 (Roizman). Additional exemplary herpes simplex virus vectors include HFEM/ICP6-LacZ disclosed in WO95/04139 (Wistar Institute), pHSVlac described in Geller (1988) Science 241:1667-1669 and in WO90/09441 and WO92/07945, HSV Us3::pgC-lacZ described in Fink (1992) 77«mαn Gene Therapy 3:11-19 and HSV 7134, 2 RH 105 and GAL4 described in EP 0453242 (Breakefield), and those deposited with the ATCC as accession numbers ATCC VR-977 and ATCC VR-260. Also contemplated are alpha virus gene therapy vectors that can be employed in this invention. Preferred alpha virus vectors are Sindbis viruses vectors. Togaviruses, Semliki Forest virus (ATCC VR-67; ATCC VR-1247), Middleberg virus (ATCC VR-370), Ross River virus (ATCC VR-373; ATCC VR-1246), Venezuelan equine encephalitis virus (ATCC VR923; ATCC VR-1250; ATCC VR-1249; ATCC VR-532), and those described in US patents 5,091,309, 5,217,879, and WO92/10578. More particularly, those alpha virus vectors described in US Serial No.08/405,627, filed March 15, 1995.W094/21792, WO92/10578, WO95/07994, US 5,091,309 and US 5,217,879 are employable. Such alpha viruses may be obtained from depositories or collections such as the ATCC in Rockville, Maryland or isolated from known sources using commonly available techniques. Preferably, alphavirus vectors with reduced cytotoxicity are used (see USSN 08/679640). DNA vector systems such as eukaryotic layered expression systems are also useful for expressing the nucleic acids of the invention. See W 095/07994 for a detailed description of eukaryotic layered expression systems. Preferably, the eukaryotic layered expression systems of the invention are derived from alphavirus vectors and most preferably from Sindbis viral vectors. Other viral vectors suitable for use in the present invention include those derived from poliovirus, for example ATCC VR-58 and those described in Evans, Nature 339 (1989) 385 and Sabin (1973) J. Biol. Standardization 1:115; rhinovirus, for example ATCC VR-1110 and those described in Arnold (1990) J Cell Biochem L401; pox viruses such as canary pox virus or vaccinia virus, for example ATCC VR-111 and ATCC VR-2010 and those described in Fisher-Hoch (1989) Proc Natl Acad Sci 86:317; Flexner (1989) Ann NY Acad Sci 569:86, Flexner (1990) Vaccine 8:17; in US 4,603,112 and US 4,769,330 and WO89/01973; SV40 virus, for example ATCC VR-305 and those described in Mulligan (1979) Nature 277:108 and Madzak (1992) J Gen Virol 73:1533; influenza virus, for example ATCC VR-797 and recombinant influenza viruses made employing reverse genetics techniques as described in US 5,166,057 and in Enami (1990) Proc Natl Acad Sci 87:3802-3805; Enami & Palese (1991)7 Virol 65:2711-2713 and Luytjes (1989) Cell 59:110, (see also McMichael (1983) NE] Med 309:13, and Yap (1978) Nature 273:238 and Nature (1979) 277:108); human immunodeficiency virus as described in EP-0386882 and in Buchschacher (1992) /. Virol.66:2731; measles virus, for example ATCC VR-67 and VR-1247 and those described in EP-0440219; Aura virus, for example ATCC VR-368; Bebaru virus, for example ATCC VR-600 and ATCC VR-1240; Cabassou virus, for example ATCC VR-922; Chikungunya virus, for example ATCC VR-64 and ATCC VR-1241; Fort Morgan Virus, for example ATCC VR-924; Getah virus, for example ATCC VR-369 and A TCC VR-1243 ; Kyzylagach virus, for example ATCC VR-927 ; M ayaro virus, for example ATCC VR-66; M ucambo virus, for example ATCC VR-580 and ATCC VR-1244; Ndumu virus, for example ATCC VR-371 ; Pixuna virus, for example ATCC VR-372 and ATCC VR-1245; Tonate virus, for example ATCC VR-925 ; Triniti virus, for example ATCC VR-469 ; Una virus, for example ATCC VR-374; W hataroa virus, for example ATCC VR-926; Y-62-33 virus, for example ATCC VR-375 ; O 'Nyong virus, Eastern encephalitis virus, for example ATCC VR-65 and ATCC VR-1242; W estern encephalitis virus, for example ATCC VR-70, ATCC VR-1251 , ATCC VR-622 and ATCC VR- 1252; and coronavirus, for example ATCC VR-740 and those described in Hamre (1966) Proc Soc Exp Biol Med 121 :190. Delivery of the compositions of this invention into cells is not limited to the above mentioned viral vectors. Other delivery methods and media may be employed such as, for example, nucleic acid expression vectors, polycationic condensed DNA linked or unlinked to killed adenovirus alone, for example see US Serial No. 08/366,787, filed December 30, 1994 and Curiel (1992) Hum Gene Ther 3 :147-154 ligand linked DNA, for example see W u (1989) J Biol Chem 264: 16985-16987, eucaryotic cell delivery vehicles cells, for example see US Serial No.08/240,030, filed M ay 9, 1994, and US Serial No. 08/404,796, deposition of photopolymerized hydrogel materials, hand-held gene transfer particle gun, as described in U S Patent 5,149,655, ionizing radiation as described in US5,206,152 and in W O92/11033, nucleic charge neutralization or fusion with cell membranes. Additional approaches are described in Philip (1994) Mol Cell Biol 14:2411 -2418 and in W offendin (1994) Proc Natl Acad Sci 91 :1581 -1585. Particle mediated gene transfer may be employed, for example see US Serial No. 60/023,867. Briefly, the sequence can be inserted into conventional vectors that contain conventional control sequences for high level expression, and then incubated with synthetic gene transfer molecules such as polymeric DNA-binding cations like polylysine, protamine, and albumin, linked to cell targeting ligands such as asialoorosomucoid, as described in W u & W u (1987) J. Biol. Chem. 262:4429-4432, insulin as described in Hucked (1990) Biochem Pharmacol 40:253-263, galactose as described in Plank (1992) Bioconjugate Chem 3:533-539, lactose or transferrin. Naked DNA may also be employed. Exemplary naked DNA introduction methods are described in W O 90/11092 and US 5,580,859. Uptake efficiency may be improved using biodegradable latex beads. DNA coated latex beads are efficiently transported into cells after endocytosis initiation by the beads. The method may be improved further by treatment of the beads to increase hydrophobicity and thereby facilitate disruption of the endosome and release of the DNA into the cytoplasm. Liposomes that can act as gene delivery vehicles are described in US 5,422,120, W 095/13796, W 094/23697, W 091/14445 and EP-524,968. As described in USSN . 60/023,867, on non-viral delivery, the nucleic acid sequences encoding a polypeptide can be inserted into conventional vectors that contain conventional control sequences for high level expression, and then be incubated with synthetic gene transfer molecules such as polymeric DNA -binding cations like polylysine, protamine, and album in, linked to cell targeting ligands such as asialoorosomucoid, insulin, galactose, lactose, or transferrin. Other delivery systems include the use of liposomes to encapsulate DNA comprising the gene under the control of a variety of tissue-specific or ubiquitously-active promoters. Further non-viral delivery suitable for use includes mechanical delivery systems such as the approach described in W offendin et al (1994) Proc. Natl. Acad. Sci. USA 91 (24):11581 -1 1585. M oreover, the coding sequence and the product of expression of such can be delivered through deposition of photopolymerized hydrogel materials. Other conventional methods for gene delivery that can be used for delivery of the coding sequence include, for example, use of hand-held gene transfer particle gun, as described in US 5,149,655 ; use of ionizing radiation for activating transferred gene, as described in US 5,206,152 and W 092/1 1033 Exemplary liposome and polycationic gene delivery vehicles are those described in US 5,422,120 and 4,762,915 ; in W O 95/13796; W 094/23697 ; and W 091/14445 ; in EP-0524968; and in Stryer, B iochemistry, pages 236-240 (1975) W .H . Freeman, San Francisco; Szoka ( 1980) Biochem Biophys Ada 600:1 ; B ayer (1979) Biochem Biophys Ada 550:464; Rivnay (1987) Meth Enzymol 149 :1 19; W ang (1987) Proc Natl Acad Sci 84:7851 ; Plant (1989) Anal Biochem 176:420. A polynucleotide composition can comprises therapeutically effective amount of a gene therapy vehicle, as the term is defined above. For purposes of the present invention, an effective dose will be from about 0.01 mg/ kg to 50 mg/kg or 0.05 mg/kg to about 10 mg/kg of the DNA constructs in the individual to which it is administered. Delivery Methods Once formulated, the polynucleotide compositions of the invention can be administered (1 ) directly to the subject; (2) delivered ex vivo, to cells derived from the subject; or (3) in vitro for expression of recombinant proteins. The subjects to be treated can be mammals or birds. Also, human subjects can be treated. Direct delivery of the compositions will generally be accomplished by injection, either subcutaneously, intraperitoneally, intravenously or intramuscularly or delivered to the interstitial space of a tissue. The compositions can also be administered into a lesion. Other modes of administration include oral and pulmonary administration, suppositories, and transdermal or transcutaneous applications (eg. see W O98/20734), needles, and gene guns or hyposprays. Dosage treatment may be a single dose schedule or a multiple dose schedule. M ethods for the ex vivo delivery and reimplantation of transformed cells into a subject are known in the art and described in eg. W 093/14778. Examples of cells useful in ex vivo applications include, for example, stem cells, particularly hematopoetic, lymph cells, macrophages, dendritic cells, or tumor cells. Generally, delivery of nucleic acids for both ex vivo and in vitro applications can be accomplished by the following procedures, for example, dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, and direct microinjection of the DNA into nuclei, all well known in the art. Polynucleotide and polypeptide pharmaceutical compositions In addition to the pharmaceutically acceptable carriers and salts described above, the following additional agents can be used with polynucleotide and/or polypeptide compositions. A. Polypeptides One example are polypeptides which include, without lim itation: asioloorosomucoid (ASOR); transferrin; asialoglycoproteins; antibodies; antibody fragments; ferritin; interleukins; interferons, granulocyte, macrophage colony stimulating factor (GM -CSF), granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M -C SF), stem cell factor and erythropoietin. Viral antigens, such as envelope proteins, can also be used. Also, proteins from other invasive organisms, such as the 17 amino acid peptide from the circumsporozoite protein of plasmodium falciparum known as RII. B .Hormones. V itamins, etc. Other groups that can be included are, for example: hormones, steroids, androgens, estrogens, thyroid hormone, or vitamins, folic acid. C .Polvalkylenes, Polysaccharides, etc. Also, polyalkylene glycol can be included with the desired polynucleotides/polypeptides. In a preferred embodiment, the polyalkylene glycol is polyethlylene glycol. In addition, mono-, di-, or polysaccharides can be included. In a preferred embodiment of this aspect, the polysaccharide is dextran or DEAE-dextran. Also, chitosan and poly(lactide-co-glycolide) D . Lipids. and Liposomes The desired polynucleotide/polypeptide can also be encapsulated in lipids or packaged in liposomes prior to delivery to the subject or to cells derived therefrom. Lipid encapsulation is generally accomplished using liposomes which are able to stably bind or entrap and retain nucleic acid. The ratio of condensed polynucleotide to lipid preparation can vary but will generally be around 1 :1 (mg DNA :micromoles lipid), or more of lipid. For a review of the use of liposomes as carriers for delivery of nucleic acids, see, Hug and Sleight (1991 ) Biochim. Biophys. Ada. 1097:1-17; Straubinger (1983) Meth. Enzymol. 101 :512-527. Liposomal preparations for use in the present invention include cationic (positively charged), anionic (negatively charged) and neutral preparations. Cationic liposomes have been shown to mediate intracellular delivery of plasmid DNA (Feigner (1987) Proc. Natl. Acad. Sci. USA 84:7413-7416); mRNA (M alone (1989) Proc. Natl. Acad. Sci. USA 86:6077-6081 ); and purified transcription factors (Debs (1990) 7. Biol. Chem. 265 :10189-10192), in functional form . Cationic liposomes are readily available. For example, N[l-2,3-dioleyloxy)propyl]-N,N,N- triethylammonium (DOTM A) liposomes are available under the trademark Lipofectin, from GIB CO B RL, Grand Island, N Y . (See, also, Feigner supra). Other commercially available liposomes include transfectace (DDAB/DOPE) and DOTAP/DOPE (B oerhinger). Other cationic liposomes can be prepared from readily available materials using techniques well known in the art. See, eg. Szoka (1978) Proc. Natl. Acad. Sci. USA 75:4194-4198; W O90/11092 for a description of the synthesis of DOTAP (l ,2-bis(oleoyloxy)-3-(trimethylammonio)propane) liposomes. Similarly, anionic and neutral liposomes are readily available, such as from Avanti Polar Lipids (B irmingham , A L), or can be easily prepared using readily available materials. Such materials include phosphatidyl choline, cholesterol, phosphatidyl ethanolam ine, dioleoylphosphatidyl choline (DOPC), dioleoylphosphatidyl glycerol (DOPG), dioleoylphoshatidyl ethanolamine (DOPE), among others. These materials can also be mixed with the DOTM A and DOTAP starting materials in appropriate ratios. M ethods for making liposomes using these materials are well known in the art. The liposomes can comprise multilammelar vesicles (M LVs), small unilamellar vesicles (SUVs), or large unilamellar vesicles (LUV s). The various liposome-nucleic acid complexes are prepared using methods known in the art. See eg. Straubinger (1983) Meth. Immunol. 101 :512-527 ; Szoka (1978) Proc. Natl. Acad. Sci. USA 75 :4194-4198 ; Papahadjopoulos (1975) Biochim. Biophys. Ada 394:483 ; W ilson (1979) Cell 17:77); Deamer & B angham (1976) Biochim. Biophys. Ada 443 :629; Ostro (1977) Biochem. Biophys. Res. Commun. 76:836; Fraley (1979) Proc. Natl. Acad. Sci. USA 76:3348); Enoch & Strittmatter (1979) Proc. Natl. Acad. Sci. USA 76:145 ; Fraley (1980) ]. Biol. Chem. (1980) 255: 10431 ; Szoka & Papahadjopoulos (1978) Proc. Natl. Acad. Sci. USA 75 :145; and Schaefer-Ridder (1982) Science 215 :166. E.Lipoproteins In addition, lipoproteins can be included with the polynucleotide/polypeptide to be delivered. Examples of lipoproteins to be utilized include: chylomicrons, HDL, ID L, LD L, and VLD L. M utants, fragments, or fusions of these proteins can also be used. Also, modifications of naturally occurring lipoproteins can be used, such as acetylated LDL. These lipoproteins can target the delivery of polynucleotides to cells expressing lipoprotein receptors. Preferably, if lipoproteins are including with the polynucleotide to be delivered, no other targeting ligand is included in the composition. Naturally occurring lipoproteins comprise a lipid and a protein portion. The protein portion are known as apoproteins. At the present, apoproteins A , B , C, D , and E have been isolated and identified. At least two of these contain several proteins, designated by Roman numerals, Al, All, AIV ; CI, CII, CIII. A lipoprotein can comprise more than one apoprotein. For example, naturally occurring chylomicrons comprises of A, B , C, and E, over time these lipoproteins lose A and acquire C and E apoproteins. VLD L comprises A, B , C , and E apoproteins, LDL comprises apoprotein B ; and HD L comprises apoproteins A , C , and E. The amino acid of these apoproteins are known and are described in, for example, Breslow (1985) Annu Rev. B iochem 54:699; Law (1986) Adv. Exp M ed. B iol. 151 :162; Chen (1986) J B iol Chem 261 :12918 ; Kane (1980) Proc Natl Acad Sci USA 77:2465 ; and Utermann (1984) Hum Genet 65:232. Lipoproteins contain a variety of lipids including, triglycerides, cholesterol (free and esters), and phospholipids. The composition of the lipids varies in naturally occurring lipoproteins. For example, chylomicrons comprise mainly triglycerides. A more detailed description of the lipid content of naturally occurring lipoproteins can be found, for example, in Meth. Enzymol. 128 (1986). The composition of the lipids are chosen to aid in conformation of the apoprotein for receptor binding activity. The composition of lipids can also be chosen to facilitate hydrophobic interaction and association with the polynucleotide binding molecule. Naturally occurring lipoproteins can be isolated from serum by ultracentrifugation, for instance. Such methods are described in Meth. Enzymol. (supra); Pitas (1980) J. Biochem. 255:5454-5460 and M ahey (1979) J Clin. Invest 64:743-750. Lipoproteins can also be produced by in vitro or recombinant methods by expression of the apoprotein genes in a desired host cell. See, for example, Atkinson (1986) Annu Rev Biophys Chem 15 :403 and Radding (1958) Biochim Biophys Ada 30: 443. Lipoproteins can also be purchased from commercial suppliers, such as B iomedical Techniologies, Inc., Stoughton, M assachusetts, USA . Further description of lipoproteins can be found in Zuckermann et al. W O98/06437. F.Polvcationic A gents Polycationic agents can be included, with or without lipoprotein, in a composition with the desired polynucleotide/polypeptide to be delivered. Polycationic agents, typically, exhibit a net positive charge at physiological relevant pH and are capable of neutralizing the electrical charge of nucleic acids to facilitate delivery to a desired location. These agents have both in vitro, ex vivo, and in vivo applications. Polycationic agents can be used to deliver nucleic acids to a living subject either intramuscularly, subcutaneously, etc. The following are examples of useful polypeptides as polycationic agents: polylysine, polyarginine, polyornithine, and protamine. Other examples include histones, protamines, human serum albumin, DNA binding proteins, non-histone chromosomal proteins, coat proteins from DNA viruses, such as (X 174, transcriptional factors also contain domains that bind DNA and therefore may be useful as nucleic aid condensing agents. Briefly, transcriptional factors such as C/CEB P, c-jun, c-fos, AP-1 , AP-2, A P-3, CPF, Prot-1 , Sp-1 , Oct-l , Oct-2, CREP, and TFIID contain basic domains that bind DNA sequences. Organic polycationic agents include: spermine, spermidine, and purtrescine. The dimensions and of the physical properties of a polycationic agent can be extrapolated from the list above, to construct other polypeptide polycationic agents or to produce synthetic polycationic agents. Synthetic polycationic agents which are useful include, for example, DEAE-dextran, polybrene. Lipofectin™, and lipofectAM INE™ are monomers that form polycationic complexes when combined with polynucleotides/polypeptides. Immunodiasnostic Assays Neisserial antigens of the invention can be used in immunoassays to detect antibody levels (or, conversely, anti-Neisserial antibodies can be used to detect antigen levels). Immunoassays based on well defined, recombinant antigens can be developed to replace invasive diagnostics methods. Antibodies to Neisserial proteins within biological samples, including for example, blood or serum samples, can be detected. Design of the immunoassays is subject to a great deal of variation, and a variety of these are known in the art. Protocols for the immunoassay may be based, for example, upon competition, or direct reaction, or sandwich type assays. Protocols may also, for example, use solid supports, or may be by immunoprecipitation. M ost assays involve the use of labeled antibody or polypeptide; the labels may be, for example, fluorescent, chemiluminescent, radioactive, or dye molecules. Assays which amplify the signals from the probe are also known; examples of which are assays which utilize biotin and avidin, and enzyme-labeled and mediated immunoassays, such as ELISA assays. Kits suitable for immunodiagnosis and containing the appropriate labeled reagents are constructed by packaging the appropriate materials, including the compositions of the invention, in suitable containers, along with the remaining reagents and materials (for example, suitable buffers, salt solutions, etc.) required for the conduct of the assay, as well as suitable set of assay instructions. Nucleic Acid Hybridisation "Hybridization" refers to the association of two nucleic acid sequences to one another by hydrogen bonding. Typically, one sequence will be fixed to a solid support and the other will be free in solution. Then, the two sequences will be placed in contact with one another under conditions that favor hydrogen bonding. Factors that affect this bonding include: the type and volume of solvent; reaction temperature; time of hybridization; agitation; agents to block the non-specific attachment of the liquid phase sequence to the solid support (Denhardt's reagent or B LOTTO); concentration of the sequences; use of compounds to increase the rate of association of sequences (dextran sulfate or polyethylene glycol); and the stringency of the washing conditions following hybridization. See Sambrook et al. [supra] Volume 2, chapter 9, pages 9.47 to 9.57. "Stringency" refers to conditions in a hybridization reaction that favor association of very similar sequences over sequences that differ. For example, the combination of temperature and salt concentration should be chosen that is approximately 120 to 200°C below the calculated Tm of the hybrid under study. The temperature and salt conditions can often be determined empirically in preliminary experiments in which samples of genomic DNA immobilized on filters are hybridized to the sequence of interest and then washed under conditions of different stringencies. See Sambrook et al. at page 9.50. Variables to consider when performing, for example, a Southern blot are (1) the complexity of the DNA being blotted and (2) the homology between the probe and the sequences being detected. The total amount of the fragment(s) to be studied can vary a magnitude of 10, from 0.1 to lμ g for a plasmid or phage digest to 10 " to 10 "8 g for a single copy gene in a highly complex eukaryotic genome. For lower complexity polynucleotides, substantially shorter blotting, hybridization, and exposure times, a smaller amount of starting polynucleotides, and lower specific activity of probes can be used. For example, a single-copy yeast gene can be detected with an exposure time of only 1 hour starting w ith 1 μ g of yeast DNA , blotting for two hours, and hybridizing for 4-8 hours with a probe of 10 8 cpm/μ g. For a single-copy mammalian gene a conservative approach would start with 10 μ g of DNA , blot overnight, and hybridize overnight in the presence of 10% dextran sulfate using a probe of greater than 10 8 cpm/μ g, resulting in an exposure time of -24 hours. Several factors can affect the melting temperature (Tm) of a DNA-DNA hybrid between the probe and the fragment of interest, and consequently, the appropriate conditions for hybridization and washing. In many cases the probe is not 100% homologous to the fragment. Other commonly encountered variables include the length and total G+C content of the hybridizing sequences and the ionic strength and formamide content of the hybridization buffer. The effects of all of these factors can be approximated by a single equation: Tm= 81 + 16.6(logιoCi) + 0.4[% (G + C)]-0.6(% formamide) - 600/n-1.5(% mismatch). where Ci is the salt concentration (monovalent ions) and n is the length of the hybrid in base pairs (slightly modified from M einkoth & W ahl (1984) Anal. Biochem. 138: 267-284). In designing a hybridization experiment, some factors affecting nucleic acid hybridization can be conveniently altered. The temperature of the hybridization and washes and the salt concentration during the washes are the simplest to adjust. As the temperature of the hybridization increases (ie. stringency), it becomes less likely for hybridization to occur between strands that are nonhomologous, and as a result, background decreases. If the radiolabeled probe is not completely homologous with the immobilized fragment (as is frequently the case in gene family and interspecies hybridization experiments), the hybridization temperature must be reduced, and background will increase. The temperature of the washes affects the intensity of the hybridizing band and the degree of background in a similar manner. The stringency of the washes is also increased with decreasing salt concentrations. In general, convenient hybridization temperatures in the presence of 50% formamide are 42°C for a probe with is 95% to 100% homologous to the target fragment, 37°C for 90% to 95% homology, and 32°C for 85% to 90% homology. For lower homologies, formamide content should be lowered and temperature adjusted accordingly, using the equation above. If the homology between the probe and the target fragment are not known, the simplest approach is to start with both hybridization and wash conditions which are nonstringent. If non-specific bands or high background are observed after autoradiography, the filter can be washed at high stringency and reexposed. If the time required for exposure makes this approach impractical, several hybridization and/or washing stringencies should be tested in parallel. Identification of the meningococcal 80-85kDa protein It was observed that various outer membrane vesicle preparations from N. meningitidis serogroup B contained a component of approximately 80-85kDa. This protein was purified from SDS-PAGE gels and N-terminal sequenced (SEQ ID 1). Antibodies raised against the SDS-PAGE purified protein cross-reacted with equivalent proteins in more than 50 N. meningitidis strains of diverse serogroups and serotypes. Cross-reactivity with N.gonorrhoeae, N.polysaccharia and N. lactamica was also observed. Post-immune sera from vaccinated patients also reacted with the protein. The complete gene was cloned from serogroup B N. meningitidis (SEQ ID 2) and the encoded protein was inferred (SEQ ID 3). By comparison with the N-terminal sequencing described above, a signal peptide (SEQ ID 4) and a mature sequence (SEQ ED 5) are inferred. Identification of corresponding genes in N.meningitidis serogroup A and N.gonorrhoeae On the basis of the serogroup B N.meningitidis sequence, the corresponding genes from N.meningitidis serogroup A and N.gonorrhoeae were cloned and sequenced. The complete gene from serogroup A N.meningitidis is shown in SEQ ID 6, with the encoded protein in SEQ ID 7. The signal peptide and mature sequence are SEQ IDs 8 and 9. The complete gene from N.gonorrhoeae is shown in SEQ ID 10, with the encoded protein in SEQ ED 11. The signal peptide and mature sequence are SEQ EDs 12 and 13. Sequence comparisons The protein sequences were compared and are highly homologous. The N.meningitidis serogroup B sequence and the N.gonorrhoeae sequence show 95.4% identity in 797 aa overlap: 10 20 30 40 50 60 orf21. pep MK KQIASALMMLGISPLALADFTIQDIRVEG QRTEPSTVFNYLPVKVGDTYNDTHGSA orf21ng . pep MK KQIASA MM GISPLAFADFTIQDIRVEGLQRTEPSTVFNYLPVKVGDTYNDTHGSA 10 20 30 40 50 60 70 80 90 100 110 120 orf21.pep IIKSLYATGFFDDVRVETADGQLLLTVIERPTIGSLNITGAKMLQNDAIKKNLESFGLAQ orf2Ing.pep IIKS YATGFFDDVRVETADGQL TVIERPTIGSLNITGAKMLQNDAIKK ESFGLAQ 70 80 90 100 110 120 130 140 150 160 170 180 orf21.pep SQYFNQATLNQAVAGLKEEYLGRGKLNIQITPKVTKLARNRVDIDITIDEGKSAKITDIE orf2Ing.pep SQYFNQATLNQAVAGLKEEYLGRGKLNIQITPKVTK ARNRVDIDITIDEGKSAKITDIE 130 140 150 160 170 180 190 200 210 220 230 240 orf21. pep FEGNQVYSDRKLMRQMS TEGGIWTWLTRSNQFNEQKFAQDMEKVTDFYQN GYFDFRIL orf2Ing . pep FEGNQVYSDRK MRQMS TEGGI TW TRSDRFDRQKFAQDMEKVTDFYQNNGYFDFRIL 190 200 210 220 230 240 250 260 270 280 290 300 orf21.pep DTDIQTNEDKTKQTIKITVHEGGRFRWGKVSIEGDTNEVPKAELEKLLTMKPGKWYERQQ orf21ng .pep DTDIQTNEDKTRQTIKITVHEGGRFR GKVSIEGDTNEVPKAE EKL TMKPGKWYERQQ 250 260 270 280 290 300 310 320 330 340 350 360 orf21. pep MTAVLGEIQNRMGSAGYAYSEISVQP PNAETKTVDFVLHIEPGRKIYVNEIHITGNNKT orf2Ing.pep MTAVLGEIQNRMGSAGYAYSEISVQPLPNAGTKTVDFVLHIEPGRKIYV EIHITGNNKT 310 320 330 340 350 360 370 380 390 400 410 420 orf21.pep RDEWRRELRQMESAPYDTSKLQRSKERVEL GYFDNVQFDAVPLAGTPDKVDLNMSLTE orf2Ing.pep RDEWRRELRQMESAPYDTSKLQRSKERVEL GYFD VQFDAVP AGTPDKVD NMS TE 370 380 390 400 410 420 430 440 450 460 470 480 orf21. pep RSTGS DLSAGWVQDTG VMSAGVSQDNLFGTGKSAA RASRSKTTLNGS SFTDPYFTA orf2Ing.pep RSTGSLDLSAGWVQDTG VMSAGVSQDNLFGTGKSAA RASRSKTT NGSLSFTDPYFTA 430 440 450 460 470 480 490 500 510 520 530 540 orf21. pep DGVΞ GYDVYGKAFDPRKASTSIKQYKTTTAGAGIRMSVPVTEYDRVNFG VAEHLTVNT orf21ng . pep DGVSLGYDIYGKAFDPRKASTSVKQYKTTTAGGGVRMGIPVTEYDRVNFGLAAEHLTVNT 490 500 510 520 530 540 550 560 570 580 590 600 orf21. pep YNKAPKHYADFIKKYGKTDGTDGSFKGW YKGTVG GR KTDSA PTRGYLTGV AEIA orf2Ing . pep YNKAPKRYADFIRKYGKTDGADGSFKG LYKGTVGWGRNKTDSAS PTRGYLTGV AEIA 550 560 570 580 590 600 610 620 630 640 650 660 orf21. pep LPGSK QYYSATHNQTWFFPLSKTFT MLGGEVGIAGGYGRTKEIPFFENFYGGGLGSVR orf21ng.pep LPGSKLQYYSATHNQTWFFPLSKTFTLMLGGEVGIAGGYGRTKEIPFFENFYGGGLGSVR 610 620 630 640 650 660 670 680 690 700 710 720 orf21.pep GYESGTLGPKVYDEYGEKISYGGNKKAJ SAELLFPMPGAKDARTVRLS FADAGSVWDG orf2Ing. pep GYESGTLGPKVYDEYGEKISYGGNKKANVSAELLFPMPGAKDARTVRLSLFADAGSV DG 670 680 690 700 710 720 730 740 750 760 770 780 orf21. pep KTYDDNSSSATGGRVQNIYGAGNTHKSTFTNELRYSAGGAVTWLSPLGPMKFSYAYPLKK orf2Ing.pep RTY TAAENGN KSVYSE-NAHKSTFTNELRYSAGGAVTWLSPLGPMKFSYAYPLKK 730 740 750 760 770 790 orf21. pep KPEDEIQRFQFQLGTTF orf2Ing.pep KPEDEIQRFQFQ GTTFX 780 790 The N.meningitidis serogroup A and B sequences show 99.9% identity in 797 aa overlap: 10 20 30 40 50 60 orf21.pep MK KQIASALMMLGISPLALADFTIQDIRVEGLQRTEPSTVFNY PVKVGDTYNDTHGSA ll lll II orf2la.pep MK KQIASA MVLGISPLALADFTIQDIRVEGLQRTEPSTVFNY PVKVGDTYNDTHGSA 10 20 30 40 50 60 70 80 90 100 110 120 orf21.pep IIKSLYATGFFDDVRVETADGQLLLTVIERPTIGSLNITGAKM QNDAIKK ESFGLAQ III IMIIIIMII I orf2la. pep IIKS YATGFFDDVRVETADGQL LTVIERPTIGSLNITGAKMLQNDAIKKNLESFGLAQ 70 80 90 100 110 120 130 140 150 160 170 180 orf21. pep SQYFNQATLNQAVAGLKEEYLGRGK NIQITPKVTKLARNRVDIDITIDEGKSAKITDIE orf2la.pep SQYFNQATLNQAVAG KEEYLGRGKLNIQITPKVTK ARNRVDIDITIDEGKSAKITDIE 130 140 150 160 170 180 190 200 210 220 230 240 orf21. pep FEGNQVYSDRK MRQMSLTEGGIWTWLTRSNQFNEQKFAQDMEKVTDFYQNNGYFDFRIL orf21a.pep FEGNQVYSDRK MRQMSLTEGGIWTWLTRSNQFNEQKFAQDMEKVTDFYQNNGYFDFRIL 190 200 210 220 230 240 250 260 270 280 290 300 orf21.pep DTDIQTNEDKTKQTIKITVHEGGRFR GKVSIEGDTNEVPKAE EK LTMKPGK YERQQ II orf21a.pep DTDIQTNEDKTKQTIKITVHEGGRFRWGKVSIEGDTNEVPKAE EK TMKPGKWYERQQ 250 260 270 280 290 300 310 320 330 340 350 360 orf21. pep MTAV GEIQNRMGSAGYAYSEISVQP PNAETKTVDFV HIEPGRKIYVNEIHITGNNKT orf2la. ep MTAVLGEIQNRMGSAGYAYSEISVQP PNAETKTVDFVLHIEPGRKIYVNEIHITGN KT 310 320 330 340 350 360 370 380 390 400 410 420 orf21.pep RDEWRRE RQMESAPYDTSKLQRΞKERVELLGYFDNVQFDAVPLAGTPDKVD NMS TE IIIIMIIIMII orf2la. pep RDEWRRELRQMESAPYDTSKLQRSKERVELLGYFDNVQFDAVPLAGTPDKVDLN SLTE 370 380 390 400 410 420 430 440 450 460 470 480 orf21.pep RSTGSLDLSAGWVQDTGLVMSAGVSQDNLFGTGKSAALRASRSKTTLNGSLSFTDPYFTA orf2la. pep RSTGS DLSAG VQDTG VMSAGVSQDN FGTGKSAA RASRSKTTLNGSLSFTDPYFTA 430 440 450 460 470 480 490 500 510 520 530 540 orf21. pep DGVSLGYDVYGKAFDPRKASTSIKQYKTTTAGAGIRMSVPVTEYDRVNFGLVAEH TVNT orf2la. pep DGVSLGYDVYGKAFDPRKASTSIKQYKTTTAGAGIRMSVPVTEYDRVNFG VAEHLTVNT 490 500 510 520 530 540 550 560 570 580 590 600 orf21.pep YNKAPKHYADFIKKYGKTDGTDGSFKGW YKGTVGWGRNKTDSA PTRGY TGV AEIA orf2la . pep YNKAPKHYADFIKKYGKTDGTDGSFKGW YKGTVGWGR KTDSAL PTRGYLTGVNAEIA 550 560 570 580 590 600 610 620 630 640 650 660 orf21.pep PGSK QYYSATHNQTWFFP SKTFT MLGGEVGIAGGYGRTKEIPFFENFYGGG GSVR orf2la. pep LPGSK QYYSATHNQTWFFPLSKTFTLMLGGEVGIAGGYGRTKEIPFFENFYGGGLGSVR 610 620 630 640 650 660 670 680 690 700 710 720 orf21.pep GYESGTLGPKVYDEYGEKISYGGNKKANVSAELLFPMPGAKDARTVRLS FADAGSVWDG orf2la.pep GYESGTLGPKVYDEYGEKISYGGNKKANVSAELLFPMPGAKDARTVRLS FADAGSV DG 670 680 690 700 710 720 730 740 750 760 770 780 orf21. pep KTYDDNSSSATGGRVQNIYGAGNTHKSTFTNELRYSAGGAVTWLSPLGPMKFSYAYPLKK orf2la . pep KTYDDNSSSATGGRVQNIYGAGNTHKSTFTNELRYSAGGAVT LSPLGPMKFSYAYP KK 730 740 750 760 770 780 790 orf21.pep KPEDEIQRFQFQLGTTF orf21a.pep KPEDEIQRFQFQLGTTFX 790 The high degree of conservation suggests that a single protein may be able to induce immune responses against a variety of Neisseriae species. Vaccines The three proteins identified above were expressed and used for immunisation. Good immune responses were observed against the proteins. Combination vaccines En addition, the proteins were each combined with antigens against other pathogenic organisms (e.g. the Chiron polysaccharide vaccine against serogroup C meningitis), and used for immunisation. Good immune responses were observed. Further NmB components Whilst it is efficacious, the protection elicited by the Norwegian OMV vaccine is restricted to the strain used to make the vaccine. The clinical trials on the vaccine obtained only 57.2% efficacy after 29 months in teenagers, although IgG responses were observed in almost 100% of patients [e.g. Rosenqvist et al. (1995) Infect. Immun. 63:4642-4652]. Surprisingly, it has been found that the addition of further defined components to the Norwegian OMV vaccine significantly broadens its efficacy. The Norwegian vaccine does not elicit protection against NmB strain 2996. Defined proteins from strain 2996 were added to the Norwegian vaccine, and it was shown that the efficacy of the vaccine was increased by a surprising degree. Furthermore, the addition of a NmC polysaccharide conjugate antigen [e.g. Costantino et al. (1992) Vaccine 10:691-698] gave excellent results. The bactericidal activities of the combinations are shown in the following table: Three different NmB antigens were used: #1 : ORF1 - e.g. example 77 of W099/24578 (see also W099/55873) #2: protein '287' - e.g. Figure 21 of WO99/57280 (also SEQ IDs 3103-3108) #3: a mixture in Al(OH) 3 of #1, #2 and protein '919' (SEQ ID 14 herein; see also WO99/57280 Figure 23 and SEQ IDs 3069-3074 therein). It can readily be seen that the inefficacy of the Norwegian OMV vaccine against strain 2996 (group 1) can be overcome by adding defined antigens from strain 2996. The results using NmB protein '287' are particularly good. The Norwegian vaccine can thus be improved without needing to prepare OMVs from a number of different strains. This vaccine also offers protection against heterologous MenB strains. The same vaccines, prepared using 2996 strain proteins, was tested against five other strains. Titres were as follows: * Controls: strains 2996, BZ133 & 1000 = OMVs prepared from homologous strain; strain BZ232 = OMVs prepared from 2996; MC58 & NGH38 = SEAM3 A second study supplemented 'Norwegian' OMVs with proteins from NmB strain 2996: - protein 919, expressed in E.coli without any fusion partner - ORF1, expressed in E.coli as a His-tagged fusion - Protein 287, expressed in E.coli as a GST fusion - A mixture of these three proteins, optionally with the NmC conjugate . The preparations were adjuvanted with Al(OH) 3 and tested against the homologous strain using the bactericidal assay. Results were as follows: * the antibodies were bacteriostatic, not bactericidal Further work with antigen 287 Combinations of Norwegian OMVs with antigen 287 were investigated further. 20μg antigen 287 was combined with Norwegian OMP vaccine (15μg OMP + Al(OH) 3 ) and used to immunise mice. The antibodies were tested in the bactericidal assay, and were effective against all strains tested. The results were as follows: In almost all cases, therefore, the combination of OMVs + protein 287 surprisingly gives better results that the OMVs alone. Recombinant OMVs E.coli were transformed to express ORFl, ORF40 and ORF46. OMVs prepared from the recombinant E.coli were able to induce bactericidal antibodies against N.meningitidis. ORFl, ORF40 and ORF46 (strain 2996) were expressed as His-tagged fusions in E.coli and were prepared either as pure proteins or in the form of OMVs. Bactericidal titres against both preparations were tested using strain 2996 as challenge: Bactericidal titres using heterologous challenge strains were also measured. ORF46 gives a titre against strain MC58 of 4096 in pure form, but this rises to 32000 when in the form of OMVs. ORFl gives a titre against NmA strain F6124 of 128 in pure form, but this rises to 512 when in the form of OMVs. ORF40 gives a titre against strain MC58 of 512 in pure form, but this doubles when in the form of OMVs. These data show that NmB antigens retain immunogenicity when prepared in E.coli as OMVs and, furthermore, that immunogenicity can actually be enhanced. It will be understood that this application describes the invention by way of example only and modifications may be made whilst remaining within the scope and spirit of the invention.

Claims: CLAIMS 1. A composition comprising (a) N.meningitidis serogroup B outer membrane preparation, and (b) an immunogenic component selected from one or more of the following: • a protein disclosed in WO99/57280, or an immunogenic fragment thereof; • a protein disclosed in WO99/36544, or an immunogenic fragment thereof; • a protein disclosed in WO99/24578, or an immunogenic fragment thereof; • a protein disclosed in WO99/66791, or an immunogenic fragment thereof; • a protein disclosed in Tettelin et al. [Science (2000) 287: 1809-1815], or an immunogenic fragment thereof; • a protein disclosed in WO97/28273, or an immunogenic fragment thereof; • a protein disclosed in WO96/29412, or an immunogenic fragment thereof; • a protein disclosed in WO95/03413, or an immunogenic fragment thereof; • a protein disclosed in WO99/31132, or an immunogenic fragment thereof; • a protein disclosed in WO99/58683, or an immunogenic fragment thereof; • a protein disclosed in WO99/55873, or an immunogenic fragment thereof; and/or • Neisseria meningitidis protein PorA, TbpA, TbpB, PilC, Op A, Omp85. 2. The composition of claim 1 , wherein component (b) is a NmB protein. 3. The composition of claim 2, wherein the NmB protein is protein 919, protein 235, protein 519, protein 225, protein ORF40, protein 287, ORFl, ORF4, or ORF46. 4. The composition of any preceding claim, wherein component (b) includes a protein from a different NmB strain from that from which component (a) is derived. 5. The composition of any preceding claim, wherein one or more of its components is adsorbed on Al(OH) . 6. The composition of any preceding claim, wherein component (a) comprises OMVs. 7. The composition of claim 6, wherein the OMVs are a deoxycholate extract from NmB. 8. The composition of any preceding claim, wherein component (a) is adsorbed on Al(OH) 3 . 9. The composition of any preceding claim, wherein further comprising one or more of the following components: • a protective antigen against Neisseria meningitidis serogroup A; • a protective antigen against Neisseria meningitidis serogroup C; • a protective antigen against Neisseria meningitidis serogroup Y; • a protective antigen against Neisseria meningitidis serogroup W; • a protective antigen against Haemophilus influenzae; • a protective antigen against pneumococcus; • a protective antigen against diphtheria; • a protective antigen against tetanus; • a protective antigen against whooping cough; • a protective antigen against Helicobacter pylori; • a protective antigen against polio; and/or • a protective antigen against hepatitis B virus. 10. The composition of any preceding claim, wherein the composition is a vaccine. 11. The composition of any preceding claim, for use as a medicament. 12. The use of composition according to any preceding claim in the manufacture of: (i) a medicament for treating or preventing infection due to Neisserial bacteria; (ii) a diagnostic reagent for detecting the presence of Neisserial bacteria or of antibodies raised against Neisserial bacteria; and/or (iii) a reagent which can raise antibodies against Neisserial bacteria. 13. A method of treating a patient, comprising administering to the patient a therapeutically effective amount of a composition according to any one of claims 1 to 10. 14. A bacterial outer membrane preparation comprising an immunogenic component selected from one or more of the following: • a protein disclosed in WO99/57280, or an immunogenic fragment thereof; • a protein disclosed in WO99/36544, or an immunogenic fragment thereof; • a protein disclosed in WO99/24578, or an immunogenic fragment thereof; • a protein disclosed in WO99/66791, or an immunogenic fragment thereof; • a protein disclosed in Tettelin et al. [Science (2000) 287:1809-1815], or an immunogenic fragment thereof; • a protein disclosed in WO97/28273, or an immunogenic fragment thereof; • a protein disclosed in WO96/29412, or an immunogenic fragment thereof; • a protein disclosed in WO95/03413, or an immunogenic fragment thereof; • a protein disclosed in WO99/31132, or an immunogenic fragment thereof; • a protein disclosed in WO99/58683, or an immunogenic fragment thereof; • a protein disclosed in WO99/55873, or an immunogenic fragment thereof; and/or • Neisseria meningitidis protein PorA, TbpA, TbpB, PilC, OpA, Omp85.

Inventors: GIULIANI MARZIA, PIZZA MARIAGRAZIA, RAPPUOLI RINO, GIULIANI, MARZIA, PIZZA, MARIAGRAZIA, RAPPUOLI, RINO
Applicants: CHIRON SPA, GIULIANI MARZIA, PIZZA MARIAGRAZIA, RAPPUOLI RINO, GIULIANI, MARZIA, PIZZA, MARIAGRAZIA, RAPPUOLI, RINO
Application year: 2001
Main classification: A61K39
IPC codes: A61K39, A61P25, A61P29, A61P31