Immunogenic polysaccharide-protein conjugates containing...

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Conjugate or complex

Reexamination Certificate

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C424S137100, C424S194100, C424S197110, C424S241100, C530S403000, C530S404000, C530S405000, C530S406000

Reexamination Certificate

active

06632437

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates, in general, to polysaccharide-protein conjugates and vaccines. In particular the present invention relates to polysaccharide-protein conjugates that elicit serum IgG and IgM antibodies to poly &agr;(2→8) NeuNAc, or to both poly &agr;(2→8) NeuNAc and poly &agr;(2→9) NeuNAc, or to poly &agr;(2→8),&agr;(2→9) NeuNAc.
2. Background Information
Neisseriae meningitidis
are a major cause of systemic infections, especially meningitis, in humans. Capsular polysaccharide (CP) vaccines are licensed for meningococcal groups A,C,Y and W135. Diseases caused by group B meningococci continue to occur in endemic and epidemic forms and remain an important health problem (Gotschlich, E. C. (1984) in Bacterial Vaccines. Ed. Germanier (Academic Press, NY) pp. 237-255; Peltola, H. (1983) Rev. Infect. Dis. 5, 71-91; Poolman, J. T. et al. (1986) Lancet, ii,555-557).
Escherichia coli
(
E. coli
) K1 is a major cause of neonatal meningitis, upper urinary tract infections and systemic infections in hospitalized patients and in domesticated and laboratory animals (Robbins, J. B. et al. (1974) N. Eng. J. Med. 290, 1216-1220; Kaijser, B. et al. (1977) Lancet i, 663-664; Cross, A. S. et al. (1984) J. Infect. Dis. 149, 184-193; Ørskov, I., & Ørskov, F. (1985) J. Hyg. Camb. 95, 551-575). Despite antibiotic treatment and supportive care, meningitis caused by these two pathogens continues to exert a high morbidity, including permanent CNS injury, and mortality (Peltola, H. (1983) Rev. Infect. Dis. 5, 71-91; Schneerson, R. (1988) in Understanding Mental Retardation. ed. Kavanagh, J. F. (Paul Brookes Publishing Co. Baltimore), pp. 237-249; Brandtzaeg, P. et al. (1989) J. Infect. Dis. 159, 195-204; McCracken, G. H., Jr. et al. (1974) Lancet, ii, 246-250).
The CP of Group B meningococci and of
E. coli
K1 are identical (poly &agr;(2→8) NeuNAc) and serve as essential virulence factors and protective antigens for both pathogens (Grados, O., & Ewing, W. H. (1970) J. Infect. Dis. 122, 100-103; Kasper, D. L. et al. (1973) J. Immunol. 110, 262-268; Bhattacharjee, A. K. et al. (1975) J. Biol. Chem. 250, 1926-1932; Robbins, J. B. et al. (1974) N. Eng. J. Med. 290, 1216-1220). Poly &agr;(2→8) NeuNAc is also a surface antigen of
Moraxella nonliquefaciens
and
Pasteurella haemolytica,
serotype A-2 (Bøvre, K. et al. (1983) NIHP Annals. 6, 65-73; Devi, S. J. N. et al. (1991) Infect. Immun. 59, 732-736; Adlam, C. et al. (1987) FEMS Microbiol. Lett. 42, 23-25). The latter is the major cause of outbreaks of pasteurellosis in young lambs which suggests that poly &agr;(2→8) NeuNAc may serve as a virulence factor for yet another bacterial species.
Attempts to induce protective immunity to group B meningococci and
E. coli
K1 have been thwarted because poly &agr;(2→8) NeuNAc, alone or complexed to outer membrane proteins, induced low and transient levels of IgM antibodies (Kasper, D. L. et al. (1973) J. Immunol. 110, 262-268; Wyle, F. A. et al. (1972) J. Infect. Dis. 126, 514-522; Zollinger, W. D. et al. (1979) J. Clin. Invest. 63, 836-842; Moreno, C. et al. (1985) Infect. Immun. 47, 527-533; Frasch, C. E. et al. (1988) J. Infect. Dis. 158, 710-718; Lifely, M. R. et al. (1991) Vaccine 9, 60-66). Covalent attachment of periodate-treated (Jennings, H. & Lugowski, C. (1981) J. Immunol. 127, 1011-1018) or acid-hydrolyzed poly &agr;(2→8) NeuNAc (Porro, M. et al. (1983) Med. Trop. 43, 129-132) to a protein also failed to elicit antibodies to this antigen. Further, this CP has been considered as a “self antigen”, because &agr;(2→8) NeuNAc is found as monomers or dimers on glycoproteins and gangliosides in adults and up to ≈11 residues in fetal tissues including N-CAMs (Finne, J. et al. (1983) Lancet, ii, 355-357; Finne, J. et al. (1987) J. Immunol. 138, 4402-4407; Soderstrom, T. et al. (1984) N. Eng. J. Med. 310, 726-727). Accordingly, investigators have studied other components, such as LPS, outer membrane proteins and iron-binding proteins, or chemically modified poly &agr;(2→8) NeuNAc, as potential vaccines (Zollinger, W. D. et al. (1979) J. Clin. Invest. 63, 836-842; Moreno, C. et al. (1985) Infect. Immun. 47, 527-533; Frasch, C. E. et al. (1988) J. Infect. Dis. 158, 710-718; Jennings, H. J. et al. (1984) Infect. Immun. 43, 407-412; Jennings, H. J. et al. (1986) J. Immunol. 137, 1708-1713; Frasch, C. E. (1989) Clin. Microbiol. Rev. 2(Suppl), S134-S138).
Most newborns and adults have bactericidal antibodies to the three major serogroups (A,B,C) of meningococci (Goldschneider, I. et al. (1969) J. Exp. Med. 129, 1307-1326); most of the bactericidal activity, including of group B meningococci, was removed by adsorption with the homologous CP (Frasch, C. E. et al. (1988) J. Infect. Dis. 158, 710-718; Brandt, B. L. et al. (1972) J. Immunol. 108, 913-920; Kasper, D. L. et al. (1973) J. Infect. Dis. 127, 378-387; Skevakis, L. et al. (1984) J. Infect. Dis. 149, 387-396). The peak incidence of disease caused by meningococci, including group B, is when the maternally-derived antibodies have waned and the adult levels have not yet developed (Gotschlich, E. C. (1984) in Bacterial Vaccines. Ed. Germanier (Academic Press, NY) pp. 237-255; Goldschneider, I. et al. (1969) J. Exp. Med. 129, 1307-1326). Rises in poly &agr;a(2→8) NeuNAc antibodies, including those of the IgG isotype, are detectable in patients convalescent from group B meningococcal meningitis (Wyle, F. A. et al. (1972) J. Infect. Dis. 126, 514-522; Zollinger, W. D. et al. (1979) J. Clin. Invest. 63, 836-842; Frasch, C. E. et al. (1988) J. Infect. Dis. 158, 710-718; Skevakis, L. et al. (1984) J. Infect. Dis. 149, 387-396; Craven, D. E. et al. (1982) Infect. Immun. 37, 132-137; Mandrell, R. E. & Zollinger, W. D. (1982) J. Immunol. 129, 2172-2178; Leinonen, M. & Frasch, C. E. (1982) Infect. Immun. 38, 1203-1207). Polyclonal and monoclonal (mAb) poly &agr;(2→8) NeuNAc antibodies were raised in animals by multiple intravenous injections of bacteria (Robbins, J. B. et al. (1974) N. Eng. J. Med. 290, 1216-1220; Moreno, C. et al. (1985) Infect. Immun. 47, 527-533; Mandrell, R. E. & Zollinger, W. D. (1982) J. Immunol. 129, 2172-2178; Allen, P. Z. et al. (1982) J. Clin. Microbiol. 15, 324-329; Craven, D. E. et al. (1979) J. Clin. Microbiol. 10, 302-307; Frosch, M. et al. (1985) Proc. Natl. Acad. Sci. (USA) 82, 1194-1198). Monoclonal antibodies to this antigen were identified in a healthy 81 year old male and from hybridoma cultures (Kabat, E. A. et al. (1986) J. Exp. Med. 164, 642-654; Kabat, E. A. et al. (1988) J. Exp. Med. 168, 699-711; Raff, H. V. et al. (1988) J. Infect. Dis. 157, 118-126). These antibodies exert biologic activities which have been correlated with protective immunity; 1) complement-dependent bacteriolysis on Group B meningococci (Gotschlich, E. C. (1984) in Bacterial Vaccines. Ed. Germanier (Academic Press, NY) pp. 237-255; Goldschneider, I. et al. (1969) J. Exp. Med. 129, 1307-1326); 2) protection against lethal infection of rodents by
E. coli
K1 (Robbins, J. B. et al. (1974) N. Eng. J. Med. 290, 1216-1220; Glode, M. P. et al. (1977) Infect. Immun. 16, 75-80; Kim, K. S. et al. (1985) Infect. Immun. 50, 734-737).
There are two other bacterial NeuNAc polymers: 1) group C
N. meningitidis
CP composed of poly &agr;(2→9) NeuNAc; most strains are variably O-acetylated at C7 or C8 (Bhattacharjee, A. K. et al. (1975) J. Biol. Chem. 250, 1926-1932). Although differing from poly &agr;(2→8) NeuNAc only by linkage, poly &agr;(2→9) NeuNAc is immunogenic and is a licensed vaccine against group C meningococci (World Health Organization Expert Committee on Biological Standardization. (1977) Technical Report Series, 610. WHO, Geneva, Switzerland); 2)
E. coli
K92 CP (
FIG. 1
) with the disaccharide repeat unit of alternating &agr;(2→8),&agr;(2→9) NeuNAc (The structure of this polysaccharide can be written as 9)-NeuNAc-&agr;-(2→8)-NeuNAc-&agr;-(2→.) (Robbins, J. B. et al. (1972) Infect. Immun. 6

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