AU2010235984B2

AU2010235984B2 - Vaccine composition

Info

Publication number: AU2010235984B2
Application number: AU2010235984A
Authority: AU
Inventors: Dominique Boutriau; Carine Capiau; Pierre Michel Desmons; Dominique Lemoine; Jan Poolman
Original assignee: GlaxoSmithKline Biologicals SA
Current assignee: GlaxoSmithKline Biologicals SA
Priority date: 2001-04-03
Filing date: 2010-10-22
Publication date: 2013-03-21
Anticipated expiration: 2022-03-28
Also published as: AU2007200116A1; AU2010201399A1; AU2010235979A1; AU2010201399B2; AU2010235984A1

Abstract

The present invention relates to new, advantageous DTP-based combination vaccine formulations, and concomitantly administered combination vaccine kits. Methods of administration of these vaccines and kits are also provided.

Description

AUSTRALIA Patents Act COMPLETE SPECIFICATION (ORIGINAL) Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Name of Applicant: GlaxoSmithKline Biologicals s.a. Actual Inventor(s): Dominique Boutriau, Carine Capiau, Pierre Michel Desmons, Dominique Lemoine, Jan Poolman Address for Service and Correspondence: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: VACCINE COMPOSITION Our Ref: 898539 POF Code: 456838/462676 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): -1 aaoeI Vaccine Composition The present application is a divisional application from Australian patent application number 2010201399 which in turn is a divisional of Australian patent application number 2007200116 which in turn is a divisional of Australian patent application number 2002310903 5 the entire disclosure of each of which are incorporated herein by reference. The present invention relates to new combination vaccine formulations. Combination vaccines (which provide protection against multiple pathogens) are very desirable in order to minimise the number of immunisations required to confer protection against multiple pathogens, to lower administration costs, and to increase acceptance and coverage rates. The 10 well-documented phenomenon of antigenic competition (or interference) complicates the development of multi-component vaccines. Antigenic interference refers to the observation that administering multiple antigens often results in a diminished response to certain antigens relative to the immune response observed when such antigens are administered individually. Combination vaccines are known which can prevent Bordetella pertussis, Clostridium 15 tetani, Corynebacterium diphtheriae, and optionally Hepatitis B virus and/or Haemophilus influenzae type b (see, for instance, WO 93/24148 and WO 97/00697). The present invention concerns the manufacture of the most ambitious multi-valent vaccines to date, the administration of which can prevent or treat infection by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, and N. 20 meningitidis, and preferably also Haemophilus influenzae, Streptococcus pneumoniae, Hepatitis A virus and/or Polio virus, wherein the components of the vaccine do not significantly interfere with the immunological performance of any one component of the vaccine. The discussion of documents, acts, materials, devices, articles and the like is included 25 in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application. Throughout the description and claims of the specification the word "comprise" and 30 variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps. Accordingly, in one aspect of the invention there is provided a vaccine kit for concomitant administration comprising two multi-valent immunogenic compositions for conferring protection in a host against disease caused by Bordetella pertussis, Clostridium I a tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and Streptococcus pneumoniae, said kit comprising a first container comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), 5 (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and (e) Inactivated polio virus, and a second container comprising: (2a) one or more conjugates of a carrier protein and a capsular 10 polysaccharide or oligosaccharide from Streptococcus pneumoniae, and wherein the first container comprises a DT content of 60-120 tg, and the second container comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197. The above immunogenic composition may further comprise one, two, three, four, five, 15 or six components selected from the following list: N. meningitidis type A polysaccharide [MenA] (preferably conjugated), N. meningitidis type W polysaccharide [MenW] (preferably conjugated), the Vi polysaccharide of Salmonella typhi, N. meningitidis (preferably serotype B) outer membrane vesicles, one or more N. meningitidis (preferably serotype B) outer membrane (surface-exposed) proteins, and killed, attenuated Hepatitis A virus (HepA 20 preferably the product known as 'HavrixTM' [SmithKline Beecham Biologicals]) without substantial interference problems for any of the antigens of the composition. In a second aspect of the invention there is provided various advantageous kits comprising two or three multi-valent immunogenic compositions, said kits being capable of conferring protection in a host against disease caused by Bordetella pertussis, Clostridium 25 tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and Streptococcus pneumoniae, and optionally also N. meningitidis, and Haemophilus influenzae. In a first embodiment of the second aspect of the invention there is provided a kit comprising two multi-valent immunogenic compositions for conferring protection in a host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium 30 diphtheriae, Hepatitis B virus, Polio virus and Streptococcus pneumoniae, and optionally also N. meningitidis, and Haemophilus influenzae. The kit comprises a first container comprising: (a) either killed whole-cell Bordetella pertussis (Pw), or two or more acellular pertussis components (Pa) [preferably the latter], 35 (b) tetanus toxoid (TT or T), (c) diphtheria toxoid (DT or D), 2 (d) Hepatitis B surface antigen (HepB or HB), and (e) Inactivated polio virus (IPV), and a second container comprising: (2a) one or more conjugates of a carrier protein and a capsular 5 polysaccharide from Streptococcus pneumoniae (where the capsular polysaccharide is preferably from a pneumococcal serotype selected from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 7P, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F). 10 In further advantageous embodiments of the above kit of the invention, the first container additionally comprises: (f) either or both conjugates of a carrier protein and a capsular polysaccharide of a bacterium selected from the group N. meningitidis type Y (MenY) and N. meningitidis type C (MenC), and (g) a conjugate of a carrier 15 protein and the capsular polysaccharide of H. influenzae type B (Hib); or the second container additionally comprises: (2b) either or both conjugates of a carrier protein and a capsular polysaccharide of a bacterium selected from the group N meningitidis type Y (MenY) and N. meningitidis type C (MenC), and (2c) a conjugate of a carrier protein and the capsular polysaccharide of H. influenzae type B (Hib); or the first 20 container additionally comprises: (f) either or both conjugates of a carrier protein and a capsular polysaccharide of a bacterium selected from the group N meningiddis type Y (MenY) and N. meningitidis type C (MenC), and the second container additionally comprises (2b) a conjugate of a carrier protein and the capsular polysaccharide of H. influenzae type B (Hib); or the first container additionally comprises (f) a conjugate of 25 a carrier protein and the capsular polysaccharide of H. influenzae type B (Hib), and the second container additionally comprises: (2b) either or both conjugates of a carrier protein and a capsular polysaccharide of a bacterium selected from the group N. meningitidis type Y (MenY) and N. meningitidis type C (MenC). 30 In a second embodiment of the second aspect of the invention there is provided a kit comprising two multi-valent immunogenic compositions for conferring protection in a host against disease caused by Bordetellapertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus, N. meningitidis, and Haemophilus influenzae. 3 The kit comprises a first container comprising: (a) either killed whole-cell Bordetella pertussis (Pw), or two or more acellular pertussis components (Pa) [preferably the latter], (b) tetanus toxoid (TT or T), 5 (c) diphtheria toxoid (DT or D), (d) Hepatitis B surface antigen (HepB or HB), and (e) Inactivated polio virus (IPV), and a second container comprising: (2a) either or both conjugates of a carrier protein and a capsular 10 polysaccharide of a bacterium selected from the group N. Meningittdis type Y (MenY) and N. meningitidis type C (MenC), and (2b) a conjugate of a carrier protein and the capsular polysaccharide oflH influenzae type B (Hib). 15 In a third embodiment of the second aspect of the invention there is provided a kit comprising three multi-valent immunogenic compositions for conferring protection in a host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and N. meningitidis, 20 Haemophilus influenzae and Streptococcuspneumoniae. The kit comprises a first container comprising: (a) either killed whole-cell Bordetella pertussis (Pw), or two or more acellular pertussis components (Pa) [preferably the latter], (b) tetanus toxoid (TT or T), 25 (c) diphtheria toxoid (DT or D), (d) Hepatitis B surface antigen (HepB or HB), and (e) Inactivated polio virus (IPV), and a second container comprising: (2a) one or more conjugates of a carrier protein and a capsular 30 polysaccharide from Streptococcus pneumoniae [where the capsular polysaccharide is preferably from a pneumococcal serotype selected from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, I1A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F], 4 and a third container comprising: (3a) either or both conjugates of a carrier protein and a capsular polysaccharide of a bacterium selected from the group N. meningitidis type Y (MenY) and N. meningitidis type C (MenC), 5 and (3b) a conjugate of a carrier protein and the capsular polysaccharide of H. influenzae type B (Hib). 10 Any or the above containers of the above kits of the invention may further comprise one, two, three, four, five; 'six or seven components selected from the following list: N meningitidis type A polysaccharide [MenAl (preferably conjugated), N. meningitidis type W polysaccharide [MenW] (preferably conjugated), the Vi polysaccharide of Salmonella typhI, N. meningitidis (preferably serotype B) outer 15 membrane vesicles, one or more N. meningitdis (preferably serotype B) outer membrane (surface-exposed) proteins, HepA (as described above), and one or more S. pneumoniae proteins (preferably surface-exposed) without substantial interference problems for any of the antigens of the composition. The containers of the kit can be packaged separately or, preferably, packed 20 together. Preferably the kit is provided with a list of instructions for administration of the vaccines in the two or more containers. ' Where a container in a kit contains a certain polysaccharide conjugate, it is preferred that the same conjugate is not present in the other containers of the kit. 25 The inventors have surprisingly found that a kit provided in the above ways advantageously presents the various antigens to a host's imunune system in an optimal manner. The kit provides a medical practitioner with an optimal method of immunising a host with one or more of the following advantages (preferably 2 or 3, and most preferably all): protective efficacy for all antigens, minimal reactogenicity, 30 minimal carrier suppression interference, minimal adjuvant/antigen interference, or minimal antigen/antigen interference. In such a way, these goals may be achieved with the minimum number (two) administrations, preferably occurring at the same visit to the practitioner. 5 5 In another embodiment the present invention relates to a vaccine kit when used for concomitant administration comprising two multi-valent immunogenic compositions conferring protection in a host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and Streptococcus pneumoniae, said kit comprising a first container comprising: 0 (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and (e) Inactivated polio virus, 5 and a second container comprising: (2a) one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, and wherein the first container comprises a DT content of 60-120 pg, and the second container comprises one or more polysaccharides or oligosaccharides conjugated to DT 0 and/or CRM197. In another embodiment the present invention relates to a multi-valent immunogenic composition comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, 5 (b) tetanus toxoid (TT), (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and (e) Inactivated polio virus; conferring protection in a host against disease caused by Bordetella pertussis, Clostridium 30 tetani, Corynebacterium diphtheriae, Hepatitis B virus, and Polio virus, when used for the manufacture of a medicament, wherein said medicament is coadministered with another immunogenic composition comprising: (2a) one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, 35 conferring protection in a host against Streptococcus pneumoniae, 5a 5 and wherein the multi-valent immunogenic composition comprises a DT content of 60-120 pg, and the other immunogenic composition comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197. In another embodiment the present invention relates to an immunogenic composition 0 comprising: (2a) one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, conferring protection in a host against Streptococcus pneumoniae, when used for the manufacture of a medicament, wherein said medicament is coadministered 5 with a multi-valent immunogenic composition comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and 0 (e) Inactivated polio virus; conferring protection in a host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, and Polio virus, wherein the immunogenic composition comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197 and the multi-valent immunogenic 5 composition comprises a DT content of 60-120 tg. In another embodiment the present invention relates to the use of a multi-valent immunogenic composition comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, 30 (b) tetanus toxoid (TT), (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and (e) Inactivated polio virus, wherein the multi-valent immunogenic composition comprises a DT content of 60-120 pig in 35 the manufacture of a medicament when used for conferring protection against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and Streptococcus pneumoniae, wherein said medicament is coadministered with 5b 5 an immunogenic composition comprising one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, which immunogenic composition comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM 197. 0 In another embodiment the present invention relates to the use of an immunogenic composition comprising one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, which immunogenic composition comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197 in the manufacture of a medicament when used for conferring protection 5 against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and Streptococcus pneumoniae, wherein said medicament is coadministered with a multi-valent immunogenic composition comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), 0 (c) diphtheria toxoid (DT), (d) Hepatitis B surface, and (e) Inactivated polio virus wherein the immunogenic composition comprises a DT content of 60-120 ptg. 5c Although in a preferred embodiment the vaccines of the first and second (and third where applicable) containers are administered concomitantly at different sites (as described later), in an alternative embodiment the inventors envision that the contents of the first and second containers may be mixed (preferably extemporaneously) before 5 administration as a single vaccine. The antigens of the invention Methods of preparing tetanus toxoid (TT) are well known in the art. For instance, TT is preferably produced by purification of the toxin from a culture of 10 Clostridium tetani followed by chemical detoxification, but is alternatively made by purification of a recombinant, or genetically detoxified analogue of the toxin (for example, as described in EP 209281). 'Tetanus toxoid' also encompasses immunogenic fragments of the full-length protein (for instance Fragment C - see EP 478602). 15 Methods of preparing diphtheria toxoid (DT) are also well known in the art. For instance, DT is preferably produced by purification of the toxin from a culture of Corynebacterium diphtheriae followed by chemical detoxification, but is alternatively made by purification of a recombinant, or genetically detoxified analogue of the toxin (for example, CRM197, or other mutants as described in US 4,709,017, US 20 5,843,711, US 5,601,827, and US 5,917,017). Acellular pertussis components (Pa) are well known in the art. Examples include pertussis toxoid (PT), filamentous haemagluttinin (FHA), pertactin (PRN) and agglutinogens.2 and 3. These antigens are partially or highly purified. Preferably 2 or more acellular pertussis components are used in the vaccine. More preferably 2, 3, 4 25 or all 5 of the above example acellular pertussis components are incorporated in the vaccine. Most preferably PT, FHA and PRN are included. PT may be produced by a variety of manners, for instance by purification of the toxin from a culture of B. pertussis followed by chemical detoxification, or alternatively by purification of a genetically detoxified analogue of PT (for example, as described in US 5,085,862). 30 Methods of preparing killed, whole-cell Bordetella pertussis (Pw) suitable for this invention is disclosed in WO 93/24148, as are suitable formulation methods for producing DT-TT-Pw-HepB and DT-TT-Pa-HepB vaccines. Inactivated Polio Virus (IPV) preferably comprises types 1, 2 and 3 as is standard in the vaccine art. Most preferably it is the Salk polio vaccine. 6 Typically the Streptococcus pneumoniae vaccine of the present invention will comprise polysaccharide antigens (preferably conjugated), wherein the polysaccharides are derived from at least four serotypes of pneumococcus chosen from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, llA, 12P, 14, 5 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F. Preferably the four serotypes include 6B, 14, 19F and 23F. More preferably, at least 7 serotypes are included in the composition, for example those derived from serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F. More preferably still more than 7 serotypes are included in the composition, for instance at least 11 serotypes. For example the composition in one embodiment 10 includes 11 capsular polysaccharides derived from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F (preferably conjugated). In a preferred embodiment. of the invention at least 13 polysaccharide antigens (preferably c6njugated) are included, although further polysaccharide antigens, for example 23 valent (such as serotypes 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 1OA, lIA, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 15 23F and 33F), are also contemplated by the invention, For elderly vaccination (for instance for the prevention of pneumonia) it is advantageous to include serotypes 8 and 12F (and most preferably 15 and 22 as well) to the preferred 11 valent antigenic composition described above to form a 13/15 valent vaccine, whereas for infants or toddlers (where otitis media is of more concern) 20 serotypes 6A and 19A are advantageously included to form a 13 valent vaccine. Coniugates The bacterial capsular polysaccharide conjugates may comprise any carrier peptide, polypeptide or protein comprising at least one T-helper epitope. Preferably 25 the carrier protein(s) used is selected from the group coriprising: tetanus toxoid, diphtheria toxoid, CRM197, recombinant diphtheria toxin (as described in any of US 4,709,017, WO 93/25210, WO 95/33481, or WO 00/48638), pneumolysin (preferably chemically detoxified, or a detoxified mutant) from S. pneumoniae, OMPC from N. meningitidis, and protein D from H. influenzae (EP 594610). Due to the known effect 30 of carrier suppression, it is advantageous if in each of the compositions of the invention the polysaccharide antigens contained therein ('n' antigens) are conjugated to more than one carrier. Thus (n-1) of the polysaccharides could be carried (separately) on one type of carrier, and 1 on a different carrier, or (n-2) on one, and 2 on two different carriers, etc. For example, in a vaccine containing 4 bacterial 7 polysaccharide conjugates, 1,.2 or all four could be conjugated to different carriers). Protein D, however, is advantageously used as a carrier in the compositions of the invention as it may be used for various (2, 3, 4 or more) polysaccharides in a composition without a marked carrier suppression effect. Most preferably Hib is 5 present as a TT conjugate, pneumococcal polysaccharides are protein D, DT or CRM197 conjugates,.and MenA, MenC, MenY and MenW are either TT or PD conjugates. Protein D is also a useful carrier as it provides a further antigen which can provide protection against H. influenzae. The polysaccharide may be linked to the carrier protein by any known method 10 (for example, by Likhite, U.S. Patent 4,372,945 and by Armor et aL., U.S. Patent 4,474,757). Preferably, CDAP conjugation is carried out (WO 95/08348). In CDAP, -the cyanylating reagent l-cyano-dimethylaminopyridinium tetrafluoroborate (CDAP) is preferably used for the synthesis of polysaccharide protein conjugates. The cyanilation reaction can be performed under relatively mild 15 conditions, which avoids hydrolysis of the alkaline sensitive polysaccharides. This synthesis allows direct coupling to a carrier protein. Properties of the immunogenic compositions of the invention The immunogenic compositions of the invention are preferably formulated as 20 a vaccine for in vivo administration to the host in such a way that the individual components of the composition are formulated such that the immunogenicity of individual components is not substantially impaired by other individual components of the composition. By not substantially impaired, it is meant that upon immunisation, an antibody titre (e.g. IgG) against each component is obtained which is more than 25 60%, preferably more than 70%, more preferably more than 80 %, still more preferably more than 90%, and most preferably more than 95-100% of the titre obtained when the antigen is administered in isolation. Interestingly, with the kit combinations described above, it is possible, upon immunisation, to obtain antibody titres against Hib capsular polysaccharide or some 30 pneumococcal polysaccharides approaching, or in excess of, 100% of the titre obtained when the antigen is administered in isolation. 8 Vaccine formulations The immunogenic compositions of the invention are preferably formulated as a vaccine for in vivo administration to the host, such that they confer an antibody titre superior to the criterion for seroprotection for each antigenic component for an 5 acceptable percentage of human subjects. This is an important test in the assessment of a vaccine's efficacy. throughout the population. Antigens with an associated antibody titre above which a host is considered to be seroconverted against the antigen are well known, and such titres are published by organisations such as WHO. Preferably more than 80% of a statistically significant sample of subjects is 10 seroconverted, more preferably more than 90%, still more preferably more than 93% and most preferably 96-100%. The immunogenic compositions of the invention are preferably adjuvanted. Suitable adjuvants include an aluminium salt such as aluminium hydroxide gel (alum) or aluminium phosphate, but may also be a salt of calcium, iron or zinc, or may be an 15 insoluble suspension of acylated tyrosine, or acylated sugars, cationically or anionically derivatised polysaccharides, or polyphosphazenes. The adjuvant may also be selected to be a preferential inducer of a THI type of response to aid the cell mediated branch of the immune response. High levels of Thl-type cytokines tend to favour the induction of cell 20 mediated immune responses to a given antigen, whilst high levels of Th2-type cytokines tend to favour the induction of humoral immune responses to the antigen. Suitable adjuvant systems which promote a predominantly Thl response include, Monophosphoryl lipid A or a derivative thereof, particularly 3-de-O-acylated monophosphoryl lipid A, and a combination of monophosphoryl lipid A, preferably 3 25 de-O-acylated monophosphoryl lipid A (3D-MPL) together with an aluminium salt. An enhanced system involves the combination of a monophosphoryl lipid A and a saponin derivative, particularly the combination of QS21 and 3D-MPL as disclosed in WO 94/00153, or a less reactogenic composition where the QS21 is quenched with cholesterol as disclosed in WO 96/33739. A particularly potent adjuvant formulation 30 involving QS21, 3D-MIPL and tocopherol in an oil in water emulsion is described in WO 95/17210. The vaccine may additionally comprise a saponin, more preferably QS21. The formulation may also comprises an oil in water emulsion and tocopherol (WO 95/17210). Ulnmethylated CpG containing oligonucleotides (WO 96/02555) are 9 also preferential inducers of a TH1 response and are suitable for use in the present invention. Aluminium salts are preferred adjuvants in the above immunogenic compositions. In particular, HepB should preferably be adsorbed onto aluminium 5 phosphate before admixing with the other components. Pertactin is preferably adsorbed onto aluminium hydroxide before admixing with the other components. In order to minimise the levels of adjuvant (particularly aluminium salts) in the compositions of the invention, the polysaccharide conjugates may be unadjuvanted. The present invention also provides a method for producing a vaccine 10 formulation comprising the step of mixing the components of the vaccine together with a pharmaceutically acceptable excipient. A particularly preferred DTPa composition of the invention (for independent use or as the contents of the first container of one of the above-described kits) 15 comprises: Tr, DT, Pa (preferably comprising PT, FRA and PRN - with PRN preferably adsorbed onto aluminium hydroxide), HepB (preferably adsorbed onto aluminium phosphate), IPV, MenC (preferably conjugated onto either protein D, TT, DT or CRM197), and, optionally, MenY (preferably conjugated onto either protein D, TT, DT or CRM197). The composition may also optionally comprise Hib (preferably 20 conjugated onto TT and/or unadsorbed onto adjuvant). Preferably the vaccine may be supplied in 2 vials, the first containing DTPa-IPV-HepB in a liquid form, and a second containing MenC (and optionally MenY and/or Hib) in a lyophilised form, preferably in the presence of an anti-caking agent such as sucrose, lactose or trehalose. The contents of the vials may be mixed extemporaneously in a single 25 container before administering to a host in a single administration/injection. This composition may also be used in a kit described above (the contents of the first container). For the purpose of kits comprising a container comprising Hib (preferably conjugated onto TT and/or unadsorbed onto adjuvant) and/or either or both of MenC 30 and MenY (preferably conjugated onto either protein D, TT, DT or CRM197 and/or unadsorbed onto adjuvant), this composition is preferably stored in a lyophilised form, preferably in the presence of an anti-caking agent such as sucrose, lactose or trehalose. 10 For the purpose of DTPa compositions of the invention (for independent use or as the contents of the first container of one of the above-described kits) comprising a container comprising DTPa and Hib and/or either or both of MenC and MenY, where the Hib and/or Men components are conjugated to TT, it is preferable to 5 balance the TT content in the vaccine such that the total content of TT in a single container is not more than a critical threshold (such as 40, 45, 50, 60, 70 or 80 pg TT) to reduce, minimise or prevent TT immune interference or carrier suppression of TT conjugated polysaccharides. Preferably this threshold is 50 pg. The inventors have found that the ratio of polysaccharide:TT may be reduced in the above conjugates to 10 1:0.5-1.5 by weight (preferably 1: 0.6-1.2, most preferably around 1:1) to be beneficial in this respect. For instance in a DTPa-HB-IPV-Hib(TT)-MenC(TT) vaccine the amount of T in DTPa should preferably be reduced below a typical standard quantity (preferably about one to three quarters, most preferably about a half of the regular amount) to, for instance, 10-30 pg TT, preferably 20-25 pg TI'. For 15 example, if the amount of TT conjugated to Rib is around 12 pg TT, and the amount conjugated to MenC is around 5 pg TT, and the amount of unconjugated TT is 24 pg, then the total TT will be about 41 4g. A particularly preferred Hib / pneumococcal polysaccharide composition (for 20 independent use or as the contents of the second container of one of the above described kits) comprises: Hib (preferably conjugated onto TI and/or unadsorbed onto adjuvant) and multiple (for instance more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11) pneumococcal polysaccharide conjugates (for instance those combinations described in the paragraph on 'the Streptococcus pneumoniae vaccine of the present invention' 25 above). Most preferably 11 polysaccharides (from serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F) are included. Preferably pneumococcal polysaccharides are conjugated onto PD, DT, CRM197 or TT. In a preferred embodiment, the Hib polysaccharide antigen is not adsorbed onto an adjuvant, particularly aluminium salts. Although the pneumococcal polysaccharide antigen(s) may be adjuvanted (preferably 30 onto aluminium phosphate), they may also be not adsorbed onto an adjuvant, particularly aluminium salts. In a particular embodiment, there is no aluminium adjuvant salt present in the composition. Further antigens may be included in the compositions of the invention (for instance N. meningitidis Type C capsular 11 polysaccharide conjugate (preferably conjugated onto either protein D, TT, DT or CRM197 and/or unadsorbed onto adjuvant]), however, in an alternative embodiment, Hib and pneumococcal polysaccharide conjugates are the only antigens present in the composition. In a further specific embodiment of the above formulations, the Hib and 5 pneumococcal polysaccharides are not conjugated to the same carrier (particularly where the carrier is CRM1 97). The vaccine may be supplied in one container (with the contents either in a liquid or lyophilised form), or in two vials, the first containing Hib (preferably lyophilised), the second containing the pneumococcal antigens (preferably in a liquid 10 form). Lyophilised compositions are preferably in the presence of an anti-caking agent such as sucrose, lactose or trehalose. The contents of the vials may be mixed extemporaneously in a single container before administering to a host in a single administration/injection. With such a formulation it is possible, upon immunisation, to obtain antibody titres against Hib capsular polysaccharide approaching, or most often 15 in excess of, 100% of the tire obtained when the antigen is administered in isolation. In preferred embodiments, no (significantly) detrimental effect occur to the pneumococcal polysaccharide conjugates (in terms of protective efficacy) in the combination as compared to their administration in isolation. This can be assessed in terms of measuring post-primary geometric mean concentrations (GMC) of anti 20 polysaccharide antibody I month after the last primary dose (primary doses being the priming administrations - usually 3 - in the first year of life). The GMC (in pg/ml) for a vaccine of the invention should be preferably over 55% (more preferably over 60, 70, 80, or 90%) of the GMC when the pneumococcal polysaccharides are administered without the Hib conjugate. Another indication that no detrimental effect 25 has occurred is if the % of subjects with antibody concentrations of no less than 0.5 ptg/ml differs by no more than 10% (preferably less than 9, 7, 5, 3 or 1%) when comparing 1 month post-primary administrations of the vaccine of the invention versus the vaccine without Hib conjugate. Although the above refers to Hib, pneumococcal and meningococcal 30 'polysaccharides' it is envisaged that the invention may be extended to Rib and pneumococcal 'sized-polysaccharides' and 'oligosaccharides' (polysaccharides reduced in size for manageability, which are still capable of inducing a protective immune response in a host) which are well known in the vaccine art (see for instance 12 EP 497525). Advantageously, MenY may be present as an oligosaccharide conjugate with the oligosaccharide 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, or 0.9 times the molecular weight of the native polysaccharide. 5 In a further aspect of the present invention there is provided an immunogenic composition or vaccine as herein described for use in a medicament. In a still further aspect of the invention there is provided a use of the immunogenic compositions of the invention in the manufacture of a medicament for the treatment or prevention of diseases caused by infection by Bordetella pertussis, 10 Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and N. meningitidis (and optionally H. influenzae). Furthermore, there is provided a use of the immunogenic compositions of the invention in the manufacture of a vaccine kit for the treatment or prevention of diseases caused by infection by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio 15 virus, Haemophilus influenzae, Streptococcwpneumoniae and N. meningitidis. Additionally, a method of immunising a human host against disease caused by Bordetella pertussis, Clostridium letani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and N. meningitidis (and optionally H. influenzae), which method comprises administering to the host an immunoprotective dose of the immunogenic 20 composition of the invention is also provided. A further aspect of the invention concerns a method of immunising a human host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterum diphtheriae, Hepatitis B virus, and Polio virus, and one or more of 25 Haemophilus influenzae, Streptococcus pneumoniae and N. meningitidis, with the kits of the invention described above, which method involves a concomitant administration schedule as defined below. Concomitant administration schedule 30 Such a schedule comprises the step of administering to a host an immunoprotective dose of an immunogenic composition of a first container of a kit (for instance one of the kits of the invention) at a different site drained by a different lymph nodes from the site at which the immunogenic composition of the second (or third) container of the kit is administered. Preferably the different sites are different 13 limbs. Preferably the administration of the vaccines occurs within 24 hours of each other, more preferably within the same day, and most preferably at the same visit of the host to the practitioner. Preferably, the host is subsequently primed with both (or all) vaccines in the same way one or more (preferably 2) further times, each time 5 separated by 2-12 weeks (preferably approximately 1 month). Often a third priming administration may be .given between 2 weeks and 7 months after the second administration. For instance, the vaccine may be administered as above according to a normal administration schedule for DTP vaccines (such as a three visit system, each visit separated by 1 month, for instance a 3, 4 and 5 month of age schedule; or a 3, 5 10 and 11; or a 3, 5 and 12 month of age schedule). Such an administration schedule allows the optimisation of the immune response against the antigens in both (or all) containers 'of the kit. A booster administration of the vaccines may be given in the same way anytime from the second year of life to adulthood. Although priming is preferably 15 done via the intramuscular route, boosting may advantageously be carried out mucosally, optionally in the presence of a mucosal adjuvant (preferably laureth 9 or Heat Labile Toxin [LT] from E. coli and mutants or fragments thereof), (for instance intranasal administration of the vaccines is easy to administer and can work extremely well especially when the host is primed parenternally), and site of administration of 20 the vaccines need not drain to different lymph nodes. The use of the immunogenic compositions of the invention within containers in a method of manufacturing a vaccine kit of the invention for concomitant administration is also envisaged. 25 Kits comprising TT in two or more containers A further aspect of the invention concerns vaccine kits for concomitant administration (as defined above) where the TT content of two or more containers are balanced to advantageously reduce, minimise or prevent TT immune interference or carrier suppression of TIT conjugated polysaccharides. TT is an extremely good 30 carrier, however it is known that it has limitations if used to excess in a vaccine composition, particularly if free Tr is also present. If used excessively, all antigens conjugated to TT exhibit reduced antibody titres. There is therefore a distinct problem in the art of how to use TT in many different areas (for instance as free antigen and as carrier for many polysaccharide antigens) within a large combination vaccine without 14 the above disadvantages. The present inventors have found an optimal method of solving this problem; that by using a kit concomitant administration schedule (as defined above), a vaccine in a first container comprising TT in a quantity not more than a critical threshold where immune interference or carrier suppression occurs can 5 be administered with a vaccine in a second (and optionally third) container comprising IT in a quantity not more than a critical threshold where immune interference or carrier suppression occurs such that the total quantity of TT concomitantly administered is above this critical threshould, and immune interference (or carrier suppression) is minimised (i.e. less than if the components had been 10 administered in one injection) and preferably does not occur at all. The critical threshold can be 40, 45, 50, 60, 70 or 80 pg TT, and is preferably about 50 pg TT.. The maximum total TT that can be administered is therefore approximately up to a quantity derived from the number of containers of the kit (two or three) multiplied by the critical threshould. 15 The present invention therefore provides a kit comprising two (or three) containers comprising two (or three) immunogenic compositions for concomitant administration each comprising TT in a free and/or conjugated form, wherein the quantity of TT in each container is not more than a critical threshold to prevent or minimise TT immune interference (or carrier suppression) effects, but the total TT in 20 all containers is more than said critical threshold. Preferably at least one of the containers should include free (unconjugated) TT, most preferably in the context of a DTPa or DTPw multivalent vaccine. Although the quantity of free TT can be present at around normal levels of approximately 42 pg, a further advantage of the invention allows lower quantities to be present (10-30 25 or 10-20 pg, for instance 10, 15, 20, 25 or 30 pg) but optimal anti-TT antibody titres may still be elicited with minimal (or no) immune interference or carrier suppression effects. Preferably at least one (but possibly 2 or 3) of the containers should include at least one (but possibly 2, 3, 4, 5, 6, 7 or more) TT conjugated polysaccharide. Where 30 free TT is present in one container, it is preferred that at least one TT-conjugated polysaccharide should be in one of the other containers of the kit. The polysaccharide may be any described in this application, preferably one or more pneumococcal polysaccharides (as described above), or MenC, MenY, or Bib. 15 Preferably the kit is any of the kits of the invention as described above. Preferably one, two, three or all the polysaccharide-TT conjugates present in the kit are such that the ratio of polysaccharide:TT is reduced (compared to standard conjugates) to 1:0.5-1.5 by weight (preferably 1: 0.6-1.2, most preferably around 1:1) 5 such that the conjugates are still immunologically functional, but TT immune interference or carrier suppression effects are facilitated in being minimised or prevented. Further provided is a method of immunising a human host using the above kit, which method comprises administering an immunoprotective dose of the 10 immunogenic composition of the first container to the host at a first site, administering an .immunoprotective dose of the immunogenic composition of the second container to. the host at a second site (and optionally administering an immunoprotective dose of the immunogenic composition of the third container to the host at a third site), wherein the first and second (and third) sites are drained by 15 different lymph nodes. Concomitant administration should be carried out as described above. Preferably the first and second (and third) sites represent different limbs of the host. Preferably the administration of the immunogenic compositions of the first and second (and third) containers occurs on the same day. Preferably the host is 20 subsequently vaccinated in the same way one or more further times, each time separated by 2-12 weeks, more preferably two further times, each time separated by approximately a period of 1-2 months. 25 Kits comprising DT or CRM197 in two or more containers A still further aspect of the invention concerns vaccine kits for concomitant administration (as defined above) where the DT content (including DT and any immunologically identical mutants such as CRM197) of two or more containers are balanced advantageously to enhance DT (or CRM197) conjugated polysaccharide 30 antibody titres whilst minimising reactogenicity (i.e. lower reactogenicity than if the components of the containers were administered in a single injection). DT and CRM197 are extremely good carriers, however it is known that DT contributes largely to the reactogenicity of vaccines containing it. The present inventors have found that by using a kit concomitant administration schedule (as defined above), a vaccine in a 16 first container comprising DT (and/or CRM197) is advantageously present in a high amount (40-150 pg, preferably 60-120 I±g, more preferably 70-100 pg, most preferably around 95 pg) where a vaccine in a second (and optionally third) container comprising a DT- or CRM197-conjugated polysaccharide is concomitantly 5 administered. The advantages- of this invention are that a) although the DT content is high in the first container it is not high enough to induce DT immune interference or carrier suppression effects, b) the DT- or CRM-197 polysaccharide conjugate is separated from the first container so that the reactogenicity of the vaccine of the first container 10 is not increased, yet c) the antibody titre against the polysaccharide conjugated to DT or CRMI 97 is not reduced and may be enhanced (greater titres compared to where the conjugate is administered separately, or compared to where lower quantities of DT are present in the first container). The present -invention therefore provides a kit comprising two (or three) 15 containers comprising two (or three) immunogenic compositions for concomitant administration (as defined above), wherein the first container comprises a DT content (DT plus CRM197; preferably free or unconjugated) which is present in a high amount (as defined above), and the second (and third) containers comprise one or more polysaccharides conjugated to DT and/or CRM197. 20 Preferably the first container should include free (unconjugated) DT, most preferably in the context of a DTPa or DTPw multivalent vaccine. The DT/CRM197 conjugated polysaccharide(s) may be any described in this application; preferably one or more from the following list: pneumococcal polysaccharides 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 25 18C, 19A, 19F, 20, 22F, 23F or 33F, MenC, MenY, or Hib. Preferably according to this invention the immune response (antibody titres) against one or more of these polysaccharides is maintained compared to administering the conjugate by itself, and is most preferably enhanced. Preferably the kit is any of the kits of the invention as described above. 30 Preferably one, two, three or all the polysaccharide-DT (or CRM197) conjugates present in the kit are such that the ratio of polysaccharide:DT/CRM197 is reduced (compared to standard conjugates) to 1:0.5-1.5 by weight (preferably 1: 0.6 1.2, most preferably around 1:1). 17 Further provided is a method of immunising a human host using the above kit, which method comprises administering an immunoprotective dose of the immunogenic composition of the first container to the host at a first site, administering an immunoprotective dose of the immunogenic composition of the 5 second container to the host at a second site (and optionally administering an immunoprotective dose of the immunogenic composition of the third container to the host at a third site), wherein the first and second (and third) sites are drained by different lymph nodes. Concomitant administration should be carried out as described above. 10 Preferably the first and second (and third) sites represent different limbs of the host. Preferably the administration of the immunogenic compositions of the first and second (and third) containers occurs on the same day. Preferably the host is subsequently vaccinated in the same way one or more father times, each time separated by 2-12 weeks, more preferably two further times, each time separated by 15 approximately a period of 1-2 months. The vaccine preparations of the present invention may be used to protect or treat a mammal susceptible to infection, by means of administering said vaccine via 20 systemic or mucosal route. These administrations may include injection via the intramuscular, intraperitoneal, intradermal or subcutaneous routes; or via mucosal administration to the oral/alimentary, respiratory (e.g. intranasal), genitourinary tracts. The amount of antigen in each vaccine dose is selected as an amount which induces an immunoprotective response without significant, adverse side effects in 25 typical vaccines. Such amount will vary depending upon which specific immunogen is employed and how it is presented. Generally, it is expected that each dose will comprise 0.1-100 pg of polysaccharide, preferably 0.1-50 pg, preferably 0.1-10 pg, of which I to 5 pg is the most preferable range. The content of protein antigens in the vaccine will typically be in the range 1 30 100pug, preferably 5-50pg, most typically in the range 5 - 25pg. Following an initial vaccination, subjects may receive one or several booster immunisations adequately spaced. 18 Vaccine preparation is generally described in Vaccine Design ("The subunit and adjuvant approach" (eds Powell M.F. & Newman MJ.) (1995) Plenum Press New York). Encapsulation within liposomes is described by Fullerton, US Patent 4,235,877. 5 EXAMPLES Examples are provided solely for the purposes of illustration and are not 10 intended to limit the scope of the invention. Example 1: Preparation of a DT-TT-Pa-IPV-HepB (DTPaIPVHepB) Vaccine This was done as described in WO 93/24148. The vaccine is commercially 15 available under the name Infanrix-PeNTaTm (SmithKline Beecham Biologicals). Example 2: Preparation of a MenC or MenC-MenY Vaccine MenC: N meningitidis type C capsular polysaccharide conjugated onto either 20 protein D or TT (using the CDAP technique) present in an amount of 5 pg of polysaccharide in the conjugate per 0.5 mL human dose. The pH was adjusted to 6.1, and was lyophilised in the presence of sucrose. MenCMenY: N. meningitidis type C capsular polysaccharide conjugated onto either protein D or TT (using the CDAP technique) and N. meningitidis type Y 25 capsular polysaccharide conjugated onto either protein D. or TT were mixed together in an amount of 5 pg of polysaccharide in each conjugate per 0.5 nL human dose. The pH was adjusted to 6.1, and was lyophilised in the presence of sucrose. 30 19 Example 3: Preparation of a DT-TT-Pa-IPV-HepB-MenC-MenY (DTPaIPVHepB/IMenCMenY) or a DT-TT-Pa-IPV-HepB-MenC (DTPaIPVHepB/MenC) Vaccine 5 The vaccines of Example 1 and Example 2 were mixed extemporaneously (on the same day) before use. Example 4: Preparation of a Hib - 11 valent pneumococcal conjugate 10 (Hib/Strep1IV) Vaccine H. influenzae type b capsular polysaccharide conjugated onto TT (10 pg of polysaccharide in the conjugate per dose) which had been lyophilised at a pH of 6.1 in the presence of lactose [HiberixTm (SmithKline Beecham Biologicals)] was 15 extemporaneously (on the same day as use) dissolved in a liquid solution of eleven valent pneumococcal capsular polysaccharide (serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F) conjugated onto PD (1 pg of polysaccharide in each conjugate per dose). The pneumococcal vaccine had previously been adsorbed onto 0.5 mg Al (as

AIPO

4 ). 20 Example 5: Clinical Trials 25 Studies on the Vaccine of Example 4 The vaccine of Example 4 and a control vaccine were administered in a three dose (3, 4, 5 months of age) schedule to German infants. The immune response results (measured 1 month after the last primary 30 administration) were as follows. 35 20 Anti pncumococcal IgG antibodies: GMC (Vg/ml) yElisa) PS GroupA GroupD Anti- Timin N S+[%) GMC N S+[%] Mc Anti-i Pl 30 100 1.23 33 100 0.99 Anti-3 PM 30 100 2.04 33 97.0 1.20 Anti-4 PM 30 100 0.98 33 100 1.03 Anti-5 PM 30 100 1.33 33 100 1.34 Anti-6B PM 30 1db 0.54 33 100 0.62 Anti-7F PM 30 100 1.60 33 100 1.33 Anti-9V PM 30 100 1.61 33 100 1.21 Anti-14 PM 30 100 2.27 33 100 2.32 Anti-18C PM 30 100 1.06 33 100 1.04 Anti-19F PM 30 100 2.05 33 100 1.92 Anti-23F PH 30 96.7 0.75 33 100 0.76 Group A- 1lPn-PD + Infanrix-HeXaM (Infanrix-Penta plus added Hib conjugate - DTPa-HB-1PV 5 Hib) Group D -i iPn-PD/Hb + Infanrix-PeNTa" (DTPa-HB-IPV) + indicates concomitant (in different limbs) rather than combined administration. Percent of subjects with antibody concentrations no less than 0.5 g/nml Group PS 13 4 5 6B 7F 7V 14 1 9F 23F D 84.8 87.9 87.9 90.9 51.5 90.9 93.9 97.0 81.8 97.0 72.7 A .7 96. 6.7 90.0 50. 93.3 90.0 90.0 80.0 96.7 66.7 10 Anti PRP antibodies: GMC (pgm1) (By Elisa) GroupD (N -34) n :1 GMC 33 100 10.75 100% of subjects had anti-PRP (Hib polysaccharide) antibody concentrations no less than 1.0 pg/ml. 15 Hiberix (unadsorbed Hib-TT conjugate) has a GMC after a similar administration schedule of about 6 pg/ml. The immune response, in terms of ELISA antibodies, of infants who received the 1lPn-PD/Hib vaccine was similar to that observed for those who received the 20 l1Pn-PD vaccine for all of the serotypes, with the exception of serotypes 1, 3 and 9V for which a trend to lower geometric mean concentrations was observed for the 1 lPn PD/Hib vaccine. However, these differences were not significant as shown by the overlapping of 95% confidence intervals. 21 The 1 lPn-PDHib vaccine induced functional (opsonophagocytic) antibodies to al 111 serotypes. Combining the Hib vaccine with the pneumococcal conjugate vaccine did not significantly interfere with the pneumococcal immune response and surprisingly 5 enhanced the anti PRP response compared to both the registered vaccines Infanrix HeXa and Hiberix. Studies on the Vaccines of Example 3, or the Concomitant Administration of the 10 Vaccines of Example 3 and Example 4 Study 1: The safety and immunogenicity of Infanrix-PeNTa mixed with MenC conjugate vaccine given with a Rib vaccine or concomitantly with 11-valent pneumococcal 15 vaccine mixed with Hiberix can be evaluated. Both PD and TT carriers can be evaluated for the MenC conjugate. The vaccines can be administered as a three dose vaccine in infants. Concomitant injection can be in different limbs, administered in the same visit to the practitioner. 20 Study 2: The safety and immunogenicity of Infanrix-PeNTa mixed with MenC-MenY conjugate vaccine given with a Hib vaccine or concomitantly with 1 1-valent pneumococcal vaccine mixed with Hiberix can be evaluated. Both PD and IT carrier can be evaluated for the MenC and MenY conjugates. The vaccines can be 25 administered as a three dose vaccine in infants. Concomitant injection can be in' different limbs, administered in the same visit to the practitioner. 22 Throughout the description and claims of the specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps. A reference herein to a patent document or other matter which is given as prior 5 art is not to be taken as an admission or a suggestion that that document or matter was, known or that the information it contains was part of the common general knowledge as at the priority date of any of the claims. 22a

Claims

1. A vaccine kit when used for concomitant administration comprising two multi-valent immunogenic compositions conferring protection in a host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, 0 Polio virus and Streptococcus pneumoniae, said kit comprising a first container comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and 5 (e) Inactivated polio virus, and a second container comprising: (2a) one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcuspneumoniae, and wherein the first container comprises a DT content of 60-120 ptg, and the second .0 container comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197.

2. The vaccine kit of claim 1 wherein the first container comprises a DT content of 70 100 tg. 5

3. The vaccine kit of claim 1 or 2, wherein the one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197 are selected from a list consisting of pneumococcal polysaccharides or oligosaccharides 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, llA, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F, meningococcal 30 polysaccharides or oligosaccharides MenC and MenY, and H influenzae type b Hib.

4. The vaccine kit of claims 1-3, wherein the second container comprises seven pneumococcal polysaccharides or oligosaccharides derived from serotypes 4, 6B, 9V, 14, 18C, 19F and 23F which are conjugated to CRM197. 35 23 5

5. The vaccine kit of claims 1-4 wherein one or more of the polysaccharides or oligosaccharides conjugated to DT and/or CRM197 have a ratio of polysaccharide or oligosaccharide: DT or CRM197 of 1:0.5-1.5 by weight.

6. The vaccine kit of claims 1-5, wherein the one or more conjugates of a carrier protein 0 and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae is derived from one or more pneumococcal serotypes selected from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 1OA, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F.

7. The vaccine kit of claims 1-6, wherein the Hepatitis B surface antigen is adsorbed 5 onto aluminium phosphate.

8. The vaccine kit of claims 1-7 wherein the first container additionally comprises: (f) either or both conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide of a bacterium selected from the group N. meningitidis type Y (MenY) and N. meningitidis 0 type C (MenC), and (g) a conjugate of a carrier protein and the capsular polysaccharide or oligosaccharide of H influenzae type B (Hib).

9. The vaccine kit of claims 1-7 wherein the second container additionally comprises: (2b) either or both conjugates of a carrier protein and a capsular polysaccharide or 5 oligosaccharide of a bacterium selected from the group N. meningitidis type Y (MenY) and N. meningitidis type C (MenC), and (2c) a conjugate of a carrier protein and the capsular polysaccharide or oligosaccharide of H influenzae type B (Hib).

10. The vaccine kit of claims 1-7 wherein the first container additionally comprises: (f) 30 either or both conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide of a bacterium selected from the group N. meningitidis type Y (MenY) and N. meningitidis type C (MenC), and the second container additionally comprises (2b) a conjugate of a carrier protein and the capsular polysaccharide or oligosaccharide of H influenzae type B (Hib). 35

11. The vaccine kit of claims 1-7 wherein the first container additionally comprises (f) a conjugate of a carrier protein and the capsular polysaccharide or oligosaccharide of H influenzae type B (Hib), and the second container additionally comprises: (2b) either or both conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide of a 24 5 bacterium selected from the group N. meningitidis type Y (MenY) and N. meningitidis type C (MenC).

12. The vaccine kit of claims 1-7 additionally comprising a third container comprising: (3a) either or both conjugates of a carrier protein and a capsular polysaccharide or ) oligosaccharide of a bacterium selected from the group N meningitidis type Y (MenY) and N. meningitidis type C (MenC), and (3b) a conjugate of a carrier protein and the capsular polysaccharide or oligosaccharide of H. influenzae type B (Hib).

13. The vaccine kit of claims 1-12 wherein two or more containers comprise TT, the 5 quantity of TT in each of said two or more containers being not more than a critical threshold of 50 tg TT to prevent or minimise TT immune interference or carrier suppression effects, but the total TT in all containers of the vaccine kit being more than said critical threshold.

14. The vaccine kit of claim 13 wherein 1, 2 or 3 of the containers includes one or more ) TT-conjugated polysaccharide or oligosaccharide selected from a list consisting of a pneumococcal polysaccharide or oligosaccharide, MenC, MenY, and Hib.

15. The vaccine kit of claim 14 wherein one or more of the TT-conjugated polysaccharides or oligosaccharides have a ratio of polysaccharide or oligosaccharide:TT of 5 1:0.5-1.5 by weight.

16. A multi-valent immunogenic composition comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), 0 (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and (e) Inactivated polio virus; conferring protection in a host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, and Polio virus, 5 when used for the manufacture of a medicament, wherein said medicament is characterised in that it is coadministered with another immunogenic composition comprising: 25 5 (2a) one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, conferring protection in a host against Streptococcus pneumoniae, and wherein the multi-valent immunogenic composition comprises a DT content of 60-120 ptg, and the other immunogenic composition comprises one or more polysaccharides or D oligosaccharides conjugated to DT and/or CRM197.

17. An immunogenic composition comprising: (2a) one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, 5 conferring protection in a host against Streptococcus pneumoniae, when used for the manufacture of a medicament, wherein said medicament is characterised in that it is coadministered with a multi-valent immunogenic composition comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), J (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and (e) Inactivated polio virus; conferring protection in a host against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, and Polio virus, 5 wherein the immunogenic composition comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197 and the multi-valent immunogenic composition comprises a DT content of 60-120 pig.

18. The use of a multi-valent immunogenic composition comprising: 0 (a) acellular pertussis components comprising pertussis toxoid and FHA, (b) tetanus toxoid (TT), (c) diphtheria toxoid (DT), (d) Hepatitis B surface antigen, and (e) Inactivated polio virus, 5 wherein the multi-valent immunogenic composition comprises a DT content of 60-120 ptg in the manufacture of a medicament when used for conferring protection against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, 26 5 Polio virus and Streptococcus pneumoniae, wherein said medicament is coadministered with an immunogenic composition comprising one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, which immunogenic composition comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197. 0

19. The use of an immunogenic composition comprising one or more conjugates of a carrier protein and a capsular polysaccharide or oligosaccharide from Streptococcus pneumoniae, which immunogenic composition comprises one or more polysaccharides or oligosaccharides conjugated to DT and/or CRM197 in the manufacture of a medicament 5 when used for conferring protection against disease caused by Bordetella pertussis, Clostridium tetani, Corynebacterium diphtheriae, Hepatitis B virus, Polio virus and Streptococcus pneumoniae, wherein said medicament is coadministered with a multi-valent immunogenic composition comprising: (a) acellular pertussis components comprising pertussis toxoid and FHA, 0 (b) tetanus toxoid (TT), (c) diphtheria toxoid (DT), (d) Hepatitis B surface, and (e) Inactivated polio virus wherein the immunogenic composition comprises a DT content of 60-120 tg. 5

20. The vaccine kit of claim 1 substantially as hereinbefore described with reference to any of the examples and/or figures.

21. An immunogenic composition according to claim 16 or 17 substantially as 30 hereinbefore described with reference to any of the examples and/or figures.

22. A use according to claim 18 or 19 substantially as hereinbefore described with reference to any of the examples and/or figures. 35 27