Title:  Multi-component vaccine comprising at least three antigens to protect against disease cased by Haemophilus influenzae

United States Patent:  6,342,232

Assignee:  Aventis Pasteur Limited (Toronto, CA)

Filed:  March 3, 1999

A multi-component immunogenic composition confers protection on an immunized host against infection caused by Haemophilus influenzae. Such composition comprises at least three different antigens of Haemophilus influenzae, two of which are adhesins. High molecular weight (HMW) proteins and Haemophilus influenzae adhesin (Hia) proteins of non-typeable Haemophilus influenzae comprise the adhesin components while the other antigen is a non-proteolytic analog of Hin47 protein. Each component does not impair the immunogenicity of the others. The Haemophilus vaccine may be combined with DTP component vaccines, which may contain inactivated poliovirus, including type 1, type 2 and/or type 3, and/or a conjugate of a capsular polysaccharide of Haemophilus influenzae and tetanus or diphtheria toxoid, including PRP-T, to provide a multi-valent component vaccine without impairment of the immunogenic properties of the other antigens.

SUMMARY OF THE INVENTION

The present invention is directed towards the provision of a multi-component vaccine comprising at least three antigens, to protect against disease caused by infection with Haemophilus influenzae, including otitis media.

In accordance with one aspect of the present invention, there is provided an immunogenic composition for conferring protection in a host against disease caused by infection by Haemophilus influenzae, including otitis media, comprising at least three different antigens of Haemophilus influenzae, at least two of which different antigens are adhesins.

One of the antigens which is an adhesin may be a high molecular weight protein (HMW) of a non-typeable strain of Haemophilus, particularly an HMW1 or HMW2 protein of the non-typeable strain, which may be produced recombinantly.

The other of the antigens which is an adhesin is a Haemophilus influenzae adhesin (Hia) protein of a non-typeable strain of Haemophilus influenzae or a Haemophilus influenzae surface fibril (Hsf) protein of a typeable strain of Haemophilus influenzae.

The antigen of Haemophilus influenzae which is not an adhesin may be a non-proteolytic heat shock protein of a strain of Haemophilus influenzae. The non-proteolytic heat shock protein of a strain of Haemophilus influenzae may be an analog of Haemophilus influenzae Hin47 protein having a decreased protease activity which is less than about 10% of that of the natural Hin47 protein.

In accordance with a preferred embodiment of the aspect of the invention, there is provided an immunogenic composition for conferring protection in a host against disease caused by Haemophilus influenzae, including otitis media, which comprises:

(a) an analog of Haemophilus influenzae Hin47 protein having a decreased protease activity which is less than about 10% of natural Hin47 protein,

(b) a Haemophilus influenzae adhesin (Hia) protein of a non-typeable strain of Haemophilus influenzae, and

(c) a high molecular weight (HMW) protein of a strain of non-typeable Haemophilus influenzae.

The analog of Hin47 protein may be one in which at least one amino acid of the natural Hin47 protein contributing to protease activity has been deleted or replaced by a different amino acid and which has substantially the same immunogenic properties as natural Hin47 protein.

Such at least one amino acid may be selected from the group consisting of amino acids 91, 121 and 195 to 207 of natural Hin47 protein. Specific mutants which may be used including serine-197 replaced by alanine, Histidine-91 replaced by alanine, lysine or arginine and Asp-121 replaced by alanine.

The HMW protein of the non-typeable strain of Haemophilus influenzae may be a HMW1 or HMW2 protein and may be recombinantly produced. The HMW1 and HMW2 proteins are derived from the respective strains of non-typeable Haemophilus influenzae and possess respective molecular weights as set forth in the following Table I:

                                                      TABLE I
                                    Non-typeable H. influenzae Strain
    Molecular Weight (kDa)  12   JoyC   K21   LCDC2   PMH1    15
    Mature Protein: HMW1     125  125.9 104.4  114.0   102.4   103.5
                             HMW2     120  400.9            111.7   103.9   121.9


The Hia protein may be produced recombinantly and may comprise the N-terminal truncated V38 rHia protein.

The immunogenic composition of the invention may be further formulated with an adjuvant. Such adjuvant for use in the present invention may include (but not limited to) aluminum phosphate, aluminum hydroxide, QS21, Quil A, derivatives and components thereof, ISCOM matrix, calcium phosphate, calcium hydroxide, zinc hydroxide, a glycolipid analog, an octadecyl ester of an amino acid, a muramyl dipeptide, polyphosphazene, ISCOPREP, DC-chol, DDBA and a lipoprotein and other adjuvants. Advantageous combinations of adjuvants are described in copending U.S. patent applications Ser. No. 08/261,194 filed Jun. 16, 1994 and 08/483,856 filed Jun. 7, 1995, assigned to the assignee hereof and the disclosures of which are incorporated herein by reference (WO 95/34308, published Nov. 21, 1995). The adjuvant preferably may comprise aluminum phosphate or aluminum hydroxide (collectively known as alum).

The components of the immunogenic composition may be present in appropriate quantities to provide the desired immune response. The components may be formulated as a vaccine for in vivo administration to the host. The vaccine composition may contain:

(a) about 25 to 100 .mu.g of the Hin47 protien,

(b) about 25 to 100 .mu.g of the Hia protien, and

(c) about 25 to 100 .mu.g of the HMW protien.

The immunogenic compositions may be formulated with other antigenic components to provide a multivalent vaccine in which the additional antigenic component(s) confer protection against disease caused by another pathogen(s). Such additional antigens should be such that and be present in quantities that the immunogenicity of the individual components of the resulting vaccine is not impaired by other individual components of the composition. Such additional antigens preferably are purified antigens in defined quantities to produce a component vaccine.

Such additional antigens may be those traditionally found in multivalent protective vaccines, such as diphtheria toxoid, tetanus toxoid and pertussis antigens, including pertussis toxoid, filamentous hemagglutinin, pertactin and/or agglutinogens.

The resulting multivalent vaccine also may contain non-virulent poliovirus, such as inactivated poliovirus, which may be type 1, type 2 and/or type 3 poliovirus. The multi-component vaccine further may comprise a conjugate of a tetanus or diphtheria toxoid and a capsular polysaccharide of Haemophilus influenzae, preferably PRP-T.

The invention extends to a method of immunizing a host against disease caused by infection by Haemophilus influenzae, including otitis media, which comprises administering to the host an immunoeffective amount of the immunogenic composition provided herein.

Advantages of the present invention include a multi-component vaccine that can confer protection against encapsulated and non-encapsulated Haemophilus influenzae diseases in a safe and efficacious manner.

Claim 1 of 20 Claims

What we claim is:

1. An immunogenic composition for conferring protection in a host against disease caused by Haemophilus influenzae, which comprises:

(a) an isolated and purified analog of Haemophilus influenzee Hin47 protein having a decreased protease activity which is less than about 10% of natural Hin47 protein,

(b) an isolated and purified Haemophilus influenzae adhesin (Hia) protein of a non-typeable strain of Haemophilus influenzae, and

(c) an isolated and purified high molecular weight (HMW) protein of a non-typeable strain of Haemophilus influenzae.

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