Proteinaceous adjuvants

a technology of proteinaceous adjuvants and adjuvants, which is applied in the direction of antibody medical ingredients, vertebrate antigen ingredients, bacteria antigen ingredients, etc., can solve the problems of ineffective influenza vaccination, inability to induce protective immune responses, and inability to use in humans,

Inactive Publication Date: 2003-04-17
AVENTIS PASTUER LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

0024] The present invention relates to avoiding the problems associated with the use of alum as an adjuvant in immunogenic compositions by employi...

Problems solved by technology

Other antigens, however, fail to induce, for example, a protective immune response or induce only a weak immune response.
Some of these adjuvants are toxic, however, and can cause undesirable side-effects, making them unsuitable for use in humans and many animals Indeed, only aluminum hydroxide and aluminum phosphate (collectively commonly referred to as alum) are routinely used as adjuvants in human and veterinary vaccines.
While the usefulness of alum is well established for some applications, it has limitations.
For example, it is ineffective for influenza vaccination and inconsistently elicits a cell mediated immune response.
The antibodies elicited by alum-adjuvanted antigens are mainly of the IgG1 isotype in the mouse, which may not be optimal for protection by some vaccinal agents.
Although the whole cell pertussis vac...

Method used

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Examples

Experimental program
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example 1

[0071] This Example describes the formulation of vaccines.

[0072] The DPT whole-cell vaccine and an experimental component acellular vaccine were produced by Connaught Laboratories Ltd. The component acellular pertussis vaccine was alum adsorbed (1.5 mg / dose) and consisted of 10 .mu.g protein nitrogen of glutaraldehyde toxoided pertussis toxin , 5 .mu.g protein nitrogen each of FHA and agglutinogens 2 and 3 and 3 .mu.g of pertactin along with 5 Lf of tetanus toxoid and 25 Lf of diphtheria toxoid per dose. The recombinant component vaccine also contained 5 .mu.g protein nitrogen each of FHA and agglutinogens 2 and 3 and 3 .mu.g protein nitrogen of pertactin in addition to 5 Lf of tetanus toxoid and 25 Lf of diphtheria toxoid per dose. However, it varied from the other acellular vaccine in that it contained 20 .mu.g protein nitrogen of the recombinant PT mutant holotoxin, K9G129, and was not alum adsorbed. The K9G129 pertussis toxin molecule as well as the purified FHA, agg 2+3 and per...

example 2

[0073] This Example describes immunization of animals.

[0074] Female BALB / c mice weighing 15 to 18 grams were obtained from Charles River Canada (St. Constant, Quebec). The mice were housed in microisolators and used in accordance with the guidelines set by the Canadian Council on Animal Care (CCAC). The animals were specific pathogen free and the housing rooms were monitored for Murine Hepatitis virus outbreaks through the use of sentinel mice. Water was provided ad libitum and the diet was ovalbumin-free. The mice were immunized on Day 0 with the vaccine formulations in groups of six. A booster dose of vaccine was administered an Day 21. On Day 28 the animals were bled via jugular vein laceration and spleneostomized. The serum samples were stored at -20.degree. C. until assayed.

example 3

[0075] This Example describes antigen specific imunuoassays.

[0076] The vaccine antigen specific IgG responses were determined by indirect EIA. The antigens of interest were pertussis toxin, pertactin, filamentous hemagglutinin, agglutinogens, as well as diphtheria and tetanus toxoids. High binding capacity microplates (Nuno) were coated with 4 .mu.g / mL of each of the above antigens in a volume of 50 uls / well of 50 mM carbonate buffer pH 9.6. After an overnight incubation, the plates were washed and successively blocked with a 0.1% solution of bovine serum albumin (sigma) for one hour at room temperature. The excess block was removed and the microplates were washed. The murine serum samples were then serially diluted in PBS-Tween 20 (0.05%) and plated out at a volume of 100 uls. The samples were incubated overnight at 4.degree. C. The antigen specific fraction of IgG antibodies was detected by a peroxidase conjugated sheep anti-mouse IgG conjugate (Jackson Laboratories). The plates w...

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Abstract

A modulated immune response to an antigen is achieved by coadministering the antigen and a genetically-detoxified pertussis holotoxin, particularly one retaining its immunogenicity, to a host. The modulated immune response enables immunogenic compositions, including multivalent pediatric vaccines, such as DTP, to be provided which produce a modulated immune response in the absence of extrinsic adjuvants, such as alum. The adjuvanting effect achieved by the genetically-detoxified pertussis holotoxin enables at least the same level of adjuvanting effect to be achieved as previously attained by alum, without the undesirable side effects thereof.

Description

REFERENCE TO RELATED APPLICATION[0001] This application is a continuation-in-part of copending U.S. patent application Ser. No. 08 / 258,228 filed Jun. 10, 1994.[0002] The present invention relates to the field of immunology and is particularly concerned with proteinaceous adjuvants., i.e. materials which modulate immune responses to an antigen.[0003] Vaccines have been used for many years to protect humans and animals against a wide variety of infectious diseases. Such conventional vaccines consist of attenuated pathogens (for example, polio virus), killed pathogens (for example, Bordetalla pertussis) or immunogenic components of the pathogen (for example, diphtheria toxoid). Some antigens are highly immunogenic and are capable alone of eliciting immune responses. Other antigens, however, fail to induce, for example, a protective immune response or induce only a weak immune response.[0004] Immunogenicity can be significantly improved if the antigens are co-administered with adjuvants...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/10A61K39/116A61K39/39A61P37/04
CPCA61K39/0005A61K39/099A61K39/39A61K2039/70A61K2039/55544A61K2039/57A61K2039/55505A61K2039/55516A61P31/00A61P31/04A61P31/12A61P35/00A61P37/04Y02A50/30
Inventor GAJEWCZYK, DIANE M.BOUX, HEATHER A.NOVAK, ANTONKLEIN, MICHEL H.
Owner AVENTIS PASTUER LTD
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