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Adjuvant compositions

a technology of adjuvant compositions and compositions, applied in the field of adjuvant compositions, can solve the problems of severe side effects of cfa, pain, abscess formation and fever, and conventional vaccines often fail to provide adequate protection against the targeted pathogen, and achieve the effect of safe and effective, enhancing immunogenicity

Inactive Publication Date: 2010-01-07
GLAXOSMITHKLINE BIOLOGICALS SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is based on the discovery that combining a type 1 interferon inducer with an antigen delivery system and / or an immunostimulatory molecule can significantly increase the immune response to a coadministered antigen compared to using either component alone. This combination can be used in a therapeutically effective amount to stimulate an immune response in a vertebrate subject. The use of these components in an adjuvant composition can lead to higher antibody titers and a more effective immunogenicity of vaccine antigens. The invention is useful for enhancing the immunogenicity of a variety of vaccine antigens and can be applied in both prophylactic and therapeutic compositions.

Problems solved by technology

Although effective as an adjuvant, CFA causes severe side-effects, including pain, abscess formation and fever, primarily due to the presence of the mycobacterial component.
Despite the presence of adjuvants, conventional vaccines often fail to provide adequate protection against the targeted pathogen.

Method used

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  • Adjuvant compositions
  • Adjuvant compositions
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Examples

Experimental program
Comparison scheme
Effect test

example 1

Production of HCV E1E2

[0211]An HCV E1E2 complex for use in the present vaccine compositions was prepared as a fusion protein as follows. In particular, mammalian expression plasmid pMH-E1E2-809 (FIG. 2, ATCC Deposit No. PTA-3643) encodes an E1E2 fusion protein which includes-amino acids 192-809 of HCV la (see, Choo et al., Proc. Natl. Acad. Sci. USA (1991) 88:2451-2455).

[0212]Chinese Hamster Ovary (CHO) cells were used for expression of the HCV E1E2 sequence from pMH-E1E2-809. In particular, CHO DG44 cells were used. These cells, described by Uraub et al., Proc. Natl. Acad. Sci. USA (1980) 77:4216-4220, were derived from CHO K-1 cells and were made dihydrofolate reductase (dhfr) deficient by virtue of a double deletion in the dhfr gene.

[0213]DG44 cells were transfected with pMH-E1E2-809. The transfected cells were grown in selective medium such that only those cells expressing the dhfr gene could grow (Sambrook et al., supra). Isolated CHO colonies were picked (˜800 colonies) into ...

example 2

Purification of HCV E1E2

[0214]Following expression, CHO cells were lysed and the intracellularly produced E1E2809 was purified by GNA-lectin affinity chromatography (GNA step), followed by hydroxyapatite (HAP) column chromatography (HAP step), DV50 membrane filtration (DV50 step), SP Sepharose HP column chromatography (SP step), Q membrane filtration (Q step) and G25 Sephadex column chromatography G25 step). At the completion of each of the processing steps, the product pool was either 0.2 μ filtered and held at 2-8° C. or processed immediately through the next purification step. At the completion of the purification process, the antigen was 0.2 μ filtered and held frozen at −60° C., or lower until filtered for formulation.

[0215]Specifically, to lyse the cells, two volumes of chilled lysis buffer (1% Triton X-100 in 100 mM Tris, pH8, and 1 mM EDTA) were added to the CHO cells at 2-8° C. The mixture was centrifuged at 5000 rpm for 45 min at 2-8° C. to remove debris. The supernatant ...

example 3

Use of dsRNA Adjuvant Compositions and HCV Antigens

[0223]The ability of dsRNA, in combination with a representative delivery system, to enhance the immunogenicity of HCV E1E2809, produced and purified as described above, was determined as follows.

[0224]The formulations used in this study are summarized in Table 3. MF59, a representative submicron oil-in-water emulsion which contains 4-5% w / v squalene, 0.5% w / v TWEEN 80™, 0.5% SPAN 85™, was produced as described previously. See, International Publication No. WO 90 / 14837; U.S. Pat. No. 6,299,884, incorporated herein by reference in its entirety; and Ott et al., “MF59—Design and Evaluation of a Safe and Potent Adjuvant for Human Vaccines” in Vaccine Design: The Subunit and Adjuvant Approach (Powell, M. F. and Newman, M. J. eds.) Plenum Press, New York, 1995, pp. 277-296.

[0225]The dsRNA used in these studies was poly[rI-rC], available from Sigma Chemical Co. (St. Louis, Mo.). The dsRNA was reconstituted in RNase-free distilled water and...

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Abstract

Adjuvant compositions comprising type 1 interferon inducers, such as double-stranded RNA, in combination with antigen delivery systems and / or immunostimulatory molecules, such as immunostimulatory nucleic acid sequences, for enhancing the immune response of a coadministered antigen, are described.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001]This application a continuation of U.S. application Ser. No. 10 / 265,083, filed Oct. 3, 2002 from which priority is claimed under 35 U.S.C. 120, which application claims the benefit under 35 U.S.C. 119(e)(1) of Provisional Patent Application No. 60 / 326,929, filed Oct. 3, 2001, and Provisional Patent Application No. 60 / 373,547, filed Apr. 17, 2002. All of the foregoing applications are incorporated herein by reference in their entireties.TECHNICAL FIELD [0002]The present invention pertains generally to adjuvant compositions. In particular, the invention relates to the use of adjuvant compositions comprising type 1 interferon inducers, such as double-stranded RNA, in combination with antigen delivery systems and / or immunostimulatory molecules, such as immunostimulatory nucleic acid sequences, for enhancing the immune response of a coadministered antigen. The adjuvant compositions will find use in both prophylactic and therapeutic composition...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/21A61K39/00A61K39/12A61K39/095A61K39/29A61P31/12A61K31/7088A61K9/107A61K31/7105A61K38/00A61K38/21A61K39/39A61K45/06A61K47/06A61K47/26A61K47/34A61K47/44A61K47/48A61K48/00A61P31/04A61P31/18A61P35/00A61P37/04C12N15/31C12N15/48
CPCA61K39/39A61K2039/55522A61K2039/55566A61K2039/55561A61K2039/55555A61P31/04A61P31/12A61P31/18A61P35/00A61P37/04A61K39/095A61K39/21A61K39/29C12N7/00C12N2740/16034C12N2770/24234
Inventor O'HAGAN, DEREKVALIANTE, NICHOLAS
Owner GLAXOSMITHKLINE BIOLOGICALS SA
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