Nanoemulsion vaccines

A technology of nanoemulsion and adjuvant, applied in pharmaceutical formulations, ester active ingredients, organic active ingredients, etc., can solve the problems of ineffective and repeated immune broad-spectrum diseases

Inactive Publication Date: 2009-06-24
RGT UNIV OF MICHIGAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Important problems inherent in existing vaccines include the need for repeated immunizations and the ineffectiveness of existing vaccine delivery systems for a broad spectrum of diseases

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0360] Materials and methods: Nanoemulsion inactivated vaccinia virus

[0361] animal. Pathogen-free, 5- to 6-week-old female Balb / c mice were purchased from Charles River Laboratories. According to the American Association for Accreditation of Laboratory Animal Care (American Association for Accreditation of Laboratory Animal Care) standards, the inoculation group was housed separately, with 5 animals in one cage. All methods involving mice were performed in accordance with the University Committee on Use and Care of Animals (UCUCA) of the University of Michigan.

[0362] Virus. Two exemplary vaccinia viruses (VV), VV WR and VV WR-Luc . VV was obtained from the American Type Culture Collection (ATCC), USA WR (NIH TC-adapted). rVV WR-Luc Expression of firefly luciferase via the p7.5 early / late promoter has been described (see, eg, Luker et al., Virology. 2005, 341(2):284-300). VV WR-Luc It was not attenuated in vitro or in vivo because the virus was constructed using...

Embodiment 2

[0381] Nasal immunization with nanoemulsion-inactivated vaccinia virus resulted in the induction of specific systemic IgG responses.

[0382] To estimate the virucidal activity of NE in vitro, a series of NE concentrations were compared with VV WR or VV WR-Luc Mix, and incubate at 37°C for 1 to 3 hours. The results of plaque reduction (PRA) and luciferase bioluminescence assays both showed NE concentration-dependent inactivation of both viruses. 10% NE can completely inactivate more than 10 6 pfu of vaccinia (see Figure 1A and B). Subsequent passage of culture supernatants from cells infected with VV (inactivated with 10% NE) showed no viable virus.

[0383] Intranasal administration of inactivated VV WR Later, complete inactivation of virus in NE formulations was further demonstrated in vivo by PCR amplification using DNA isolated from post-administration mouse lungs. In any treated mice (see Figure 1C No viral DNA was detected in ), whereas the control PCR (incorpo...

Embodiment 3

[0387] Subjects administered nanoemulsion-killed vaccinia virus have mucosal immunity against vaccinia virus

[0388] Mucosal immunity was determined by VV-specific secretory IgA antibodies in bronchoalveolar lavage (BAL). In from using 10 3 / NE or 10 5 Anti-VV IgA was detected in the BAL of / NE immunized animals. Animals vaccinated with formulations containing formalin-killed virus (whether diluted in saline or diluted in NE) did not produce a measurable mucosal response despite the presence of serum anti-VV IgG (see Figure 2B ). Accordingly, the invention provides that a composition comprising NE-killed VV produces mucosal immunity in a subject (e.g., as evidenced by the presence of VV-specific secretory IgA antibodies in the subject's BAL), However, compositions that do not contain NE-killed VV (eg, formalin-killed VV) cannot generate mucosal immunity against VV.

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Abstract

The present invention provides methods and compositions for the stimulation of immune responses. Specifically, the present invention provides methods and compositions for the use of nanoemulsion compounds as mucosal adjuvants to induce immunity against environmental pathogens. Accordingly, in some embodiments, the present invention provides nanoemulsion vaccines comprising a nanoemulsion and an inactivated pathogen or protein derived from the pathogen. The present invention thus provides improved vaccines against a variety of environmental and human-released pathogens.

Description

[0001] This application claims priority to US Provisional Patent Application Serial Nos. 60 / 791,800, 60 / 791,759, and 60 / 791,758, all filed April 13, 2006, each of which is incorporated herein by reference in its entirety. [0002] This invention was made with Government support under contract MDA 972-97-1-0007 awarded by the Defense Advanced Research Projects Agency and contract U54 AI57153-02 awarded by the National Institutes of Health. The Government has certain rights in this invention. field of invention [0003] The present invention provides methods and compositions for stimulating an immune response. In particular, the invention provides methods of inducing an immune response to an agent (eg, a bacterium or virus) in a subject (eg, a human subject) and compositions (eg, comprising a bacterium or virus or its constituent nanoemulsions). The compositions and methods of the invention find use in clinical (eg, therapeutic and preventive medicine (eg, vaccination)) and re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/22A61K31/225
Inventor 小J·R·贝克尔A·比林斯卡A·麦斯
Owner RGT UNIV OF MICHIGAN
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