Recombinant intracellular pathogen vaccines and methods for use

A pathogenic and pathogenic technology, applied in the direction of chemical instruments and methods, chlamydia antigen components, drug combinations, etc., can solve the problems of no indication of prevention of mammalian intracellular pathogenic diseases, no disclosure, etc.

Inactive Publication Date: 2003-08-20
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disclosed vaccine was used to induce an immune response in a non-human animal model to produce antibodies thereto, but was not shown to prevent intracellular pathogenic

Method used

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  • Recombinant intracellular pathogen vaccines and methods for use
  • Recombinant intracellular pathogen vaccines and methods for use
  • Recombinant intracellular pathogen vaccines and methods for use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0060] To determine the effect of pathogen strain variation on vaccination, various embodiments of the invention were prepared using two different BCG strains: BCG Tice and BCG Connaught. Wild-type M. bovis BCG Tice was purchased from Organon, and wild-type M. bovis BCG Connaught was obtained from Connaught Laboratories, Toronto, Canada. The strains were grown in 7H9 medium pH 6.7 (Difco) at 37°C with 5% CO 2 A -95% air atmosphere was maintained as a non-shaking culture. Cultures were sonicated for 5 min in a sonication water bath once or twice a week to reduce bacterial colony clumps.

[0061] Recombinant BCG TICE (rBCG30 Tice) expressing the 30 kDa major extracellular non-fusion protein of M. tuberculosis was prepared as follows. The recombinant construct plasmid pMTB30 of a kind of Escherichia coli / mycobacterium shuttle plasmid pSMT3 prepares as the method that the inventor introduces previously in following document: Harth, G., B.-Y.Lee and M.A.Horwitz.1997, in fast Hig...

Embodiment 2

[0112] Example 2 demonstrates that using mycobacteria closely related to BCG but unable to replicate in mammalian hosts, in vivo expression of the immunogens of the invention does not induce significant levels of protection against challenge with M. tuberculosis. Examples 3 and 4 demonstrate that slow release of the immunogen of the invention by synthetic vaccine microcarriers also fails to induce significant levels of protection against challenge with M. tuberculosis.

[0113] Thus, the following examples serve to emphasize that the intracellular pathogen vaccines of the present invention represent a completely unexpected and significant advance in the field of infectious disease immunology.

Embodiment 1

[0116] Immunization with BCG plus recombinant Mycobacterium tuberculosis 30kDa major extracellular protein (r30) Guinea pigs fail to induce high levels of protection against challenge with Mycobacterium tuberculosis

[0117] We have previously immunized guinea pigs with a highly potent adjuvant (SAF, Syntex adjuvant formulation) of BCG plus r30. The r30 protein (100 μg per immunization) was intradermally administered three times. This induces a strong cutaneous delayed-type hypersensitivity (C-DTH) response to r30 ( Figure 5 ). In fact, the C-DTH response was comparable to that induced by recombinant BCG expressing r30. Nevertheless, immunization with both BCG and r30 did not induce high levels of protection against challenge with M. tuberculosis (Table 5). CFU levels in the lungs and spleens of animals immunized with both BCG and r30 were not lower than those of animals immunized with BCG alone. This result is in direct contrast to the above results in which animals ...

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Abstract

Vaccines and immunotherapeutics for preventing intracellular pathogen diseases in mammals are provided that consist of recombinant attenuated intracellular pathogens that have been transformed to express recombinant immunogenic antigens of the same r other intracellular pathogens. Exemplary vaccines and immunotherapeutics include attenuated recombinant Mycobacteria expressing the major extracellular non-fusion proteins of Mycobacterial and/or other intracellular pathogens. These exemplary vaccines are shown to produce surprisingly potent protective immune response in mammals that surpass those of any previously known anti-mycobacterium vaccine. More specifically, a recombinant BCG expressing the 30 kDa major extracellular non-fusion protein of Mycobacterium tubercolosis is provided. Additionally, methods for preventing and treating diseases caused by intracellular pathogens are provided. The methods of treating and preventing intracellular pathogen diseases utilize the described surprisingly efficacious vaccines and immunotherapeutics.

Description

[0001] About Government Rights [0002] This invention was made with US Government support under Grant No. AI31338 from the Department of Health and Human Services. The US Government has certain rights in this invention. field of invention [0003] The present invention generally relates to immunotherapeutics and vaccines against intracellular pathogenic organisms such as bacteria, protozoa, viruses and fungi. More specifically, unlike prior art vaccines and immunotherapeutics based on pathogen subunits, inactivated pathogens, and attenuated natural pathogens, the present invention uses immunogens that express and secrete selected pathogens that stimulate an effective immune response in the mammalian host Recombinant attenuated pathogens or closely related species of sex determinants. The immunostimulatory vaccines and immunotherapeutics of the present invention are derived from recombinant attenuated intracellular pathogens or closely related species expressing immunogenic ...

Claims

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Application Information

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IPC IPC(8): A61K38/00A61K39/02A61K39/04A61K39/108A61K39/116A61K39/118A61P31/06A61P37/04C07K14/35
CPCA61K2039/522A61K2039/523A61K38/00A61K39/04A61P31/02A61P31/04A61P31/06A61P33/02A61P37/04
Inventor M·A·霍尔维茨G·哈斯
Owner RGT UNIV OF CALIFORNIA
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