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Vaccine against microbial pathogens

A technology of microorganisms and pathogens, applied in the direction of antibody medical components, bacterial antigen components, protozoa antigen components, etc., can solve the problem of unproposed endogenous hsp peptide complex vaccine and other problems

Inactive Publication Date: 2003-10-29
IMMUNOBIOLOGY LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although pathogenic HSPs have been widely used as antigens and adjuvants, to date, the use of pathogen-derived endogenous HSP-peptide complexes (hspC) has not been proposed as vaccines

Method used

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  • Vaccine against microbial pathogens
  • Vaccine against microbial pathogens

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Experimental program
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Embodiment 1

[0062] Embodiment 1: the preparation of heat shock microorganism

[0063] M. vaccae strain NCTC11659 was grown to saturation in Sauton's medium, diluted in fresh medium and grown overnight to provide a log-phase culture, which was then heat-shocked at 42°C for 3 hours, or at 39°C. ℃ heat shock for 5 hours, and incubated overnight. Cells are then washed in culture medium followed by saline and either lyophilized in separate vaccine aliquots or directly immunized into experimental animals. For comparison, isolated endogenous SP-peptide complexes were prepared as described in PCT Application NoGB00 / 03228. Briefly, the washed stress-induced cells were resuspended in a homogeneous buffer such as PBS with 5% Tween, and then the cells were cycled through freeze-thaw or using a cell disruptor (e.g. Bead Shaker, France )broken. Cell lysates are then processed by centrifugation, typically at 3-5000 g for 5 minutes to remove nuclei and cell debris, followed by a high speed centrifugat...

Embodiment 2

[0067] Immunization with intact stress-induced microorganisms, and especially lyophilized microorganisms, provided better immunity than that induced by isolated endogenous SP-peptide complexes, including the production of IFN- Gamma and antibodies increased. Example 2: Preparation and use of constitutive hsp mutants

[0068] Microbes that constitutively produce hsp can be constructed by knocking out transcriptional regulator repressor genes such as hspR, MerR (mercury resistance operon regulatory protein), HmrR (heavy metal regulatory protein) and their homologues or the transcriptional control gene rho and sigma. Such genes are readily identified by scanning genome sequence databases with typical test sequences. A typical test sequence of this kind is, for example, the hsp gene of Mycobacterium tuberculosis (see Figure 1). The suppressor genes identified as homologous to the hspR gene are shown in Figure 2.

[0069] A M. bovis strain that constitutively produces hspC was ...

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Abstract

The present invention provides a vaccine comprising a microbial pathogen, wherein the microbial pathogen is subjected to a stress inducing stimuli. The stress inducing stimuli can be heat or osmotic stress, through preferably the microbial pathogen is genetically modified such that at least one repressor gene for a heat shock protein gene is inactivated, thus allowing the constitutive expression of heat shock proteins. In particular the use of heat shock protein repressor mutant bacteria is shown to be effective for inducing immunity when comprised within vaccines of the present invention. The present invention further provides a method for producing a vaccine comprising stressed induced microbial pathogens and further the use of the heat shock protein repressor deletion mutant microbes as vaccine vectors which can be additionally allow the expression of heterologous antigen fragments.

Description

[0001] The present invention relates to a vaccine and a method of producing the vaccine. More specifically, the invention provides a vaccine comprising a microbial pathogen and a method of producing the vaccine. Background of the invention [0002] An important part of any human immune response is the presentation of antigens to T cells by antigen-presenting cells (APCs) such as macrophages, B cells or dendritic cells. Peptide fragments of foreign antigens are presented on the surface of macrophages in combination with major histocompatibility complex (MHC) molecules, associated with accessory molecules such as CD4 and CD8 molecules. Antigenic peptide fragments presented in this manner are recognized by the T cell receptors of T cells. The interaction of the antigenic peptide fragment with the T cell receptor results in the proliferation of antigen-specific T cells and the secretion of lymphokines by the T cells. The nature of the antigenic peptides presented by APCs is crit...

Claims

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Application Information

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IPC IPC(8): C12N15/09A61K39/002A61K39/02A61K39/04A61K39/106A61K39/112A61P31/04A61P37/04
CPCA61K39/04A61K2039/523A61K2039/6043A61P31/04A61P37/04
Inventor 卡米洛·安东尼·利奥·塞尔温·科拉索
Owner IMMUNOBIOLOGY LTD