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Use of panton-valentine leukocidin for treating and preventing staphylococcus infections

A technology for staphylococcal infection and staphylococcus, applied in antibacterial drugs, immunoglobulins, antibody medical components, etc., can solve problems such as uncontrolled IVIG systemic immune regulation effects

Inactive Publication Date: 2012-10-03
GLAXOSMITHKLINE BIOLOGICALS SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the authors did not demonstrate that the reported activity was due to the anti-PVL antibody itself and did not control for the systemic immunomodulatory effects of IVIG

Method used

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  • Use of panton-valentine leukocidin for treating and preventing staphylococcus infections
  • Use of panton-valentine leukocidin for treating and preventing staphylococcus infections
  • Use of panton-valentine leukocidin for treating and preventing staphylococcus infections

Examples

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Effect test

example 1

[0115] Example 1. Generation of rLukF-PV and rLukS-PV wild type clones

[0116]A strain of Staphylococcus aureus deposited from the ATCC under accession number 49775 (prototype of PVL producing high levels of PVL) was obtained by using a protocol according to the manufacturer (Promega) with minor modifications (addition of lysostaphin to the resuspension buffer) strains) to isolate genomic DNA.

[0117] The published PVL gene sequence (GenBank accession numbers X72700 and AB006796) was used to design oligonucleotide primers to classify the LukF-PV and LukS-PV genes, respectively. The forward primer was designed to eliminate a putative signal peptide and incorporate an Ncol site. The ATG at the Ncol site was designed to serve as an initiation codon for translation, avoiding the addition of the N-terminal amino acid encoded by the vector. The reverse primer was designed to incorporate a BamHI site just downstream of the stop codon. The luks-PV and lukf-PV genes were amplifi...

example 2

[0118] Example 2. Generation of rLukF-PV and rLukS-PV fusion protein clones

[0119] The PCR cloning technique was used to construct the PVL fusion protein, which is a human engineered protein encoded by the nucleotide sequence obtained by splicing together the lukf-PV and luks-PV genes. The PVL subunits were covalently linked by a short amino acid linker with the configuration rLukF-PV-aa linker-rLukS-PV or rLukS-PV-aa linker-rLukF-PV. This fusion protein is not cytotoxic because the two subunits cannot assemble and / or interact in the manner required to exhibit toxicity (eg, in the manner required for proper insertion into the leukocyte membrane). Fusion proteins are suitable for stimulating antibodies in the host to both subunits of PVL (eg, LukS-PV and LukF-PV) and to PVL toxin as a whole.

example 3

[0120] Example 3. Generation of rLukF-PV and rLukS-PV mutant clones

[0121] Using the protocol described by the manufacturer (Stratagene) and using pTrcHisBLukF-PV and / or pTrcHisBLukS-PV as templates, the following mutants were constructed using the QuickChange Mutagenesis Kit:

[0122] rLukF-PV mutants: ΔI124-S129; E191A; N173A; R197A; W176A and Y179A.

[0123] rLukS-PV mutants: ΔD1-I17; ΔF117-S122; T28D; T28F; T28N and T244A.

[0124] Those of ordinary skill in the art of genetics understand this nomenclature as standard terminology. That is, "ΔI124-S129" means a region deleted ("Δ") terminated by isoleucine at position 124 and terminated by serine at position 129 in the rLukF-PV mutant. Similarly, "ΔD1-I17" indicates that residues 1 (aspartic acid) to 17 (isoleucine) are missing in the rLukS-PV mutant. Likewise, the skilled artisan knows that "E191A" means that the glutamic acid at position 191 is replaced by alanine, and that "N173A" indicates that the rLukF-PV mutan...

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Abstract

The present invention relates to compositions and methods for treating Staphylococcus aureus (S. aureus) infections. In particular, the present invention provides vaccines comprising a Panton- Valentine Leukocidin (PVL) antigen, antibodies which bind a PVL antigen and compositions containing the same, methods of making such compositions and methods for treating S. aureus infections, including those that are community acquired methicillin-resistant infections. The present invention also provides PVL antibodies, including PVL antibodies specific for a single PVL subunit, and PVL antigens, including conjugated and mutated PVL antigens.

Description

[0001] This application is a divisional application with an application date of June 13, 2006, an application number of 200680024681.9, and an invention name of PANTON-VALENTINE leukocidin for the treatment and prevention of Staphylococcus infection. [0002] Cross References to Related Patent Applications [0003] This application claims priority to US Provisional Application Serial No. 60 / 689,526, filed June 13, 2005 and incorporated herein by reference. technical field [0004] The present invention generally relates to the treatment and prevention of bacterial infections. In particular, disclosed herein is the use of compositions comprising the Panton-Valentine leukocidin (PVL) antigen or an antibody that specifically binds thereto for the treatment and prevention of Staphylococcus aureus (S. aureus) infections, including community-acquired Compositions and methods for methicillin-resistant Staphylococcus aureus (CA-MSRA) infection). The invention also generally relates...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/085A61K45/00A61P31/04
CPCC07K16/1271A61K39/085A61K2039/505C07K2316/96C07K14/31C07K2317/76A61P31/04C07K16/12A61K39/395
Inventor 金伯利·路易斯·泰勒阿里·易卜拉欣·法托姆
Owner GLAXOSMITHKLINE BIOLOGICALS SA
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