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Streptococcus pneumoniae pilus antigens

A technology of Streptococcus pneumoniae and fimbriae, applied in the field of peptides, can solve problems such as unclear adhesion mechanism

Inactive Publication Date: 2010-09-29
NOVARTIS AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Streptococcus pneumoniae infection can initiate invasive disease through initial colonization of the nasopharynx, but mechanism of adhesion is unclear

Method used

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  • Streptococcus pneumoniae pilus antigens
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  • Streptococcus pneumoniae pilus antigens

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0224] In some aspects, methods for preparing antigen-adsorbed microparticles are provided. The method comprises: (a) providing an emulsion by dispersing a mixture comprising (i) water, (ii) a detergent, (iii) an organic solvent, and (iv) a biodegradable polymer selected from : Poly(alpha-hydroxy acids), polyhydroxybutyrates, polycaprolactones, polyorthoesters, polyanhydrides and polycyanoacrylates. The concentration of the polymer in the mixture is generally about 1%-30% relative to the organic solvent, and the weight ratio of soil release agent-polymer in the mixture is generally about 0.00001:1-0.1:1 (more typically about 0.0001 : 1-0.1: 1, about 0.001: 1-0.1: 1, or about 0.005: 1-0.1: 1); (b) removing the organic solvent in the emulsion; and (c) allowing the antigen to be adsorbed on the surface of the microparticle . In some embodiments, the concentration of the biodegradable polymer is about 3%-10% relative to the organic solvent.

[0225] In some embodiments, the mic...

Embodiment 1

[0496] Example 1: Detection of pili II islands (INV104B) in genomic DNA

[0497] The presence of S. pneumoniae pili II islands (INV104B) shown in Table 1 was determined on purified genomic DNA or directly on bacteria (colony PCR). Table 2 lists the four oligonucleotides used for amplification.

[0498] Table 2 PCR oligonucleotides

[0499] logo

direction

sequence

1008 Forward

5’→3’

GCTGGATCGAGTTTGAAACCAGAA (SEQ ID

NO: 24)

1009 reverse

3’→5’

TAAGGATCACCAAAGTCCAAGGCA (SEQ ID

NO: 25)

Int - reverse

3’→5’

TTTCAGTGTATGTTTTAGTGCTTCA (SEQ ID

NO: 26)

Int - Forward

5’→3’

ATGGCTTCAGGGGCTATGTTCGGTG (SEQ ID

NO: 27)

[0500] Three diagnostic PCRs were performed on each of the strains listed in Table 1 using the following PCR oligonucleotide combinations: 1008 forward-1009 reverse; 1008 forward-int reverse; and int forward-1009 reverse. 1008 forward-int reverse and in...

Embodiment 2

[0503] Example 2: Sequences of 23F, INV200 and OXC141

[0504] 1. Sequence download and assembly

[0505] The initial sequences of the four S. pneumoniae strains were downloaded from the Sanger website (see World Wide Web "sanger.ac.uk / Projects / Microbes / ”). The Sanger sequences consisted of varying numbers of non-contiguous contigs. Details of the downloaded sequences are listed in Table 3:

[0506] Table 3. Download sequence

[0507] strain

serotype

ST

number of contigs

total bp

23F

23F

81

21

2225211

INV104B

1

227

68

1986609

NV200

14

9

167

2022487

[0508] strain

serotype

ST

number of contigs

total bp

OXC141

3

180

120

1962139

[0509] To identify the likely sequence of the contig, this sequence was aligned with the complete TIGR4 sequence using MUMmer3.19 to form a single pseudomolecule. To separate the ...

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Abstract

Polypeptides from Streptococcus pneumoniae are described. In some aspects the polypeptides include pili polypeptides from a second pili island (pilus II island (INVl 04B)) identified in Streptococcus pneumoniae isolate INVl 04. In other aspects the polypeptides include pili polypeptides and non-pilus polypeptides from Streptococcus pneumoniae strains 23F, INV200, and OXC141 that are absent from Streptococcus pneumoniae isolate INV104. The polypeptides, including fragments and variants thereof, may be used in immunogenic compositions for prophylactic or therapeutic immunization against Streptococcus pneumoniae. The polypeptides are also disclosed to be used in compositions useful for the production of antibodies and immunostimulants. Also presented are methods of inhibiting Streptococcus pneumoniae, methods of treating Streptococcus pneumoniae infection, methods of identifying inhibitors of Streptococcus pneumoniae and methods for diagnosing / detecting Streptococcus pneumoniae infection.

Description

technical field [0001] The present invention relates to polypeptides, including pili proteins from Streptococcus pneumoniae (S. pneumoniae), including fragments and variants thereof, and methods of use thereof in the treatment and immunization of S. pneumoniae infection. Background technique [0002] The gram-positive bacterium Streptococcus pneumoniae (also known as Spn or pneumococcus) is a leading cause of morbidity and mortality worldwide, and together with HIV, malaria, and tuberculosis, is one of the four major infectious disease killers (Bruyn, G.A.W. and van Furth, R. (1991) Eur.J.Clin.Microbiol.Infect.Dis.10, 897-910; Ryan, M.W. and Antonelli, P.J. (2000) Laryngoscope 110, 961-964; Cutts, F.T., Zaman, S.M., Enwere, G., Jaffar, S., Levine, O.S., Okoko, C. Oluwalana, A., Vaughan, S., Obaro, A., Leach, A. et al., (2005) Lancet 365, 1139-1 146; Swiatlo, E., Champlin, F.R., Holman, S.C., Wilson, W.W. & Watt, J.M. (2002) Infect. Immun. 70, 412-415; Sandgren, A., Albiger,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/315C12N9/10C07K16/12A61K39/09G01N33/569
CPCA61K2039/53G01N2333/315C12N9/52C07K14/3156G01N33/56944G01N2469/10A61K2039/505A61K39/00A61P31/04A61P37/04
Inventor C·多纳蒂V·马西格纳尼M·莫拉M·莫斯切奥尼
Owner NOVARTIS AG
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