Streptococcus pneumoniae antigens and vaccines

a technology of streptococcus pneumoniae and antigens, applied in the field of streptococcus pneumoniae antigens, can solve the problems of provoking meningitis-like symptoms, unable to answer questions about its virulence, and unable to prove its efficacy,

Inactive Publication Date: 2005-08-18
HUMAN GENOME SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In spite of the vast number of publications on S. pneumoniae many questions about its virulence are still unanswered, and this pathogen remains a major causative agent of serious human disease, especially community-acquired pneumonia.
Partially purified neuraminidase was observed to induce meningitis-like symptoms in mice; however, the reliability of this finding has been questioned because the neuraminidase preparations used were probably contaminated with cell wall products.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Expression and Purification of S. pneumoniae Polypeptides in E. coli

[0163] The bacterial expression vector pQE10 (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, Calif., 91311) is used in this example for cloning of the nucleotide sequences shown in Table 1 and for expressing the polypeptides identified in Table 1. The components of the pQE10 plasmid are arranged such that the inserted DNA sequence encoding a polypeptide of the present invention expresses the polypeptide with the six His residues (i.e., a “6×His tag”)) covalently linked to the amino terminus.

[0164] The DNA sequences encoding the desired portions of the polypeptides of Table 1 are amplified using PCR oligonucleotide primers from either a DNA library constructed from S. pneumoniae, such as the one deposited by the inventors at the ATCC™ for convenience, ATCC™ Deposit No. 97755, or from DNA isolated from the same organism such as the S. pneumoniae strain deposited with the ATCC™ as Deposit No. 55840. A list of PCR prime...

example 2

Immunization and Detection of Immune Responses Methods

Growth of Bacterial Innoculum, Immunization of Mice and Challenge with S pneumoniae.

[0172] Propagation and storage of, and challenge by S. pneumoniae are preformed essentially as described in Aaberge, I. S. et al., Virulence of Streptococcus pneumoniae in mice: a standardized method for preparation and frozen storage of the experimental bacterial inoculum, Microbial Pathogenesis, 18:141 (1995), incorporated herein by reference.

[0173] Briefly, Todd Hewitt (TH) broth (Difco laboratories, Detroit, Mich.) with 17% FCS, and horse blood agar plates are used for culturing the bacteria. Both broth and blood plates are incubated at 37° C. in a 5% CO2 atmosphere. Blood plates are incubated for 18 hr. The culture broth is regularly 10-fold serially diluted in TH broth kept at room temperature and bacterial suspensions are kept at room temperature until challenge of mice.

[0174] For active immunizations C3H / HeJ mice (The Jackson Laborat...

example 3

Detection of Streptococcus mRNA Expression

[0179] Northern blot analysis is carried out using methods described by, among others, Sambrook et al., supra. to detect the expression of the S. pneumoniae nucleotide sequences of the present invention in animal tissues. A cDNA probe containing an entire nucleotide sequence shown in Table 1 is labeled with 32p using the rediprime™ DNA labeling system (Amersham Life Science), according to manufacturer's instructions. After labeling, the probe is purified using a CHROMA SPIN-100™ column (Clontech Laboratories, Inc.), according to manufacturer's protocol number PT1200-1. The purified labeled probe is then used to detect the expression of Streptococcus mRNA in an animal tissue sample.

[0180] Animal tissues, such as blood or spinal fluid, are examined with the labeled probe using ExpressHyb™ hybridization solution (Clontech) according to manufacturer's protocol number PT 1190-1. Following hybridization and washing, the blots are mounted and ex...

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Abstract

The present invention relates to novel vaccines for the prevention or attenuation of infection by Streptococcus pneumoniae. The invention further relates to isolated nucleic acid molecules encoding antigenic polypeptides of Streptococcus pneumoniae. Antigenic polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same. The invention additionally relates to diagnostic methods for detecting Streptococcus nucleic acids, polypeptides and antibodies in a biological sample.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a divisional of U.S. application Ser. No. 08 / 961,083, filed Oct. 30, 1997, which claims benefit of U.S. Provisional Application No. 60 / 029,960, filed Oct. 31, 1996; this application is also a divisional of U.S. application Ser. No. 09 / 765,271, filed Jan. 22, 2001, which is a continuation of U.S. application Ser. No. 09 / 536,784, filed Mar. 28, 2000 (now U.S. Pat. No. 6,573,082, issued Jun. 3, 2003), which is a continuation of U.S. application Ser. No. 08 / 961,083, filed Oct. 30, 1997, which claims benefit of U.S. Provisional Application No. 60 / 029,960, filed Oct. 31, 1996; U.S. application Ser. No. 09 / 765,271 is also a continuation of U.S. application Ser. No. 08 / 961,083, filed Oct. 30, 1997, which claims benefit of U.S. Provisional Application No. 60 / 029,960, filed Oct. 31, 1996.REFERENCE TO SEQUENCE LISTING ON COMPACT DISC [0002] This application refers to a “Sequence Listing” which is provided as an electronic docum...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01K67/027A61K38/00A61K39/00A61K39/09A61K48/00G01N33/53A61P31/04C07K14/315C07K16/12C12N1/15C12N1/19C12N1/21C12N5/00C12N5/10C12N5/18C12N15/09C12N15/31C12P21/02C12Q1/68G01N33/50G01N33/569G06F17/30
CPCA61K38/00C07K14/3156C07K14/315A61K39/00A61P31/04A61P37/04
Inventor CHOI, GIL H.KUNSCH, CHARLES A.BARASH, STEVEN C.DILLON, PATRICK J.DOUGHERTY, BRIANFANNON, MICHAEL R.ROSEN, CRAIG A.
Owner HUMAN GENOME SCI INC
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