Compounds and methods for treatment and diagnosis of chlamydial infection

A technology of chlamydia and antigen, applied in chemical instruments and methods, botany equipment and methods, biochemical equipment and methods, etc., can solve problems such as infertility

Inactive Publication Date: 2003-08-20
CORIXA CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, by the time a woman seeks medical care for PID, irreversible damage may have already occurred and lead to infertility

Method used

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  • Compounds and methods for treatment and diagnosis of chlamydial infection
  • Compounds and methods for treatment and diagnosis of chlamydial infection
  • Compounds and methods for treatment and diagnosis of chlamydial infection

Examples

Experimental program
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Embodiment Embodiment 1

[0141] EXAMPLES Example 1: Cloning of DNA sequences encoding Chlamydia antigens

[0142] The Chlamydia antigens of the present invention are isolated by expression cloning of a Chlamydia trachomatis LGV II genomic DNA library substantially as described in Sanderson et al., J. Exp. Med., 182:1751-1757, 1995, which demonstrates immunoreactivity. Induction of PBMC proliferation and IFN-γ production in T cell lines.

[0143] Chlamydia-specific T cell lines were generated by stimulating PBMCs from normal individuals without a history of Chlamydia genital tract infection with primary bodies of Chlamydia trachomatis LGV II. This T cell line, called TCL-8, was found to recognize monocyte-derived dendritic cells infected with C. trachomatis and C. pneumoniae.

[0144] A randomly sheared genomic library of Chlamydia trachomatis LGV II was constructed in λZAP (Stratagene, La Jolla, CA), and the amplified library was plated in 96-well microtiter plates at a densit...

Embodiment 2

[0157] Additional Chlamydia trachomatis antigens were identified by serological expression cloning. As mentioned above, these studies used pooled sera from several Chlamydia-infected individuals, but used IgA and IgM antibodies as secondary antibodies in addition to IgG. Clones selected by this method have enhanced detection of antigens recognized by the early immune response to Chlamydia infection, the mucosal humoral immune response. The following immunoreactive clones were identified and the inserts containing Chlamydia genes were sequenced: CTL2gam-1 (SEQ ID NO: 290), CTL2gam-2 (SEQ ID NO: 289), CTL2gam-5 (SEQ ID NO: 288), CTL2gam-6-3' (SEQ ID NO: 287, the second genome sequence determined, representing the 3' end), CTL2gam-6-5' (SEQ ID NO: 286, the first genome sequence determined, representing the 5 ' end), CTL2gam-8 (SEQ ID NO: 285), CTL2gam-10 (SEQ ID NO: 284), CTL2gam-13 (SEQ ID NO: 283), CTL2gam-15-3' (SEQ ID NO: 282, The second determined genome sequence represent...

Embodiment 3

[0166] Peptides can be synthesized using HPTU (O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate) activated FMOC chemistry on a Millipore 9050 peptide synthesizer. A Gly-Cys-Gly sequence can be attached to the amino terminus of the peptide to provide a means of conjugating or labeling the peptide. Peptides can be cleaved from the solid support using the following cleavage mixture: trifluoroacetic acid:ethanedithiol:thioanisole:water:phenol (40:1:2:2:3). After 2 hours of cleavage, the peptide can be precipitated in cold methyl tert-butyl ether. The peptide pellet was then dissolved in water containing 0.1% trifluoroacetic acid (TFA) and lyophilized prior to purification by C18 reverse phase HPLC. Peptides can be eluted using a gradient of 0-60% acetonitrile (with 0.1% TFA) in water (with 0.1% TFA). After lyophilization of the pure fractions, the peptides can be identified using electrospray mass spectrometry and amino acid analysis. Example 4: Isolation and Ch...

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Abstract

The present invention discloses compounds and methods for the diagnosis and treatment of Chlamydia infection. Compounds provided by the invention include polypeptides comprising at least one antigenic portion of a Chlamydia antigen and DNA sequences encoding these polypeptides. The present invention also provides pharmaceutical compositions and vaccines comprising these polypeptides or DNA sequences, as well as antibodies against these polypeptides. Diagnostic kits comprising these polypeptide or DNA sequences and suitable detection agents can be used to detect Chlamydia infection in patients and biological samples.

Description

technical field [0001] In general, the present invention relates to the detection and treatment of Chlamydia infection. In particular, the present invention relates to polypeptides comprising Chlamydia antigens, and the use of these polypeptides for serodiagnosis and treatment of Chlamydia infection. Background of the invention [0002] Chlamydia are intracellular bacterial pathogens responsible for a variety of important infections in humans and animals. Chlamydia trachomatis, one of the most common causes of sexually transmitted diseases, can cause pelvic inflammatory disease (PID), leading to blocked fallopian tubes and infertility. Chlamydia trachomatis may also play a role in male infertility. The cost of treating PID in the United States was estimated at $4 billion in 1990. Trachoma, caused by ocular infection with Chlamydia trachomatis, is the leading cause of preventable blindness worldwide. Chlamydia pneumoniae is a major cause of acute res...

Claims

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Application Information

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IPC IPC(8): C12N15/31C07K14/295C12N15/62C07K16/12A61K38/16A61K39/118A61K48/00G01N33/569C12Q1/68G01N33/53A61K38/00A61K39/00A61K39/39A61K39/395A61P31/04C07K19/00C12N1/19C12N1/21C12N5/10C12N15/09C12P21/08G01N33/566G01N33/571
CPCC07K14/295C07K2319/00A61K2039/51A61K48/00A61K38/00A61K39/00A61P31/04
Inventor P·普罗布斯特A·巴蒂亚Y·A·N·斯凯基S·P·福林J·舒勒
Owner CORIXA CORP
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