Strings of epitopes useful in diagnosing and eliciting immune responses to sexually transmitted infections

Patent metadata

Field	Value
Publication number	US-10386369-B2
Application number	US-201815877635-A
Country	US
Kind code	B2
Filing date	Jan 23, 2018
Priority date	Mar 13, 2013
Publication date	Aug 20, 2019
Grant date	Aug 20, 2019

Abstract

The invention provides methods and compositions for detecting and diagnosing sexually transmitted infections using a string of epitopes (SOE) specific for detection of causative microorganisms. The antigenic epitopes may be single epitope sequences, a plurality of epitope sequences joined by repeats of glycine (-GG-) and/or lysine (-KK-) to form a series of epitopes (SOE), or nucleotide sequences encoding one or more SOEs and host cells harboring said SOE nucleotide sequences. SOEs specific for highly immunogenic regions of proteins from Trichomonas, Treponema and Neisseria species are provided. SOEs to detect the presence of trichomonas species comprise regions from Trichomonas-sptciric aldolase, GAPDH, a-enolase and a-actinin proteins. Pharmaceutical compositions comprising SOEs can also be used as vaccines or to elicit an immune response to specific microorganisms.

First claim

What is claimed is:

1. A method for detecting the presence of one or more Trichomonas vaginalis microorganisms in a biological sample of a subject, comprising the steps of:
combining said biological sample with a polypeptide including a series of epitopes (SOE) which includes at least a plurality of epitopes selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO:82, and SEQ ID NO: 137, said epitopes being arranged as a linear array with each of said epitopes being connected by an amino acid linker consisting of a repeat of an amino acid, wherein said combining is performed under conditions whereby antigen-antibody complexes are permitted to form; and
detecting formation of at least one antigen-antibody complex as an indication of a presence of at least one microorganism of said one or more Trichomonas vaginalis microorganisms in said biological sample.

2. The method of claim 1 , wherein said Trichomonas vaginalis microrganisms are selected from the group consisting of T. vaginalis isolates T016, T068-II, UT40, and VB102.

3. The method of claim 1 , wherein said detecting step is performed using an immunoassay.

4. The method of claim 3 , wherein said immunoassay is an enzyme-linked immunosorbent assay (ELISA).

5. The method of claim 1 , wherein said biological sample is selected from the group consisting of serum, plasma, blood, saliva, semen, cerebrospinal fluid, semen, prostatic fluid, urine, sputum, joint fluid, body cavity fluid, whole cells, cell extracts, tissue, biopsy material, aspirates, exudates, vaginal washings, pap smear samples, pap smear preparations, slide preparations, fixed cells, and tissue sections.

6. The method of claim 1 , wherein said subject is selected from the group of human, non-human primate, dog, cat, cattle, sheep, swine, horse, bird, mouse and rat.

7. A polypeptide including a series of epitopes (SOE) which includes at least a plurality of epitopes selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO:82, and SEQ ID NO: 137 said epitopes being arranged as a linear array with each of said epitopes being connected by an amino acid linker consisting of a repeat of an amino acid.

8. A kit for detecting the presence of one or more Trichomonas vaginalis microorganisms comprising at least one polypeptide of claim 7 and assay reagents or media for detecting formation of at least one antigen-antibody complex comprising said at least one polypeptide.

9. A method of eliciting an immune response to a Trichomonas vaginalis microorganism in a subject, comprising the steps of
preparing a pharmaceutical composition comprising
at least one SOE, which includes at least a plurality of epitopes selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO:82, and SEQ ID NO: 137, said epitopes being arranged as a linear array with each of said epitopes being connected by an amino acid linker consisting of a repeat of an amino acid , and a suitable carrier and adjuvant, and

administering said pharmaceutical composition to said subject in an amount sufficient to stimulate formation of antibodies to said SOE by the immune system of said subject.

Assignees

• Univ Washington State

Inventors

• Alderete John F

Classifications

• C07K16/20

  from protozoa · CPC title

• G01N2333/20

  from Spirochaetales (O), e.g. Treponema, Leptospira · CPC title

• C07K2319/40

  containing a tag for immunodetection, or an epitope for immunisation · CPC title

• C07K14/22

  from Neisseriaceae (F) · CPC title

• G01N2333/22

  from Neisseriaceae (F), e.g. Acinetobacter · CPC title