Identification of CD8 T-lymphocyte epitopes in OmpB of Rickettsia conorii

Zhen Li, C. Marcela Díaz-Montero, Gustavo Valbuena, Xue Jie Yu, Juan Olano, Hui Min Feng, David Walker

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The 1.2-kb DNA fragment of the Rickettsia conorii outer membrane protein B gene (OmpB451-846) was subcloned using site-specific PCR primers and expressed as six smaller fragments: OmpB458-652, OmpB595-744, OmpB595-654, OmpB654-692, OmpB689-744, and OmpB739-848. NCTC cells transfected with a mammalian expression vector expressing the fragments OmpB689-744 and OmpB739-848 stimulated immune anti-R. conorii CD8 T lymphocytes, suggesting the presence of CD8 T-lymphocyte-stimulating epitopes on these fragments. In order to further characterize the CD8 T-lymphocyte-stimulatory elements, CD8 T-lymphocyte epitopes on OmpB689-744 and OmpB739-848 were mapped by overlapping synthetic peptides. The ability of these synthetic peptides to stimulate immune CD8 T lymphocytes was determined by gamma interferon (IFN-γ) production and cell proliferation after incubation with simian virus 40-transformed murine vascular endothelial cells in the presence of a 20 μM solution of each synthetic peptide. Five synthetic peptides, SKGVNVDTV (OmpB708-716), ANVGSFVFN (OmpB735-743), IVSGTVGGQ (OmpB749-757), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), induced secretion of IFN-γ at significantly higher levels than the controls. Three of these five peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), also stimulated the proliferation of immune CD8 T lymphocytes. Significantly higher levels of specific cytotoxic T-lymphocyte killing were observed with the same three synthetic peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820).

Original languageEnglish (US)
Pages (from-to)3920-3926
Number of pages7
JournalInfection and Immunity
Volume71
Issue number7
DOIs
StatePublished - Jul 1 2003

Fingerprint

Rickettsia conorii
T-Lymphocyte Epitopes
Peptides
T-Lymphocytes
Simian virus 40
Cytotoxic T-Lymphocytes
Interferons
Interferon-gamma
Membrane Proteins
Endothelial Cells
Cell Proliferation
Polymerase Chain Reaction
DNA

ASJC Scopus subject areas

  • Immunology

Cite this

Identification of CD8 T-lymphocyte epitopes in OmpB of Rickettsia conorii. / Li, Zhen; Díaz-Montero, C. Marcela; Valbuena, Gustavo; Yu, Xue Jie; Olano, Juan; Feng, Hui Min; Walker, David.

In: Infection and Immunity, Vol. 71, No. 7, 01.07.2003, p. 3920-3926.

Research output: Contribution to journalArticle

Li, Zhen ; Díaz-Montero, C. Marcela ; Valbuena, Gustavo ; Yu, Xue Jie ; Olano, Juan ; Feng, Hui Min ; Walker, David. / Identification of CD8 T-lymphocyte epitopes in OmpB of Rickettsia conorii. In: Infection and Immunity. 2003 ; Vol. 71, No. 7. pp. 3920-3926.
@article{3548529cc2884a0b9175cb67c440d892,
title = "Identification of CD8 T-lymphocyte epitopes in OmpB of Rickettsia conorii",
abstract = "The 1.2-kb DNA fragment of the Rickettsia conorii outer membrane protein B gene (OmpB451-846) was subcloned using site-specific PCR primers and expressed as six smaller fragments: OmpB458-652, OmpB595-744, OmpB595-654, OmpB654-692, OmpB689-744, and OmpB739-848. NCTC cells transfected with a mammalian expression vector expressing the fragments OmpB689-744 and OmpB739-848 stimulated immune anti-R. conorii CD8 T lymphocytes, suggesting the presence of CD8 T-lymphocyte-stimulating epitopes on these fragments. In order to further characterize the CD8 T-lymphocyte-stimulatory elements, CD8 T-lymphocyte epitopes on OmpB689-744 and OmpB739-848 were mapped by overlapping synthetic peptides. The ability of these synthetic peptides to stimulate immune CD8 T lymphocytes was determined by gamma interferon (IFN-γ) production and cell proliferation after incubation with simian virus 40-transformed murine vascular endothelial cells in the presence of a 20 μM solution of each synthetic peptide. Five synthetic peptides, SKGVNVDTV (OmpB708-716), ANVGSFVFN (OmpB735-743), IVSGTVGGQ (OmpB749-757), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), induced secretion of IFN-γ at significantly higher levels than the controls. Three of these five peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), also stimulated the proliferation of immune CD8 T lymphocytes. Significantly higher levels of specific cytotoxic T-lymphocyte killing were observed with the same three synthetic peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820).",
author = "Zhen Li and D{\'i}az-Montero, {C. Marcela} and Gustavo Valbuena and Yu, {Xue Jie} and Juan Olano and Feng, {Hui Min} and David Walker",
year = "2003",
month = "7",
day = "1",
doi = "10.1128/IAI.71.7.3920-3926.2003",
language = "English (US)",
volume = "71",
pages = "3920--3926",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "7",

}

TY - JOUR

T1 - Identification of CD8 T-lymphocyte epitopes in OmpB of Rickettsia conorii

AU - Li, Zhen

AU - Díaz-Montero, C. Marcela

AU - Valbuena, Gustavo

AU - Yu, Xue Jie

AU - Olano, Juan

AU - Feng, Hui Min

AU - Walker, David

PY - 2003/7/1

Y1 - 2003/7/1

N2 - The 1.2-kb DNA fragment of the Rickettsia conorii outer membrane protein B gene (OmpB451-846) was subcloned using site-specific PCR primers and expressed as six smaller fragments: OmpB458-652, OmpB595-744, OmpB595-654, OmpB654-692, OmpB689-744, and OmpB739-848. NCTC cells transfected with a mammalian expression vector expressing the fragments OmpB689-744 and OmpB739-848 stimulated immune anti-R. conorii CD8 T lymphocytes, suggesting the presence of CD8 T-lymphocyte-stimulating epitopes on these fragments. In order to further characterize the CD8 T-lymphocyte-stimulatory elements, CD8 T-lymphocyte epitopes on OmpB689-744 and OmpB739-848 were mapped by overlapping synthetic peptides. The ability of these synthetic peptides to stimulate immune CD8 T lymphocytes was determined by gamma interferon (IFN-γ) production and cell proliferation after incubation with simian virus 40-transformed murine vascular endothelial cells in the presence of a 20 μM solution of each synthetic peptide. Five synthetic peptides, SKGVNVDTV (OmpB708-716), ANVGSFVFN (OmpB735-743), IVSGTVGGQ (OmpB749-757), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), induced secretion of IFN-γ at significantly higher levels than the controls. Three of these five peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), also stimulated the proliferation of immune CD8 T lymphocytes. Significantly higher levels of specific cytotoxic T-lymphocyte killing were observed with the same three synthetic peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820).

AB - The 1.2-kb DNA fragment of the Rickettsia conorii outer membrane protein B gene (OmpB451-846) was subcloned using site-specific PCR primers and expressed as six smaller fragments: OmpB458-652, OmpB595-744, OmpB595-654, OmpB654-692, OmpB689-744, and OmpB739-848. NCTC cells transfected with a mammalian expression vector expressing the fragments OmpB689-744 and OmpB739-848 stimulated immune anti-R. conorii CD8 T lymphocytes, suggesting the presence of CD8 T-lymphocyte-stimulating epitopes on these fragments. In order to further characterize the CD8 T-lymphocyte-stimulatory elements, CD8 T-lymphocyte epitopes on OmpB689-744 and OmpB739-848 were mapped by overlapping synthetic peptides. The ability of these synthetic peptides to stimulate immune CD8 T lymphocytes was determined by gamma interferon (IFN-γ) production and cell proliferation after incubation with simian virus 40-transformed murine vascular endothelial cells in the presence of a 20 μM solution of each synthetic peptide. Five synthetic peptides, SKGVNVDTV (OmpB708-716), ANVGSFVFN (OmpB735-743), IVSGTVGGQ (OmpB749-757), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), induced secretion of IFN-γ at significantly higher levels than the controls. Three of these five peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820), also stimulated the proliferation of immune CD8 T lymphocytes. Significantly higher levels of specific cytotoxic T-lymphocyte killing were observed with the same three synthetic peptides, SKGVNVDTV (OmpB708-716), ANSTLQIGG (OmpB789-797), and IVEFVNTGP (OmpB812-820).

UR - http://www.scopus.com/inward/record.url?scp=0037634239&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037634239&partnerID=8YFLogxK

U2 - 10.1128/IAI.71.7.3920-3926.2003

DO - 10.1128/IAI.71.7.3920-3926.2003

M3 - Article

VL - 71

SP - 3920

EP - 3926

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 7

ER -