Methods of MHC class II epitope mapping, detection of autoimmune T cells and antigens, and autoimmune treatment

a technology of autoimmune t cells and antigens, which is applied in the field of mhc class ii epitope mapping, can solve the problems of labor and time-consuming process, and achieve the effect of determining the efficacy or effectiveness of the therapeutic agen

Inactive Publication Date: 2006-10-26
BENAROYA RES INST AT VIRGINIA MASON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cells are then expanded and assayed for MHC restriction and peptide specificities by screening clones with individual peptides which cover the antigen, a labor intensive and time consuming process.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0113] In this example, a panel of 60 overlapping HSV-2 VP16 peptides were analyzed to identify MHC class II epitopes in HSV-2 VP16.

Methods and Materials

[0114] Generation of DRA1*0101 / DRB1*0401 and DRA1*0101 / DRB1*0404 Tetramers

[0115] Soluble human MHC class II α and β chains were expressed using a Drosphila Schneider S-2 cell system. The MHC class II α and β chains were expressed from separate expression vectors, pRMHa-2 alpha and pRMHa-3 beta, which are derived from the Cu-inducible pRmHa-3 Drosophila expression vector. The alpha chain was expressed from an expression cassette that includes the metallothionen promoter upstream of soluble MHC class II α chain which is fused to a leucine zipper coding region. The beta chains was expressed from a similar expression cassette that includes the metallothionen promoter upstream of the MHC class II β chain, which is fused to a leucine zipper coding region and a biotinylation coding sequence. These expression cassettes containing the co...

example 2

[0139] In this example, a comparison is made of T cell epitopes as selected by the TEPITOPE program and those identified by the TGEM method.

[0140] Computer-Implemented Identification of Candidate Epitopes.

[0141] Human subjects infected with HSV-2 mount T cell responses to viral antigens, including CD4+ T cell responses to peptides from the VP16 tegument protein (see Koelle et al., J. Virol. 72:7476-83 (1998)). Using methods according to the present invention, peripheral blood T cells from HSV-2 infected individuals carrying HLA DRB1*0101, DRB1*0401, DRB1*0402, DRB1*0404, DRB1*1104 and DRB1*1501 MHC class II alleles were tested for their ability to bind specific MHC-VP16 epitope tetramers. VP16 T cell epitopes identified by methods of the present invention were compared with those epitopes predicted by the TEPITOPE program to bind that same MHC class II molecules. Table II lists the VP16 epitopes selected by the TEPITOPE program at a threshold level of 3%. The TEPITOPE program pred...

example 3

[0145] A T cell line that is directed against an epitope of prostate specific antigen, PSA 64-78, as been generated, establishing PSA 64-78 as an antigenic epitope. To evaluate whether PSA 64-78-loaded tetramers can be used to stain PSA specific T cells, HLA-DR0401 transgenic mice were immunized with the PSA peptide in the presence of CFA. Mice were sacrificed on day 7, T cells from the inguinale lymph nodes were isolated and stained with the DR041 / PSA 64-78 tetramers. The data show that DR041 / PSA 64-78 tetramers stain T cells from the PSA 64-78-immunized DR-4-1-IE mice. In contrast, T cells were not stained by the DR0401 / PSA 64-78 tetramers in mice immunized with hemagglutinin peptide 307-319

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Abstract

The present invention provides of using multimeric MHC class II / peptide complexes. In one aspect, methods provided for identifying MHC class II-restricted immune epitopes of a predetermined polypeptide antigen. Methods for identifying an immunostimulatory epitope for a predetermined polypeptide antigen are provided. In a related aspect, methods for screening a therapeutic polypeptide agent for an MHC class II epitope are provided. In other aspects, methods for modulating T cells and for determining or monitoring an MHC class II-restricted immune status of a patient are also provided.

Description

CONTINUITY [0001] This application is a divisional of U.S. patent application Ser. No. 10 / 116,846, filed Apr. 5, 2002, which claims the benefit of U.S. Provisional Patent Application Nos. 60 / 282,328, filed Apr. 5, 2001, and 60 / 308,962, filed Jul. 30, 2001, the entire disclosures of which are incorporated by reference herein.[0002] This work was supported by U.S. Government grant numbers AI-44443 and AI-30731, awarded by the National Institutes of Health. The U.S. Government may have certain rights in the invention.BACKGROUND OF THE INVENTION [0003] The T cell-mediated response to complex antigens involves recognition of selected peptide epitopes presented in the context of MHC molecules expressed on antigen presenting cells. The choice of these immunogenic epitopes from among the often hundreds or thousands of amino acids comprising an antigenic protein depends significantly on the binding properties of a given MHC type and the interactions of specific amino acids with a T cell rece...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/14C40B30/06C40B40/10A61K39/00G01N33/50G01N33/564G01N33/569
CPCA61K39/0008A61K2039/5156G01N33/505G01N2800/24G01N33/56977G01N2800/042G01N33/564
Inventor KWOK, WILLIAMNEPOM, GERALDGEBE, JOHNREIJONEN, HELENALIU, ANDREW
Owner BENAROYA RES INST AT VIRGINIA MASON
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