T-CELL VACCINE
An improved T-cell vaccine and methods of making the vaccine are described. The vaccine may be made by stimulating T-cells with all epitopes of an antigenic polypeptide that may be capable of stimulating autoreactive T cells.
1 . A method of identifying an epitope within a polypeptide antigen for an autoreactive T-cell, comprising:
(a) providing a sample comprising T cells isolated from a host;
(b) adding one or more different peptides to a plurality of portions of the sample, wherein the sequences of the peptides collectively comprise a portion of the sequence of the polypeptide antigen; and
(c) identifying a portion of the sample comprising activated autoreactive T cells,
wherein a peptide that activates the autoreactive T cells identified by (c) comprises the epitope.
2 . The method of claim 1 wherein the sequences of the peptides collectively comprise the complete sequence of the polypeptide antigen.
3 . The method of claim 1 wherein the polypeptide antigen is MBP, PLP, MOG or a combination thereof.
4 . The method of claim 1 wherein the different peptides comprise overlapping sequence of 8-12 or 4-19 amino acids.
5 . The method of claim 1 wherein the different peptides comprise about 12-16 or 8-20 amino acids.
6 . The method of claim 1 wherein the Stimulation Index of the sample comprising activated autoreactive T cells is above a predetermined value.
7 . A method of preparing a T cell vaccine, comprising
(a) providing a sample comprising T cells isolated from a patient;
(b) contacting the T-cells with one or more different peptides, whereby autoreactive T-cells are activated;
(c) expanding activated autoreactive T cells; and
(d) attenuating the autoreactive T cells;
wherein the one or more different peptides comprise all epitopes of an antigenic polypeptide capable of stimulating autoreactive T-cells with a stimulation index above a predetermined value.
8 . The method of claim 7 wherein the antigenic polypeptide is MBP, PLP, MOG or a combination thereof.
9 . A method of preparing a T cell vaccine, comprising
(a) providing a sample comprising T cells isolated from a patient;
(b) contacting the T-cells with one or more different peptides comprising an epitope identified by the method of claim 1 , whereby autoreactive T-cells are activated;
(c) expanding activated autoreactive T cells; and
(d) attenuating the autoreactive T cells;
wherein the one or more different peptides comprise all epitopes of an antigenic polypeptide capable of stimulating autoreactive T-cells with a stimulation index above a predetermined value.
10 . A method of detecting epitope shift in a T cell mediated disease, comprising:
(a) identify the epitopes of an autoreactive antigen according to the method of claim 1 ; and
(b) comparing the epitopes of (a) to epitopes at a control,
wherein epitope shift has occurred if the epitopes are different.
11 . A method of diagnosing an epitope shift, comprising:
(a) detecting an epitope shift according to the method of claim 10 ,
wherein detection of epitope shift indicates that epitope shift has occurred.
12 . A method of monitoring an epitope shift, comprising:
(a) detecting an epitope shift according to the method of claim 10 ; and
(b) comparing the epitopes to a previous time,
wherein epitope shift has occurred if the epitopes are different.
13 . A T cell vaccine comprising T cells that are specific to an antigenic polypeptide wherein each epitope of the antigenic polypeptide capable of producing a stimulation index above a predetermined value is recognized by autoreactive T cells present in the vaccine.
14 . The T cell vaccine of claim 13 wherein the antigenic polypeptide is MBP, PLP, MOG or a combination thereof.
15 . The T cell vaccine of claim 13 wherein the autoreactive T cells present in the vaccine comprise less than 50% T cells that recognize the epitopes of the antigenic polypeptide capable of producing a stimulation index of less than a predetermined value.
16 . The T cell vaccine of claim 13 wherein the autoreactive T cells present in the vaccine are positive for cell makers selected from one the following: CD3, CD4, CD8, CD25, TCR αβ, TCR γδ, HSP60 or a combination thereof.