Cleavage of transaldolase by granzyme B causes the loss of enzymatic activity with retention of antigenicity for multiple sclerosis patients.

Multiple sclerosis (MS) is an autoimmune demyelinating disease of the CNS resulting from a progressive loss of oligodendrocytes. Transaldolase (TAL) is expressed at selectively high levels in oligodendrocytes of the brain, and postmortem sections show concurrent loss of myelin basic protein and TAL from sites of demyelination. Infiltrating CD8(+) CTLs are thought to play a key role in oligodendrocyte cell death.

Measurement of polyclonal and antigen-specific cytotoxic T cell function.

Measurement of in vitro cytotoxic function of human T cells can be accomplished by polyclonal stimulation of T cell effectors using anti-CD3 antibody, which stimulates all cytolytic effector cells, or with a specific stimulating antigen. Accordingly, two sets of assays of cytolytic T cell function are described in this unit, one for measuring anti-CD3-mediated cytotoxicity and the other for measuring antigen-specific cytotoxicity. Although the calcein release assay (CARE-LASS) described here is for use with antigen-activated cytotoxic T lymphocytes (CTL) as well as natural killer (NK) or lymphokine-activated killer (LAK) cells, minor changes in the protocols that address polyclonal T cell activation are described that make them suitable for use with calcein-labeled target cells.

Peptide binding motifs for MHC class I and II molecules.

This overview discusses the use of peptide-binding motifs to predict interaction with a specific MHC class I or II allele, and gives examples for the use of MHC binding motifs to predict T-cell recognition.

Generation of continuously growing B cell lines by epstein-barr virus transformation.

Primary cultures of cells tend to have a limited life span, which in turn limits the availability of a consistent population of cells to study. In this unit, Epstein-Barr virus produced by a marmoset cell line is used to transform B cells into a continuously growing cell line that produces no virus. These cell lines can be used as a constant source of cells for further study.