Immune Tolerance: from Gene Expression to Drug Discovery. Therapeutic Immunology Group (Prof. H. Waldmann) Therapeutic Antibody Centre (Prof. G. Hale). Immune Tolerance and Therapy. Therapy to reverse breakdown of self tolerance in autoimmune diseases
Therapeutic Immunology Group
(Prof. H. Waldmann)
Therapeutic Antibody Centre
(Prof. G. Hale)
What are the approaches to find new therapeutics:
Random screening of chemical libraries in a “surrogate assay” (eg. suppression of antigen specific proliferation in vitro).
Look for monoclonal antibodies that modulate a function (eg. same assay).
Targeted chemical design/antibodies against specific protein structures.
How to identify the most relevant/specific target proteins on possibly rare cells? Ideally we want targets expressed ONLY on target cells to avoid potential toxicity against other tissues.
Look for genes that are specifically expressed in the functional cell type of interest – in our example, Th1 but NOT T regulatory (Treg) cells.
Analysis of known genes (RT-PCR/Antibodies/Protein Gels):
There are >1000 interesting “immunological” genes and probably many more important but unidentified genes. How to choose?
Differential Display and Gene Cloning:
Clones genes over-expressed in one cell compared to one other, but these may be shared with other cells and you don’t know what you are working with until you have it cloned and sequenced. How to choose?
Can identify patterns of expression from many (10,000+) genes and multiple samples. Genes must already have been cloned (<1/3 of genome?), it is quick, but not very sensitive (or reliable?), and currently expensive.
SAGE (Serial Analysis of Gene Expression)
Can identify almost the entire pattern of gene expression (the “transcriptome”) with no a priori knowledge of the gene sequences. Multiple samples are directly comparable as a database. Sensitivity depends on the number of tags sequenced: this can be labour intensive.
CD4+ T cell clones/lines against DBY-Ek male antigen
Clone Source Polarised in Type Cytokines
R2.2 A1(M)xRAG-1-/- IL-2 Th1 IFN-g
R2.4 A1(M)xRAG-1-/- IL-4 Th2 IL-4, IL-10
Tr1D1 A1(M)xRAG-1-/- IL-10 Tr1/Treg IL-10 (IL-4)
(naïve) (aCD3 cloned)
A1MP A1(M) naïve Anti-CTLA4 Treg IL-10 (IL-4)
SKA A1(M) + male DBY-Ek peptide Tskin/Treg IL-10
line skin graft CD4+
AE = Nla-III
TE = BsmF1
T cell clone cluster
Spleen CD4 cell cluster
Clustered Expression Chart of approx. 300 known genes (CD antigens, cytokines and receptors)
A close up of a T antigens, cytokines and receptors)reg
cluster of known genes
Real Time PCR Machine antigens, cytokines and receptors)
TM4 antigens, cytokines and receptors)
Ratio of tolerant to rejecting skin graft expression
Fluorescence Activated Cell Scanner skin grafts
Th2 skin grafts
CD25 -> skin grafts
The Therapeutic Antibody Centre skin grafts
expressed by effector but not regulatory T cells
Therapeutic antibody purification molecules
Clinical Trials of Therapeutic Antibody molecules
Or go to the Pathology Web site:
And click on “Herman Waldmann”