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Immune system's bare essentials used to speedily detect drug targets
Date:10/31/2010

the antigen into its visible, identifiable protein parts.

In the first set of experiments, the team mixed chemical solutions of each antigen with the five key proteins and used mass spectrometry an electron-beaming device that can measure the exact make-up of molecules to determine the best-fitting peptide based on precisely which segment of the antigen appeared as mass peaks. Peaks would indicate that HLA-DR had successfully bound to the antigen at a likely epitope.

Next, researchers confirmed their mass spectrometry findings by injecting mice bred to produce human HLA-DR with each antigen to trigger a standard immune response and collecting samples of the resulting T cells. The T cells were then grown in the lab and exposed to various peptides, including the suspect epitopes, to identify and confirm that only one triggered the greatest chemical response from the cultured T cells. The scientists knew that if they could match a peak highlighted by mass spectrometry to the peptide that produced the greatest T cell reaction, they had found the most heavily favored epitope.

When both tests were performed on any of the four disease antigens, researchers were able to narrow the suspect binding sites to one "immunodominant" epitope each for Texas strain of the flu, type II collagen, avian flu and malaria.

Kim, a postdoctoral research fellow at Johns Hopkins, says designing both experiments and completing the verification study took some seven years, noting that adding HLA-DM, which she calls a protein editor, was the pivotal factor in making the initial epitope-selection process work.

Researchers say their next steps are to broaden and refine their chemical mixture for selecting and identifying possible epitopes for other kinds of HLA because the current set of experiments analyzes only one of the most common HLA-type molecules in whites.


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Contact: David March
dmarch1@jhmi.edu
410-955-1534
Johns Hopkins Medical Institutions
Source:Eurekalert

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