Vaccine and Infectious Disease Division

An ORFeome approach to identifying T-cell immune responses

Knowing which viral proteins produce the best immune response for fighting disease is essential in viral vaccine design.  However, identifying such immune response-inducing proteins in the laboratory can be extremely time consuming and expensive because of the large number of proteins expressed by some viruses.  VIDI affiliate investigator Dr. David Koelle and colleagues designed a new method of identifying reactive proteins using arrays to increase the assay efficiency.  With this method, Koelle’s team used bacteria (E. coli) to produce protein from all 200 open reading frames (ORFs) of the Vaccinia virus genome, and then pooled these proteins in arrays to detect immune responses in blood samples from people who were vaccinated with Vaccinia.  The proteins are expressed inexpensively and are biologically active at extreme dilutions, thus increasing throughput and decreasing cost.   This method will benefit future studies focused on pathogens containing a large number of proteins.

ORFeome approach to the clonal, HLA allele-specific CD4 T-cell response to a complex pathogen in humans.  Jing L et al.  J Immunol Methods. 2009 347(1-2):36-45.