The protocols being used or under development by our Technology Development Component are listed below.  Click on the title for access to the decsription of the protocol.  We will revise and add protocols as needed.

Collection of Cell Types from Whole Blood Each 50 ml bleed will be processed in the same manner to isolate 4 cell populations needed for the studies (NK , pDC, mDC and T cells). The first step is a ficoll purification of the leukocyte rich buffy coat.

Microneutralization Assays This assay is under development. We will use reverse genetics technology to create an eight segmented influenza virus which has it’s HA gene segment replaced with a GFP gene maintaining the replication, transcription and packaging signals of the HA.

Transcriptome Fingerprinting Microarrays allow study of many transcripts simultaneously, although they have low sample throughput and limited accuracy.  Accurately profiling the gene responses to a large number of chimeric viruses at multiple time points would not be possible with microarrays.  In contrast to microarrays, massively parallel high throughput quantitative PCR provides high accuracy and sample throughput. The use of stringent assay standardization and robotic instrumentation make possible accurate measurement of hundreds of transcripts in thousands of samples at low cost/data point.