Protective Immunity Project

1
Protective Immunity Project

• Chris Larsen, MD, DPhil,
• PI
• Christine Martens, Ph.D.
• Project Manager
• Vicki Hertzberg, Ph.D.
• Director of Data Management and Analysis Core

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3 studies

• 2 human
• New renal transplants (n60) and 20 controls,
  followed for 2 years
• Existing renal transplants (n60) and 30
  controls, receiving flu vaccine, followed for
  short time
• 1 macaque study, 30 macaques randomized to 6
  groups

3
Longitudinal studies
ID Time 1 Time 2 Time t
1

n
4
Measurements

• At each time point for each subject obtain
  measurements from
• Flow cytometry
• ELISPOT/ELISA
• M-array

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Flow cytometry

• For each subject at each time, take several blood
  samples.
• For each blood sample, label all of the PBMCs
  with 8-10 markers (a panel). Cells will fluoresce
  according to the amount of each marker present.
• Apply 6-8 panels for each subject at each time.
• Each panel usually has 2 markers that are common
  to all other panels

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What to the data look like?

• X-Y graph for fluoresence intensity for marker X
  vs marker Y.
• Gates are drawn that
• Separate present from absent
• Isolate cells of different types

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Issues?

• How to analyze?
• Can we deal with fluorescence intensity rather
  than presence or absence?
• How do we institute QC for the gating?