The Institute for Immunity, Transplantation and Infection at Stanford

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The Institute for Immunity, Transplantation and
Infection at Stanford
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• In order to fulfill the mission of the ITI, the
  following
• goals were set
• (1) To help recruit faculty whose interests are
  in ITI based diseases,
• and whose research is aimed toward moving
  discoveries from the bench to
• bedside or bedside to bench.
• To develop and fund young clinical faculty in the
  area of
• clinical trial development and technology
  transfer in order to both understand the
• pathophysiology as well as the treatment of ITI
  based diseases.
• To establish a Postdoctoral Fellowship Program
  for MD
• or MD PhD fellows who will do a project either in
  basic research aimed toward
• pre-clinical models or clinical trials.

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• Continued Goals of the ITI
• To establish new interactions among the ITI
  faculty
• through seminars, retreats and educational
  initiatives
• (5) To support high school summer students who
  will
• spend at least eight weeks studying in a lab
  oriented toward ITI based disease
• pathophysiology or therapy.
• (established Dr P.J. Utz, Director)
• (6) To develop an Immune Monitoring Unit.
• This unit will allow innovative techniques in
  monitoring the immune system
• developed at Stanford and elsewhere to be applied
  to patient care.
• (7) To Develop Innovative Curriculum, a new
  program has been
• developed to teach clinical approaches to PhD
  students from multiple disciplines.

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A Clinical Immunology Summer School for High
School Students P.J. Utz, M.D.
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• Summer Student Model
• Background
• Initiated in summer 2000 with 10 students
• Funded by donors and the NCAF
• Expanded to 25 students in 2001 - 2004
• 15, 42, 141, and 193 applications
• 2003 statistics Mean GPA 4.06, SAT 1363,
• s3 valedictorians, 10 1 in class
• Students from public and private schools
• 80 women

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• Goals for High School and Undergraduate Program
• Interest students in careers in biomedical
  research
• Improve teaching skills of graduate students
  and
• postdoctoral fellows
• Encourage interactions among faculty members
• Community outreach
• University
• Medical School
• Industry

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• Existing MODEL
• Program
• Lab selection by student from faculty
• 15 lectures on basic immunology
• 300 page syllabus with assigned reading
• Lectures by individual faculty
• Lectures on poster assembly and presentation
• Intensive research experience
• Poster presentation
• Longitudinal study of program graduates is
  ongoing

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• This model has been exported nationally to
• 10 of 30 FOCIS Centers of Excellence
• Centralized educational materials
• Syllabus
• PowerPoint lectures
• All administrative materials
• Advertisements
• Application forms
• Letters of acceptance/rejection/waitlist
• Organizational materials
• Lecture schedules
• Public relations materials
• Information for participating mentors/labs

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Immune Monitoring Core at Stanford

• Needed to support new therapies and innovative
  trials.
• Current facilities are in multiple labs
• A centralized facility will provide better
  patient care
• Advantages of proposed immune monitoring core
• Provide a resource to the community
• Designed to allow novel techniques in genomics
  and proteomics to be integrated as information
  based medicine by new bioinformatics technology
• Substantial opportunity currently exists to build
  on existing expertise in research and to increase
  both clinical investigation and delivery of novel
  therapies.

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• Potentially Useful Multiplex Proteomics Assays
• FACS/Phospho FACS
• Autoantibody Profiling
• Cytokine Profiling
• Bead-Based
• Cleavable Tags
• Planar Arrays
• Signaling Molecule Assays
• FACS-Based Techniques
• Planar Capture Arrays
• Cleavable Tags
• Lysate Arrays
• Serum Proteome Analysis by Mass Spectroscopy

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Activated vs Static Signaling Proteomics in
Autoimmunity.
Garry P. Nolan, Ph.D. Stanford University Dept.
of Microbiology Immunology
Signaling Phenotypes in single immune cells
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Why use Flow Cytometry to Measure Signaling?
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Revealing Lupus (SLE) Immune Deficiencies
requires stimulation of cells
SLE prone animals were given a drug iv
and isolated cells assayed before and following
activation ex vivo
T cells
Autoimmune
Normal
Peter Krutzik Matt Hale
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Proteomic Assays P.J. Utz, M.D. and Bill
Robinson, MD. Ph.D. Stanford University
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Produce arrays using a robotic microarrayer
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The 1152 Feature CTD Chip
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• Details of Current Arrays
• Coated Glass Slides
• 150 Slides Per Print Run
• Approximately 4,000-5,000 Spots/Features Per
  Slide
• Features are Duplicated
• 200uM Feature, 200pg Antigen
• Protein, Peptide, Nucleic Acid, RNP, Complex
• Monoclonal Antibody or Complex Mixtures
• 30ul Volume
• Fluorescent Detection (Cy3, Cy5, BoDipy)
• Secondary Antibody vs Competition
• Sensitive and Specific

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• Potential Applications
• Multiplex Diagnostic Test
• Epitope Spreading
• Follow Response to Therapy
• Guide Selection of Therapy
• Discovery Tool

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Reverse Phase Protein Lysate Microarrays P.J.
Utz, M.D.
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Protein MicroarraysDetection of Phosphorylated
Proteins
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• In Vitro Studies Using Lysate Arrays
• Analysis of Rare Cell Populations
• FACS-Sorted Cells
• Laser Capture Microdissection
• Antigen-Specific Cells (e.g., Tetramers)
• Correlation With Transcript Profiling (genomics)
  Experiments

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A Cellular MicroArray
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MHC-Cytokine Arrays
Secondary cytokine Detection antibody Conjugated
to a flurophore
Co-spotted Cytokine Capture antibody
Cytokine secreted by T cell after recognition
of Peptide/MHC
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aCD8 Co-Spots MART1/A2
Co-Spots
aCD8 brightfield
MART1/A2 brightfield
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Functional T Cell Responses to Peptide Vaccines
Key
of Blocks Denotes gp100 Specific Activity
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What is now possible

• Identifying disease subsets by proteomic
  signatures
• Correlating proteomic signatures with clinical or
  therapy induced outcome in disease
• DIRECT build-out of patient-specific proteomic
  maps and mechanistic inferences
• Observation of rare subsets that would be missed
  by all other signaling analyses (mass spec,
  chromatography, elisa, bead)

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Introduction to Medicine for PhD
studentsImmunology 230

• How to learn about a disease using a prototype
  (diabetes)
• Betsy Mellins MD

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Course Goals

• Understand how medical knowledge is organized
• Appreciate the concerns and tools of the various
  scientific fields and clinical disciplines within
  medicine
• Identify quality sources of medical information
• Gain some familiarity with medical language
• Learn specific information about human physiology
  and pathophysiology
• Identify opportunities to apply your primary
  discipline to unsolved medical problems

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Focus on one disease

• This models the situation that may be confronted
  in the future
• The idea is that a guided learning experience
  about one disease will teach skills that can be
  applied to learning about other diseases
• Focus on Diabetes
• Multi-system disorder Many branches of medicine
  needed to understand it
• Relatively common
• Partially understood many ?s that will draw on
  your expertise

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Projects

• Project goals
• To work in inter-disciplinary teams on a medical
  problem that draws on each students primary
  expertise
• To practice learning about medicine in the
  context of a focused effort to solve a problem
• To demonstrate your acquisition of medical
  knowledge in the project report
• Project teams
• 3-4 students from different disciplines with a
  coach who is a graduate of last years course

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Design of Human Microarray Experiment for
Identifying Genes Associated with Insulin
Resistance

• Presented by
• Su-In Lee, Mechanical Engineering
• Kevin Pan, Biophysics

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Our Commitment to medical research and
education and their role in serving the public go
to the very core of what the University is
about. John Hennessy,
President