Educational Level:

Roadmap Objectives:

• • Article: Cell Interactions in the Induction of Tolerance, The role of Thymic Lymphocytes (Gershon, R.K. and Kondo, K., Immunology, 1970)
  • Content area/major concepts: First paper to describe the discovery of lymphocytes (Tregs) that induce immunological tolerance.
    
    Antigen recognition (protein shape, bonds used, peptide configuration).
    Cell signaling (cross talk, proliferative and survival signals).
    Antibody production and circulation.
    Lymphocyte development and associated organs.
    
  • Methods or technology used to obtain data: Thymectomy of mice, Irradiation of mice, Working with cells (General cell culture technique, Cell counting for viability, Titration of cells), Retro-orbital blood collection, Hemagglutination test, Student T-Test
  • How the CREATE strategy was used:
  • Biggest teaching challenge: While the techniques are very “low” tech in this paper from 1970, there will many things to cartoon. I will also have to make sure to provide more background methodology information, as it is not very “complete” in the steps needed to get the big picture of two of the experimental techniques used. Also the graphs are a bit challenging at first glance, BUT this will make it great to be able to re-graph the data in a more reader friendly way.
• • Article: Organ-Specific Autoimmune Diseases Induced in Mice by Elimination of T Cell Subset (Sakagushi, S., et. al., J. Exp. Med., 1985)
  • Content area/major concepts: Evidence that a subset of T cells (Tregs) participate in natural self-tolerance, as their removal is correlated to the induction of autoimmunity.
    
    Inoculation.
    Soluble immune complex formation.
    MHC (importance and function).
    Graft vs. host disease.
    
  • Methods or technology used to obtain data: Histological examination (Preparation of tissue samples, Hematoxylin and Eosin staining, Immunofluorescence staining), Adoptive transfer of disease in mice, Use of Knock out mice, Production of anti-sera, C-dependent Cytotoxicity Test
  • How the CREATE strategy was used:
  • Biggest teaching challenge: This paper is pretty straight forward, but the C-dependent cytotoxicity test will be challenging to explain.
• • Article: GIVEN as HW (REVIEW ARTICLE) Role of Cytokines in Determining T-Lymphocyte Function (O’Garra, A. and Murphy, K., Curr. Opin. Immunol., 1994)
  • Content area/major concepts: First insight into the characterization of the cytokine profile of a subset of CD4+ T cells (Tregs) with immunomodulatory function.
    
    
  • Methods or technology used to obtain data: ELISA, ELISpot
  • How the CREATE strategy was used:
  • Biggest teaching challenge:
• • Article: Autoimmune Disease as a Consequence of Developmental Abnormality of a T Cell Subpopulation (Masanao, A., Sakagushi, S., et. al., J. Exp. Med., 1996)
  • Content area/major concepts: First discovery of the CD25 cell surface marker for T regs and its purpose. Set the stage of its use as a way to characterize T regs (CD4+CD25+).
    
    Clusters of Differentiation use for characterizing cells.
    Co-receptor activation mechanisms.
    Cytokines and chemokines.
    Transcription.
    Translation.
    
  • Methods or technology used to obtain data: Flow cytometry, FACS sorting, Isolation of lymphocytes from Spleen and Lymph nodes, RT-PCR
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Explaining FACS and FACS sorting
• • Article: A CD4+ T-cell Subset Inhibits Antigen-specific T-cell Responses and Prevents Colitis (Groux, H., O’Garra, A., et. al., Nature, 1997)
  • Content area/major concepts: Article that establishing that CD4+CD25+ (Tregs) are indeed a unique NEW subset of T cells.
    
    Immunological environments induce immune cell differentiation and proliferation.
    Immunological environments promote or inhibit inflammation.
    
  • Methods or technology used to obtain data: Cloning. Proliferation assay.
  • How the CREATE strategy was used:
  • Biggest teaching challenge: Cloning and intracellular cytokine signaling techniques might be difficult to explain
• • Article: CD4+CD25+ T Cells Inhibit Bothe the Induction and Effector Functon of Autoreactive T Cells and Represent a Unique Lineage of Immunoregulatory Cells (Suri-Payer, E., et. al., 1998)
  • Content area/major concepts: Paper that establishes an accepted function role of T regs as inhibitors of autoreactive T cells.
    
    Experimental design for characterizing the physical and functional aspects of immunological cells.
  • Methods or technology used to obtain data: Preparation of microscopes. Preparation of H/K ATPase expressing cell membranes.
  • How the CREATE strategy was used:
  • Biggest teaching challenge:
• • Article: Disruption of a New Forkhead/Winged-helix Protein, Scurfin, Results in the Fatal Lymphoproliferative Disorder of the Scurfy Mouse (Brunkow, M.E., et. al., Nature Genetics, 2001)
  • Content area/major concepts: Discovery of the FoxP3 transcription factor and association with T regs.
    
    Transcription.
    Translation.
    
  • Methods or technology used to obtain data: cDNA library building and use. Northern blots.
  • How the CREATE strategy was used:
  • Biggest teaching challenge:
• • Article: FoxP3 Programs the Development and Function of CD4+CD25+ Regulatory T Cells (Fontenot, J.D., et. al., Nature Immunol., 2003)
  • Content area/major concepts: Complete characterization of the functional role of FoxP3 transcription factor for Tregs.
    
    Cellular signaling.
    Gene expression and regulation.
    
  • Methods or technology used to obtain data: Generation of knock out mice. Retrovivral transfection of cells. MACS sorting. Quantitative RT-PCR.
  • How the CREATE strategy was used:
  • Biggest teaching challenge:

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