Molecular Regulation of the Production of Highly Pathogenic Autoantibodies
Focus:
The papers that I have selected all relate to the study of the molecular regulation of the production of highly pathogenic autoantibodies, particularly as related to the autoimmune disorder Systemic Lupus Erythematosus (SLE). The connection of the selected journal articles is not focused on the work from one laboratory but rather based on the understanding of the function of Activation-Induced Cytidine Deaminase (AID). AID belongs to a family of proteins termed APOBEC (Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like) proteins that function as cytidine deaminases. In the context of this module, we will focus on the role of AID in antibody diversification (i.e. somatic hypermutation and class switch recombination).
The progression of the journal articles moves from the demonstration of the overexpression of AID and its association with increased production of highly pathogenic autoantibodies to understanding the hormonal modulation (i.e. estrogen) and molecular regulation of the protein, particularly in the context of the female dominated autoimmune disorder SLE.
Overview:
I have selected three journal articles and, as an alternative to a fourth journal article, I have chosen to assign a commentary article (Women, autoimmunity, and cancer: a dangerous liaison between estrogen and activation-induced deaminase?) published in the same volume of the Journal of Experimental Medicine to highlight Paper 2. I would also assign sections of a review article (Hormonal modulation of B cell development and repertoire selection) that would serve to provide background and clarify some of the complex topics (i.e. autoimmunity, SLE, B cell development, selection and activation, estrogen receptors and estrogen response elements) within the journal articles selected for the module. The use of a variety of CREATE tools will be used to explore both the commentary and review article in greater depth.
Applicable for Courses:
Educational Level:
Upper-levelRoadmap Objectives:
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- Article: Women, autoimmunity, and cancer: a dangerous liaison between estrogen and activation-induced deaminase? (2009) Robert M. Maul and Patricia J. Gearheart. Journal of Experimental Medicine. 206(1): 99-111
- Content area/major concepts: To call attention to the medical fact that women are more susceptible to autoimmune diseases and the new finding that the hormone estrogen may play an important regulatory role.
- Methods or technology used to obtain data:
- How the CREATE strategy was used:
- Biggest teaching challenge:
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- Article: Hormonal modulation of B cell development and repertoire selection. (2005) Grimaldi et al. Molecular Immunology 42:811-820
- Content area/major concepts: To provide background and clarify some of the complex topics (i.e. autoimmunity, SLE, B cell development, selection and activation, estrogen receptors and estrogen response elements) within the journal articles selected for the module.
- Methods or technology used to obtain data:
- How the CREATE strategy was used:
- Biggest teaching challenge:
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- Article: Overexpression of Activation-Induced Cytidine Deaminase in B Cells is Associated with Production of Highly Pathogenic Autoantibodies. (2007) Hsu et al. Journal of Immunology 178: 5357-5365
- Content area/major concepts: cell biology, B cell development and biology (maturation, activation, somatic hypermutation, receptor editing, class switch recombination), T cell biology (maturation, activation, regulation), molecular biology (mutation)
- Methods or technology used to obtain data: work with mice, work with murine and human cell lines and primary cells, generation of hybridomas, cloning and sequencing, RT-PCR, immunofluorescence, immunohistochemistry, FACS (fluorescence activated cell sorting), flow cytometry, stimulation and proliferation assays, construction of rAd (recombinant adenoviral) vectors, ELISA (enzyme linked immunosorbent assay), ELISPOT
- How the CREATE strategy was used:
- Biggest teaching challenge: jargon of immunology and molecular biology, complicated experiments and techniques, substantial number of concepts and experiments
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- Article: Estrogen directly activates AID transcription and function. (2009) Pauklin et al. (lab of S.K. Petersen-Mahrt) Journal of Experimental Medicine 206(1): 99-111
- Content area/major concepts: B cell development and biology (maturation, somatic hypermutation, class switch recombination), molecular biology (eukaryotic regulation of gene expression)
- Methods or technology used to obtain data: work with murine cell lines and primary cells, RT-PCR, qRT-PCR, promoter analysis, EMSA (Electromobility Shift Assay), ChIP (chromatin immunoprecipitation)
- How the CREATE strategy was used:
- Biggest teaching challenge: jargon of immunology and molecular biology, more complicated experiments and techniques, substantial number of concepts and experiments
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- Article: A role for the RNA pol II-associated PAF complex in AID-induced immune diversification. (2012) Willmann et al. (lab of S.K. Petersen-Mahrt) Journal of Experimental Medicine 209(11): 2099-2111
- Content area/major concepts: B cell development and biology (maturation, somatic hypermutation, class switch recombination), molecular biology (eukaryotic regulation of gene expression), cell biology
- Methods or technology used to obtain data: work with murine cell lines and primary cells, flow cytometry, real-time qRT-PCR, chromatin isolation, immunoprecipitation, mass spectrometry, western blotting, in vitro translation, shRNA knockdown, ChIP
- How the CREATE strategy was used:
- Biggest teaching challenge: jargon of immunology and molecular biology, even more complicated experiments and techniques, substantial number of concepts and experiments