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Pennsylvania State University College of Medicine

9 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Directed Differentiation of Induced Pluripotent Stem Cells towards T Lymphocytes

Fengyang Lei¹, Rizwanul Haque¹, Xiaofang Xiong¹, Jianxun Song¹

¹Department of Microbiology and Immunology, Pennsylvania State University College of Medicine

Generation of T lymphocytes from induced pluripotent stem (iPS) cells gives an alternative approach of using embryonic stem cells for T cell-based immunotherapy. The method shows that by utilizing either in vitro or in vivo induction system, iPS cells are able to differentiate into both conventional and antigen-specific T lymphocytes.

Immunology and Infection

Accelerated Type 1 Diabetes Induction in Mice by Adoptive Transfer of Diabetogenic CD4+ T Cells

Gregory Berry¹, Hanspeter Waldner¹

¹Department of Microbiology & Immunology, Pennsylvania State University College of Medicine

We provide a reproducible method to induce type 1 diabetes (T1D) in mice within two weeks by the adoptive transfer of islet antigen-specific, primary CD4+ T cells.

Immunology and Infection

Development of Stem Cell-derived Antigen-specific Regulatory T Cells Against Autoimmunity

Mohammad Haque¹, Kristin Fino¹, Praneet Sandhu¹, Jianxun Song¹

¹Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine

We present here a method to develop functional antigen (Ag)-specific regulatory T cells (Tregs) from induced pluripotent stem cells (iPSCs) for immunotherapy of autoimmune arthritis in a murine model.

Neuroscience

A Free-breathing fMRI Method to Study Human Olfactory Function

Jianli Wang¹, Sebastian Rupprecht¹, Xiaoyu Sun¹, Diana Freiberg¹, Courtney Crowell¹, Emma Cartisano¹, Megha Vasavada¹, Qing X. Yang^1,2

¹Center for NMR Research, Department of Radiology, Pennsylvania State University College of Medicine, ²Department of Neurosurgery, Pennsylvania State University College of Medicine

We present the technical challenges and solutions for obtaining reliable functional magnetic resonance imaging (fMRI) data from the human central olfactory system. This includes special considerations in olfactory fMRI paradigm design, descriptions of fMRI data acquisition with an MRI-compatible olfactometer, odorant selection, and a special software tool for data post-processing.

Immunology and Infection

Lung microRNA Profiling Across the Estrous Cycle in Ozone-exposed Mice

Nathalie Fuentes¹, Patricia Silveyra^1,2

¹Pulmonary, Immunology and Physiology Laboratory, Department of Pediatrics, Pennsylvania State University College of Medicine, ²Department of Biochemistry and Molecular Biology, Pennsylvania State University College of Medicine

Here we describe a method to assess lung expression of miRNAs that are predicted to regulate inflammatory genes using mice exposed to ozone or filtered air at different stages of the estrous cycle.

Genetics

HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries

Huacheng Luo¹, Amin Sobh², Christopher D. Vulpe², Edmond Brewer¹, Sinisa Dovat¹, Yi Qiu³, Suming Huang¹

¹Department of Pediatrics, Pennsylvania State University College of Medicine, ²Department of Physiological Sciences, University of Florida, ³Department of Anatomy and Cell Biology, University of Florida

A CRISPR/sgRNA library has been applied to interrogating protein-coding genes.However, the feasibility of a sgRNA library to uncover the function of a CTCF boundary in gene regulation remains unexplored. Here, we describe a HOX loci specific sgRNA library to elucidate the function of CTCF boundaries in HOX loci.

Immunology and Infection

Stem Cell-Derived Viral Ag-Specific T Lymphocytes Suppress HBV Replication in Mice

Xiaofang Xiong¹, Fengyang Lei², Mohammad Haque¹, Jianxun Song¹

¹Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, ²Department of Ophthalmology, Harvard Medical School

Presented here is a protocol for the effective suppression of hepatitis B virus (HBV) replication in mice by utilizing adoptive cell transfer (ACT) of stem cell-derived viral antigen (Ag)-specific T lymphocytes. This procedure may be adapted for potential ACT-based immunotherapy of HBV infection.

A Xenograft Mouse Model to Assess Efficacy of Therapeutic Agents for Human Acute Leukemia

Chandrika Gowda^*1, Charyguly Annageldiyev^*2,3, Pavan Kumar Dhanyamraju¹, Morgann Klink¹, Sinisa Dovat¹, Mark Kester^4,5, Thomas P. Loughran, Jr^4,6, David Claxton^2,3, Arati Sharma^1,2,7

¹Department of Pediatrics, Pennsylvania State University College of Medicine, ²Penn State Hershey Cancer Institute, Pennsylvania State University College of Medicine, ³Departments of Medicine, Division of Hematology and Oncology, Pennsylvania State University College of Medicine, ⁴University of Virginia Cancer Center, ⁵nanoSTAR Institute, University of Virginia, ⁶Division of Hematology and Oncology, Department of Medicine, University of Virginia School of Medicine, ⁷Department of Pharmacology, Pennsylvania State University College of Medicine

Mouse (Mus Musculus) models are being widely used to develop xenografts using human leukemia cells. These models provide a comparable biological system to study drug efficacy, pharmacodynamics, and pharmacokinetics. Modeling acute myeloid leukemia in immunocompromised mice is described in detail using the U937 cell line xenograft as an example.

Biology

3D Culturing of Organoids from the Intestinal Villi Epithelium Undergoing Dedifferentiation

Christina Li¹, Jeel Shah¹, Kylee Wrath¹, Dahlia Matouba¹, Connor Mills¹, Kishore Punnath², Ansu Perekatt¹

¹Department of Biology and Chemical Biology, Stevens Institute of Technology, ²Department of Pharmacology, Pennsylvania State University College of Medicine

The procedure describes isolation of the villi from the mouse intestinal epithelium undergoing dedifferentiation to determine their organoid forming potential.