Nebraska Center for Virology

3 ARTICLES PUBLISHED IN JoVE

Biology

A Technique to Simultaneously Visualize Virus-Specific CD8+ T Cells and Virus-Infected Cells In situ

Qingsheng Li¹, Pamela J. Skinner², Lijie Duan¹, Ashley T. Haase¹

¹Department of Microbiology, Medical School, University of Minnesota, ²Department of Veterinary and Biomedical Sciences, University of Minnesota

A technique combining in situ tetramer staining and in situ hybridization (ISTH) enables visualization, mapping and analysis of the spatial proximity of virus-specific CD8+ T cells to their virus-infected targets, and determination of the quantitative relationships between these immune effectors and targets to infection outcomes.

Immunology and Infection

In Situ Detection of Autoreactive CD4 T Cells in Brain and Heart Using Major Histocompatibility Complex Class II Dextramers

Chandirasegaran Massilamany¹, Arunakumar Gangaplara¹, Ting Jia¹, Christian Elowsky², Qingsheng Li³, You Zhou², Jay Reddy¹

¹School of Veterinary Medicine and Biomedical Sciences, University of Nebraska, Lincoln, ²Center for Biotechnology, University of Nebraska, Lincoln, ³Nebraska Center for Virology and School of Biological Sciences, University of Nebraska, Lincoln

The protocol to detect self-reactive CD4 T cells in brain and heart by direct staining with major histocompatibility complex class II dextramers has been described in this report. For comprehensive analysis, a reliable method to enumerate the frequencies of antigen-specific CD4⁺ T cells in situ is also devised.

Biochemistry

A Double Humanized BLT-mice Model Featuring a Stable Human-Like Gut Microbiome and Human Immune System

Lance Daharsh^1,2, Jianshui Zhang^1,2, Amanda Ramer-Tait³, Qingsheng Li^1,2

¹Nebraska Center for Virology, ²School of Biological Sciences, University of Nebraska-Lincoln, ³Department of Food Science and Technology, University of Nebraska-Lincoln

We describe a novel method for generating double humanized BLT-mice that feature a functional human immune system and a stable engrafted human-like gut microbiome. This protocol can be followed without the need for germ-free mice or gnotobiotic facilities.