The Zhu lab utilizes cutting edge tools to better understand the spatial-temporal relationship between the tissue-based immune response and HSV in humans.

We have developed novel techniques to define the mechanisms by which CD8+ TRM contain virus. The lab uses in situ immunofluorescence staining combined with laser capture microdissection (LCM) on patient biopsies to define immune control mechanisms, taking into consideration their anatomic site and native physiological state. We also use high-throughput sequencing of the T cell receptor (TCR) to determine the breadth and clonality of the TCR repertoire in locally affected skin/mucosa tissue and in blood. These tools help us understand how the quantity, quality and diversity of CD8+ TRM impacts antiviral activity in barrier tissues and disease outcomes in individuals with HSV-2.

Multi-parameter in situ staining & high-dimensional imaging analysis

To gain the most from limited human samples, we have continued to expand the capabilities of immunofluorescence staining so that we are able to obtain both qualitative and quantitative data.

Qdot conjugated pMHC multimers for in situ detection of antigen-specific T cells in tissue

In addition to assessing total T cell responses in the tissue, it is critical to also consider the antigen-specific response. We have developed a system to use Quantum dots (Qdots, fluorescent nanoscaled crystalline probes) conjugated to peptide–MHC multimers (Qdot multimers) in order to provide higher sensitivity and photostability than conventional tetramers for identifying antigen-specific T cells. Using this method we have been able to carefully track HSV-specific CD8 T cell localization and function in genital tissue.

Cell-Type-Specific Laser Capture Microdissection (LCM)

A primary advantage of using tissue sections with immunofluorescence imaging is the spatial information we are able to gather. LCM is a method of isolating individual cells from tissue sections after rapid staining and allows us to gain valuable transcriptome information in conjugation with cell location.

Transcriptional profiling

Using LCM and single cell RNA sequencing, we are able to bring together spatial orientation with T cell function and how these dynamic processes change over time.

Skin-on-chip & microfluidic platforms

The organ-on-chip field has grown from the need for alternatives to animal and human research for ethical reasons and in order to do larger scale studies that are also financially sustainable. We have pursued the development of a skin-on-chip model to study immune dynamics of HSV infection as well as to test novel anti-viral drugs.