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Research areas
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T cell receptor (TCR) and immunoglobulin (IG) repertoires in health and disease
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Autoimmunity
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Hematopoietic stem cell transplantation
Main objectives
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To develop new technologies for the efficient deep profiling of immune repertoires
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To understand better the ensemble function of the adaptive immune system
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To rationally improve therapies that modulate immune function
Content of research
Every individual’s adaptive immune system is composed of tens of millions or more different T and B lymphocyte clones, each of which respectively expresses a distinct T cell receptor or antibody molecule. This tremendous diversity allows the immune system to generate receptors for a broad spectrum of antigens and to thereby ensure an efficient, targeted immune response. After the immune system has eliminated pathogens or diseased tissues, expanded pools of antigen-specific cells can subsequently persist for many years as memory cells, and these may therefore be used to track an individual‘s history of infections, vaccinations, and possibly even autoimmune and anti-tumor responses. Next generation sequencing (NGS) techniques are capable of reading millions of short DNA fragments, and thus allow deep analysis of the diversity of antibodies and T cell receptors within an individual repertoire. My team employs NGS to study adaptive immunity and, in parallel, also works on advanced molecular technologies and specialized software solutions that collectively allow us to minimize quantitative biases, eliminate input bottlenecks and efficiently correct PCR and sequencing errors to ensure accurate and quantitative analysis of T cell receptor and antibody repertoires.
1. Ratiometric genetically encoded fluorescent sensors for measurement of intracellular pH values
Supervisor: Dmitriy Chudakov, Ph.D., DSc.
Annotation:
Intracellular pH influences a lot of processes in cellular cytoplasm and intracellular compartments. [1-10]. Fluorescent proteins have many advantages compared to chemical fluorescent dyes in living cell imaging, including less invasive measurement and possibility of tethering the reporter to any cellular organelle or compartment. Sensitivity to pH has been used to develop genetically engineer encodable pH indicators [11]. Genetically encoded fluorescent pH reporters enable to measure spatial and temporal changes of pH in dynamics. To date, in spite of development of different geneticaly encoded fluorescent-based pH indicators, both single-color and ratiometric pH sensors have been ultimately used rather to detect the direction of pH changes, than to measure exact pH value in live cell microscopy. The project will be dedicated to creation of ratiometric pH sensors based on pH-sensitive GFP mutants and highly pH-independent red fluorescent protein which can be used for precise imaging of pH value for a great variety of biological events and signals.
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