Michael A. Lynes, Ph.D- Director

michael.lynes@uconn.edu

My lab group brings a variety of backgrounds and expertise to the study of the immune system and the genetic, biochemical and cellular processes that serve to regulate normal and aberrant immune activities.  Our main biological research theme is the study of the role metallothionein (MT, an important cell stress protein) plays in immune regulation.  We have shown that MT can have dramatic influences on the progression of both innate and adaptive immunity.  These influences probably occur as a result of several different contributions that MT can make to mammalian physiology, but one of the more surprising recent findings from our lab group is the observation that extracellular MT can initiate chemotactic cell movement.  This suggests that the pool of MT that exists outside of cells may influence the trafficking patterns of leukocytes toward sites of cellular stress.  A logical consequence of this observation is that in cases where MT is synthesized as a consequence of stresses not associated with a beneficial immune response, the trafficking patterns of leukocytes may be altered in detrimental ways.  We have shown that manipulations of MT using genetic tools or antibody reagents can alter the immune capacity of the organism.  Our current studies include experiments to identify the targets for MT interactions with leukocytes, and investigations of the consequences of MT manipulation to the course of autoimmune and infectious disease.  Our collaborators on these studies include groups from the Wadsworth Center of the NY Department of Health, the Jackson Laboratory, and Ghent University (Belgium).
            We are also involved in the development of novel research tools.  In collaboration with the Knecht laboratory, we have developed an automated chemotactic assay that measures cell movement in real time.  Using this assay, one can more readily assess the chemotactic potential of new agents, and the therapeutic potential of new drugs in a more objective manner.  This technology is now commercially available from Applied Biophysics, Inc.
            Finally, we are engaged in collaborative studies with Ciencia, Inc.,  the NY Department of Health, the Center for Excellence in Vaccine Research (CEVR at UCONN), the Plum Island Animal Disease Center, and the University of Massachusetts Medical Center to develop Grating-coupled Surface Plasmon Resonance Imaging (GCSPRI) and Grating-coupled fluorescence plasmonics (GCFP)-based instruments for the detection and characterization of molecular and cellular targets.  The foundation GCSPRI platform developed by Ciencia is now commercially available from GE Healthcare.

Current projects ongoing in our lab:
                       

1. The molecular basis of metallothionein influences on  in vivo and in vitro cell movement.
2. GCSPRI and GCFP-based mapping of transcription factor activation
3. Metallothionein-mediated effects on macrophage function (Gregory Marusov, Cell Biology Ph.D. student)
4. Signal transduction cascades altered by metallothionein, and the potential consequences of these changes on transcription factor activation
5. GCSPRI and GCFP phenotypic and functional monitoring of CD4 and CD8 T cells in Type 1 diabetes.
6. GCSPRI and GCFP-based diagnostics for infectious disease
7. Metallothionein binding proteins
8. Biomarker signatures of biological, chemical and psychological stress.
9. GCFP-based characterization of molecular and cellular characteristics of rheumatoid arthritis
10. Pathogen biosensors

Greg Marusov, M.S.- Ph.D student

gregory.marusov@uconn.edu

My research focuses on the effects of metallothionein (MT) expression on macrophage function.  Preliminary experiments show that the innate immune response to Listeria infection is significantly altered in MT overexpressing and MT knockout mice, and macrophages play a critical role in this innate response.  I have been optimizing methods for the purification of primary peritoneal macrophages from mice for in vitro analysis.  I have also been designing custom immunoassays for capturing activated macrophages on a chip and analyzing their functional responses using GCSPRI.  This involves development of a custom SPR chip for the multiplexed analysis of cytokines, cell surface markers, and other biomarkers of cellular activation.  Future work will use flow cytometry to compare ROS and RNS production and intracellular levels of labile Zn2+ in congenic strains of mice that differ only in MT expression levels.

Jamie Rice, M.S.- Ph.D Student

jamesmrice1@gmail.com

My research is focused on the isolation and quantification of reactive T cells as a diagnostic method for the pre-symptomatic detection of Type I Diabetes. I am currently designing and optimizing an immunoassay using recombinant peptide/MHC complexes on GCSPRI and GCSPEF detection platforms to capture, quantify, and characterize reactive T cells from peripheral blood. Other research interests include extending the application of SPR based technology to isolate, activate, and characterize populations of cells in real time.

Kathryn Pietrosimone - Ph.D student

kathryn.pietrosimone@uconn.edu

My research focuses on the function of bacterial metallothionein (smtA) in response to immune cells. This research aims to uncover the role of smtA in bacterial infection, specifically when exposed to reactive oxygen
species (ROS) produced by immune cells to fight infection.
I am also using the GCSPRI technology to identify bacterial pathogens.
This involves identifying antibodies specific to particular pathogens, as
well as groups of bacteria, and capturing the pathogens with these
antibodies.

Clare Melchiorre - research technician

Phil Gorecki- University Scholar

I am currently working on my University Scholar project in Dr. Lynes’s lab, which integrates aspects of both immunology and economics.  My research in the lab focuses on the development of a set of diagnostic tools using grating coupled surface plasmon resonance imaging (GCSPRI) to test for early molecular indicators of agricultural disease in swine.  Through testing of inactivated prototype samples of infectious agents, I will determine how the presence of certain cytokines, antibodies, and other immune cells in the biosignature of a swine are indicative of certain classes of disease.  After determining how the presence of certain immune components in the biosignature such as cytokines IL-2, IL-4, IL-6, IFN-γ, and TNF-α correspond to certain disease states, samples from animals with an unknown health history could be evaluated for those same analytes in the biosignature to determine whether the sample came from an infected animal.  The aim is to be able to determine the characteristic biosignatures of several agricultural diseases affecting swine including foot and mouth disease virus (FMDV), classical swine fever virus (CSFV), and vesicular stomatitis virus (VSV).

The other portion of the University Scholar project will look at the economic significance of these tools in preventing the financial losses caused by outbreaks of agricultural disease.

Stephanie Davis - University Scholar

My research is focused on the bacterial homolog of the mammalian metallothionein, SmtA, and its function over the course of mammalian infection. I am currently exploring the role of SmtA in bacteria exposed to oxidative stress, specifically reactive oxygen species known to be released during the respiratory bursts of mammalian neutrophils and macrophges. These students my elucidate an important role for SmtA in bacterial pathogenesis and may be a valuable target for theurapeutic intervention.

Will Page - Post-Baccalaureate

william.d.page@uconn.edu

My present research is focused on the discovery and characterization of polypeptides that bind to metallothionein and UC1MT. These peptides are being selected for by means of a phage display kit. Briefly, this technique exposes a plate coated with MT or UC1MT to phage particles engineered to have a highly variable region, so that every possible permutation (of exactly seven amino acids) is expressed. After several round of panning and amplification, a small number of high-affinity interactions can be selected. An understanding of the chemical natures of these MT binding peptides could provide a insight into which specific cell receptors bind MT, as well as an approach for MT-dependent immunosuppressive therapies.

Chana Rich - University Scholar

My research focuses on the characteristic biomarker signatures produced
from a set of diverse stressors including psychological, biological and
chemical stress. A biomarker signature is a distinctive biological
indicator of a specific condition. I am using enzyme-linked immunosorbent
assay (ELISA) and grated-coupling surface plasmon resonance imaging
(GCSPRI) will be used to categorize these unique endogenous responses to
each form of stress. By identifying the different biomarker signatures
that are associated with each stressful condition, better diagnosis could
be made and the effects of different stresses on physiological processes
may be better predicted and controlled.

Chelsea Bleckwehl - Undergraduate Researcher

Monalissa Rodrigues - M.S student

Peter Reinhold - M.S. Student

Ryan Molony - University Scholar

My University Scholar Project deals with the investigation of
transcription factor biosignatures in the three strains of mice with
varying Metallothionein expression. I will be conducting experiments using
both GCSPR and SPEF technologies, making use of both antibodies and
consensus oligonucleotide sequences as bait molecules. I will additionally
expose cells to Cadmium or LPS to induce stress, potentially resulting in
the expression of more prominent transcription factor biosignatures.

Meaghan Roy-O'Reilly - Undergraduate Researcher

My research focuses on the lymphoproliferative effects of metallothionein
(MT). I am currently optimizing an assay that uses a novel tetrazolium
compound (MTS)to quantify cell proliferation. In the future, I will
attempt to block the interaction between MT and leukocytes that stimulates
this proliferation with various competitors including 2-mercaptoethanol,
pertussis toxin, cholera toxin, UC1MT and UC2 (anti-MT antibodies),
bacterial metallothionein (smtA) and MT binding peptide motifs. The
blockade patterns of these competitors will help me to elucidate the
mechanism by which MT binds to leukocyte plasma membranes and stimulates
lymphoproliferation.

Rachel Wood - Undergraduate

Priya Ranade - Undergraduate

Carol Norris, Ph.D- Facility Scientist

Flow Cytometry and Confocal Microscopy Facility of Uconn Biotech Center

carol.norris@uconn.edu

Krishna Patel- undergraduate researcher

Jacklyn Kubinski- undergraduate researcher

Doug Donaldson, B.S.

Xiuyun Yin, Ph.D- postdoctoral researcher

Jennifer Joukhadar- undergraduate researcher

Gabriel Pilar- affiliated researcher

"Sasha" Alexandra Isenberg- undergraduate researcher

Lorena Nunez- undergraduate researcher

Darryn Unfricht, Ph.D.

Emilia Falkowska - Doctoral Student at Yeshiva University

Kirsten Frostad

Michele Barber, Ph.D. - Assistant Professor of Biology and Chair of the Science Department at Norwalk Community College

Nacima Hadjout, Ph.D. - Postdoctoral Fellow (Canada)

Warren Brooks, Ph.D. - Associate Director of Medical Affairs at Regeneron Pharmaceuticals

Peter C. Lai

Jacqueline Samson Lagoy - Medical Student at UConn Medical Center

Sara Colpitts, Ph.D. - Postdoctoral Fellow Uconn Medical Center