PENN STATE CMIID LABS
Dr. K Sandeep Prabhu Lab
Research description
Selenium is a unique micronutrient present in a
variety of foods, which is charged onto a selenocysteine-specific tRNAsec
(Trsp) and incorporated into 25
selenoproteins with the aid of various proteins, such as SECIS binding
protein-2 (Sbp2). Only a few of these
25 selenoproteins have been functionally characterized with the majority
exhibiting redox functionality. Other functions include, but are not limited
to, thyroid metabolism, Se transport and storage, removal of damaging
peroxides, and protein folding. Studies in our laboratory are focused around
various cellular and molecular mechanisms by which selenium acting through
selenoproteins, and other products of natural origin, alter the host response
and immune function when responding to oxidative stress caused by environmental
toxicants and pathogens.
Current projects in the
lab:
1.
Modulation
of inflammatory signaling pathways by selenium
2.
Modulation
of steady state and stress hematopoiesis by selenium
3.
Effect
of endogenous and exogenous fatty acid metabolites on leukemic stem cells
4.
Epigenetic
regulation by antioxidant
Techniques
- ·
Flow cytometry (intra- and extracellular) and fluorescence-activated cell sorting
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Western blot
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Real-time PCR
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Semi-quantitative PCR
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Electrospray
ionization mass spectrometry (small molecules/peptides: 80-1500 daltons)
- ·
High performance liquid chromatography (HPLC)
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Cell culturing (bone marrow derived macrophages,
various stem cell populations and primary cell lines)
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Transfection/transduction (plasmids and sh- and
siRNA knockdown)
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Bone marrow transplantation
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Collecting various mouse tissues
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Mouse breeding
Animal Strains (Mice)
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C57Bl6 – Taconic
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GPX1 whole body KO – C57Bl6
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TRSP homozygous floxed – C57Bl6
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TRSP macrophage specific KO (homo TRSP
floxed/hetero LysM Cre+) – C57Bl6
- ·
SBP2 homozygous floxed – C57Bl6
- ·
SBP2 macrophage specific KO (homo SBP2
floxed/homo or hetero LysM Cre+) – C57Bl6
- ·
TRSP hematopoietic stem cell specific KO (homo
TRSP floxed/hetero VAV Cre+) – C57Bl6
Dr. Margherita Cantorna
Research Description
It has long been known that vitamins are important in a
variety of physiological processes. The
Cantorna lab studies the how essential micronutrients regulate immune responses
during homeostasis, pregnancy, infection and autoimmunity, mainly focusing on
mucosal immunology. We examine how
vitamin D and the vitamin D receptor (VDR) modulate antigen specific B cell
responses, regulate intestinal immunity during inflammation and maintain maternal
tolerance during pregnancy. Another part
of the lab studies how other micronutrients (vitamin A and selenium) regulate
mucosal and systemic immunity.
Techniques
- ·
Flow cytometry (surface and intra-cellular)
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Cell
sorting
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Cell culture
- ·
Models of intestinal inflammation
o
DSS colits
o
Anti-CD40 colits
o
C.
rodentium infections
- ·
Experimental autoimmune encephalomyelitis (EAE –
mouse model for multiple sclerosis)
- ·
ELISA
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LC/MS
- ·
GC/MS
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RT-PCR
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Western blotting
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Mucosal cell isolations
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Timed pregnancies
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OVA immunizations
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Bone marrow transplantations
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Germ free mouse work
Animal strains (Mice)
- C57Bl6 – Jackson Labs
- Balb/c – Jackson Labs
- CD45.1 – C57Bl6
- Rag KO – C57Bl6
- Cyp27B1 KO – C57Bl6
- LCK-cre – C57Bl6
- LCK-cre TRSP– C57Bl6 KO
- LCK-cre
VDR KO – C57Bl6
- CD19 cre-VDR KO – C57Bl6
- VDR – IL10 double KO – C57Bl6
- Villin – cre – C57Bl6
- Villin cre-VDR KO – C57Bl6
- Dominant negative Retinoic Acid Receptor (dnRAR Floxed) – C57Bl6
- Villin cre- dnRAR – C57Bl6
Dr. Girish Kirimanjeswara
Lab
Research Description
Tularemia is a disease caused by
the facultative intracellular bacterium, Francisella
tularensis. The CDC classifies F.
tularensis as a tier one potential biothreat, because a miniscule number of
these bacteria can cause life-threatening disease in humans. The Kirimanjeswara
(Girish) lab is working towards advancing our understanding of tularemia by
studying the contributions of the host, pathogen, and environment to the
establishment, progression and outcome of infection. Part of the lab’s work
will be conducted at biosafety level (BSL)-3 following CDC certification of the
new Eva J. Pell Laboratory for Advanced Biological Research.
One focus of the lab involves
identifying and characterizing virulence factors that operate at various stages
of the pathogen’s intracellular lifecycle- i.e. factors that allow the
bacterium to attach to and enter host cells, survive within and escape from the
phagosome, replicate in they cytosol, or evade killing by intracellular innate
immune mechanisms. Other lab members are studying the relevance of the
micronutrient selenium to the host response to F. tularensis. The impact of selenium status is being evaluated at
the molecular, cellular and systemic levels, with an additional project
focusing specifically on the contribution of selenium to antigen presentation
by B cells. Another current project is addressing differences between the
immune responses raised against intradermal versus pulmonary F. tularensis challenge. Most studies
focus on pneumonic tularemia as this is most relevant to a potential bioterror
attack, however any approved vaccine is likely to be administered via
injection, and additionally the most prevalent route of natural exposure to F. tularensis is via skin-piercing
arthropod vectors.
Techniques
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Cloning (bacterial gene deletion,
complementation, overexpression, ectopic gene expression)
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PCR, RT-PCR and qPCR
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Flow cytometry (intracellular cytokine assays,
cell surface markers, endocytosis assays, calcium flux assays, antigen
presentation assays)
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Fluorescence microscopy
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Tissue culture (primary cells and cell lines)
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Intranasal, intradermal, intraperitoneal animal
infections
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Bacterial burden enumeration by organ
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Histopathology
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ELISA
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Bacterial culture
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In vitro
infection assays
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Western Blot
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Metabolomics
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In vitro
Antigen Presentation assay
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BSL-2 and BSL-3 containment
Animal Strains (Mice)
- ·
C57BL/6J
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Balb/c
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IL10eGFP (C57BL/6J background)
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IL1α KO(C57BL/6J background)
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IL1β KO(C57BL/6J background)
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IL1RKO(C57BL/6J background)
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TrspFlox/Flox C57BL/6J background)
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TrspM (C57BL/6J background – Trsp fl/fl
LysMWT/Cre)
- ·
CD19 cre (C57BL/6J)
Dr. Adam Glick
Research Description
The Glick Lab primarily studies the
signaling pathways and immune phenotypes associated with benign and malignant
skin cancer. Our lab examines signal transduction pathways that commonly cause
malignant phenotypes in many developing tumors, such as RAS and TGFb.
Three major avenues we study are: The roles of the adaptive immune system in
orchestrating an immune response to a developing skin tumor, the roles of ER
stress in malignant phenotypes of transformed keratinocytes, and the importance
of antigen-presenting cell function in the response to ultraviolet-induced skin
damage. Using a variety of techniques, we hope to elucidate the mechanisms that
lead to skin tumor progression, and identify potential therapeutic targets.
Techniques
- ·
Flow Cytometry
- ·
Immunohistochemistry
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Western Blotting
- ·
RT-qPCR
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shRNA knockdown
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Plasmid preparation
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Cell sorting
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Primary cell culture
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Fluorescence Microscopy
Animal Strains (Mice)
- ·
TetoRas- FVB/n
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Involucrin tTA- FVB/n
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K14 rTA- FVB/n
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Rag1-/- - FVB/n
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K5 rTA – FVB/n
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TetoRas- C57B6J
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Involucrin tTA- C57B6J
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K14 rTa- C57B6J
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CD11cDN – C57B6J
Dr. Troy Ott Lab
Research Description
Physiology of
Reproduction/Reproductive Immunology
Early embryonic mortality in livestock occurs at a rate of
30-40 percent in cattle, sheep and swine, and exceeds 50 percent in horses. It
is also a significant problem in human reproductive medicine.
The primary focus of research in the lab is the molecular and
immunological mechanisms regulating fertility and infertility in domestic farm
animals. We hypothesize that the function of endometrial immune cells is
altered during maternal recognition of pregnancy to facilitate survival of
embryo and development of placenta. Cross talk between the developing conceptus
and the mother is essential to promote tolerance to paternal alloantigens and
to bring about uterine remodeling.
We currently focus on studying these changes in normal early
pregnancy in order to understand the reasons for growing infertility in cattle.
Unraveling these mechanisms would help in devising therapeutic measures to
increase fertility and thereby economically benefit the dairy industry.
Techniques
- ·
Real time PCR
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Immunofluorescence- Paraffin, Frozen
- ·
Flow cytometry
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Western Blot
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ELISA
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Large animal –Blood collection
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Uterine immune cell isolation
Animal Strains
- ·
Large animal model- Holstein dairy heifers and
cows
Dr. Joy Pate Lab
Research description
The long term goal of this research
is to increase our understanding of the function of the bovine corpus luteum to
enhance reproductive efficiency of dairy cattle. The decline in reproductive
efficiency of dairy cows is well-documented and results in substantial economic
loss within the agricultural industry each year. Progesterone production by the
corpus luteum is essential for the establishment of pregnancy in all mammals.
Therefore, understanding the mechanisms that regulate luteal development and
survival will lead to enhanced reproductive efficiency and lower the costs of
food production.
A focus of our research has been to
understand the regulatory interactions that exist between parenchymal
(steroidogenic and endothelial) cells and immune cells that result in normal
luteal function. Compelling evidence exists that immune cells and the cytokines
they produce regulate luteal function and facilitate luteolysis. However,
immune cells, and all components necessary to activate those cells, are present
within luteal tissue long before the onset of luteolysis. Specifically our
research is focused on luteal cell secreted extracellular vesicles (exosomes
and microvesicles), micro RNA (miRNA), and macrophages that regulate normal
luteal function.
Techniques
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Cell culture
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Enzymatic tissue dissociation
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Western blots
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Flow cytometry (Guava, Flowsight…etc)
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Immunohistochemistry
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PCR/RT-qPCR
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Electrophoresis
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Corpora lutea collection via culpotomy
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Cell sorting (autoMACS)
Animal Strains
- ·
Large animal model- Holstein dairy heifers and
cows
Dr. Anthony Schmitt Lab
Research description
Our lab
works on the assembly and budding of paramyxoviruses-enveloped, non-segmented,
negative sense single stranded RNA viruses.
Viruses from this group include many important human and animal
pathogens such as mumps virus, measles virus, Newcastle disease virus, and
Henipavirus. We are studying assembly
and budding using a prototypic model, parainflusnza virus 5 (PIV5) that is
non-pathogenic in humans. We can also
study assembly and budding using virus-like particles (VLPs) that lack
infection genome. VLPs are used in our
lab to safely study mechanisms of pathogenic viruses like mumps virus, Nipah
virus and human parainfluenza virus 2.
VLPs can be prepared by expression a subset of viral proteins necessary
for particle formation. Using this and
other techniques, we study viral protein-protein interactions that are
necessary for particle formation. Since
viruses hijack a variety of host proteins during their life cycle, we are also
looking into viral-host protein interactions.
Detailed information about these mechanisms can be of potential use in
creating antiviral or other inhibitors of virus replications.
Techniques
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Fluorescence microscopy
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Virus-like particle production
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Transfection of mammalian cells
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Ultracentrifugation
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Generation of recombinant viruses
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Cloning
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Plaque assay
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35S labeling of proteins for use in
pulse chase and other experiments
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Co-immunopreciptation assays
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Affinity purification of proteins by FPLC
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Western Blots
Cell Lines
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HEK 293T
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Vero
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BHK
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MDBK
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MDCK
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A549
