Research Project

Saurabh Dixit

301-***-**** *******.*****@***.********://www.linkedin.com/in/saurabh-dixit-phd-44184616/

Immunology Expert

Experienced scientist in discovery research and transitional biology of infectious disease, vaccine development, immunology, inflammation, host pathogen interactions.

I have led projects in collaborative environment to develop cell based assay optimization and validation, gene silencing, animal model development and Immunoassays.

Proficient in use of FACS analysis software (FloJo and BD Diva)

Education:

Year

Degree

School

March 2007

Ph. D. (Life Science)

Central Drug Research Institute (CDRI), Lucknow, India

March 2000

Master (Microbiology)

Dr. Ram Manohar Lohia Avadh University, Faizabad, India

Research Skills:

Perform and design immunization and drug screening studies.

Measuring immune response using different immunological and molecular biological techniques and instruments, such as ELISA, Bioplex, Antibody ELISA from serum, Transfection, siRNA, Western blotting, Flow cytometry, Southern blotting and Immuno-fluorescent microscopy.

Isolation and culture of primary cells and cell lines. T cell purification from spleen and nodes.

Thymidine and CFSE based T cell proliferation assay.

Isolate and differentiate bone marrow cells in to bone marrow derived macrophage and dendritic cells from mice.

3D cell culture on biodegradable scaffold for skin culture using mouse COCA fibroblast, human keritanocytes, fibroblast and KYOU induced pluripotent stem cells.

Extensive experience working on mouse model and nonhuman primate models. Animal model development, necropsy of small animal and transgenic parasite.

Protein expression and purification. Express and purify chlamydia outer membrane peptide M278 and MOMP using Ni-NTA column

Nucleic acid extraction (DNA and RNA) from primary cells, cell lines, blood and tissue. cDNA synthesis, Primer probe design, one step and 2 step qPCR, RT-PCR and PCR array

Maintaining animal facility knowledge of regulation and disposal of biohazard waste.

GLP and Laboratory management.

Grants writing, SOPs (experience (live link) online laboratory protocol management system) and manuscript writing

Data analysis PRISM, EXCEL, Word and Power point presentation.

Standardized all the protocols and wrote SOPs (standard operating procedure) with help of live link.

Trained for working in high safety area BSL2-3 and regulated biological materials.

Professional appointments

National Institute of Environmental Health Science NIEHS/ NIH, RTP, NC 2016 -Present

Scientist/Contractor

Project focus: Effect of Human COX-2 Single Nucleotide Polymorphisms on T cells Differentiation

oNaïve T cells purified from whole blood were cultured and differentiated in to different T cell subset (TH1, TH2, TH9, TH17, TH22 and Treg).

oMy task is to analyze complex FACS data collected form peripheral blood and cultured T cells from all 125 patients belonging to three SNP groups and extract mRNA from T cells.

oQuantify effector cytokine (IFN-γ, IL4, IL9, IL17A, IL22 and transcription factor (FOXP3) by FACS (BD Diva software) and qPCR.

oInterpret data how single nucleotide polymorphism in human COX-2 effecting T cell differentiation.

Alabama State University, Center for Nanobiotechnology Research, Montgomery, AL 2012 -2016

Postdoctoral Associate

Project focus: Evaluation of Nano encapsulate Outer membrane protein of Chlamydia muridarum as vaccine and mechanism of antigen presentation by dendritic cells

oPerformed immunogenicity testing in mouse model by challenging mouse with EBs.

oMeasuring cell mediated and humoral responses (determined concentration of coating antigen and dilution of serum by checker board) by ELISA.

oDCs serve as the main protagonist of T-cell activation; as such we employed bone marrow-derived DCs (BMDCs) as APCs to assess their interaction with nano-vaccines for receptor-mediated recognition, uptake, processing, and T-cell activation to produce Th1 protective immune responses.

oCell surface activation markers including MHC class I/II molecules, CD11b/CD205 for regulation of Th1/Th2 responses, and CD80/CD86/CD40 co-stimulatory/maturation markers were evaluated by (FACS).

oPrimed DCs presented to CD4+ and CD8+ T-cells were evaluated for lymphoproliferation (CFSE), quantifying Th1/Th2/TH17 cytokines, and memory T-cell phenotypes.

oStaining of costimulatory molecules CD80, 40 and 86, surface marker MHC I and MHCII,CD3, CD4 and CD8 intracellular cytokines and memory T cells, effector T cells and naïve T cells by probing for CD62L and CD44 surface receptors for memory and naïve T cell markers. Data collected on BD Canto and LSR II and analyzed by (Flow Jo).

National Institute of Allergy and Infectious diseases NIH/NIAID, Rockville, MD 2008 -2012

Visiting Fellow

Project focus: Development of nonhuman primate model for malaria CSP vaccine testing/and new drugs

oI have generated transgenic P. knowlesi (PkPfCSP) in which P. knowlesi CSP (PkCSP) gene was replaced by PfCSP (P. falciparumCSP) by homologues integration. Schizont stage of malarial parasites were purified from blood using magnetic column. Transfected with gene of interest by electroporation.

oTransfected parasites were replicated in rhesus host under drug selection.

oBlood stages of parasites were checked for integrated insert by PCR and Southern blotting.

oWe evaluated transmission blocking activities by membrane feeding assay of anti-Pfs25 human plasma samples from the Pfs25/ISA51 Phase I trial, against P. falciparum infectious stages from patients in two different geographical regions of the world, Thailand and Burkina Faso.

oDNA isolated from human blood was genotyped for Pfs25 gene seven microsatellites. The results indicate that despite different genetic backgrounds among parasite isolates Pfs25 is highly conserved.

Tulane Primate Research Center, Covington, LA 2007 -2008

Research Specialist

Project focus: Immune signaling pathways regulated by Borrelia burgdorferi (agent of Lyme disease)

To determine how Lyme bacteria, evade host innate immune system by regulating inflammatory response. We looked at parasite sensing receptors and cellular pathway involved in regulation of inflammation with main focus on TLRs and SOCS.

oUsed siRNA and electroporation technique to knock down TLR’s, MyD88 and SOCS in mouse and human macrophage cell lines.

oQuantified knock down efficiency using lipid reagents for TLR2, SOCS’s and MyD88 by FACS and qPCR

oDifferent cytokine mediators and transcription factors effected by knock down by using biochemical and molecular technique (ELISA, Western Blotting, FACS and qPCR).

Research Publication: N= 24 Publications, Kindly check google scholar citation link

https://scholar.google.com/citations?user=E79zep0AAAAJ&hl=en

Patent

1.Indian patent No. IN201000774-I1; Title: Process for preparation of bio-active sub-fraction from flowers of Simarouba glauca active against filariasis.

Workshops / Training Courses Attended:

June 13th- June 14th 2017

Attended two-day workshop by Duke University Department of Office of Regulatory Affairs and Quality (ORAQ) Investigational New Drug (IND) and Investigational Device Exemption (IDE).

July 10th - August 21st 2017

Attended six-day annual training program by The Duke Office of Regulatory Affairs and Quality (ORAQ) for IND and IDE

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