Cellular immunology: Multicolor Flow cytometry, cell sorting, flow jo analysis. Intracellular cytokine and Cell surface Staining (ICS). PBMCs Isolation. Tissue cells isolation from muscles, liver, lung, spleen and stem cells from bone marrow (BM), Cell based ELISA like EliSpot .
Humoral and viral Immunology: Antibody purifications and quantification, ELISA (RIA), Western blot, Virus TCID50 detection, viral neutralization technique.
Genetics: Gene cloning, vector construction, qPCR, Cell transduction and transfection with viral vectors and DNA.
Protein assay: Recombinant protein in bacterial and mammalian cells system; Isolated and purified proteins, function assay; Immunoassay (IHC, ELISA, Western blot etc.).
Tissue and cells culture: T and B cell line creation. McAbs hybridoma skill. Embryo culture, Stem cells (BMC, Mesenchymal cells like CD271 cell).
In vivo and In vitro bioluminescence skill.
- Full time
- Part time
- One time
Harvard Medical School/Forsyth Collaboration
January 2011 - January 2011
Projects: Conducted researches on bacterial vaccine preclinical trials. 1). Created DNA, Adenovirus vector based vaccine candidates. 2). Characterized the role of cytokine responses by intracellular staining in different tissues. 3). Explored the adjuvant effect in vaccine induced mucosal immunogenicity. 4). Defined the host cell in Mtb Infection by creating engineered Mtb-GFP strain.
Results shown that 1).TB antigen with a novel epitope elicited strong immunogenicity in CD8-T cell against TB challenge. This strong immune response mainly occurred in the lung tissue following TB infection and vaccine immunization. 2). Following vaccine inoculation, strong IL-17 response happened earlier in the lung and last a short while, however, IFN-g response peaked relatively later compared to IL-17 response but last longer in the lung tissue. 3). Natural Treg peaked parallelly with the Th17 response in lung tissue. 4). By administration of the purified TB protein antigen plus CTB adjuvant intranasally, an enhanced CD4 and CD8 IL-17 expression in both the lung and spleen, particularly in the lung CD8-T cells.
5). Used genetic recombined technique, we created the GFP-Mtb strain, inoculated i.v in mice, isolated cells from tissues in the lung, spleen and bone marrow to detect the distribution of the infected cells. Two biomarkers were used, CD11b for monocytes and CD271 the stem cell. During the latent stage of Mtb infection (12 weeks following inoculation), two populations, CD11b and CD271, still being detected GFP-positive, indicated a novel finding for bacterial latent reservoir.
Skills: Mice handling and inoculation with i.v and i.m. DNA and Adv vaccine construction and cell transfection and transduction. DNA subclone and purification. In vitro protein expression, purification and quantification. Western blot. Tissue (lung, spleen and bone marrow) cell isolation. Antibody ELISA titration.
Senior Research Fellow at MGH
Harvard Medical School
January 2008 - January 2010
Projects and Results: 1). Responsible for the new research lab set-up and management. Created SOPs used for cellular immune response study including human PBMCs and liver T cells isolations, freezing, thawing, human T cell surface markers immune staining and cytokine intracellular staining. New lab member orientation. 2). Conducted study on human T-cell inhibitory receptors (PD-1, Tim-3, Lag-3, 2B4 and CD160) to understand the relevance of HCV persistence and evidenced that HCV persistence was correlated with increased PD-1, CD160, Lag-3 and decreased 2B4 expression on human CD8-T cell. 3). Defined the role of CD161-T cell population, and found CD161 was related HCV persistence by expression IL-17 both in CD4- and CD8-T cells in the early stage of HCV infection.
Skills: Human blood PBMCs and liver infiltrated T cell isolation, freezing, thawing. Cell culture including created the CD4- and CD8-T cell lines. Multicolor flow cytometry, flow jo analysis. Cytokine intracellular stainning. Cell based ELISA such as EliSpot.
BIDMC, Harvard Medical School
January 2005 - January 2008
Project and Results: Focused on cellular immunology and vaccine immunology.
1. Defined the adjuvant plasmid GM-CSF to enhance the HIV DNA vaccine immunogenicity. Constructed HIV gp120 plasmid, i.m inoculation mice to understand local innate immune response how correlated the systemic immune response, and found adjuvant mediated local muscle cytokine response, particularly type I interferon expression and macrophage migration can lead to the change of adaptive immune responses induced by HIV DNA vaccination.
2. Conducted the research on the immunoregulation effect for nTreg in vaccine immunized animals. Balb/c mice pre-depleted natural Treg by inoculation neutralizing anti-CD25 antibody, 3 days later following nTreg depletion. Mice were inoculated with HIV plasmid gp120, by detection of the binding of HIV gp120-specificific tetramer on CD8-T cell. Enhanced immunogenicity both from primary and memory response, ICS results showed increased polyfunctional IFN-g and IL-2 response from both CD4- and CD8-T cells.
Skills: Gene cloning, subcloning, vector construction, mice immunization i.m. eye retrobleeding, PBMCs isolation, muscle immune cell isolation, muscle homogenization and cytokine quantification. FACS and in vivo, in vitro bioluminescence measurement. ELISA Antibody subtypes titration, EliSpot.
Qualifications & Certifications
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