Sushmita Kumari

PEDV neutralization assay development & HEV culture

• Validating neutralization assays to quantify Porcine Epidemic Diarrhea Virus (PEDV) neutralizing antibodies in serum from vaccinated animals to support vaccine evaluation.
• Establishing culture systems for hepatitis E virus (HEV) infectivity studies in human and swine-derived cell lines and organoids to investigate HEV infection mechanisms in intestinal cells and effect of bile enzymes on the infectivity.

Influenza Vaccine Development in Mouse, Chicken, and Pig Models

• Optimized lipid nanoparticle (LNP) formulations for antigenic DNA encapsulation; characterized encapsulation efficiency, stability, and transfection efficacy in host-specific cell lines.
• Assessed post-vaccination immune responses in animals, demonstrating strong antibody production, T cell activation, and protection against influenza virus in controlled laboratory studies.
• Collaborated with Dr. Hinh Li (University of Minnesota) to evaluate a recombinant viral vector-based influenza vaccine (pichinde virus), which showed robust immune activation and sterile protection in vivo.

Monoclonal Antibody Production and B Cell Engineering

• Generated and maintained multiple monoclonal antibody clones from antigen-stimulated murine splenocytes using standard hybridoma techniques.
• Adapted protocols for swine hybridomas, successfully creating and optimizing porcine monoclonal antibody clones.
• Labeled antigens with fluorophores for flow cytometric sorting of antigen-specific B cells. Isolated single B cells post-vaccination, optimized in vitro proliferation, and recovered IgG genes for future cloning and recombinant antibody production.

Primary Cell Culture and Tissue-Derived Isolation

• Established and maintained primary cultures of: Mouse bone marrow-derived macrophages Swine PBMC-derived macrophages Swine tracheal, lung, and blood-derived fibroblasts Chicken embryo fibroblasts

Collaborative Projects

• Mentored colleagues in immunological techniques and supported manuscript preparation for publication.
• Assisted in the isolation of swine peritoneal and alveolar macrophages; guided mouse macrophage isolation techniques. Enabled successful evaluation of PRRSV susceptibility in swine macrophages, a population previously thought to be resistant.

Innate and Adaptive Immune Mechanisms in IBD and Colon Biology

• Investigated target molecule expression across immune and tissue-resident cells in mouse (bone marrow, macrophages, dendritic cells, splenocytes, MLN, colon explants) and human (PBMCs, monocytes, whole blood).
• Assessed pathway activation by stimulating cells with LPS, LTA, MDP, nigericin, ATP, TNF-α, and DSS to understand inflammatory signaling relevant to IBD pathology.

Immunological Assay Development and Drug Screening

• Designed and optimized functional in vitro assays in murine thioglycolate-elicited peritoneal macrophages and bone marrow-derived dendritic cells.
• Screened compound libraries targeting immune pathways; quantified cytokine and chemokine suppression (e.g., IL-6, IL-8, IL-12p40, IFN-γ, IL-1β, TNF-α, GLP-1, PGE2) using ELISA.