Neha M.

Fremont, California, United States Contact Info
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With over 6 years of experience in scientific/clinical research and medical writing, I am…

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  • Illumina

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Publications

  • Characterization of an umbilical cord blood sourced product suitable for allogeneic applications

    Mukta S Sane, Huiyuan Tang, Neha Misra, Xinzhu Pu, Sara Malara, Christopher D Jones, Soumyajit Banerjee Mustafi

    Aim: Umbilical cord blood (UCB) sourced allografts are promising interventions for tissue regeneration. As applications of these allografts and regulations governing them continue to evolve, we were prompted to identify parameters determining their quality, safety and regenerative potential. Materials & methods: Flow-cytometry, mass-spectrometry, protein multiplexing, nanoparticle tracking analysis and standard biological techniques were employed. Results: Quality attributes of a uniquely…

    Aim: Umbilical cord blood (UCB) sourced allografts are promising interventions for tissue regeneration. As applications of these allografts and regulations governing them continue to evolve, we were prompted to identify parameters determining their quality, safety and regenerative potential. Materials & methods: Flow-cytometry, mass-spectrometry, protein multiplexing, nanoparticle tracking analysis and standard biological techniques were employed. Results: Quality attributes of a uniquely processed UCB-allograft (UCBr) were enumerated based on identity (cell viability, immunophenotyping, proteomic profiling, and quantification of relevant cytokines); safety (bioburden and microbiological screening), purity (endotoxin levels) and potency (effect of UCBr on chondrocytes and mesenchymal stem cells derived exosomes). These attributes were stable up to 24 months in cryopreserved UCBr. Conclusion: We identified a comprehensive panel of tests to establish the clinical efficacy and quality control attributes of a UCB-sourced allograft.

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  • Cytokines in umbilical cord blood-derived cellular product: a mechanistic insight into bone repair

    Regenerative medicine

    Aim: Umbilical cord blood (UCB) finds frequent applications in regenerative medicine. We evaluated the role of cytokines present in a uniquely processed, UCB-derived cellular allograft product (UCBp). Materials & methods: Luminex multiplex assay and standard cell biology methods were employed. Results: Study with allografts from 33 donors identified 44 quantifiable cytokines in the UCBp derived conditioned media (CM). The UCBp-CM elevated proliferation and migration rates of mesenchymal stem…

    Aim: Umbilical cord blood (UCB) finds frequent applications in regenerative medicine. We evaluated the role of cytokines present in a uniquely processed, UCB-derived cellular allograft product (UCBp). Materials & methods: Luminex multiplex assay and standard cell biology methods were employed. Results: Study with allografts from 33 donors identified 44 quantifiable cytokines in the UCBp derived conditioned media (CM). The UCBp-CM elevated proliferation and migration rates of mesenchymal stem cells (MSCs) and bone marrow stromal cells. Moreover, UCBp-CM induced secretion of VEGF-A and osteoprotegerin, which promoted angiogenesis of endothelial cells and positively influenced the osteogenic differentiation of MSCs, respectively. Conclusion: Cytokines in UCBp stimulate cellular processes important for bone regeneration, making UCBp an excellent candidate for potential applications in orthopedic procedures like bone non-union and spinal fusion.

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  • Biochemical characterization of pure dehydrated binate amniotic membrane: role of cytokines in the spotlight.

    Abstract AIM: Placental allografts used for tissue regeneration differ in membrane compositions and processing techniques. A uniquely folded dehydrated binate amniotic membrane (DBAM) was biochemically characterized to evaluate its potential role in wound healing. METHODS: Histology, Luminex-based immunoassay and standard in vitro cell biology techniques were employed.
    RESULTS: Histological staining confirmed that the DBAM was chorion free with epithelial cell layer of the respective amnion…

    Abstract AIM: Placental allografts used for tissue regeneration differ in membrane compositions and processing techniques. A uniquely folded dehydrated binate amniotic membrane (DBAM) was biochemically characterized to evaluate its potential role in wound healing. METHODS: Histology, Luminex-based immunoassay and standard in vitro cell biology techniques were employed.
    RESULTS: Histological staining confirmed that the DBAM was chorion free with epithelial cell layer of the respective amnion membranes facing outward. DBAM had quantifiable levels of relevant cytokines that induced proliferation and migration while bolstering secretory activity of the cells. DBAM retained biological efficacy at a broad range of temperatures. CONCLUSION: Cytokines in DBAM stimulate bone marrow stromal and stem cells that may lead to tissue regeneration and wound healing in a clinical setup.

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  • Immunogenicity of a Staphylococcus aureus-cholera toxin A2/B vaccine for bovine mastitis.

    Abstract
    Staphylococcus aureus causes a chronic, contagious disease of the udder, or mastitis, in dairy cows. This infection is often refractory to antibiotic treatment, and has a significant economic impact on milk production worldwide. An effective vaccine to prevent S. aureus mastitis would improve animal health, reduce antibiotic dependence and inform human vaccine approaches. The iron-regulated surface determinant A (IsdA) and clumping factor A (ClfA) are conserved S. aureus…

    Abstract
    Staphylococcus aureus causes a chronic, contagious disease of the udder, or mastitis, in dairy cows. This infection is often refractory to antibiotic treatment, and has a significant economic impact on milk production worldwide. An effective vaccine to prevent S. aureus mastitis would improve animal health, reduce antibiotic dependence and inform human vaccine approaches. The iron-regulated surface determinant A (IsdA) and clumping factor A (ClfA) are conserved S. aureus extracellular-matrix adhesins and target vaccine antigens. Here we report the results of two bovine immunogenicity trials using purified IsdA and ClfA-cholera toxin A2/B chimeras (IsdA-CTA2/B and ClfA-CTA2/B). Cows were intranasally inoculated with IsdA-CTA2/B + ClfA-CTA2/B at dry off and followed for 70 days. Trial 1 utilized three groups with one or two booster doses at a total concentration of 600 or 900 μg. Trial 2 utilized two groups with one booster at a total concentration of 1200 μg. Humoral immune responses in serum and milk were examined by ELISA. Responses in serum were significant between groups and provide evidence of antigen-specific IgG induction after vaccination in both trials. Cellular proliferation was detected by flow cytometry using antigen-stimulated PBMCs from day 60 of Trial 2 and revealed an increase in CD4+ T cells from vaccinated cows. IsdA and ClfA stimulation induced IL-4 expression, but not IFN-γ or IL-17, in PBMCs from day 60 as determined by cytokine expression analysis. Opsonophagocytosis of S. aureus confirmed the functional in vitro activity of anti-IsdA antibodies from Trial 2 serum and milk. The vaccine was well tolerated and safe, and results support the potential of mucosally-delivered CTA2/B chimeras to protect cows from mastitis caused by S. aureus.

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  • Immunoproteomics to identify Staphylococcus aureus antigens expressed in bovine milk during mastitis.

    Abstract
    Staphylococcus aureus is an opportunistic pathogen affecting both human and animal species. An effective vaccine to prevent S. aureus bovine disease and transmission would have positive effects on animal well-being, food production, and human health. The objective of this study was to identify multiple antigens that are immunoreactive during udder colonization and disease for exploration as vaccine antigens to prevent bovine mastitis. Staphylococcus aureus produces several cell…

    Abstract
    Staphylococcus aureus is an opportunistic pathogen affecting both human and animal species. An effective vaccine to prevent S. aureus bovine disease and transmission would have positive effects on animal well-being, food production, and human health. The objective of this study was to identify multiple antigens that are immunoreactive during udder colonization and disease for exploration as vaccine antigens to prevent bovine mastitis. Staphylococcus aureus produces several cell wall-anchored and surface-associated virulence factors that play key roles in the pathogenesis of mastitis. Many of these proteins are conserved between different strains of S. aureus and represent promising vaccine candidates. We used an immunoproteomics approach to identify antigenic proteins from the surface of S. aureus. The expression of cell wall and surface proteins from S. aureus was induced under low iron conditions, followed by trypsin extraction and separation by 2-dimensional electrophoresis. The separated proteins were blotted with antibodies from mastitic bovine milk and identified by liquid chromatography-mass spectrometry. Thirty-eight unique proteins were identified, of which 8 were predicted to be surface exposed and involved in S. aureus virulence. Two surface proteins, iron-regulated surface determinant protein C (IsdC) and ESAT-6 secretion system extracellular protein (EsxA), were cloned, expressed, and purified from Escherichia coli for confirmation of immune reactivity by ELISA. A PCR of 37 bovine S. aureus isolates indicated that the presence of esxA and isdC is conserved, and amino acid alignments revealed that IsdC and EsxA sequences are highly conserved. The immunoproteomics technique used in this study generated reproducible results and identified surface exposed and reactive antigens for further characterization.

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  • Expression, immunogenicity and variation of iron-regulated surface protein A from bovine isolates of Staphylococcus aureus.

    FEMS Microbiology Letters

    Staphylococcus aureus iron-regulated surface protein A (IsdA) is a fibrinogen and fibronectin adhesin that also contributes to iron sequestration and resistance to innate immunity. IsdA is conserved in human isolates and has been investigated as a human vaccine candidate. Here we report the expression of isdA, the efficacy of anti-IsdA responses, and the existence of IsdA sequence variants from bovine Staphylococcus. Clinical staphylococci were obtained from U.S. dairy farms and assayed by PCR…

    Staphylococcus aureus iron-regulated surface protein A (IsdA) is a fibrinogen and fibronectin adhesin that also contributes to iron sequestration and resistance to innate immunity. IsdA is conserved in human isolates and has been investigated as a human vaccine candidate. Here we report the expression of isdA, the efficacy of anti-IsdA responses, and the existence of IsdA sequence variants from bovine Staphylococcus. Clinical staphylococci were obtained from U.S. dairy farms and assayed by PCR for the presence and expression of isdA. isdA positive species from bovines included S. aureus, S. haemolyticus and S. chromogenes. Immunoassays on bovine milk and serum confirmed the induction and opsonophagocytic activity of anti-IsdA humoral responses. The variable region of isdA was sequenced and protein alignments predicted the presence of two main variants consistent with those from human S. aureus. Mouse antibodies against one IsdA variant reduced staphylococcal binding to fibronectin in vitro in an isotype-dependent manner. Purified IsdA variants bound distinctly to fibronectin and fibrinogen. Our findings demonstrate that variability within the C-terminus of this adhesin affects immune reactivity and binding specificity, but are consistent with the significance of IsdA in bovine disease and relevant for vaccine development.

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  • Assessment of mineral phosphate-solubilizing properties and molecular characterization of zinc-tolerant bacteria

    Journal of Basic Microbiology

    Plant growth-promoting bacteria with the ability to tolerate heavy metals have importance both in sustainable agriculture and phytoremediation. The present study reports on the isolation and characterization of mineral phosphate-solubilizing (MPS) bacteria associated with the Achyranthes aspera L. plant (prickly chaff, flower plant). Out of 35 bacterial isolates, 6 isolates, namely RS7, RP23, EPR1, RS5, RP11 and RP19, with high MPS activity were selected and subjected to the assessment of MPS…

    Plant growth-promoting bacteria with the ability to tolerate heavy metals have importance both in sustainable agriculture and phytoremediation. The present study reports on the isolation and characterization of mineral phosphate-solubilizing (MPS) bacteria associated with the Achyranthes aspera L. plant (prickly chaff, flower plant). Out of 35 bacterial isolates, 6 isolates, namely RS7, RP23, EPR1, RS5, RP11 and RP19, with high MPS activity were selected and subjected to the assessment of MPS activity under various stress conditions, viz. ZnSO4 (0.30–1.5 M), NaCl and temperature. MPS activity by the selected isolates was observed at concentrations of as high as >1.2 M ZnSO4. Significant improvement in plant growth was observed on bacterization of seeds (pearl millet) with all of the six selected isolates. Plant growth was measured in terms of root length, shoot length, fresh weight and % increase in root biomass. The molecular diversity among the phosphate-solubilizing bacteria was studied employing enterobacterial repetitive intergenic sequence-PCR (ERIC-PCR). Representative strains from each ERIC type were identified, on the basis of a partial sequence of the 16S rRNA gene, as members of the genera Pseudomonas, Citrobacter, Acinetobacter, Serratia, and Enterobacter. Among all the isolates, RP19 was the best in terms of phosphate-solubizing activity and its response to various stresses. The ability of RP19 and other isolates to exhibit MPS activity at high ZnSO4 concentrations suggests their potential as efficient biofertilizer for growing plants in metal (ZnSO4)-contaminated soil.

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Courses

  • Biophysics

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  • Drug Discovery

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  • Flow Cytometry

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  • Immunology

    -

  • Microbiology

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  • Molecular Biology

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  • Plant Biotechnology

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  • Recombinant DNA technology

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  • Vaccinology

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Projects

  • Detection of antibiotic response of uro-pathogens by selective media

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Honors & Awards

  • Recipient of Distinguished Doctoral Dissertation Award, Boise State University

    Boise State University

  • Awarded ISV Presidential Trainee Award at International Society of Vaccine conference.

    International society of Vaccines Conference at Boston

    Was one of 8 students to win a $500 Trainee Award at the International Society of Vaccines (ISV) conference in Boston, MA. Oct 2016

  • Recipient of Sigma Xi research grant award

    Sigma Xi

    Awarded grant money for my research proposal submitted to the Sigma Xi Committee Grants-in-Aid

Languages

  • English

    Native or bilingual proficiency

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