Enrichment and culture of myeloid cells for immunogenicity assays and study of inflammation

Dendritic cell-based assays for vaccine screening
 

Collaboration between academic institutes, pharmaceutical companies, and CROs is accelerating the vaccine development process today as never before.

At the base of the pre-clinical phase of vaccine development is the need for standardized antigen-presentation assays that can be used to evaluate safety and immunogenicity of candidate antigens. Such immuno-assays are often based on co-culture of monocyte-derived dendritic cells (Mo-DCs) and T cells, for which Miltenyi Biotec can offer the following supporting tools and protocols:'
 

• Monocytes and T cells can be quickly enriched using StraightFrom Blood products
• Premium grade MACS® Cytokines, such as Human IL-4 and Human GM-CSF, guarantee standardized differentiation and activation of monocyte-derived dendritic cells
• PepTivator Peptide Pools can be efficiently presented by Mo-DCs and enable virus-specific activation and stimulation of CD4+ and CD8+ T cells. PepTivator Peptide Pools are  available for SARS-CoV-2.
• MACS Antibodies and the MC CD14 Monocyte Cocktail allow optimal phenotyping of Mo-DC  and T cells
• For other products related to enrichment and phenotyping of T cells, please visit Solutions for stimulation, enrichment, and analysis of rare virus-specific T cells 
   

Study of inflammatory myeloid cells to understand COVID-19 pathology

During highly inflammatory states, such as in COVID-19-related pneumonia, myeloid cells release excessive amounts of chemokines, which result in cytokine storm syndrome (CSS). Recent RNA-sequencing data suggest that alveolar macrophages become hyperactive in the severely injured lungs of patients with SARS-Co-V2, leading to CSS, respiratory distress syndrome, and multi-organ dysfunction.  

Moreover, some researchers have already reported that inhibition of viral endocytosis by resident macrophages might help in reducing inflammation during  SARS-CoV-2 infection.  For this reason, studying myeloid cell dysregulation in COVID-19 patients is currently of great interest.  

Miltenyi Biotec can support such endeavors with: 

• The CD163 MicroBead Kit for isolation of inflammatory or steady state monocytes/macrophages from human peripheral blood mononuclear cell suspension or single-cell suspensions from tissues (e.g. lung tissues, BALF)
• CD11b MicroBeads to enrich inflammatory or steady state myeloid cells from human peripheral blood mononuclear cells or single-cell suspensions from tissues (e.g. lung, BALF). Also available for non-human primates
• The StraightFrom Whole Blood CD14 MicroBead Kit for isolation of monocytes from peripheral  blood, and CD14 MicroBeads UltraPure to enrich tissue-resident and tissue-infiltrating monocytes
• Products for fast isolation of granulocytes directly from blood, such as the MACSxpress Whole Blood Neutrophil Isolation Kit, the MACSxpress Whole Blood Eosinophil Isolation Kit, StraightFrom Whole Blood CD66b MicroBeads and CD15 MicroBeads
• The CD304 (BDCA-4/Neuropilin-1) MicroBead Kit for isolation of Neuropilin-1 positive cells such as plasmacytoid dendritic cells from peripheral blood, cord blood, or bone marrow
   

References

• The landscape of lung bronchoalveolar immune cells in COVID-19 revealed by single-cell RNA sequencing.; Liao et al.
• Insights from nanomedicine into chloroquine efficacy against COVID-19; Hu et al.; Nat Nanotechnol
• Monocytes and Macrophages as Viral Targets and Reservoirs; Nikitina et al.; Int J Mol Sci (PMID: 30231586)
• Hydroxychloroquine, a less toxic derivative of chloroquine, is effective in inhibiting SARS-CoV-2 infection in vitro; Liu et al.; (PMID: 32194981)
• Macrophage expression and prognostic significance of the long pentraxin PTX3 in COVID-19.; Enrico Brunetta et al.; Nature Immunology (PMID: 33208929) 
• Altered bioenergetics and mitochondrial dysfunction of monocytes in patients with COVID-19 pneumonia.; Lara Gibellini et al.; EMBO Molecular Medicine (PMID: 33078545) 
• Neuropilin-1 facilitates SARS-CoV-2 cell entry and infectivity.; Ludovico Cantuti-Castelvetri et al.; Science (PMID: 33082293)
• Neuropilin-1 is a host factor for SARS-CoV-2 infection.; James L Daly et al.; Science (PMID: 33082294)