Neha Mishra
Current Organizations / Institutes
The organizations/institutes from the scholar's latest article
Research Subjects
Neha Mishra has a total of 4 co-authors. Here are the top co-authors by number of shared publications:
- Almut Meyer-Bahlburg
University Medicine Greifswald 21 Department of Pediatrics and Adolescent Medicine Greifswald
1 shared publication - Ann-Kathrin Behrendt
Pediatric Rheumatology and Immunology, Department of Pediatrics, University Medicine Greifswald, 17475 Greifswald, Germany
1 shared publication - Lukas Bossaller
Section of Rheumatology, Department of Medicine A, University Medicine Greifswald Greifswald Germany
1 shared publication - Nico Wittmann
Section of Pediatric Rheumatology and Immunology, Department of Pediatric and Adolescent Medicine, University Medicine Greifswald Greifswald Germany
1 shared publication
Latest Publications
- Open Access
Instructions for Flow Cytometric Detection of ASC Specks as a Readout of Inflammasome Activation in Human Blood
Inflammasome activation is linked to the aggregation of the adaptor protein ASC into a multiprotein complex, known as the ASC speck. Redistribution of cytosolic ASC to this complex has been widely used as a readout for inflammasome activation and precedes the downstream proteolytic release of the proinflammatory cytokines, IL-1β and IL-18. Although inflammasomes are important for many diseases such as periodic fever syndromes, COVID-19, gout, sepsis, atherosclerosis and Alzheimer’s disease, only a little knowledge exists on the precise and cell type specific occurrence of inflammasome activation in patient samples ex vivo. In this report, we provide detailed information about the optimal conditions to reliably identify inflammasome activated monocytes by ASC speck formation using a modified flow cytometric method introduced by Sester et al. in 2015. Since no protocol for optimal sample processing exists, we tested human blood samples for various conditions including anticoagulant, time and temperature, the effect of one freeze–thaw cycle for PBMC storage, and the fast generation of a positive control. We believe that this flow cytometric protocol will help researchers to perform high quality translational research in multicenter studies, and therefore provide a basis for investigating the role of the inflammasome in the pathogenesis of various diseases.
Open AccessInstructions for Flow Cytometric Detection of ASC Specks as a Readout of Inflammasome Activation in Human Blood
Inflammasome activation is linked to the aggregation of the adaptor protein ASC into a multiprotein complex, known as the ASC speck. Redistribution of cytosolic ASC to this complex has been widely used as a readout for inflammasome activation and precedes the downstream proteolytic release of the proinflammatory cytokines, IL-1β and IL-18. Although inflammasomes are important for many diseases such as periodic fever syndromes, COVID-19, gout, sepsis, atherosclerosis and Alzheimer’s disease, only a little knowledge exists on the precise and cell type specific occurrence of inflammasome activation in patient samples ex vivo. In this report, we provide detailed information about the optimal conditions to reliably identify inflammasome activated monocytes by ASC speck formation using a modified flow cytometric method introduced by Sester et al. in 2015. Since no protocol for optimal sample processing exists, we tested human blood samples for various conditions including anticoagulant, time and temperature, the effect of one freeze–thaw cycle for PBMC storage, and the fast generation of a positive control. We believe that this flow cytometric protocol will help researchers to perform high quality translational research in multicenter studies, and therefore provide a basis for investigating the role of the inflammasome in the pathogenesis of various diseases.