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Characterization of Immunological Niches within Peyer’s Patches by ex vivo Photoactivation and Flow Cytometry Analysis

Authors: Adi Biram and Ziv Shulman, date: 03/20/2020, view: 227, Q&A: 0

[Abstract] T follicular helper (Tfh) cells regulate B cell selection for entry into the germinal center (GC) reaction or for differentiation into antibody forming cells. This process takes place at the border between the T and B zones in lymphoid organs and involves physical contacts between T and B cells. During these interactions, T cells endow the B cells ...

Analyzing the Quenchable Iron Pool in Murine Macrophages by Flow Cytometry

Authors: Michael Riedelberger and Karl Kuchler, date: 03/20/2020, view: 164, Q&A: 0

[Abstract] Tissue-resident macrophages are pivotal for a tightly-regulated iron metabolism at a cellular and systemic level, since subtle iron alterations increase the susceptibility for microbial infections or drive multiple diseases. However, research on cellular iron homeostasis in macrophages remains challenging due to the limited amount of available ...

Method for Measuring Mucociliary Clearance and Cilia-generated Flow in Mice by ex vivo Imaging

Authors: Eric Song and Akiko Iwasaki, date: 03/20/2020, view: 203, Q&A: 0

[Abstract] Ex vivo biophysical measurements provide valuable insights into understanding both physiological and pathogenic processes. One critical physiological mechanism that is regulated by these biophysical properties is cilia-generated flow that mediates mucociliary clearance, which is known to provide protection against foreign particles and ...

Optogenetic Tuning of Ligand Binding to The Human T cell Receptor Using The opto-ligand-TCR System

Authors: O. Sascha Yousefi, Maximilian Hörner, Maximilian Wess, Vincent Idstein, Wilfried Weber and Wolfgang W. A. Schamel, date: 03/05/2020, view: 500, Q&A: 0

[Abstract] T cells are one major cell type of the immune system that use their T cell antigen receptor (TCR) to bind and respond to foreign molecules derived from pathogens. The ligand-TCR interaction half-lives determine stimulation outcome. Until recently, scientists relied on mutating either the TCR or its ligands to investigate how varying TCR-ligand ...

Zebrafish Bacterial Infection Assay to Study Host-Pathogen Interactions

Authors: Faiza Basheer, Clifford Liongue and Alister C Ward, date: 03/05/2020, view: 286, Q&A: 0

[Abstract] The study of host-pathogen interactions has improved our understanding of both pathogenesis and the response of the host to infection, including both innate and adaptive responses. Neutrophils and macrophages represent the first line of innate host defense against any infection. The zebrafish is an ideal model to study the response of these cells ...

Skin Transplantation and Lymphoid Organ Analysis in Mice

Authors: Julie Ruer-Laventie, Rajesh Jayachandran, Mathias Schmaler, Simona W. Rossi and Jean Pieters, date: 02/20/2020, view: 446, Q&A: 0

[Abstract] Skin transplantation in mice is an important procedure to evaluate immune responses generated against heterologous grafts, especially given its highly immunogenic nature. In fact, skin is one of the most challenging organs in terms of allograft retention. In this protocol, we provide a detailed procedure for skin grafting using the tail skin as ...

Transcervical Mouse Infections with Chlamydia trachomatis and Determination of Bacterial Burden

Authors: Karthika Rajeeve and Rajeeve Sivadasan, date: 02/05/2020, view: 351, Q&A: 0

[Abstract] Chlamydia trachomatis is an obligate human pathogen. It infects the genital tract of humans ascending into the fallopian tube, exacerbated by chronic pelvic pain, pelvic inflammatory disease, and fallopian tube scaring resulting in infertility and other malignancies. The major hurdle in controlling chlamydial spread is that the ...

Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro

Authors: Nicholas D. Condon, Jennifer L. Stow and Adam A. Wall, date: 01/20/2020, view: 407, Q&A: 0

[Abstract] Cell surface protrusions include F-actin rich, wave-like ruffles that are erected transiently in response to stimuli and during cell migration. Macrophages are innate immune cells that ruffle constitutively and more dramatically in cells activated by pathogens. Dorsal ruffles and their resulting macropinosomes are key sites for environmental ...

Quantifying HIV-1-Mediated Gut CD4⁺ T Cell Death in the Lamina Propria Aggregate Culture (LPAC) Model

Authors: Stephanie M. Dillon, Kejun Guo, Moriah J. Castleman, Mario L. Santiago and Cara C. Wilson, date: 01/20/2020, view: 471, Q&A: 0

[Abstract] Gut CD4 T cells are major targets of HIV-1 and are massively depleted early during infection. To better understand the mechanisms governing HIV-1-mediated CD4 T cell death, we developed the physiologically-relevant Lamina Propria Aggregate Culture (LPAC) model. The LPAC model is ideal for studying CD4 T cell death induced by clinically-relevant ...

Isolation and Stimulation of Peritoneal Macrophages with Apoptotic Jurkat Cells to Produce IL-10

Authors: Mei Song and Xiaojing Ma, date: 12/20/2019, view: 623, Q&A: 0

[Abstract] Clearance of apoptotic cells by macrophages is critical to ensuring cellular homeostasis and suppression of autoimmunity. Macrophage recognition of apoptotic cells triggers an anti-inflammatory response, which is mediated by the release of IL-10, TGF-β etc. with concurrent inhibition of pro-inflammatory cytokines (such as TNFα, IL-12, ...

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