Isolation of Mononuclear Cells From Tonsillar Tissue

Johnston, Andrew; Sigurðardóttir, Sigrún; Ryon, Judith J.

doi:10.1002/0471142735.im0708s00

Cited by 6 publications

(8 citation statements)

References 6 publications

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“…Human PBMCs were isolated from buffy coats by density gradient centrifugation using Ficoll‐Paque PLUS (GE Healthcare). Mononuclear cells (MCs) from human tonsils were isolated from patients undergoing tonsillectomy as previously described . After density gradient centrifugation using Ficoll‐Paque PLUS, ILCs were enriched by using magnetic cell depletion of CD3 + T cells with CD3 microbeads and LD columns (Miltenyi) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies^*

et al. 2017

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International audienceThe classical model of hematopoiesis established in the mouse postulates that lymphoid cells originate from a founder population of common lymphoid progenitors. Here, using a modeling approach in humanized mice, we showed that human lymphoid development stemmed from distinct populations of CD127(-) and CD127(+) early lymphoid progenitors (ELPs). Combining molecular analyses with in vitro and in vivo functional assays, we demonstrated that CD127(-) and CD127(+) ELPs emerged independently from lympho-mono-dendritic progenitors, responded differently to Notch1 signals, underwent divergent modes of lineage restriction, and displayed both common and specific differentiation potentials. Whereas CD127(-) ELPs comprised precursors of T cells, marginal zone B cells, and natural killer (NK) and innate lymphoid cells (ILCs), CD127(+) ELPs supported production of all NK cell, ILC, and B cell populations but lacked T potential. On the basis of these results, we propose a "two-family" model of human lymphoid development that differs from the prevailing model of hematopoiesis

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Section: Methodsmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies^*

et al. 2017

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“…Written informed consent was obtained from all patients prior to sample acquisition and experiments have been approved by the Ethics Committee of the Charité Medical University, Berlin (EA2‐078‐16, EA1/149/1). Mononuclear cells (MCs) from human tonsils were isolated from patients undergoing tonsillectomy as previously described . After mashing and density gradient centrifugation using Ficoll‐Paque PLUS, ILCs were enriched by using magnetic cell depletion of CD3 + T cells with CD3 mAb microbeads and LD columns (Miltenyi) according to the manufacturer's instructions.…”

Section: Flow Cytometric Phenotyping Of Cells Across Species and Tissuesmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

et al. 2019

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These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer‐reviewed by leading experts in the field, making this an essential research companion.

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“…Cell populations from human tonsils were isolated from patients undergoing tonsillectomy, as previously described (Ryon, 2001). Gut LP cells were isolated from CRC or IBD patients undergoing colectomy, by using a modified version of the protocol of Bull and Bookman (Bull and Bookman, 1977).…”

Section: Experimental Procedures Cell Isolation and Flow Cytometrymentioning

confidence: 99%

RORγt+ Innate Lymphoid Cells Acquire a Proinflammatory Program upon Engagement of the Activating Receptor NKp44

et al. 2013

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RORγt⁺ innate lymphoid cells (ILCs) are crucial players of innate immune responses and represent a major source of interleukin-22 (IL-22), which has an important role in mucosal homeostasis. The signals required by RORγt⁺ ILCs to express IL-22 and other cytokines have been elucidated only partially. Here we showed that RORγt⁺ ILCs can directly sense the environment by the engagement of the activating receptor NKp44. NKp44 triggering in RORγt⁺ ILCs selectively activated a coordinated proinflammatory program, including tumor necrosis factor (TNF), whereas cytokine stimulation preferentially induced IL-22 expression. However, combined engagement of NKp44 and cytokine receptors resulted in a strong synergistic effect. These data support the concept that NKp44⁺ RORγt⁺ ILCs can be activated without cytokines and are able to switch between IL-22 or TNF production, depending on the triggering stimulus.

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Isolation of Mononuclear Cells From Tonsillar Tissue

Cited by 6 publications

References 6 publications

Guidelines for the use of flow cytometry and cell sorting in immunological studies^*

Guidelines for the use of flow cytometry and cell sorting in immunological studies^*

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

RORγt+ Innate Lymphoid Cells Acquire a Proinflammatory Program upon Engagement of the Activating Receptor NKp44

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Isolation of Mononuclear Cells From Tonsillar Tissue

Cited by 6 publications

References 6 publications

Guidelines for the use of flow cytometry and cell sorting in immunological studies*

Guidelines for the use of flow cytometry and cell sorting in immunological studies*

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

RORγt+ Innate Lymphoid Cells Acquire a Proinflammatory Program upon Engagement of the Activating Receptor NKp44

Contact Info

Product

Resources

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Guidelines for the use of flow cytometry and cell sorting in immunological studies^*

Guidelines for the use of flow cytometry and cell sorting in immunological studies^*