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Flow Cytometry Analysis of mTOR Signaling in Antigen-Specific B Cells

Qizhao Huang1, Haoqiang Wang1, Lifan Xu1

  • 1Institute of Immunology, Third Military Medical University, Chongqing, 400038, China.

Methods in Molecular Biology (Clifton, N.J.)
|February 2, 2018
PubMed

Insights

This study presents protocols to measure mTOR activity in B cells. Evaluating this kinase is crucial for understanding B cell responses to pathogens.

Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • B lymphocytes produce antibodies to combat pathogens via the B cell antigen receptor (BCR).
  • BCR activation triggers signaling pathways, including mTOR, which regulates cell proliferation and metabolism.
  • mTOR signaling downstream of PI3K/Akt is presumed essential for B cell function.

Purpose of the Study:

  • To introduce two experimental protocols for assessing mTOR activity.
  • To focus on B cells expressing BCR specific to the 4-hydroxy-3-nitrophenylacetyl (NP)-hapten antigen.

Main Methods:

  • The study details two distinct protocols.
  • These methods are designed for evaluating mTOR kinase activity.
  • The focus is on B cells with NP-hapten specific BCR.

Main Results:

  • Protocols for evaluating mTOR activity in specific B cells are presented.
  • The methods allow for the assessment of a key signaling pathway in B cell activation.

Conclusions:

  • The presented protocols offer a means to investigate mTOR's role in BCR-mediated signaling.
  • Understanding mTOR activity is vital for deciphering B cell physiology and immune responses.

Keywords:
Antigen specific B cellB cell receptorEx-vivoFlow cytometryIn vitroNP-haptenmTOR

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