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Biology
Aislamiento de CD4+ las células de T y análisis de circulación folicular T Helper (cTf...
Aislamiento de CD4+ las células de T y análisis de circulación folicular T Helper (cTf...
JoVE Journal
Biology
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JoVE Journal Biology
Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Aislamiento de CD4+ las células de T y análisis de circulación folicular T Helper (cTfh) subconjuntos de la célula del uso de sangre periférica citometría de flujo de 6 colores

Full Text
12,757 Views
07:39 min
January 7, 2019

DOI: 10.3791/58431-v

Elliot Byford1, Matthew Carr1, Lucia Piñon2, Matthew J. Ahearne1, Simon D. Wagner1

1Leicester Cancer Research Centre and Ernest and Helen Scott Haematology Research Institute,University of Leicester, 2MRC Toxicology Unit,University of Leicester

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article presents a protocol for isolating and characterizing CD4 + T-cell subsets from human peripheral blood. It emphasizes the use of flow cytometry to analyze the proportions of T-follicular helper cell subsets in different health states.

Key Study Components

Research Area

  • Immunology
  • T-cell biology

Background

  • CD4 + T-cells play a crucial role in the adaptive immune response.
  • T-follicular helper cells are vital for B-cell responses and antibody production.

Methods Used

  • Flow cytometry for cell subset analysis
  • Human peripheral blood as the biological system
  • Batch processing technique for sample analysis

Main Results

  • Identification of differences in T-follicular helper cell subsets between healthy and diseased individuals.
  • Proven correlation of T-follicular helper cell subsets with clinical disease markers.
  • Verification of method efficacy and practicality in clinical settings.

Conclusions

  • This study demonstrates a reliable method for evaluating T-follicular helper subsets in various health statuses.
  • The findings are significant for understanding disease progression and potential biomarkers.

Frequently Asked Questions

What are CD4 + T-cells?
CD4 + T-cells are a type of white blood cell that plays an essential role in the immune system, particularly in helping other cells respond to infections.
How does flow cytometry work?
Flow cytometry is a technology used to analyze the physical and chemical characteristics of cells or particles as they flow in a fluid stream.
Why are T-follicular helper cells important?
T-follicular helper cells assist in the activation of B-cells and the production of antibodies, which are critical for immune defense.
Can this method be used in clinical settings?
Yes, the protocol developed in this study is practical for use in clinical laboratories for assessing disease states.
What advantages does this technique offer?
The method is fast, allows batch processing of samples, and maintains cell viability for further analyses.
How are the results of T-follicular helper subsets utilized?
Results can be used as biomarkers for disease progression and severity in patients.
Who contributed to this research?
Dr. Lucia Pi has contributed significantly to demonstrating this procedure.

Aquí, un protocolo para el aislamiento y la caracterización de CD4+ T-cell se describe subconjuntos de sangre periférica humana. Purificada de CD4+ las células T son analizadas por citometría de flujo para determinar proporciones de subconjuntos de células helper T-folicular.

Este método puede ayudar a demostrar las diferencias entre los subconjuntos de células auxiliares T-foliculares circulantes en estados sanos y enfermos. Las principales ventajas de esta técnica son que es rápida, que las muestras se pueden procesar por lotes y que las celdas permanecen activas para el análisis funcional posterior. Esta técnica facilita la evaluación de subconjuntos auxiliares T-foliculares circulantes como biomarcadores para la progresión de la enfermedad, ya que se correlacionan con marcadores de gravedad clínica. Ayudar con la demostración de este procedimiento será la Dra. Lucia Pi

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