Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester

Overview

This protocol provides a sensitive method for measuring proliferating CD4 + T cells in response to antigenic proteins or peptides. It allows for the detection and isolation of rare antigen-specific T cells, which is crucial for studying autoimmune conditions.

Key Study Components

Area of Science

• Immunology
• Cell Biology
• Autoimmunity

Background

• CD4 + T cells play a critical role in the immune response.
• Detection of rare antigen-specific T cells is often challenging.
• This assay can be adapted for cloning specific T cells.
• It is particularly relevant for studying autoimmune diseases like type one diabetes.

Purpose of Study

• To develop a protocol for measuring CD4 + T cell responses.
• To facilitate the detection of weak antigen-specific T cell populations.
• To provide a method for isolating these cells for further study.

Methods Used

• Peripheral blood sample collection from healthy donors.
• Dilution of blood in PBS and density gradient centrifugation.
• Isolation of white blood cells while avoiding red blood cell contamination.
• Washing of cells in fresh PBS for further analysis.

Main Results

• Successful isolation of CD4 + T cells from peripheral blood.
• Ability to detect rare antigen-specific T cell populations.
• Demonstrated sensitivity of the assay for weak immune responses.
• Potential for cloning antigen-specific T cells for research.

Conclusions

• This protocol enhances the ability to study CD4 + T cell responses.
• It provides a valuable tool for research in autoimmune diseases.
• The method can be adapted for various antigenic stimuli.

Frequently Asked Questions