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Spatial mapping of affinity changes for the integrin LFA-1 during cell migration using clusters identified based on

Henrik Persson¹, Wojciech Potrzebowski², Katarzyna Potrzebowska¹

¹Section of Immunology, Department of Experimental Medical Science, Lund University, Lund, Sweden.

Journal of Biophotonics

|September 30, 2018

Summary

Protein clustering of leukocyte function-associated antigen 1 (LFA-1) varies by affinity state in migrating T-cells. This clustering may influence cell adhesion to ICAM-1, impacting immune responses.

Keywords:

DBSCAN LFA-1 STORM T-lymphocyte cluster analysis integrins localization microscopy

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Area of Science:

Cellular Biology
Immunology
Biophysics

Background:

Leukocyte function-associated antigen 1 (LFA-1) is a key integrin involved in T-cell adhesion.
The role of LFA-1 clustering in modulating its avidity to ICAM-1 remains debated.
Understanding integrin clustering is crucial for deciphering cell-substrate adhesion mechanisms.

Purpose of the Study:

To investigate the clustering behavior of different affinity forms of LFA-1 in migrating T-cells.
To determine if LFA-1 clustering contributes to cell adhesion dynamics on ICAM-1.
To apply advanced cluster analysis to localization microscopy data for quantitative insights.

Main Methods:

Utilized Stochastic Optical Reconstruction Microscopy (STORM) for high-resolution imaging of LFA-1.
Applied the Density-Based Spatial Clustering of Applications with Noise (DBSCAN) algorithm to analyze molecular coordinates.
Quantified cluster parameters for various LFA-1 affinity states in migrating T-cells.

Main Results:

Observed distinct clustering patterns for different LFA-1 affinity forms.
Pan-LFA-1 formed clusters throughout the cell, with higher density in the mid and rear areas on ICAM-1.
Intermediate and high-affinity LFA-1 forms showed increased clustering in the mid-cell area, while high-affinity LFA-1 also clustered at the front and rear.

Conclusions:

LFA-1 clustering varies significantly depending on its conformational state.
Protein clustering, alongside LFA-1 conformation, likely plays a critical role in regulating T-cell adhesion to ICAM-1.
DBSCAN analysis of STORM data provides a robust method for identifying and quantifying integrin clusters.