Effect of SLAMF6 on T cell calcium signalling in response to antigen stimulation
(2026) KEML10 20261Department of Chemistry
- Abstract
- SLAMF6 is an immunoregulatory molecule studied in cancer research as a possible target for immunotherapy in acute myeloid leukaemia (AML). The absence of SLAMF6 has been shown to elevate T cell activation and improve anti-tumour responses (1). The regulatory mechanism of SLAMF6 is not fully understood. The aim of this thesis was to investigate the effect of SLAMF6 on T cell signalling using fluorescence microscopy. The experimental system was developed stepwise. First a mobile SLB containing relevant proteins (including CD58, SLAMF6, and peptide loaded major histocompatibility complex (pMHC)) was created. Cells were then added to the SLB to analyse binding patterns. The T cells were subsequently activated by pMHC binding T cell preceptors... (More)
- SLAMF6 is an immunoregulatory molecule studied in cancer research as a possible target for immunotherapy in acute myeloid leukaemia (AML). The absence of SLAMF6 has been shown to elevate T cell activation and improve anti-tumour responses (1). The regulatory mechanism of SLAMF6 is not fully understood. The aim of this thesis was to investigate the effect of SLAMF6 on T cell signalling using fluorescence microscopy. The experimental system was developed stepwise. First a mobile SLB containing relevant proteins (including CD58, SLAMF6, and peptide loaded major histocompatibility complex (pMHC)) was created. Cells were then added to the SLB to analyse binding patterns. The T cells were subsequently activated by pMHC binding T cell preceptors (TCRs) on the T cells, and a dose-response curve of the number of activated cells vs pMHC density was generated. Finally, SLAMF6 was added to the system to investigate how it affects T cell signalling. Fluorescence recovery after photobleaching (FRAP) analysis confirmed the formation of a mobile SLB with proteins successfully incorporated in the bilayer. T cells adhered to the SLB through CD58 mediated interactions and showed accumulation of SLAMF6, UCHT1-Fab and pMHC in the cell-SLB contact at the cell surface. T cell activation was observed through calcium signalling, measured using the calcium sensitive dye Fluo-4am. The fraction of signalling cells increased with increasing concentration pMHC, enabling the creation of a dose-response curve. Lastly the addition of SLAMF6 reduced the fraction of signalling cells, suggesting that SLAMF6 regulates T cell activation in this system. (Less)
- Popular Abstract
- Our immune system protects us from things that can make us sick, such as viruses, bacteria, and other germs. One important part of the immune system is T cells which are a type of white blood cells. They act like a control system, checking whether cells in the body look normal. If they detect abnormal cells, they can destroy them before they become a problem and make us sick.
Our body is constantly renewing cells by making copies of them. Sometimes this cell replication goes wrong and abnormal cells start to grow and multiply uncontrollably. If the immune system detects these abnormal cells early, it can help prevent cancer. However, if it doesn’t, cancer may develop and the body might need medical help to fight it.
Acute myeloid... (More) - Our immune system protects us from things that can make us sick, such as viruses, bacteria, and other germs. One important part of the immune system is T cells which are a type of white blood cells. They act like a control system, checking whether cells in the body look normal. If they detect abnormal cells, they can destroy them before they become a problem and make us sick.
Our body is constantly renewing cells by making copies of them. Sometimes this cell replication goes wrong and abnormal cells start to grow and multiply uncontrollably. If the immune system detects these abnormal cells early, it can help prevent cancer. However, if it doesn’t, cancer may develop and the body might need medical help to fight it.
Acute myeloid leukaemia (AML) is a type of cancer where the bone marrow starts producing abnormal white blood cells. This leads to a weaker immune system, which has a harder time fighting infections. Many cancer treatments can be stressful for the body, and there is ongoing research to improve them. One common treatment is immunotherapy, which works by strengthening the immune system so it can better recognize and attack cancer cells. A potential target for immunotherapy that has been researched is signalling lymphocytic activation molecule family member 6 (SLAMF6). How it affects the immune system and contributes to the elimination of cancer cells is still not fully understood and is therefore being actively studied. In this thesis an experimental system was developed and optimised to study on a physiochemical level how SLAMF6 affects signalling in T cells. To reach this goal, several subgoals were defined:
1) Creating a supported lipid bilayer (SLB) with proteins of interest to mimic a cell membrane. SLBs are models of cell membranes consisting of a lipid bilayer deposited on a glass slide, that have a controlled composition and can be favourably studied with fluorescence microscopy.
2) Binding T cells to the SLB using different relevant receptor-ligand pairs, including CD58-CD2, SLAMF6-SLAMF6 and peptide loaded major histocompatibility complexes (pMHC) binding to T cell receptors (TCRs). CD58 and CD2 are adhesion molecules whereas TCR is a receptor on T cells that initiates signalling upon binding pMHC molecules displaying parts of a pathogen.
3) Inducing and studying calcium signalling in the T cells. The release of calcium ions can be measured, by adding a fluorescent dye that blinks once it detects calcium in the cell. This was done both with TCR binding pMHC in the SLB as well as via antibodies targeting the TCR.
4) The concentration of TCR-pMHC pairs was varied and a dose-response curve for signalling was obtained.
5) The final step was to investigate how SLAMF6 affects the dose-response curve. Will it make the T cell more or less sensitive? This was investigated by comparing how many T cells signalled at a fixed number of pMHC molecules, with and without SLAMF6. The results indicate that the concentration of trigger molecules influences cell signalling and that SLAMF6 lowers T cell activation. Which suggests that it can reduce T cell activation in this system. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/student-papers/record/9229588
- author
- Börjesson, Oliwia LU
- supervisor
- organization
- course
- KEML10 20261
- year
- 2026
- type
- M2 - Bachelor Degree
- subject
- keywords
- Physical chemistry, Cell signaling, Fluorescence microscopy, SLAMF6, T cell activation
- language
- English
- id
- 9229588
- date added to LUP
- 2026-06-01 14:22:37
- date last changed
- 2026-06-01 14:22:37
@misc{9229588,
abstract = {{SLAMF6 is an immunoregulatory molecule studied in cancer research as a possible target for immunotherapy in acute myeloid leukaemia (AML). The absence of SLAMF6 has been shown to elevate T cell activation and improve anti-tumour responses (1). The regulatory mechanism of SLAMF6 is not fully understood. The aim of this thesis was to investigate the effect of SLAMF6 on T cell signalling using fluorescence microscopy. The experimental system was developed stepwise. First a mobile SLB containing relevant proteins (including CD58, SLAMF6, and peptide loaded major histocompatibility complex (pMHC)) was created. Cells were then added to the SLB to analyse binding patterns. The T cells were subsequently activated by pMHC binding T cell preceptors (TCRs) on the T cells, and a dose-response curve of the number of activated cells vs pMHC density was generated. Finally, SLAMF6 was added to the system to investigate how it affects T cell signalling. Fluorescence recovery after photobleaching (FRAP) analysis confirmed the formation of a mobile SLB with proteins successfully incorporated in the bilayer. T cells adhered to the SLB through CD58 mediated interactions and showed accumulation of SLAMF6, UCHT1-Fab and pMHC in the cell-SLB contact at the cell surface. T cell activation was observed through calcium signalling, measured using the calcium sensitive dye Fluo-4am. The fraction of signalling cells increased with increasing concentration pMHC, enabling the creation of a dose-response curve. Lastly the addition of SLAMF6 reduced the fraction of signalling cells, suggesting that SLAMF6 regulates T cell activation in this system.}},
author = {{Börjesson, Oliwia}},
language = {{eng}},
note = {{Student Paper}},
title = {{Effect of SLAMF6 on T cell calcium signalling in response to antigen stimulation}},
year = {{2026}},
}