Functional Characterization of Natural Killer Cell Responses in Endometriosis

Background

Endometriosis is a chronic inflammatory disease characterized by the presence of endometrial-like tissue outside the uterine cavity. It affects approximately 10% of women of reproductive age and is associated with pelvic pain, infertility, and reduced quality of life. Despite its high prevalence, the immunological mechanisms that permit ectopic endometrial tissue to persist remain incompletely understood.

Natural Killer (NK) cells are central components of the innate immune system and play a critical role in immune surveillance by eliminating aberrant or misplaced cells. In healthy individuals, NK cells can recognize and kill stressed, transformed, or foreign cells without prior sensitization. Their cytotoxic activity is regulated by a balance between activating and inhibitory receptors, which determine whether target cells are eliminated.

Impaired NK cell function has been proposed as a contributing factor in endometriosis, potentially allowing ectopic endometrial cells to evade immune clearance. However, the precise mechanisms governing NK cell-endometriosis cell interactions remain poorly defined.

Understanding how NK cells recognize and respond to endometriotic cells may reveal novel immunological mechanisms underlying disease persistence and could inform future therapeutic strategies.

Preliminary Data

Preliminary in vitro experiments demonstrate that NK cells degranulate in response to primary endometriosis-derived cells. Furthermore, blocking studies suggest that specific activating receptor-ligand interactions may play a dominant role in mediating this response.

However, the relative contribution of different receptor pathways and the overall cytotoxic capacity of NK cells toward endometriotic cells require further systematic investigation.

Project Description

The aim of this Master’s project is to functionally characterize NK cell responses toward endometriosis-derived cells and to define the receptor-ligand interactions that regulate NK cell-mediated cytotoxicity.

The student will:

·       Establish and optimize real-time NK cell cytotoxicity assays using the IncuCyte live-cell imaging system

·       Investigate NK cell degranulation and killing of endometriosis-derived cells

·       Evaluate the role of specific activating and inhibitory receptor interactions using blocking antibodies

·       Process and handle primary patient samples from individuals with endometriosis

This project combines method development, functional immunology, and translational patient-based research.

Methods

Patient Material

• Tissue preparation and cell isolation from donors
• NK cell purification using magnetic separation or flow cytometric sorting
• Culture of primary endometriosis-derived cells

Method Development: IncuCyte Cytotoxicity Assay

• Optimization of tumor/endometriosis cell labeling
• Establishment of effector-to-target ratios
• Real-time live-cell imaging to quantify NK cell-mediated killing
• Development of standardized analysis pipeline

Functional NK Cell Assays

• CD107a degranulation assays (flow cytometry)
• Cytokine production analysis (e.g., IFN-γ)

Receptor–Ligand Interaction Studies

• Blocking antibodies targeting activating and/or inhibitory NK receptors
• Functional assessment of changes in degranulation and cytotoxicity
• Analysis of ligand expression on endometriosis-derived cells

Data Analysis

• Statistical comparison of NK cell responses
• Correlation of receptor expression with functional activity
• Evaluation of donor variability

Expected Outcomes

This project will:

·       Establish a robust live-cell imaging platform for studying NK cell cytotoxicity in endometriosis

·       Define key receptor-ligand interactions regulating NK cell responses

·       Provide insight into immune evasion mechanisms in endometriosis

The results may contribute to improved understanding of immune dysregulation in endometriosis and potentially identify novel immunomodulatory targets.

Requirements

·       Background in biotechnology, biomedicine, or related field

·       Basic laboratory skills

·       Interest in immunology and translational research

·       Willingness to work with primary patient samples

Supervision

The project will be carried out in the Tumor Immunology (TIMM) Lab at Sahlgrenska Center for Cancer Research at Sahlgrenska Academy, University of Gothenburg, under the supervision of Elin Bernson.