2026 Theses Doctoral

Balancing adhesion and migration: The role of CXCR4-CXCL12 in human natural killer cell motility and development

Eisman, Shira E.

Natural killer (NK) cells are an essential component of the innate immune system and play a critical role in controlling viral infection and malignancy. As they mature, NK cells interact with various microenvironments; however, much about these interactions and how they shape NK cell development is still unknown.

This thesis examines the complex interactions between NK cells and their niches and specifically how the CXCR4-CXCL12 chemokine axis regulates human NK cell behavior across development, focusing on its role in migration, adhesion, and interaction with stromal cells. Building on prior work defining the developmental synapse between NK cells and supportive stromal cells, I investigated how CXCR4 shapes these contacts.

Using in vitro differentiation, I showed that treatment with CXCR4 antagonist AMD3100 impairs the generation of NK cells from CD34+ precursors. To understand the underlying migratory phenotypes, I applied the quantitative cell-tracking pipeline, cellPLATO, which I refined and expanded as part of this thesis to enable transparent and reproducible analysis of single-cell motility. CXCR4 blockade consistently altered key migratory features, including reduced arrest and increased displacement, suggesting that CXCR4 helps balance migration and adhesion to enable productive stromal engagement.

High-resolution imaging further demonstrates crosstalk between CXCR4 and integrins, providing a potential mechanism for chemokine-dependent modulation of adhesion. Completing these mechanistic studies, I also present cord blood NK phenotyping, highlighting the developmental trends in CXCR4, integrins, and effector molecules across early human NK subsets, as well as data from WHIM CXCR4 gain-of-function patient cells, illustrating the clinical impact of dysregulated CXCR4 signaling.

Together, these studies identify the CXCR4-CXCL12 axis as a central regulator of NK cell migration, adhesion, and development. By integrating developmental models, quantitative imaging, and human immunophenotyping, this work advances our understanding of how chemokine cues shape human NK cell development and informs strategies to improve NK cell-based immunotherapies and address CXCR4-related immunodeficiencies.