Biology subjects

King, L. A.

Publications and source records attributed to King, L. A..

4 recordsLinked to original sources

bioRxiv2026

Systematic Evaluation of Nasal Immune Cell Sampling and Antigen-specific T cell Detection using Cryopreserved Nasal Swabs

The upper respiratory tract is a key entry point for pathogens, yet local tissue-resident memory T cells (Trm) remain underexplored compared to peripheral blood. We systematically compared nasal curettes and 8 different swab types for immune cell collection, assessing yield, operator variability, and T cell phenotypes across the three turbinates and nasopharynx. The use of flocked swabs yielded higher immune cell numbers while being similarly tolerated, especially with reduced sampling duration. Nasal Trm subsets were consistent across the turbinates, whereas nasopharyngeal Trm displayed a more recently recruited phenotype. Multiple cryopreservation media were evaluated and all demonstrated high viability after thawing. Antigen-specificity was assessed using the activation induced marker (AIM) assay, peptideHLA tetramers and bulk TCR-sequencing following expansion. Notably, influenza-specific T cell frequencies were reliably detected by AIM and correlated between fresh and cryopreserved nasal samples. Downregulation of the CD3/TCR complex was observed in nasal samples. These findings establish a robust approach for nasal Trm profiling, demonstrating that cryopreservation preserves functional antigen-specific T cells. This work enables centralized, minimally invasive nasal T cell analysis for multicenter studies, including mucosal vaccination trials and controlled human infection models.

immunology↗

bioRxiv2026

Ageing of the Upper Airway Epithelial Niche Limits Tissue-Resident T Cell Immunity

Respiratory tract infections are a major cause of morbidity and mortality among older adults worldwide. Yet, how ageing shapes protective immunity within the upper respiratory tract (URT) remains poorly understood. Here, we profiled the nasal immune landscape of young adults and older adults with and without frailty, using high-dimensional cytometry, proteomics, single-cell transcriptomics and T cell receptor (TCR) sequencing. Ageing was associated with a selective reduction of tissue-resident memory T (Trm) cells, independent of effector T cell numbers, inflammageing levels or frailty and was accompanied by a decline in T cell receptor repertoire stability. Trm cells from older adults also showed decreased steady-state IFN{gamma} expression, accompanied by reduced antiviral transcriptional programs in the mucosa. Mechanistically, in vitro PBMC-epithelium co-culture models revealed that older adults have a reduced capacity for Trm differentiation, which was associated with a diminished epithelial TGF-{beta} secretion, driven by reduced expression in ciliated epithelial cells. Together, these findings reveal that URT immunity deteriorates with age due to disrupted epithelial-immune crosstalk. This identifies the epithelial niche as a key regulator of mucosal immune ageing and suggests that enhancing TGF{beta} signaling may improve the effectiveness of mucosal airway vaccination strategies in older populations.

immunology↗

bioRxiv2023

Isolation and expansion of pure and functional γδ T cells

{gamma}{delta} T cells are important components of the immune system due to their ability to elicit a fast and strong response against infected and transformed cells. Because they can specifically and effectively kill target cells in an MHC independent fashion, there is great interest to utilize these cells in anti-tumor therapies where antigen presentation may be hampered. Since only a small fraction of T cells in the blood or tumor tissue are {gamma}{delta} T cells, they require extensive expansion to allow for fundamental, preclinical and ex vivo research. Although expansion protocols can be successful, most are based on depletion of other cell types rather than {gamma}{delta} T cell specific isolation, resulting in unpredictable purity of the isolated fraction. Moreover, the primary focus only lies with expansion of V{delta}2+ T cells, while V{delta}1+ T cells likewise have anti-tumor potential. Here, we investigated whether {gamma}{delta} T cells directly isolated from blood could be efficiently expanded while maintaining function. {gamma}{delta} T cell subsets were isolated using MACS separation, followed by FACS sorting, yielding >99% pure {gamma}{delta} T cells. Isolated V{delta}1+ and V{delta}2+ T cells could effectively expand immediately after isolation or upon freeze/thawing and reached expansion ratios between 200 to 2000-fold starting from varying numbers using cytokine supported feeder stimulations. After expansion, potential effector functions of {gamma}{delta} T cells were demonstrated by IFN-{gamma}, TNF- and granzyme B production upon PMA/ionomycin stimulation and effective killing capacity of multiple tumor cell lines was confirmed in killing assays. In conclusion, pure {gamma}{delta} T cells can productively be expanded while maintaining their anti-tumor effector functions against tumor cells. Moreover, {gamma}{delta} T cells could be expanded from low starting numbers suggesting that this protocol may even allow for expansion of cells extracted from tumor biopsies.

immunology↗

bioRxiv2019

Head Stabilization During Standing in People with Persisting Symptoms after Mild Traumatic Brain Injury

Increased postural sway is often observed in people with mild traumatic brain injury (mTBI), but our understanding of how individuals with mTBI control their head during stance is limited. The purpose of this study was to determine if people with mTBI exhibit increased sway at the head compared with healthy controls. People with persisting symptoms after mTBI (n = 59, 41 women) and control participants (n = 63, 38 women) stood quietly for one minute in four conditions: eyes open on a firm surface (EO-firm), eyes closed on a firm surface (EC-firm), eyes open on a foam pad (EO-foam), and eyes closed on foam (EC-foam). Inertial sensors at the head, sternum, and lumbar region collected tri-axial accelerations. Root-mean-square (RMS) accelerations in anteroposterior (AP) and mediolateral (ML) directions. Sway ratios between the head and sternum, head and lumbar, and sternum and lumbar region, were compared between groups. Temporal coupling of anti-phase motion between the upper and lower body angular accelerations was assessed with magnitude squared coherence and cross-spectral phase angles. People with mTBI demonstrated greater sway than controls across conditions and directions. During foam-surface conditions, the control group, but not the mTBI group, reduced ML sway at their head and trunk relative to their lumbar by increasing the expression of an anti-phase hip strategy within the frontal plane. These results are consistent with suggestions of inflexible or inappropriate postural control in people with mTBI.

neuroscience↗