bioRxiv Science⌕ Search

Biology subjects

Niesner, R. A.

Publications and source records attributed to Niesner, R. A..

4 recordsLinked to original sources

Anomalous diffusion analysis reveals cooperative locomotion of adult parasitic nematodes in sex-mixed groups.

Parasitic worms are widespread throughout the world and causing agents of chronic infections in humans and animals. The majority of these pathogens parasitize the gut of the infected hosts, however very little is known about the locomotion of the worms dwelling the gut. We studied the movement of Heligmosomoides bakeri (previously Heligmosomoides polygyrus), a natural infection of mice and usually used as animal model to study the roundworm infections. We investigated the locomotion of H. bakeri in simplified environments mimicking key physical features of the intestinal lumen, i.e. various medium viscosities, and a periodical intestinal villi topography. The non-periodical nematode motion in these settings could be described by anomalous diffusion theory. Fascinatingly, an oriented, super-diffusive locomotion of nematodes in sex-mixed groups were detected, in the sense of a cooperative, but not collective (swarm-like) migration stimulated by mating and reproduction, while individual nematodes moved randomly, following a diffusive motion regime. High mucus-like medium viscosity and villi topography, representing physical constraints of nematode locomotion, slowed down but did not prevent this cooperative migration. Additionally, the mean displacement rate of nematodes in sex-mixed groups of 6{middle dot}10-4 mm/s in viscous mucus-like medium are in good agreement with estimates of nematode migration velocities between 10-4 to 10-3 mm/s in the gut. Thus, our data indicate the intestinal nematodes motion to be non-periodic and random but triggered to be oriented by kin of the different sex.

biophysics↗

In vivo optimized three-photon imaging of intact mouse tibia links plasma cell motility to functional states in the bone marrow.

Intravital multi-photon imaging of the bone marrow is crucial to the study of cellular dynamics, communication with the microenvironment and functions, however, imaging of deep tissue areas is challenging and minimally invasive methods for deep-marrow imaging in intact long bones are needed. We developed a high pulse energy 1650 nm laser prototype, which permits to surpass >100 {micro}m thick cortical bone and to perform three-photon microscopy (3PM) in more than 400 {micro}m depth in the marrow cavity of intact mouse tibia in vivo. Its unique 3 and 4 MHz laser repetition rates allowed us to analyze motility patterns of rare cells over large fields of view deep within the unperturbed marrow. In this way, we found a bi-modal migratory behavior of marrow plasma cells. Besides, the analysis of third harmonics generation (THG) in the tibia identified this signal to be a label-free indicator of the abundance of cellular organelles, in particular the endoplasmic reticulum, reflecting protein biosynthesis capacity. We found that only one third of the plasma cells in the tibia marrow of adult mice have a strong THG signal and, thus, a high protein synthesis capacity, while the other two thirds of plasma cells display a low THG signal. Finally, we identified an inverse link between migratory behavior and THG signal strength in marrow plasma cells. As in these cells, the protein biosynthesis capacity indicated by a strong THG signal is mainly associated with antibody secretion, we could relate motility to functional states of plasma cells in vivo. Our 3PM method retains the ability to connect cellular dynamics to protein biosynthesis capacity in various marrow cell types beyond plasma cells, as THG is a ubiquitous signal, opening new perspectives on understanding how tissue microenvironment impacts on cellular functions in the bone marrow.

biophysics↗

Intravital quantification of absolute cytoplasmic B cell calcium reveals dynamic signaling across B cell differentiation stages

Development, function and maintenance of lymphocytes largely depend upon the cellular mobilization and storage of Ca2+ ions. In B lymphocytes, the absolute amount of calcium mobilized and retained after cell signaling remains unknown, athough it is a crucial part of their selection within germinal centers and differentiation into plasma cells. Here, we introduce the novel reporter mouse strain YellowCaB that expresses the genetically encoded calcium indicator TN-XXL in CD19+ lymphocytes. The construct consists of the electrondonor fluorophore eCFP and the acceptor citrine, linked by a calcium sensitive domain. Its conformation and therefore donor quenching is directly linked to cytosolic calcium concentrations. By combining intravital two-photon fluorescence lifetime microscopy with our numerical approach for phasor-based analysis, we are able to extract absolute cytoplasmic calcium concentrations in activated B cells for the first time in vivo. We show that calcium concentrations in B cells are highly dynamic and fluctuations persist in extrafollicular B cells with functional relevance.

immunology↗

Low-Density Granulocytes are a novel immunopathological feature in both Multiple Sclerosis and Neuromyelitis optica spectrum disorder.

ObjectiveTo investigate whether low-density granulocytes (LDGs) are a immunophenotypic feature of patients with multiple sclerosis (MS) and/or neuromyelitis optica spectrum disorder (NMOSD).\n\nMethodsBlood samples were collected from 26 patients with NMOSD and 20 patients with MS, as well as from 18 patients with Systemic Lupus Erythematosus (SLE) and 23 Healthy Donors (HD). We isolated peripheral blood mononuclear cells (PBMCs) with density gradient separation and stained the cells with antibodies against CD14, CD15, CD16, and CD45, and analysed the cells by flow cytometry or imaging flow cytometry. We defined LDGs as CD14-CD15high and calculated their share in total PBMC leukocytes (CD45+) as well as the share of CD16hi LDGs. Clinical data on disease course, medication, and antibody status were obtained.\n\nResultsLDGs were significantly more common in MS and NMOSD than in HDs, comparable to SLE samples (median values HD 0.2%, MS 0.9%, NMOSD 2.1%, SLE 4.3%). 0/23 of the HDs, but 17/20 NMOSD and 11/17 MS samples as well as 13/15 SLE samples had at least 0.7 % LDGs. NMOSD patients without continuous immunosuppressive treatment had significantly more LDGs compared to their treated counterparts. LDG nuclear morphology ranged from segmented to rounded, suggesting a heterogeneity within the group.\n\nConclusionLDGs are a feature of the immunophenotype in some patients with MS and NMOSD.

immunology↗