Effective interface conditions for continuum mechanical models describing the invasion of multiple cell populations through thin membranes

This paper describes an advanced on-chip whole human blood fractionation and cell isolation process combining an aqueous two-phase system to create complex separation layers with a centrifugal microfluidic platform to control and automate the assay.

Download Full-text

Gamma Interferon Mediates Experimental Cerebral Malaria by Signaling within Both the Hematopoietic and Nonhematopoietic Compartments

Infection and Immunity ◽

10.1128/iai.01035-16 ◽

2017 ◽

Vol 85 (11) ◽

Cited By ~ 5

Author(s):

Ana Villegas-Mendez ◽

Patrick Strangward ◽

Tovah N. Shaw ◽

Ivana Rajkovic ◽

Vinko Tosevski ◽

...

Keyword(s):

Endothelial Cells ◽

Cerebral Malaria ◽

Myeloid Cell ◽

Gamma Interferon ◽

Cell Populations ◽

Experimental Cerebral Malaria ◽

Cerebral Pathology ◽

Multiple Cell ◽

Ifn Γ

ABSTRACT Experimental cerebral malaria (ECM) is a gamma interferon (IFN-γ)-dependent syndrome. However, whether IFN-γ promotes ECM through direct and synergistic targeting of multiple cell populations or by acting primarily on a specific responsive cell type is currently unknown. Here, using a panel of cell- and compartment-specific IFN-γ receptor 2 (IFN-γR2)-deficient mice, we show that IFN-γ causes ECM by signaling within both the hematopoietic and nonhematopoietic compartments. Mechanistically, hematopoietic and nonhematopoietic compartment-specific IFN-γR signaling exerts additive effects in orchestrating intracerebral inflammation, leading to the development of ECM. Surprisingly, mice with specific deletion of IFN-γR2 expression on myeloid cells, T cells, or neurons were completely susceptible to terminal ECM. Utilizing a reductionist in vitro system, we show that synergistic IFN-γ and tumor necrosis factor (TNF) stimulation promotes strong activation of brain blood vessel endothelial cells. Combined, our data show that within the hematopoietic compartment, IFN-γ causes ECM by acting redundantly or by targeting non-T cell or non-myeloid cell populations. Within the nonhematopoietic compartment, brain endothelial cells, but not neurons, may be the major target of IFN-γ leading to ECM development. Collectively, our data provide information on how IFN-γ mediates the development of cerebral pathology during malaria infection.

Download Full-text

Tail formation as a continuation of gastrulation: the multiple cell populations of the Xenopus tailbud derive from the late blastopore lip

Development ◽

10.1242/dev.119.4.991 ◽

1993 ◽

Vol 119 (4) ◽

pp. 991-1004 ◽

Cited By ~ 32

Author(s):

L.K. Gont ◽

H. Steinbeisser ◽

B. Blumberg ◽

E.M. de Robertis

Keyword(s):

Cell Populations ◽

Fate Map ◽

Stages Of Development ◽

Gene Markers ◽

Distinct Cell ◽

Multiple Cell ◽

Organizer Activity ◽

Dorsal Cells ◽

Late Stages ◽

Lines Of Evidence

Three lines of evidence suggest that tail formation in Xenopus is a direct continuation of events initiated during gastrulation. First, the expression of two gene markers, Xbra and Xnot2, can be followed from the blastopore lip into distinct cell populations of the developing tailbud. Second, the tip of the tail retains Spemann's tail organizer activity until late stages of development. Third, lineage studies with the tracer DiI indicate that the cells of the late blastopore are fated to form specific tissues of the tailbud, and that intercalation of dorsal cells continues during tail elongation. In particular, the fate map shows that the tip of the tail is a direct descendant of the late dorsal blastopore lip. Thus, the tailbud is not an undifferentiated blastema as previously thought, but rather consists of distinct cell populations which arise during gastrulation.

Download Full-text

Evidence for distinct lymphocytic and monocytic populations in a patient with terminal transferase--positive acute leukemia

Blood ◽

10.1182/blood.v51.6.1051.1051 ◽

1978 ◽

Vol 51 (6) ◽

pp. 1051-1056 ◽

Cited By ~ 55

Author(s):

R Mertelsmann ◽

B Koziner ◽

R Ralph ◽

D Filippa ◽

S McKenzie ◽

...

Keyword(s):

Stem Cell ◽

Acute Leukemia ◽

Clinical Course ◽

Initial Therapy ◽

Cell Populations ◽

Cell Marker ◽

Terminal Transferase ◽

Distinct Cell ◽

Multiple Cell ◽

Marker Studies

Abstract Two distinct cell populations with lymphoblastic and monocytic characteristics were separated and characterized by multiple cell markers in a patient with terminal transferase-positive acute acute leukemia. The clinical course and sequential cell marker studies were consistent with the interpretation of a defect at the level of a common stem cell giving rise to a terminal transferase--positive lymphoblastic cell population at diagnosis and, following initial therapy, a terminal transferase--negative monocytic population.

Download Full-text

Probe-Seq enables transcriptional profiling of specific cell types from heterogeneous tissue by RNA-based isolation

10.1101/735738 ◽

2019 ◽

Author(s):

Ryoji Amamoto ◽

Mauricio D. Garcia ◽

Emma R. West ◽

Jiho Choi ◽

Sylvain W. Lapan ◽

...

Keyword(s):

Transcriptional Profiling ◽

Cell Types ◽

Cell Populations ◽

Specific Cell ◽

Rna Profiling ◽

Complementary Method ◽

Dissociated Cells ◽

Multiple Cell ◽

Heterogeneous Tissue

ABSTRACTRecent transcriptional profiling technologies are uncovering previously-undefined cell populations and molecular markers at an unprecedented pace. While single cell RNA (scRNA) sequencing is an attractive approach for unbiased transcriptional profiling of all cell types, a complementary method to isolate and sequence specific cell populations from heterogeneous tissue remains challenging. Here, we developed Probe-Seq, which allows deep transcriptional profiling of specific cell types isolated using RNA as the defining feature. Dissociated cells are labelled using fluorescent in situ hybridization (FISH) for RNA, and then isolated by fluorescent activated cell sorting (FACS). We used Probe-Seq to purify and profile specific cell types from mouse, human, and chick retinas, as well as the Drosophila midgut. Probe-Seq is compatible with frozen nuclei, making cell types within archival tissue immediately accessible. As it can be multiplexed, combinations of markers can be used to create specificity. Multiplexing also allows for the isolation of multiple cell types from one cell preparation. Probe-Seq should enable RNA profiling of specific cell types from any organism.

Download Full-text

Phylogenetic analyses of melanoma reveal complex patterns of metastatic dissemination

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1508074112 ◽

2015 ◽

Vol 112 (35) ◽

pp. 10995-11000 ◽

Cited By ~ 88

Author(s):

J. Zachary Sanborn ◽

Jongsuk Chung ◽

Elizabeth Purdom ◽

Nicholas J. Wang ◽

Hojabr Kakavand ◽

...

Keyword(s):

Primary Tumor ◽

Integration Site ◽

Phylogenetic Analyses ◽

Initial Response ◽

Experimental Models ◽

Cell Populations ◽

Primary Tumors ◽

Multiple Tumor ◽

Tumor Virus ◽

Multiple Cell

Melanoma is difficult to treat once it becomes metastatic. However, the precise ancestral relationship between primary tumors and their metastases is not well understood. We performed whole-exome sequencing of primary melanomas and multiple matched metastases from eight patients to elucidate their phylogenetic relationships. In six of eight patients, we found that genetically distinct cell populations in the primary tumor metastasized in parallel to different anatomic sites, rather than sequentially from one site to the next. In five of these six patients, the metastasizing cells had themselves arisen from a common parental subpopulation in the primary, indicating that the ability to establish metastases is a late-evolving trait. Interestingly, we discovered that individual metastases were sometimes founded by multiple cell populations of the primary that were genetically distinct. Such establishment of metastases by multiple tumor subpopulations could help explain why identical resistance variants are identified in different sites after initial response to systemic therapy. One primary tumor harbored two subclones with different oncogenic mutations in CTNNB1, which were both propagated to the same metastasis, raising the possibility that activation of wingless-type mouse mammary tumor virus integration site (WNT) signaling may be involved, as has been suggested by experimental models.

Download Full-text

Evidence for distinct lymphocytic and monocytic populations in a patient with terminal transferase--positive acute leukemia

Blood ◽

10.1182/blood.v51.6.1051.bloodjournal5161051 ◽

1978 ◽

Vol 51 (6) ◽

pp. 1051-1056

Author(s):

R Mertelsmann ◽

B Koziner ◽

R Ralph ◽

D Filippa ◽

S McKenzie ◽

...

Keyword(s):

Stem Cell ◽

Acute Leukemia ◽

Clinical Course ◽

Initial Therapy ◽

Cell Populations ◽

Cell Marker ◽

Terminal Transferase ◽

Distinct Cell ◽

Multiple Cell ◽

Marker Studies

Two distinct cell populations with lymphoblastic and monocytic characteristics were separated and characterized by multiple cell markers in a patient with terminal transferase-positive acute acute leukemia. The clinical course and sequential cell marker studies were consistent with the interpretation of a defect at the level of a common stem cell giving rise to a terminal transferase--positive lymphoblastic cell population at diagnosis and, following initial therapy, a terminal transferase--negative monocytic population.

Download Full-text

Integrative transcriptomic analysis of SLE reveals IFN-driven cross-talk between immune cells

10.1101/2020.04.27.065227 ◽

2020 ◽

Author(s):

Bharat Panwar ◽

Benjamin J. Schmiedel ◽

Shu Liang ◽

Brandie White ◽

Enrique Rodriguez ◽

...

Keyword(s):

Gene Expression ◽

T Cells ◽

Autoimmune Disease ◽

Cross Talk ◽

Immune Cell ◽

Cell Types ◽

Cell Populations ◽

Multiple Cell ◽

Classical Monocytes ◽

Multiple Immune Cell

ABSTRACTThe systemic lupus erythematosus (SLE) is an incurable autoimmune disease disproportionately affecting women and may lead to damage in multiple different organs. The marked heterogeneity in its clinical manifestations is a major obstacle in finding targeted treatments and involvement of multiple immune cell types further increases this complexity. Thus, identifying molecular subtypes that best correlate with disease heterogeneity and severity as well as deducing molecular cross-talk among major immune cell types that lead to disease progression are critical steps in the development of more informed therapies for SLE. Here we profile and analyze gene expression of six major circulating immune cell types from patients with well-characterized SLE (classical monocytes (n=64), T cells (n=24), neutrophils (n=24), B cells (n=20), conventional (n=20) and plasmacytoid (n=22) dendritic cells) and from healthy control subjects. Our results show that the interferon (IFN) response signature was the major molecular feature that classified SLE patients into two distinct groups: IFN-signature negative (IFNneg) and positive (IFNpos). We show that the gene expression signature of IFN response was consistent (i) across all immune cell types, (ii) all single cells profiled from three IFNpos donors using single-cell RNA-seq, and (iii) longitudinal samples of the same patient. For a better understanding of molecular differences of IFNpos versus IFNneg patients, we combined differential gene expression analysis with differential Weighted Gene Co-expression Network Analysis (WGCNA), which revealed a relatively small list of genes from classical monocytes including two known immune modulators, one the target of an approved therapeutic for SLE (TNFSF13B/BAFF: belimumab) and one itself a therapeutic for Rheumatoid Arthritis (IL1RN: anakinra). For a more integrative understanding of the cross-talk among different cell types and to identify potentially novel gene or pathway connections, we also developed a novel gene co-expression analysis method for joint analysis of multiple cell types named integrated WGNCA (iWGCNA). This method revealed an interesting cross-talk between T and B cells highlighted by a significant enrichment in the expression of known markers of T follicular helper cells (Tfh), which also correlate with disease severity in the context of IFNpos patients. Interestingly, higher expression of BAFF from all myeloid cells also shows a strong correlation with enrichment in the expression of genes in T cells that may mark circulating Tfh cells or related memory cell populations. These cell types have been shown to promote B cell class-switching and antibody production, which are well-characterized in SLE patients. In summary, we generated a large-scale gene expression dataset from sorted immune cell populations and present a novel computational approach to analyze such data in an integrative fashion in the context of an autoimmune disease. Our results reveal the power of a hypothesis-free and data-driven approach to discover drug targets and reveal novel cross-talk among multiple immune cell types specific to a subset of SLE patients. This approach is immediately useful for studying autoimmune diseases and is applicable in other contexts where gene expression profiling is possible from multiple cell types within the same tissue compartment.

Download Full-text

High-Resolution Estimation of Multiple Cell Populations in Tissue Using Confocal Stereology

Fluorescence Microscopy ◽

10.1016/b978-0-12-409513-7.00012-9 ◽

2014 ◽

pp. 171-184 ◽

Cited By ~ 2

Author(s):

Daniel A. Peterson

Keyword(s):

High Resolution ◽

Cell Populations ◽

Multiple Cell ◽

Resolution Estimation

Download Full-text