scholarly journals Technical validation of a new microfluidic device for enrichment of CTCs from large volumes of blood by using buffy coats to mimic diagnostic leukapheresis products

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
R. Guglielmi ◽  
Z. Lai ◽  
K. Raba ◽  
G. van Dalum ◽  
J. Wu ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
White Blood Cells ◽  
High Volume ◽  
Tumor Cell Lines ◽  
Label Free ◽  
Cell Recovery ◽  
Liquid Biopsies ◽  
Technical Validation ◽  
Similar Enrichment ◽  
Blood Volumes

AbstractDiagnostic leukapheresis (DLA) enables to sample larger blood volumes and increases the detection of circulating tumor cells (CTC) significantly. Nevertheless, the high excess of white blood cells (WBC) of DLA products remains a major challenge for further downstream CTC enrichment and detection. To address this problem, we tested the performance of two label-free CTC technologies for processing DLA products. For the testing purposes, we established ficollized buffy coats (BC) with a WBC composition similar to patient-derived DLA products. The mimicking-DLA samples (with up to 400 × 106 WBCs) were spiked with three different tumor cell lines and processed with two versions of a spiral microfluidic chip for label-free CTC enrichment: the commercially available ClearCell FR1 biochip and a customized DLA biochip based on a similar enrichment principle, but designed for higher throughput of cells. While the samples processed with FR1 chip displayed with increasing cell load significantly higher WBC backgrounds and decreasing cell recovery, the recovery rates of the customized DLA chip were stable, even if challenged with up to 400 × 106 WBCs (corresponding to around 120 mL peripheral blood or 10% of a DLA product). These results indicate that the further up-scalable DLA biochip has potential to process complete DLA products from 2.5 L of peripheral blood in an affordable way to enable high-volume CTC-based liquid biopsies.

XMODULATION IN MICEAND MEN: IL-10 PRODUCING CELLS INBLOOD AND LYMPHOID TISSUE

2008 ◽  
Vol 31 (4) ◽  
pp. 3
Author(s):  
L Barrett ◽  
M Grant ◽  
R Liwski ◽  
K West
Keyword(s):  
Immune System ◽  
Peripheral Blood ◽  
Lymphoid Tissue ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
White Blood Cells ◽  
Interleukin 10 ◽  
Healthy Individuals ◽  
Human Immune System ◽  
Healthy Humans

Background: The human immune system provides remarkable protection from a plethora of pathogens, but can cause damage when activated for a prolonged time (as inpersistent infections) or against self (autoimmunity). Therefore, mechanisms of immune system downregulation and control are imperative. There is little data on how the immune system is controlled in healthy individuals. We recently described a novel population of white blood cells that constitutively produce the immunomodulatory cytokine interleukin-10 (IL-10). Our objective was to further delineate the distribution of these cells in human and mouse models, as well as potential triggers for interleukin-10 production in vitro. Methods: Human and animal protocols were reviewed and approved by the institutional ethics board and animal care facilities, and informed consent was obtained from all human donors. The ex vivo percentage of peripheral blood CD36^+IL-10^+ mononuclear cells was assessed by intracellular flow cytometry in 10 healthy individuals. IL-10 production after exposure to twoCD36 ligands, thrombospondin and oxidized low density lipoprotein (oxLDL) was measured at 8 hours. Peripheral blood mononuclear cells and splenocytes from BL/6 (n=5) and Balb/c (n=1) micewere assessed for CD36^+IL-10^+ cells ex vivo as well. Results: The percentage of CD36^+IL-10^+ cells in peripheral blood fromhealthy individuals ranges between 0.1% and 0.9%. The percentage was similar in mouse peripheral blood, with a range of 0.4%-1.1%. These cells were also found in mouse spleen at a higher frequency than peripherally (1.1-1.5%). Human CD36^+IL-10^+ cells have more IL-10 when exposed to thrombospondin, oxLDL. Conclusions: Our novel population of IL-10 producing cells is found not only in healthy humans, but also in lymphoid tissue and blood from pathogen free mice. This highlights the evolutionary conservation of the cell across species, and suggests an important homeostatic function. The physiologic ligands for CD36 are ubiquitous in circulation, and ourin vitro data suggests a link between CD36 ligation and IL-10 production. IL-10 is a known immune system modulator, and its production by these cells may help maintain homeostaticcontrol of the immune system.

Abstract MP243: Detection Of Mesenchymal Stem Cells In Mobilized Autologous Peripheral Blood Transplantation From Patients With Ischemic Heart Disease

2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Hadyanto Lim ◽  
Umar Zein ◽  
Ilham Hariaji
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Heart Disease ◽  
Ischemic Heart Disease ◽  
Peripheral Blood ◽  
Autologous Transplantation ◽  
White Blood Cells ◽  
Significant Rise ◽  
Ischemic Heart ◽  
Post Transplantation

Background: Mesenchymal stem cells (MSCs) improve the cardiac function and remodeling in patients with ischemic heart disease. However, their presence in the circulating peripheral blood and post-transplantation has not been fully elucidated. We aimed to investigate the effects of intravenous transplantation of mobilized autologous peripheral blood on the number of MSCs in patients with ischemic heart disease. Methods: Granulocyte-colony stimulating factor (G-CSF, 5.0 μg/kg/day) was given subcutaneously once a day for five days to 7 patients (4 women and 3 men, aged 54-69 years) with ischemic heart disease. Leukapheresis procedure was started on the day 5 of G-CSF using the Spectra Optia cell separator. Circulating and intravenous transplantation of autologous MSCs after leukapheresis were analyzed by flow cytometry. MSCs were identified on the basis of dual positive cells (CD73 + /CD105 + or CD90 + /CD73 + or CD90 + /CD105 + ) and detected as MSCs if a cluster of at least 10 cells could be found. Results: MSCs in the circulating peripheral blood and after transplantation were detected in 2 (28%) and 6 (85%) patients, respectively. The frequency of intravenous peripheral blood MSCs increased significantly after transplantation (from 32.57 ± 22.76 x10 -4 % to 58.57 ± 28.49 x 10 -4 % , p<0.001). Moreover, there were significant rise in the total white blood cells count (from 10.25 ± 4.86 x 10 3 /μl to 35.81 ± 7.07 x 10 3 /μl, p<0.001) and the levels of CD34 + cells (from 1.17 ± 0.93 cells/μL to 138.30 ± 11.26 cells/μL, p<0.001) after the infusion. Conclusions: The results show that intravenous transplantation of mobilized autologous peripheral blood increases the number of MSCs in patients with ischemic heart disease. Leukapheresis product of peripheral blood MSCs could therefore be a potential source for autologous transplantation in ischemic heart disease.

scholarly journals Adult Mice With Targeted Mutation of the Interleukin-11 Receptor (IL11Ra) Display Normal Hematopoiesis

Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2148-2159 ◽  
Author(s):  
Harshal H. Nandurkar ◽  
Lorraine Robb ◽  
David Tarlinton ◽  
Louise Barnett ◽  
Frank Köntgen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Bone Marrow Cells ◽  
White Blood Cells ◽  
Null Mutation ◽  
Wild Type ◽  
Normal Numbers ◽  
Interleukin 11 ◽  
Marrow Cells

Abstract Interleukin-11 (IL-11) is a pleiotropic growth factor with a prominent effect on megakaryopoiesis and thrombopoiesis. The receptor for IL-11 is a heterodimer of the signal transduction unit gp130 and a specific receptor component, the α-chain (IL-11Rα). Two genes potentially encode the IL-11Rα: the IL11Ra and IL11Ra2 genes. The IL11Ra gene is widely expressed in hematopoietic and other organs, whereas the IL11Ra2 gene is restricted to only some strains of mice and its expression is confined to testis, lymph node, and thymus. To investigate the essential actions mediated by the IL-11Rα, we have generated mice with a null mutation of IL11Ra (IL11Ra−/−) by gene targeting. Analysis of IL11Ra expression by Northern blot and reverse transcriptase-polymerase chain reaction, as well as the absence of response of IL11Ra−/− bone marrow cells to IL-11 in hematopoietic assays, further confirmed the null mutation. Compensatory expression of the IL11Ra2 in bone marrow cells was not detected. IL11Ra−/− mice were healthy with normal numbers of peripheral blood white blood cells, hematocrit, and platelets. Bone marrow and spleen contained normal numbers of cells of all hematopoietic lineages, including megakaryocytes. Clonal cultures did not identify any perturbation of granulocyte-macrophage (GM), erythroid, or megakaryocyte progenitors. The number of day-12 colony-forming unit-spleen progenitors were similar in wild-type and IL11Ra−/− mice. The kinetics of recovery of peripheral blood white blood cells, platelets, and bone marrow GM progenitors after treatment with 5-flurouracil were the same in IL11Ra−/− and wild-type mice. Acute hemolytic stress was induced by phenylhydrazine and resulted in a 50% decrease in hematocrit. The recovery of hematocrit was comparable in IL11Ra−/− and wild-type mice. These observations indicate that IL-11 receptor signalling is dispensable for adult hematopoiesis.

scholarly journals Tumor-specific CD8+ T lymphocytes derived from the peripheral blood of prostate cancer patients byin vitro stimulation with autologous tumor cell lines

2002 ◽  
Vol 98 (1) ◽  
pp. 57-62 ◽  
Author(s):  
Franck Housseau ◽  
Daniel A. Langer ◽  
Samuel D. Oberholtzer ◽  
Anitha Moorthy ◽  
Hyam I. Levitsky ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
T Lymphocytes ◽  
Tumor Cell ◽  
Cancer Patients ◽  
Cell Lines ◽  
Peripheral Blood ◽  
Autologous Tumor Cell ◽  
Autologous Tumor ◽  
Tumor Cell Lines ◽  
Cd8 T Lymphocytes

Composition of peripheral blood leukocytes and immunocompetent organs of white char in the lake Kronotskoye

2021 ◽  
pp. 42-49
Author(s):  
Ilya Ivanovich Gordeev Ilya Ivanovich Gordeev
Keyword(s):  
Fish Species ◽  
Peripheral Blood ◽  
White Blood Cells ◽  
Head Kidney ◽  
Stress Factors ◽  
Lake Basin ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Immunocompetent Organs ◽  
Blood Smears

The composition of leukocytes reflects the species features of fish. The ratio of individual cell types reflects the functional state of the organism and the nature of the influence of biotic and abiotic stress factors. Kronotskoye Lake is located in the Far East on the territory of the Kronotsky State Natural Biosphere Reserve (Kamchatka). Due to the peculiarities of the hydrological regime and the relief of the Kronotskaya River basin, anadromous migration along it is limited. Therefore, anadromous fish do not return to the lake after the marine period of their life cycle. The lake basin is inhabited by at least six reproductively isolated forms (species) of chars. White char, Salvelinus albus Glubokovsky, 1977, at the early stages of ontogenesis feeds on invertebrates, and as it grows up, it goes on to a predatory lifestyle. White char migrates around the lake: spawning occurs in the tributaries, and feeding takes place in the lake itself. This work is devoted to the study of the intensity of leukopoiesis and leukocyte composition of the blood, head kidney and spleen of the white char of Lake Kronotskoye. Seven individuals caught using gill nets from July 8 to August 14, 2011 were studied. Peripheral blood was sampled from the tail vein. Sampling also included spleen and head kidney tissues, blood smears and organ-imprinted smears of kidney and spleen. The leukocyte formula and the frequency of occurrence of leukocytes in a peripheral blood smears were calculated using light microscope. The composition of white blood cells in the white char is similar to that of other fish species (lymphocytes, monocytes, neutrophils, and blast cell forms). No basophils or eosinophils were found. In organ-imprinted smears of immunocompetent organs (kidney, spleen), the percentage of lymphocytes is lower and the percentage of blast forms is higher than in the blood leukogram. The abundance index differed from the data obtained in the study of other fish species. English version of the article is available at URL: https://panor.ru/articles/composition-of-peripheral-blood-leukocytes-and-immunocompetent-organs-of-the-white-char-of-lake-kronotskoye-kamchatka/68317.html

Development of a robust algorithm for detection of nuclei of white blood cells in peripheral blood smear images

2019 ◽  
Vol 78 (13) ◽  
pp. 17879-17898 ◽  
Author(s):  
Roopa B. Hegde ◽  
Keerthana Prasad ◽  
Harishchandra Hebbar ◽  
Brij Mohan Kumar Singh
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Blood Smear ◽  
White Blood Cells ◽  
Peripheral Blood Smear ◽  
Robust Algorithm

P1800RITUXIMAB ENHANCES THE EXPRESSION OF INTERLEUKIN-6 VIA TOLL-LIKE RECEPTOR 3 SIGNALING IN HUMAN PODOCYTE

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Shojiro Watanabe ◽  
Koji Hirono ◽  
Tomomi Aizawa ◽  
Koji Tsugawa ◽  
Hiroshi Tanaka
Keyword(s):  
Nephrotic Syndrome ◽  
B Cell ◽  
Peripheral Blood ◽  
Sustained Remission ◽  
Dependent Manner ◽  
Toll Like Receptor ◽  
Cell Recovery ◽  
Viral Dsrna ◽  
Polycytidylic Acid ◽  
Tlr3 Signaling

Abstract Background and Aims The etiology of idiopathic nephrotic syndrome (INS) in children remains unclear, but it is well known that viral infections often cause relapses of INS. Since toll-like receptor 3 (TLR3), a ligand of viral dsRNA exists in podocytes, dysregulation of TLR3 signaling in podocytes may be involved in the pathogenesis of INS. Recently, rituximab (RTX) a specific antibody to human CD20, has been successfully used to treat children with intractable nephrotic syndrome (frequent relapse nephrotic syndrome, FRNS, and steroid dependent nephrotic syndrome, SDNS). It has been reported that depletion of B cell is main mechanism of RTX treatment. However, some patients experienced recurrence before peripheral blood B cell recovery, whereas selected patients have sustained remission even after peripheral blood B cell recovery. Considering that dramatic effects of RTX in the treatment of INS, it is speculated that some B cell-independent mechanisms of RTX exist. In this context, some recent studies have reported a direct effect of RTX on podocyte, targeting sphingomyelinase phosphodiesterase acid-like 3b (SMPDL3b), but detailed mechanisms remain to be determined. In this study, we examined whether RTX influences TLR3 signaling in cultured human podocyte. Method Immortalized human podocytes in culture were treated with polyinosinic-polycytidylic acid (poly IC), a synthetic analogue of viral dsRNA. RTX were added before or after the treatment of poly IC. Then, expression of interleukin (IL)-6 were measured using real time RT-PCR and ELISA. Direct binding of RTX to podocyte were confirmed by immunofluorescence. Results Poly IC induced the expression of IL-6 in cultured human podocyte in a concentration and time-dependent manner. IL-6 expression induced by poly IC were enhanced in both mRNA and protein level by pretreatment of RTX. RTX itself were apparently stained on podocyte by immunofluorescence. Conclusion RTX binds directly to cultured human podocyte and enhances the IL-6 expression induced by poly IC. Since IL-6 from podocyte was reported to be engaged in regulating glomerular inflammation by cross-talk with endothelial cells, exploring the role of enhanced IL-6 from podocyte by RTX, at least in a part, suggests a direct mechanism of RTX in treating intractable nephrotic syndrome in children.

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