CD105-FITC (clone 2H6F11)

An article published this year in “BIORXIV” using our CD105-FITC (clone 2H6F11), by our customers from Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Santander, Spain, in the analysis of Anoxia-Induced VEGF-Release from Rat Cardiomyocites Promotes Vascular Differentiation of Human Mesenchymal Stem Cells. Congrats and Thanks.

Summary:

Background. Albeit several studies show that cellular therapy with bone marrow mesenchymal stem cells (BM-hMSCs) improves cardiac function after myocardial infarction (MI), the underlying mechanism is subject of controversy. Here we hypothesized that soluble factors, including VEGF, secreted by cardiomyocytes and BM-hMSCs under a low oxygen environment promote vascular differentiation of BM-hMSCs. Methods. Human BM-MSCs were isolated and expanded in vitro by the same procedure we employed to prepare cells suitable for cardiac cell therapy. BM-hMSCs were characterized by flow cytometry and functional analysis. Vascular differentiation was induced by VEGF or by conditioned media from neonate rat ventricular cardiomyocytes (NRVCs) cultured in anoxia and confirmed by immunostaining, tube formation over matrigel and cell migration across transwells. Presence of VEGF in conditioned media was determined by ELISA and activation of VEGF signaling by Western blot. Results. BM-hMSCs used in this study met the criteria recommended by International Society for Cellular Therapy for defining mesenchymal stromal cells. Von Willebrand factor (vWF) expression and tube formation in matrigel indicate that these cells had the capacity to differentiate into the endothelial lineage, which was further enhanced by VEGF and conditioned media from NRVCs cultured in anoxia. Furthermore, condition media and VEGF stimulated cell migration across transwells, which demonstrates the migratory capacity of BM-hMSCs. Finally, when VEGF signaling was blocked by neutralizing anti-VEGF, vascular differentiation of BM-hMSCs was reduced to basal levels. Conclusions. Soluble VEGF released in the culture media after exposure to low oxygen conditions is responsible for endothelial differentiation of BM-hMSCs.

Reference:

http://biorxiv.org/content/early/2017/02/22/110908

Product link:

http://www.immunostep.com/antibodies/2792-cd105.html

CD105-PE (clone 2H6F11)

An article published this year in “PLOS ONE” using our CD105-PE (clone 2H6F11), by our customers from Unit of Blood Diseases and Stem Cells Transplantation, Department of Clinical and Experimental Sciences, University of Brescia, ASST Spedali Civili di Brescia, Italy, in the analysis of how Mesenchymal stromal cells (MSCs) induce ex vivo proliferation and erythroid commitment of cord blood haematopoietic stem cells (CB-CD34+ cells). Congrats and Thanks.

Summary:

A human bone marrow-derived mesenchymal stromal cell (MSCs) and cord blood-derived CD34+ stem cell co-culture system was set up in order to evaluate the proliferative and differentiative effects induced by MSCs on CD34+ stem cells, and the reciprocal influences on gene expression profiles. After 10 days of co-culture, non-adherent (SN-fraction) and adherent (AD-fraction) CD34+ stem cells were collected and analysed separately. In the presence of MSCs, a significant increase in CD34+ cell number was observed (fold increase = 14.68), mostly in the SN-fraction (fold increase = 13.20). This was combined with a significant increase in CD34+ cell differentiation towards the BFU-E colonies and with a decrease in the CFU-GM. These observations were confirmed by microarray analysis. Through gene set enrichment analysis (GSEA), we noted a significant enrichment in genes involved in heme metabolism (e.g. LAMP2, CLCN3, BMP2K), mitotic spindle formation and proliferation (e.g. PALLD, SOS1, CCNA1) and TGF-beta signalling (e.g. ID1) and a down-modulation of genes participating in myeloid and lymphoid differentiation (e.g. PCGF2) in the co-cultured CD34+ stem cells. On the other hand, a significant enrichment in genes involved in oxygen-level response (e.g. TNFAIP3, SLC2A3, KLF6) and angiogenesis (e.g. VEGFA, IGF1, ID1) was found in the co-cultured MSCs. Taken together, our results suggest that MSCs can exert a priming effect on CD34+ stem cells, regulating their proliferation and erythroid differentiation. In turn, CD34+ stem cells seem to be able to polarise the BM-niche towards the vascular compartment by modulating molecular pathways related to hypoxia and angiogenesis.

Reference:

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0172430

Product link:

http://www.immunostep.com/antibodies/2793-cd105.html

FITC Annexin V Apoptosis Detection Kit with PI.

An article published this year in “ONCOTARGET” using our FITC Annexin V Apoptosis Detection Kit with PI, by our customers from Department of Medicine, Hematology and Clinical Immunology Section, University of Padova, Padova, Italy, in the analysis of how Inactivation of CK1α in multiple myeloma empowers drug cytotoxicity by affecting AKT and β-catenin survival signaling pathways. Congrats and Thanks.

Summary:

Recent evidence indicates that protein kinase CK1α may support the growth of multiple myeloma (MM) plasma cells. Here, by analyzing a large cohort of MM cases, we found that high CK1α mRNA levels are virtually associated with all MM patients. Moreover, we provided functional evidence that CK1α activity is essential for malignant plasma cell survival even in the protective niche generated by co-cultures with bone marrow stromal cells. We demonstrated that CK1α inactivation, while toxic for myeloma cells, is dispensable for the survival of healthy B lymphocytes and stromal cells. Disruption of CK1α function in myeloma cells resulted in decreased Mdm2, increased p53 and p21 and reduced expression of β-catenin and AKT. These effects were mediated partially by p53 and caspase activity. Finally, we discovered that CK1α inactivation enhanced the cytotoxic effect of both bortezomib and lenalidomide. Overall, our study supports a role for CK1α as a potential therapeutic target in MM in combination with proteasome inhibitors and/or immunomodulatory drugs.

Reference:

https://www.researchgate.net/publication/312328422_Inactivation_of_CK1a_in_multiple_myeloma_empowers_drug_cytotoxicity_by_affecting_AKT_and_b-catenin_survival_signaling_pathways

http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path%5B%5D=14654

Product link:

http://www.immunostep.com/apoptosis-tools/3727-anxvkf-100t.html

CF-Blue™ - anti Human, Cross-Reactivity CD3 - UCHT1

An article published this year in “PLOS ONE” using our Anti-CD3 CF-Blue™ (clone UCHT-1), by our customers from Immunology Service, University Hospital Marqués de Valdecilla-IDIVAL, Santander, Spain, Tissue Typing Laboratory, University Hospital Marqués de Valdecilla, Santander, Spain, in the Improvement in the definition of anti-HLA antibody profile in highly sensitized patients. Congrats and Thanks.

Summary:

The definition of anti-HLA antibody profile in highly sensitized patients on a waiting list is crucial when virtual crossmatch is used in organ allocation systems, but also when used to identify the true deleterious anti-HLA antibodies. Here we propose different levels of risk based on the results of anti-HLA antibody testing in neat serum (N) and after sera dilution (DIL) and C1q test in 18 highly sensitized patients. This group was heterogeneous in terms of anti-HLA antibody titers and their ability to fix complement. After dilution, 15 out of 18 patients (83.3%) showed a reduction of positive bead counts whereas 4 patients showed a prozone effect and complement fixation was demonstrated. The high dilution of sera and ascertaining the complement fixation allow the accurate definition of risk anti-HLA antibody profiles in highly sensitized patients, as demonstrated in 5 of the sensitized patients who were transplanted.

Reference:

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0171463

Product link:

http://www.immunostep.com/inicio/3779-cd3.html?search_query=UCHT-1&results=6

CD14-APC, CD16-PE, anti-HLA-DR-FITC

An article published this year in “JOURNAL OF IMMUNOLOGY” using our allophycocyanin-conjugated anti-human CD14, PE-conjugated anti-human CD16 (B73.1) and FITC-conjugated anti–HLA-DR, by our customers from Tumor Immunology Laboratory, Hospital La Paz Institute for Health Research, La Paz University Hospital, Madrid, Spain, in the study of how Circulating Monocytes Exhibit an Endotoxin Tolerance Status after Acute Ischemic Stroke: Mitochondrial DNA as a Putative Explanation for Poststroke Infections. Congrats and Thanks.

Summary:

Patients with acute ischemic stroke (AIS) suffer from infections associated with mortality. The relevance of the innate immune system, and monocytes in particular, has emerged as an important factor in the evolution of these infections. The study enrolled 14 patients with AIS, without previous treatment, and 10 healthy controls. In the present study, we show that monocytes from patients with AIS exhibit a refractory state or endotoxin tolerance. The patients were unable to orchestrate an inflammatory response against LPS and expressed three factors reported to control the evolution of human monocytes into a refractory state: IL-1R–associated kinase-M, NFkB2/p100, and hypoxia-inducible factor-1α. The levels of circulating mitochondrial DNA (mtDNA) in patients with AIS correlated with impaired inflammatory response of isolated monocytes. Interestingly, the patients could be classified into two groups: those who were infected and those who were not, according to circulating mtDNA levels. This finding was validated in an independent cohort of 23 patients with AIS. Additionally, monocytes from healthy controls, cultured in the presence of both sera from patients and mtDNA, reproduced a refractory state after endotoxin challenge. This effect was negated by either a TLR9 antagonist or DNase treatment. The present data further extend our understanding of endotoxin tolerance implications in AIS. A putative role of mtDNA as a new biomarker of stroke-associated infections, and thus a clinical target for preventing poststroke infection, has also been identified.

Reference:

http://www.jimmunol.org/content/early/2017/01/20/jimmunol.1601594

Product link:

http://immunostep.com/antibodies/1765-cd14.html

http://immunostep.com/antibodies/2555-cd16.html

http://immunostep.com/antibodies/1907-hla-dr.html

Cell Cycle Analysis (PI / RNASE) solution.

An article published this year in “JOURNAL OF CELL BIOLOGY” using our Cell Cycle Analysis (PI / RNASE) solution, by our customers from State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China, in the study of how Phosphoglycerate mutase 1 regulates dNTP pool and promotes homologous recombination repair in cancer cells. Congrats and Thanks.

Summary:

Glycolytic enzymes are known to play pivotal roles in cancer cell survival, yet their molecular mechanisms remain poorly understood. Phosphoglycerate mutase 1 (PGAM1) is an important glycolytic enzyme that coordinates glycolysis, pentose phosphate pathway, and serine biosynthesis in cancer cells. Herein, we report that PGAM1 is required for homologous recombination (HR) repair of DNA double-strand breaks (DSBs) caused by DNA-damaging agents. Mechanistically, PGAM1 facilitates DSB end resection by regulating the stability of CTBP-interacting protein (CtIP). Knockdown of PGAM1 in cancer cells accelerates CtIP degradation through deprivation of the intracellular deoxyribonucleotide triphosphate pool and associated activation of the p53/p73 pathway. Enzymatic inhibition of PGAM1 decreases CtIP protein levels, impairs HR repair, and hence sensitizes BRCA1/2-proficient breast cancer to poly(ADP-ribose) polymerase (PARP) inhibitors. Together, this study identifies a metabolically dependent function of PGAM1 in promoting HR repair and reveals a potential therapeutic opportunity for PGAM1 inhibitors in combination with PARP inhibitors.

Reference:

http://jcb.rupress.org/content/early/2017/01/24/jcb.201607008

Product link:

http://immunostep.com/solutions-buffers-chemicals/1289-pi-rnase-solution.html