cd44 high  (Miltenyi Biotec)

Journal: NPJ Vaccines

Article Title: Preserved efficacy of lyophilized SARS-CoV-2 mRNA vaccine incorporating novel ionizable lipids after one year at 25 °C

doi: 10.1038/s41541-025-01201-1

Figure Lengend Snippet: a Immunization scheme and sample collection schedule. Mice ( n = 5) were immunized intramuscularly at day 0 (prime) and 21(boost) with 1 µg of mRNA/animal. Blood samples were obtained at week 3 (prior to boost) and 6, and specific antibody levels were determined in serum. T-cell response was evaluated in splenocytes 3 weeks post-boost. b Reciprocal endpoint titers of antigen-specific IgG antibodies in serum samples determined by ELISA using RBD recombinant protein from wild-type variant. c Neutralization titers in serum samples were determined by neutralization assay using Spike-pseudotyped lentivirus and infection in HEK293T-ACE2-TMPRSS2 cells. NT50 titers refer to the dilution of a serum sample at which 50% of the pseudovirus infection is inhibited. d IFN-γ-secreting splenocytes quantification by ELISPOT after O.N. stimulation with SARS-CoV-2 Spike peptide pool. e IFN-γ quantification in supernatants of splenocytes after overnight stimulation with SARS-CoV-2 Spike peptide pool determined by ELISA. f T-lymphocyte CD4 and CD8 cell frequencies in spleen analyzed by flow cytometry. g Tfh and GC B cells analysis. Mice ( n = 5) were immunized intramuscularly with 5 µg of mRNA/animal. Inguinal lymph nodes were extracted on day 7. h Tfh (CD45R - , CD4 + , CD44hi, CXCR5 + , PD-1 + ) and GC B cell (CD19 + , CD95 + , GL7 + ) frequencies were determined by flow cytometry analysis. Graphs are represented as mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 as determined by two-way ANOVA ( a ) and one-way ANOVA ( c – h ) with Tukey post-test.

Article Snippet: Tfh cells were defined as CD45R - CD4 + , CD44 high , CXCR5 + , PD-1 + , and the following antibodies were used for surface staining: CD45R (B220)-APC-Vio770 (Miltenyi, 130-110-849), CD4-VioBright FITC (Miltenyi, 130-118-692), CD44-PerCP-Vio700 (Miltenyi, 130-128-625), CD185 (CXCR5)-APC (Miltenyi, 130-119-129), and CD279 (PD1)-PE (Miltenyi, 130-111-953).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Variant Assay, Neutralization, Infection, Enzyme-linked Immunospot, Flow Cytometry

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: Potentiation of 17-allylamino-17-demethoxygeldanamycin (17-AAG) cytotoxicity by nonsteroidal anti-inflammatory drugs (NSAIDs). (A) Cell surface expression of CD44 in CD44 high and parental K562 cells was quantified by flow cytometry after labeling both cells with anti-CD44 antibody. (B) CD44 high K562 cells were treated with serial doses of 17-AAG in the presence or absence of celecoxib (CCB). The percentage of cell survival was determined after 96 h of incubation using MTT assay. Each bar represents the mean ± SD of triplicate experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Expressing, Flow Cytometry, Labeling, Incubation, MTT Assay

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: (A) Enhancement of 17-AAG-induced apoptosis by NSAIDs. CD44 high K562 cells were treated with 17-AAG in the absence (w/o) or presence of CCB or ibuprofen (IBU) for 24 h, and the percentage of apoptotic cells was quantified using FACS. The upper right quadrants contain late apoptotic cells (positive for both PI and annexin V), and the lower right quadrants represent early apoptotic cells (annexin V + and PI). Images shown are representative of three independent experiments. (B) Bar graph shows mean percentage of apoptotic cells, and values represent the means ± SD. * p < 0.05, ** p < 0.01.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques:

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: Induction of autophagy and downregulation of stemness-related markers by NSAIDs, and CCB-mediated reduction in CD44 cell surface expression. CD44 high K562 cells were treated with serial doses of CCB (A), IBU (B) for 24 h, and the changed levels of autophagy (LC3B-1/II and p62) and expression of stemness-related markers (mutp53, c-Myc, CD44, and Oct4) were determined by Western blot analysis. Actin was used as a loading control. (C) CD44 high K562, CEM/VLB 100 , and MCF7-MDR cells were treated with serial doses of CCB for 24 h, and the change in CD44 cell surface expression of each cell was determined by FACS.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Expressing, Western Blot, Control

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: Comparison of autophagic-inducing effect of CCB derivatives and rapamycin, and autophagic degradation of stemness-related markers by CCB. CD44 high K562 cells were treated with CCB or 2,5-dimethyl-celecoxib (DMC) (A) or OSU-03012 or rapamycin (B) for 24 h. (C) The cells were treated with or without 25 μM CCB for 24 h and were collected at 0, 3, and 6 h after, following treatment with 20 μg/ml cycloheximide (CHX), and the levels of stemness-related marker proteins were determined by Western blot analysis.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Comparison, Marker, Western Blot

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: Crosstalk between CCB-induced apoptotic and autophagic cell death. (A) CD44 high K562 cells were treated with 25 μM CCB in the absence or presence of 10 mM 3-methyladenine (3-MA, left) or 5 mM chloroquine (CQ, right) for 24 h, and levels of p62 and stemness-related markers were determined by Western blot analysis. (B) CD44 high K562 cells were treated with CCB in the absence (w/o) or presence of Z-DEVD-FMK, and the percentages of apoptotic cells were quantified by FACS for annexin V and PI staining. Images shown are representative of three independent experiments (left). Bar graph shows mean percentage of early and late apoptotic cells, and values represent the means ± SD. *** p < 0.001.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Western Blot, Staining

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: NSAID-induced activation of AMPK and inhibition of Akt/mTOR signaling and disruption of Beclin-1/Mcl-1 complex via inhibition of STAT3 signaling pathway. (A) CD44 high K562 cells were treated with increasing doses of CCB for 36 h, and then levels of indicated molecules were determined by Western blot analysis. (B) CD44 high K562 cells were treated with increasing doses of CCB or IBU for 24 h, and the levels of STAT3 and Mcl-1 were determined by Western blot analysis. (C) Mcl-1 or Beclin-1 was immunoprecipitated from CD44 high K562 cells treated with 25 μM CCB for 24 h and analyzed for the presence of Mcl-1 or Beclin-1.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Activation Assay, Inhibition, Disruption, Western Blot, Immunoprecipitation

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: Enhancement of 17-AAG-induced autophagy, acceleration of autophagic degradation of CSC marker, and PARP activation by NSAID. CD44 high K562 cells were treated with 17-AAG in the presence or absence of 25 μM CCB (A) or 400 μM IBU (B) for 24 h. The changes in autophagy induction and levels of stemness-related markers, and cleaved PARP (cle PARP) were determined by Western blot analysis.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Marker, Activation Assay, Western Blot

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: Downregulation of HSF1/Hsps and suppression of 17-AAG-mediated Hsp70 by NSAID. (A) CD44 high K562 cells were treated with serial doses of CCB or IBU. (B) CD44 high K562 cells were treated with 17-AAG in the presence or absence of 25 μM CCB or 400 μM IBU for 24 h. The changed levels of HSF1/Hsps were determined by Western blot analysis.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Western Blot

Journal: Oncology Research

Article Title: Nonsteroidal Anti-inflammatory Drugs Sensitize CD44-Overexpressing Cancer Cells to Hsp90 Inhibitor Through Autophagy Activation

doi: 10.3727/096504019X15517850319579

Figure Lengend Snippet: A proposed model of molecular targets of NSAIDs in enhancing Hsp90 inhibitor cytotoxicity of CD44 high K562 cells. Inhibition of Hsp90 leads to disruption of regulatory complexes of Hsp90 with HSF1 and client proteins involving mutp53, thereby causing HSF1-mediated induction of Hsp70, which is responsible for the resistance of CD44 high K562 cells to Hsp90 inhibitor. NSAIDs inhibit phosphorylation of STAT3 that can upregulate HSF-1, resulting in reduced resistance against Hsp90 inhibitor in the cells. NSAIDs can promote autophagic cell death through inhibition of Akt/mTOR/p70S6K pathway and downregulation of Bcl-2/Mcl-1, which trigger autophagy-mediated degradation of multiple stemness-related marker proteins.

Article Snippet: Moreover, cell surface expression of CD44 in CEM/VLB 100 and MCF7/MDR cells as well as CD44 high K562 cells was significantly decreased by CCB, dose dependently.

Techniques: Inhibition, Disruption, Phospho-proteomics, Marker

Journal: PLoS ONE

Article Title: Anthocyanin Extracted from Black Soybean Seed Coats Prevents Autoimmune Arthritis by Suppressing the Development of Th17 Cells and Synthesis of Proinflammatory Cytokines by Such Cells, via Inhibition of NF-κB

doi: 10.1371/journal.pone.0138201

Figure Lengend Snippet: (A) Spleens from each mouse were stained for CD4 + p-STAT3(Y705) + , CD4 + p-STAT3(S727) + , CD4 + STAT3 + , and CD4 + IL-17 + T cells using antibodies specific for CD4, STAT3Y705, STAT3S727, and IL-17. The cell populations were analyzed via laser confocal microscopy (original magnification ×400). (B) The numbers of T cells positive for CD4 + STAT3Y705 + , CD4 + STAT3S727 + , CD4 + STAT3 + , and CD4 + IL-17 + in each mouse were visually counted at a higher magnification (after projection of fields onto a screen) and mean values are shown. (C) Spleens from each mouse were stained for CD4 + p-STAT5(Y694) + , CD4 + STAT5 + , and CD4 + CD25 + Foxp3 + . (D) The numbers of T cells positive for CD4 + p-STAT5(Y694) + , CD4 + STAT5 + , and CD4 + CD25 + Foxp3 + in each mouse were visually counted at a higher magnification (after projection of fields onto a screen) and mean values are shown. (Number of AEBS treated mice = 5, Number of vehicle treated mice = 5) Data are expressed as means ± SDs. * P < 0.05, ** P < 0.01. Each experiment was performed 3 times.

Article Snippet: Naïve cells were collected from this population by selecting for CD4 + CD62L high CD25 low CD44 low cells (>97% purity; Dako Cytomation, Glostrup, Denmark) and were also obtained using CD4 + CD62L + magnetic beads (Miltenyi Biotec).

Techniques: Staining, Confocal Microscopy

Journal: Oncology Letters

Article Title: Expression and clinical significance of colorectal cancer stem cell marker EpCAM high /CD44 + in colorectal cancer

doi: 10.3892/ol.2014.1907

Figure Lengend Snippet: Expression of EpCAM/CD44 in colorectal cancer.

Article Snippet: In 2008, Marhaba et al proposed that EpCAM high /CD44 + cells are a marker of colorectal cancer stem cells , and Dalerba et al found that the EpCAM high /CD44 + phenotype of colorectal cancer cells has stem cell-like properties ( ).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Prostate Cancer Stem Cell-Targeted Efficacy of a New-Generation Taxoid, SBT-1214 and Novel Polyenolic Zinc-Binding Curcuminoid, CMC2.24

doi: 10.1371/journal.pone.0069884

Figure Lengend Snippet: ( A ) Representative FACS analyses of the different cell surface markers expression in unsorted PPT2 cells grown for 4 weeks on type I collagen in MSGB medium. Each dotted square represents the population of cells expressing moderate/high levels of a particular marker conjugated with different fluorescent tags. Note a long-term retention of the CD133-APC, standard (CD44-PE) and variant (CD44v6-FITC) CD44 and CD49f-APC. In contrast, only small fractions of the PPT2 cells expressed a marker of basal cells, p63, a marker of differentiated cells, pan-keratin, CK18, AR and CXCR4. ( B ) Immunohistochemical analysis shows that, in contrast to parental tumor tissue, purified CD133 + PPT2 cells do not express PSA in vitro ; however, PPT2-induced NOD/SCID tumor xenografts are weakly PSA-positive. ( C ) Immunocytochemical analysis shows uniform expression of vimentin and nestin, with especially high levels of these markers of neural stem cells in large MNCs. ( D ) Western blot analysis shows expression of the pluripotency markers in nuclear and cytoplasmic fractions of the CD133 + PPT2 cells. Both the nuclear and cytoplasmic fractions expressed c-Myc and were negative for p53 and p21; only the nuclear fraction expressed Oct-4 and Sox-2.

Article Snippet: In order to obtain prostate cancer spheroids, MACS-CD133 and MACS-CD44 (Miltenyi Biotec, CA) or FACS sorted CD133 high /CD44 high cells were seeded at low numbers on ULA plates (1,000 cells per 6-well plate or 5-10,000 cells per 75 mm flasks) in MSCGM or SPCM.

Techniques: Expressing, Marker, Variant Assay, Immunohistochemical staining, Purification, In Vitro, Western Blot

Journal: PLoS ONE

Article Title: Prostate Cancer Stem Cell-Targeted Efficacy of a New-Generation Taxoid, SBT-1214 and Novel Polyenolic Zinc-Binding Curcuminoid, CMC2.24

doi: 10.1371/journal.pone.0069884

Figure Lengend Snippet: Phenotypic profiling of the PPT2 and PC3MM2 cells with FACS analysis*.

Article Snippet: In order to obtain prostate cancer spheroids, MACS-CD133 and MACS-CD44 (Miltenyi Biotec, CA) or FACS sorted CD133 high /CD44 high cells were seeded at low numbers on ULA plates (1,000 cells per 6-well plate or 5-10,000 cells per 75 mm flasks) in MSCGM or SPCM.

Techniques: Marker

Journal: PLoS ONE

Article Title: Prostate Cancer Stem Cell-Targeted Efficacy of a New-Generation Taxoid, SBT-1214 and Novel Polyenolic Zinc-Binding Curcuminoid, CMC2.24

doi: 10.1371/journal.pone.0069884

Figure Lengend Snippet: Representative FACS analysis shows a dose-dependent increase in the ratios of CD133 high PPT2 ( A ) and PC3MM2 ( B ) cells 24 hours after treatment with SBT-1214 and Ptx. Longer treatment (for 72 hours) with SBT-1214 (10 nM-10 µM) induced up to 65% cell death in PPT2 cells ( C ) and up to 60% in PC3MM2 cells ( D ; lower line). In contrast, treatment with the same doses of Ptx did not suppress proliferation of the tumorigenic prostate cancer cells ( C, D ; upper line). Cells were incubated with indicated drug concentrations, and data was obtained with standard MMT assay based on three independent experiments with four repeats in each treatment group. Values are the means±SD.

Article Snippet: In order to obtain prostate cancer spheroids, MACS-CD133 and MACS-CD44 (Miltenyi Biotec, CA) or FACS sorted CD133 high /CD44 high cells were seeded at low numbers on ULA plates (1,000 cells per 6-well plate or 5-10,000 cells per 75 mm flasks) in MSCGM or SPCM.

Techniques: Incubation

Journal: PLoS ONE

Article Title: Prostate Cancer Stem Cell-Targeted Efficacy of a New-Generation Taxoid, SBT-1214 and Novel Polyenolic Zinc-Binding Curcuminoid, CMC2.24

doi: 10.1371/journal.pone.0069884

Figure Lengend Snippet: NOD/SCID mice were ectopically implanted with 3,000 of CD133 + PPT2 and PC3MM2 cells on the flanks. Three weeks after injection, mice were treated with weekly i.v. injections of SBT-1214 (x4: 40, 20, 20, 20 mg/kg). This treatment modality induced dramatic reduction in tumor size in the majority of the PPT2- and PC3MM2-induced tumors ( A - D ). Representative cases are shown on B and D. Values are the means ±SD; p ≤0.0009 for PPT2-induced tumors (SBT-treated versus untreated controls; n=6), and p ≤0.0018 for PC3MM2-induced tumors (n=6). Histopathological analysis of the residual tumors showed the presence of viable cells and accumulation of large multinucleated cells in 2 of 6 PPT2 tumors and 3 of 6 PC3MM2 tumors (representative H&E staining of untreated and SBT-1214-treated PPT2 tumor tissues is shown on E, F ). Ex vivo death of the drug-treated cells in culture ( G ). Control untreated tumor cells retained profound clonogenic and sphere-forming capacities during serial passaging ( H ).

Article Snippet: In order to obtain prostate cancer spheroids, MACS-CD133 and MACS-CD44 (Miltenyi Biotec, CA) or FACS sorted CD133 high /CD44 high cells were seeded at low numbers on ULA plates (1,000 cells per 6-well plate or 5-10,000 cells per 75 mm flasks) in MSCGM or SPCM.

Techniques: Injection, Staining, Ex Vivo, Control, Passaging

Journal: PLoS ONE

Article Title: Prostate Cancer Stem Cell-Targeted Efficacy of a New-Generation Taxoid, SBT-1214 and Novel Polyenolic Zinc-Binding Curcuminoid, CMC2.24

doi: 10.1371/journal.pone.0069884

Figure Lengend Snippet: ( A , B ) The CMC2.24 as a single agent induced bi-phasic effects on prostate cancer cell proliferation: lower concentrations of it promoted proliferation, whereas higher ones were cytotoxic. ( C ) In contrast to SBT-1214, treatment with CMC2.24 did not induce an increase in the ratio of CD133 + cells (black dotted areas), but similarly to SBT-1214, increased expression of the differentiation marker pan-keratin (red dotted areas) and shifted the entire cell population toward differentiation (areas with asterisks). A combination of the two agents induced more significant cell death of the CD133 + PPT2 ( D ) and PC3MM2 ( E ) cells compared to each compound as a single agent. Data were obtained with standard MMT assay ( A , B , D , E ). Values are the means ±SD of the three independent experiments with 4 repeats for each drug concentration.

Article Snippet: In order to obtain prostate cancer spheroids, MACS-CD133 and MACS-CD44 (Miltenyi Biotec, CA) or FACS sorted CD133 high /CD44 high cells were seeded at low numbers on ULA plates (1,000 cells per 6-well plate or 5-10,000 cells per 75 mm flasks) in MSCGM or SPCM.

Techniques: Expressing, Marker, Concentration Assay

Journal: PLoS ONE

Article Title: Prostate Cancer Stem Cell-Targeted Efficacy of a New-Generation Taxoid, SBT-1214 and Novel Polyenolic Zinc-Binding Curcuminoid, CMC2.24

doi: 10.1371/journal.pone.0069884

Figure Lengend Snippet: ( A ) Multiple stem cell-relevant transcription factors are up-regulated in the CD133+ cell population compared to their differentiated counterparts. ( B ) Down-regulation of the up-regulated transcription factors after treatment with 1 µM SBT-1214 and 10 µM CMC2.24 for 24 hr (PCR array assay; SABiosciences; PAHS 501). Western blot analysis confirmed significant down-regulation of key pluripotency transcription factors, c-Myc and Sox-2 in CD133 + cells ( C ). Histon H1 was used as a loading control.

Article Snippet: In order to obtain prostate cancer spheroids, MACS-CD133 and MACS-CD44 (Miltenyi Biotec, CA) or FACS sorted CD133 high /CD44 high cells were seeded at low numbers on ULA plates (1,000 cells per 6-well plate or 5-10,000 cells per 75 mm flasks) in MSCGM or SPCM.

Techniques: Western Blot, Control

Journal: PLoS ONE

Article Title: Prostate Cancer Stem Cell-Targeted Efficacy of a New-Generation Taxoid, SBT-1214 and Novel Polyenolic Zinc-Binding Curcuminoid, CMC2.24

doi: 10.1371/journal.pone.0069884

Figure Lengend Snippet: ( A ) Previously absent pro-apoptotic/tumor suppressor proteins, p21 and p53 were induced by single treatment with SBT-1214/CMC2.24 combination for 24 hr in both CD133 + and bulk PPT2 cells. Such “gene wake-up” led to significant increase in the sensitivity of the CD133 + cells to this drug combination (B, C). Pre-treatment [SBT-1214 (1 µM) + CMC2.24 (10µM)]; second treatment [SBT-1214 (1 µM) + CMC2.24 variable]. Data were obtained with standard MMT assay.

Article Snippet: In order to obtain prostate cancer spheroids, MACS-CD133 and MACS-CD44 (Miltenyi Biotec, CA) or FACS sorted CD133 high /CD44 high cells were seeded at low numbers on ULA plates (1,000 cells per 6-well plate or 5-10,000 cells per 75 mm flasks) in MSCGM or SPCM.

Techniques: