cd14 cells depletion  (Miltenyi Biotec)

Journal: Experimental hematology

Article Title: CLL-like monoclonal B cell lymphocytosis displays an increased inflammatory signature that is reduced in early stage chronic lymphocytic leukemia

doi: 10.1016/j.exphem.2020.12.007

Figure Lengend Snippet: Flow cytometry gating strategy. FVS510 was used to select viable cells (1). After gating for singlets (2), CD45+CD14+CD4+ cells (monocytes) were selected (3 and 4) and linear mean fluorescence intensity (MFI) values of HLA-DR were registered (5a). The absence of fluorescence for PE-Cy7 on monocytes was confirmed employing isotype control (5b). On the other hand, CD3+CD4+ cells (CD4+ T cells) were gated (6). The percentage of CD4+ T cells that were positive for CXCR3 (7a) and PD1 or negative for CD27 was assessed employing isotype controls (7b), whereas the percentage of CD4+ T cells that were positive for perforin (8a) and granzyme B was evaluated using fluorescence-minus-one (FMO) controls (8b). In the plots, fluorescence intensity values are transformed into logarithmic scale.

Article Snippet: In line with this, CD14 + cell depletion from PBMCs led to decreased tumor cell viability in CLL (median: 88.7 vs. 71.6%, P -value=0.043) and a similar trend was observed in MBL (median: 85.8 vs. 75.6%, P -value=0.075) ( ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 5. caption a7 Tumor cell survival analysis.

Techniques: Flow Cytometry, Fluorescence, Control, Transformation Assay

Journal: Experimental hematology

Article Title: CLL-like monoclonal B cell lymphocytosis displays an increased inflammatory signature that is reduced in early stage chronic lymphocytic leukemia

doi: 10.1016/j.exphem.2020.12.007

Figure Lengend Snippet: Tumor cell survival analysis. All cell cultures were incubated for 24h. To evaluate the viability of tumor cells, clonal B cells were cultured alone (A). The effect of monocytes on tumor viability was assessed as the viability of tumor cells cultured with CD14-enriched cells minus the viability of tumor cells cultured alone (B). For each subject, variations in tumor viability between the two previous conditions are shown (C), as well as the viability of tumor cells within total PBMCs or CD14-depleted PBMCs (D).

Article Snippet: In line with this, CD14 + cell depletion from PBMCs led to decreased tumor cell viability in CLL (median: 88.7 vs. 71.6%, P -value=0.043) and a similar trend was observed in MBL (median: 85.8 vs. 75.6%, P -value=0.075) ( ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 5. caption a7 Tumor cell survival analysis.

Techniques: Incubation, Cell Culture

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) CD4 staining of isolated CD4 + T cells. (b) IL-2 ELISA of CD4 + T cells stimulated with or without anti-CD3/CD28 beads. (c) CD8 staining or isolated CD8 + T cells. (d) IFNγ ELISPOT assay of PBMC and purified CD8 + T cells incubated with and without EBV derived peptide AVFDRKSDAK. (e) CD56 staining of isolated NK cells. (f) NK cell degranulation assay—CD107a staining of NK cells incubated without (left panel) or with (right panel) MHC class I deficient. 221 cells at a 10:1 effector to target ratio. (g) CD303 staining of isolated pDC. (h) CD1c staining of isolated mDC. The x-axis in each flow cytometry plot indicates fluorescent intensity. The left hand peak in each flow cytometry plot indicates control staining with an irrelevant antibody. Representative white-light microscopy images of each of the purified cell populations used in Raman spectroscopy experiments are also shown.

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques: Staining, Isolation, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Purification, Incubation, Derivative Assay, Degranulation Assay, Flow Cytometry, Light Microscopy, Raman Spectroscopy

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) Mean standard Raman spectra of CD4 + , CD8 + and CD56 + NK cells. (b)-(d) Pairwise comparison of the WMRS spectra obtained from purified lymphocyte subsets. (b) Mean spectra of CD4 + versus CD8 + T cells. (c) Mean spectra of CD4 + T cells versus CD56 + NK cells. (d) Mean spectra of CD8 + T cells versus CD56 + NK cells. Solid spectra lines represent mean of each subset, with shadow lines representing the standard deviation. Shaded vertical bands indicated regions of significant difference, estimated by student’s T test at level of p<10 –7 .

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques: Purification, Standard Deviation

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) CD4 + , CD8 + T cells and CD56 + NK cells. (b) CD4 + and CD8 + T cells. (c) CD4 + T cells and CD56 + NK cells. (d) CD8 + T cells and CD56 + NK cells. (3D rotating views of these plots are available to view in the supplementary information).

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques:

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) CD4 + T cells. (b) CD8 + T cells. (c) CD56 + NK cells. (3D rotating views of these plots are available to view in the supplementary information).

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques:

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) CD4 staining of isolated CD4 + T cells. (b) IL-2 ELISA of CD4 + T cells stimulated with or without anti-CD3/CD28 beads. (c) CD8 staining or isolated CD8 + T cells. (d) IFNγ ELISPOT assay of PBMC and purified CD8 + T cells incubated with and without EBV derived peptide AVFDRKSDAK. (e) CD56 staining of isolated NK cells. (f) NK cell degranulation assay—CD107a staining of NK cells incubated without (left panel) or with (right panel) MHC class I deficient. 221 cells at a 10:1 effector to target ratio. (g) CD303 staining of isolated pDC. (h) CD1c staining of isolated mDC. The x-axis in each flow cytometry plot indicates fluorescent intensity. The left hand peak in each flow cytometry plot indicates control staining with an irrelevant antibody. Representative white-light microscopy images of each of the purified cell populations used in Raman spectroscopy experiments are also shown.

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques: Staining, Isolation, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Purification, Incubation, Derivative Assay, Degranulation Assay, Flow Cytometry, Light Microscopy, Raman Spectroscopy

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) Mean standard Raman spectra of CD4 + , CD8 + and CD56 + NK cells. (b)-(d) Pairwise comparison of the WMRS spectra obtained from purified lymphocyte subsets. (b) Mean spectra of CD4 + versus CD8 + T cells. (c) Mean spectra of CD4 + T cells versus CD56 + NK cells. (d) Mean spectra of CD8 + T cells versus CD56 + NK cells. Solid spectra lines represent mean of each subset, with shadow lines representing the standard deviation. Shaded vertical bands indicated regions of significant difference, estimated by student’s T test at level of p<10 –7 .

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques: Purification, Standard Deviation

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) CD4 + , CD8 + T cells and CD56 + NK cells. (b) CD4 + and CD8 + T cells. (c) CD4 + T cells and CD56 + NK cells. (d) CD8 + T cells and CD56 + NK cells. (3D rotating views of these plots are available to view in the supplementary information).

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques:

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: Confusion matrix for CD4 + , CD8 + and CD56 + cell subsets.

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques:

Journal: PLoS ONE

Article Title: The Use of Wavelength Modulated Raman Spectroscopy in Label-Free Identification of T Lymphocyte Subsets, Natural Killer Cells and Dendritic Cells

doi: 10.1371/journal.pone.0125158

Figure Lengend Snippet: (a) CD4 + T cells. (b) CD8 + T cells. (c) CD56 + NK cells. (3D rotating views of these plots are available to view in the supplementary information).

Article Snippet: Cells were isolated using Dynabeads (Life Technologies) untouched human CD4 T cell kit (depleting antibodies comprising anti-CD8, CD14, CD16a, CD16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human CD8 T cell kit (depleting antibodies comprising anti-CD4, CD14, CD16a, Cd16b, CD19, CD36, CD56, CD123 and CD235a), Dynabeads untouched human NK cell kit (depleting antibodies comprising anti-CD3, CD14, CD36, HLA Class II, CD123 and CD235a).

Techniques: