Isolation of Human B Cell Populations

Sims, Gary P.; Lipsky, Peter E.

doi:10.1002/0471142735.im0705s75

Cited by 6 publications

(2 citation statements)

References 28 publications

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“…Non-B cells, for example, T cells, NK cells, dendritic cells, monocytes, granulocytes, and erythroid cells, are indirectly magnetically labeled by using a cocktail of biotin-conjugated antibodies against CD2, CD14, CD16, CD36, CD43, CD235a (Glycophorin A), and Anti-Biotin Micro-Beads. Isolation of highly pure B cells is achieved by depletion of magnetically labeled cells using MACS® columns (Miltenyi Biotec GmbH) [36][37] [38]. Cells were then incubated at a concentration of 2x10 6 cells/ml in RPMI 1640 medium with 10% Fetal Bovine Serum (FBS) (Invitrogen) at 5% CO2 and 37°C overnight, before NPs testing was started.…”

Section: Methodsmentioning

confidence: 99%

B-Chronic Lymphocytic Leukemia Autophagyc Cell Death by the Use of Manganese Doped Zinc Oxide Nanoparticles and Photo-Dynamic Therapy.

Luengas

Marín

Nieto

et al. 2015

IJDDT

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B-Chronic Lymphocytic Leukemia (B-CLL) usually follows an adverse, relentless clinical course by slowly developing drug resistance to fludarabine and other chemotherapeutic agents, as well as by acquiring new different genetic abnormalities. As B-CLL cells spontaneously produce high amounts of Reactive Oxygen Species (ROS) having an altered redox state in relation to that of normal B lymphocytes, we decided to probe different metal Zinc nanoparticles (ZnNPs) and quantify the levels of Singlet Oxigen (SO) to see if variations of its intracellular concentrations could execute and accelerate deadly programs in leukemic cells rather than in normal B lymphocytes, when applied with Photodynamic Therapy (PDT). In this way, we developed and tested a variety of metal ZnNPs of which one made of 0.5% Manganese Doped Zinc Oxide (ZnO:Mn) was finally selected for further testing as it had the best fludarabine resistant B-CLL cells in vitro killing activity, specially when combined with PDT. An interesting and rapidly dying process of B-CLL cells, known as autophagy, was always seen under Transmission Electronic Microscopy (TEM) when incubated with these 0.5% Mn doped ZnO NPs. This phenomenon correlated well with those intracellular increases of SO when PDT was administered, and measured by a novel method first described by us. As this therapy seems to be very specific to fludarabine resistant B-CLL cells, producing almost no damage to normal lymphocytes, it could surely contribute in the near future as a new innovative targeted strategy to be delivered in the clinical setting for the definitive benefit of these bad prognostic patients.

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Section: Methodsmentioning

confidence: 99%

B-Chronic Lymphocytic Leukemia Autophagyc Cell Death by the Use of Manganese Doped Zinc Oxide Nanoparticles and Photo-Dynamic Therapy.

Luengas

Marín

Nieto

et al. 2015

IJDDT

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“…B cells are classified in these two subpopulations according to their expression of CD5: a marker that is present on B‐1 B cells yet absent on B‐2 B cells . In contrast to this, a general B cell marker expressed by all the B cell subsets is CD19 . A further distinction of the B cell subpopulations could be attained by detection of the surface marker CD27, also known as the general memory B cell marker .…”

Section: Introductionmentioning

confidence: 99%

Ro52- and Ro60-specific B cell pattern in the salivary glands of patients with primary Sjögren's syndrome

Aqrawi¹,

Skarstein²,

Øijordsbakken³

et al. 2013

Clinical and Experimental Immunology

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SummaryPrimary Sjögren's syndrome (pSS) is characterized by the presence of autoantibodies against the ribonucleoprotein (RNP) particles Ro/SSA and La/SSB, and mononuclear cell infiltration of exocrine tissues, especially salivary and lachrymal glands. Low numbers of autoantigen-specific memory B cells and elevated levels of plasma cells have been detected previously in the peripheral blood (PB) of pSS patients compared to controls. As both Ro52 and Ro60-specific cells have been detected in the salivary glands (SG) of pSS patients, we aimed to characterize the SSA-specific B cell pattern in SG biopsies. A series of double immunohistochemical stainings were performed on paraffin-embedded tissue from 10 well-characterized pSS patients for each Ro52 and Ro60 along with CD19, CD5, CD20 or CD27, respectively. Ro52 and Ro60-specific cells detected in SG tissue were found to be CD19 + B cells located outside the CD19 + /CD20 + B cell zones (BCZ) and also interstitially. These SSA-specific cells were also quantified. No SSA-specific cells were CD5 + , indicating that they do not belong to the B-1 B cell subset. Furthermore, no SSA-specific cells were observed within the CD20 + BCZ. Hence, no SSA-specific memory B cells were detected in these individuals. Contrary to this, SSA-specific cells were found to be CD19 + /CD27 ++ , demonstrating that they are differentiating short or long-lived plasma cells. Taken together, our findings suggest that these lower levels of SSA-specific memory B cells in PB and absence of SSA-specific memory B cells in SG of pSS patients could result from activation of these cells into plasma cells at the site of inflammation.

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Isolation of primary human B lymphocytes from tonsils compared to blood as alternative source for ex vivo application

Helm

Riedl

Gollner³

et al. 2021

Journal of Chromatography B

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Isolation of Human B Cell Populations

Cited by 6 publications

References 28 publications

B-Chronic Lymphocytic Leukemia Autophagyc Cell Death by the Use of Manganese Doped Zinc Oxide Nanoparticles and Photo-Dynamic Therapy.

B-Chronic Lymphocytic Leukemia Autophagyc Cell Death by the Use of Manganese Doped Zinc Oxide Nanoparticles and Photo-Dynamic Therapy.

Ro52- and Ro60-specific B cell pattern in the salivary glands of patients with primary Sjögren's syndrome

Isolation of primary human B lymphocytes from tonsils compared to blood as alternative source for ex vivo application

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