Effect of the interaction between fibronectin and VLA-4 on the proliferation of human B cells, especially a novel human B-cell line, OPM-3

In the present paper we report that the ROHA -9 cell line, an Epstein-Barr virus (EBV)-transformed human B cell line with accessory cell capabilities, constitutively secretes a soluble factor with the biochemical and biological characteristics of human monocyte-derived IL-1. The IL-1 derived from ROHA -9 augmented murine thymocyte proliferation and enhanced the proliferative response of human T lymphocytes to concanavalin A (Con A). The ROHA -9-derived IL-1 activity eluted from Sephacryl S-200 in two peaks, at 15- 18K and 32- 35K mol wt, eluted from DEAE-Sephacel at 50-80 and 110-130 mM NaCl, and showed charge heterogeneity with peaks at pI 7.3, 6.1, and 4.1 on isoelectrofocusing (IEF). These findings suggest that B cells may elaborate an IL-1-like activity. During the logarithmic growth of ROHA -9 cells, a inhibitory factor that inhibited the response of mouse thymocytes to IL-1 was also produced. This factor had a mol wt of 95K on Sephacryl S-200, eluted at 150 mM NaCl on DEAE-Sephacel and showed a peak of pI 4.7 on preparative IEF. The inhibitory factor appeared to be selective in its effects on IL-1 responses, since it did not inhibit the activity of IL-2 on mouse thymocytes or on the growth of the IL-2-dependent CT6 cell line. This "contra-IL-1" inhibited the response of murine thymocytes to suboptimal (1 microgram/ml) but not optimal (10 micrograms/ml) doses of Con A and the response of human peripheral blood lymphocytes to streptolysin O ( SLO ) or to alloantigens. Moreover, the factor could be absorbed by mouse thymocytes but not by CT6 cells, and such thymocytes pretreated with contra-IL-1 failed to response to IL-1. Although this inhibitor is the product of a transformed B cell line, it may be representative of regulatory substances that normally control IL-1 activities either at the extracellular or intracellular level.

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The kinetic study on cell surface antigens of a hybridoma between human B cells and a mouse B-cell line

Clinical Immunology and Immunopathology ◽

10.1016/0090-1229(86)90107-8 ◽

1986 ◽

Vol 41 (2) ◽

pp. 236-246 ◽

Cited By ~ 1

Author(s):

Teruaki Hamano ◽

Yoshinori Yasuda ◽

Tadakuni Yamasaki ◽

Yasuharu Murata ◽

Kiyoyasu Nagai

Keyword(s):

B Cells ◽

Cell Surface ◽

B Cell ◽

Kinetic Study ◽

Cell Line ◽

Surface Antigens ◽

Cell Surface Antigens ◽

Human B Cells

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Naive and memory human B cells have distinct requirements for STAT3 activation to differentiate into antibody-secreting plasma cells

Journal of Experimental Medicine ◽

10.1084/jem.20130323 ◽

2013 ◽

Vol 210 (12) ◽

pp. 2739-2753 ◽

Cited By ~ 100

Author(s):

Elissa K. Deenick ◽

Danielle T. Avery ◽

Anna Chan ◽

Lucinda J. Berglund ◽

Megan L. Ives ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Plasma Cells ◽

Cell Function ◽

Memory B Cells ◽

B Cell Differentiation ◽

Loss Of Function ◽

Human B Cells ◽

Human B Cell

Long-lived antibody memory is mediated by the combined effects of long-lived plasma cells (PCs) and memory B cells generated in response to T cell–dependent antigens (Ags). IL-10 and IL-21 can activate multiple signaling pathways, including STAT1, STAT3, and STAT5; ERK; PI3K/Akt, and potently promote human B cell differentiation. We previously showed that loss-of-function mutations in STAT3, but not STAT1, abrogate IL-10– and IL-21–mediated differentiation of human naive B cells into plasmablasts. We report here that, in contrast to naive B cells, STAT3-deficient memory B cells responded to these STAT3-activating cytokines, differentiating into plasmablasts and secreting high levels of IgM, IgG, and IgA, as well as Ag-specific IgG. This was associated with the induction of the molecular machinery necessary for PC formation. Mutations in IL21R, however, abolished IL-21–induced responses of both naive and memory human B cells and compromised memory B cell formation in vivo. These findings reveal a key role for IL-21R/STAT3 signaling in regulating human B cell function. Furthermore, our results indicate that the threshold of STAT3 activation required for differentiation is lower in memory compared with naive B cells, thereby identifying an intrinsic difference in the mechanism underlying differentiation of naive versus memory B cells.

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Characterization of Ryanodine Receptor–mediated Calcium Release in Human B Cells

Anesthesiology ◽

10.1097/00000542-200606000-00014 ◽

2006 ◽

Vol 104 (6) ◽

pp. 1191-1201 ◽

Cited By ~ 17

Author(s):

Leslie C. McKinney ◽

Thomas Butler ◽

Shawn P. Mullen ◽

Michael G. Klein

Keyword(s):

B Cells ◽

Malignant Hyperthermia ◽

B Cell ◽

Cell Lines ◽

Single Cells ◽

Human B Cells ◽

Malignant Hyperthermia Susceptible ◽

Increased Sensitivity ◽

Normal Human ◽

Human B Cell

Background Mutations in the ryanodine type 1 receptor (RyR1) are causative for malignant hyperthermia. Studies in human B lymphocytes have shown that measurement of RyR1-mediated intracellular Ca(2+) (Ca(2+)(i)) release can differentiate between normal and malignant hyperthermia-susceptible individuals. The authors have further developed the B-cell assay by pharmacologically characterizing RyR1-mediated Ca release in two normal human B-cell lines and demonstrating increased sensitivity of lymphocytes to the RyR1 agonist 4-chloro-m-cresol (4-CmC) in the porcine model of MH. Methods Ca(2+)(i) was measured fluorometrically using fura-2 in populations of cells in suspension or with fluo-4 in single cells using confocal microscopy. The Dakiki and PP normal human B cell lines were used, as well as lymphocytes obtained from normal and malignant hyperthermia-susceptible pigs. 4-CmC was used to elicit RyR1-mediated Ca release; all experiments were performed in the absence of external Ca(2+). Results EC(50) values for 4-CmC were 0.98 and 1.04 mm for Dakiki and PP cells, respectively, demonstrating reproducibility. The 4-CmC-induced increase in Ca(2+)(i) was eliminated by thapsigargin and was unaffected by xestospongin C. The Ca(2+)(i) increase was separable from mitochondrial stores and was inhibited by azumolene. Caffeine did not induce Ca(2+)(i) release, but ryanodine depleted intracellular stores by 50%. Lymphocytes from pigs carrying the Arg614Cys mutation in RyR1 showed increased sensitivity to 4-CmC (EC(50) = 0.47 vs. 0.81 mm for cells derived from normal animals). Conclusions RyR1-mediated Ca(2+) signals can be pharmacologically distinguished from other intracellular sources in human B cells, and alterations of RyR1 function can be successfully detected using Ca(2+) release from intracellular stores as an end point.

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Comprehensive analyses of B-cell compartments across the human body reveal novel subsets and a gut-resident memory phenotype

Blood ◽

10.1182/blood.2019002782 ◽

2020 ◽

Vol 136 (24) ◽

pp. 2774-2785 ◽

Cited By ~ 2

Author(s):

Nadine M. Weisel ◽

Florian J. Weisel ◽

Donna L. Farber ◽

Lisa A. Borghesi ◽

Yufeng Shen ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Surface Expression ◽

Gene Signature ◽

Organ Donors ◽

Cell Compartments ◽

Human B Cells ◽

Donor Variability ◽

And Function ◽

Human B Cell

Abstract Although human B cells have been extensively studied, most reports have used peripheral blood as a source. Here, we used a unique tissue resource derived from healthy organ donors to deeply characterize human B-cell compartments across multiple tissues and donors. These datasets revealed that B cells in the blood are not in homeostasis with compartments in other tissues. We found striking donor-to-donor variability in the frequencies and isotype of CD27+ memory B cells (MBCs). A comprehensive antibody-based screen revealed markers of MBC and allowed identification of novel MBC subsets with distinct functions defined according to surface expression of CD69 and CD45RB. We defined a tissue-resident MBC phenotype that was predominant in the gut but absent in blood. RNA-sequencing of MBC subsets from multiple tissues revealed a tissue-resident MBC gene signature as well as gut- and spleen-specific signatures. Overall, these studies provide novel insights into the nature and function of human B-cell compartments across multiple tissues.

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G protein subunit G alpha 16 expression is restricted to progenitor B cells during human B-cell differentiation

Blood ◽

10.1182/blood.v85.7.1836.bloodjournal8571836 ◽

1995 ◽

Vol 85 (7) ◽

pp. 1836-1842 ◽

Cited By ~ 21

Author(s):

MY Mapara ◽

K Bommert ◽

RC Bargou ◽

C Leng ◽

C Beck ◽

...

Keyword(s):

Cell Differentiation ◽

B Cells ◽

B Cell ◽

Cell Line ◽

Cell Lines ◽

B Cell Lymphoma ◽

Cell Phenotype ◽

Low Grade ◽

B Cell Differentiation ◽

Human B Cell

Recently G alpha 16, a new guanosine triphosphate (GTP) binding protein alpha subunit has been described to be specifically expressed in human hematopoietic cells. Expression of G alpha 16 was observed in human cell lines of myelomonocytic and T-lymphocytic origin, but not in human B-cell lines Raji and IM9. We studied the expression of G alpha 16 in human B cells corresponding to different stages of B-cell differentiation by means of reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. The human Burkitt's lymphoma cell lines Raji, Ramos, BJAB, the lymphoblastoid cell line SKW6.4, and the plasmocytoma cell line U266 were devoid of G alpha 16. In contrast, G alpha 16 was detected in the human progenitor B cell lines Reh and Nalm-6. Using the mu+, k-cell line BLIN-1 (pre-B cell phenotype) and its derived subclone 1E8 (surface mu+, k+; B-cell phenotype) G alpha 16 expression was found to disappear on transition from pre-B to B-cell differentiation stage. The analysis of a broad panel of human neoplastic B lymphocytes ranging from progenitor B-acute lymphatic leukemia (pre-pre-B-ALL), common acute leukemias (cALL), pre-B-ALL, mature B-ALL to low grade B-cell lymphoma (chronic lymphocytic leukemia of B-cell type, leukemic centrocytic non-Hodgkins lymphoma [NHL], hairy cell leukemia) showed that G alpha 16 expression is limited to progenitor and pre-B-ALL cells. Therefore, we conclude that within B-cell differentiation, G alpha 16 is expressed solely during early B cell ontogeny and downregulated during differentiation. Thus, G alpha 16 might be an important regulator involved in signaling processes in progenitor B cells.

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Human B cell precursors proliferate and express CD23 after CD40 ligation.

Journal of Experimental Medicine ◽

10.1084/jem.178.1.113 ◽

1993 ◽

Vol 178 (1) ◽

pp. 113-120 ◽

Cited By ~ 81

Author(s):

S Saeland ◽

V Duvert ◽

I Moreau ◽

J Banchereau

Keyword(s):

B Cells ◽

B Cell ◽

Surface Membrane ◽

Cd40 Ligation ◽

Gamma Receptor ◽

Human B Cells ◽

Normal Human ◽

B Lineage ◽

B Cell Precursors ◽

Human B Cell

The CD40 surface membrane molecule plays an important role in the activation of mature human B cells, but its role in earlier stages of B lineage development is unknown. Here, we have investigated the effects of triggering the CD40 antigen on B cell precursors (BCP) by crosslinking with anti-CD40 antibody presented by Fc gamma-receptor type II-transfected murine Ltk- cells (CD40 system). CD10+ surface immunoglobulin negative (sIg-) BCP, freshly isolated from fetal bone marrow or precultured on stromal cells, proliferated in the CD40 system. This effect required the presence of IL-3, which acted as a specific cosignal among a panel of cytokines examined. The association of IL-10 and IL-7 potentiated the observed IL-3 and CD40-dependent BCP proliferation, demonstrating that IL-10 can act on early B lineage cells. CD40-dependent activation of fetal BCP did not favor maturation to sIg+ B cells, but resulted in the induction of high levels of surface membrane CD23. The emerging CD23+ BCP lacked sIg and CD10, and represented an important proportion of the cycling cells in the CD40-dependent cultures. Taken together, our data demonstrate that stimulation of the CD40 antigen induces expression of the CD23 gene, and regulates cell proliferation during normal human B cell ontogeny.

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