scholarly journals B Cell–attracting Chemokine 1, a Human CXC Chemokine Expressed in Lymphoid Tissues, Selectively Attracts B Lymphocytes via BLR1/CXCR5

1998 ◽  
Vol 187 (4) ◽  
pp. 655-660 ◽  
Author(s):  
Daniel F. Legler ◽  
Marcel Loetscher ◽  
Regula Stuber Roos ◽  
Ian Clark-Lewis ◽  
Marco Baggiolini ◽  
...  
Keyword(s):  
Amino Acids ◽  
T Lymphocytes ◽  
B Cell ◽  
B Lymphocytes ◽  
Human Blood ◽  
Dna Sequences ◽  
Chemokine Receptors ◽  
Expressed Sequence Tag ◽  
Lymphoid Tissues ◽  
Cxc Chemokine

Although most leukocytes, T lymphocytes in particular, respond to several different chemokines, there is virtually no information on chemokine activities and chemokine receptors in B lymphocytes. A putative chemokine receptor, BLR1, that is expressed in Burkitt's lymphoma cells and B lymphocytes was cloned a few years ago. Deletion of the gene for BLR1 yielded mice with abnormal primary follicles and germinal centers of the spleen and Peyer's patches, reflecting the inability of B lymphocytes to migrate into B cell areas. By screening expressed sequence tag DNA sequences, we have identified a CXC chemokine, termed B cell–attracting chemokine 1 (BCA-1), that is chemotactic for human B lymphocytes. BCA-1 cDNA encodes a protein of 109 amino acids with a leader sequence of 22 residues. The mature protein shares 23–34% identical amino acids with known CXC chemokines and is constitutively expressed in secondary lymphoid organs. BCA-1 was chemically synthesized and tested for activity on murine pre–B cells 300-19 transfected with BLR1 and on human blood B lymphocytes. In transfected cells, BCA-1 induced chemotaxis and Ca2+ mobilization demonstrating that it acts via BLR1. Under the same conditions, no activity was obtained with 10 CXC and 19 CC chemokines, lymphotactin, neurotactin/fractalkine and several other peptide ligands. BCA-1 was also a highly effective attractant for human blood B lymphocytes (which express BLR1), but was inactive on freshly isolated or IL-2–stimulated T lymphocytes, monocytes, and neutrophils. In agreement with the nomenclature rules for chemokine receptors, we propose the term CXCR5 for BLR1. Together with the observed disturbance of B cell colonization in BLR1/ CXCR5-deficient mice, the present results indicate that chemotactic recruitment by locally produced BCA-1 is important for the development of B cell areas of secondary lymphoid tissues.

Molecular Cloning of DOCK10/Zizimin3, a Novel Cdc42/Rac-Interacting Protein Specifically Induced by IL4 in B Lymphocytes.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 939-939
Author(s):  
Estefania Yelo ◽  
Lourdes Gimeno ◽  
Maria Victoria Bernardo ◽  
Maria Juliana Majado ◽  
Maria Rocio Alvarez ◽  
...  
Keyword(s):  
Amino Acids ◽  
T Lymphocytes ◽  
Lymph Nodes ◽  
B Cell ◽  
B Lymphocytes ◽  
Expression System ◽  
Interleukin 4 ◽  
B Cell Differentiation ◽  
Nucleotide Exchange

Abstract Interleukin-4 (IL4) induces proliferation, differentiation and survival of B lymphocytes. IL4 protects CLL B cells from death by apoptosis. Gene expression analysis suggest that IL4 pathways are activated in CLL cells. We have identified DOCK10/Zizimin3 as an IL4-induced gene in CLL cells, and have obtained its full length sequence after cloning 1960 bp at its 5′ terminus by RACE-PCR. The human DOCK10/ZIZ3 sequence coded for a protein with 2180 amino acids and a predicted Mr of 250K. DOCK10/ZIZ3 shared homology with the other two members of the Zizimin family, and is the largest among them: DOCK9/ZIZ1 (2069 amino acids) and DOCK11/ZIZ2 (2073 amino acids) are 52% and 50% identical, respectively, to DOCK10/ZIZ3, and 58% identical between them. DOCK10 was predominantly expressed in hematopoietic tissues, particularly in peripheral blood (PB), but also in lymph nodes, thymus and spleen. Among the PB subpopulations, DOCK10 was expressed in B and T lymphocytes and, at lower levels, in monocytes. DOCK10 was also expressed in several non-hematopoietic tissues, most significantly in brain and kidney. Its homologue DOCK9, compared to DOCK10, was predominantly expressed in placenta, and less significantly in hematopoietic tissues, particularly in B lymphocytes and monocytes. DOCK11, like DOCK10, was predominantly expressed in PB. Compared to DOCK10, DOCK11 was expressed more prominently in placenta, thyroid and PB monocytes, and less significantly in brain and lymph nodes. Therefore, each of the Zizimin family members had a specific tissue distribution. Among the three genes, only DOCK10 was induced by IL4 in CLL cells in vitro. Induction of DOCK10 by IL4 was a common event in CLL, since it was observed in 10 out of 10 cases. IL4 also induced DOCK10 expression in normal PB B lymphocytes, suggesting that DOCK10 induction by IL4 in CLL cells may be normal, rather than pathological. Western blot analysis using a polyclonal antibody raised against a peptide which mapped at the N terminus of DOCK10, detected a band of the expected size of 250K. Interestingly, IL4 did not induce DOCK10 expression in CD4 or CD8 T lymphocytes in vitro. Expression of DOCK10 was also studied in 4 B-ALL, 2 T-ALL, and 1 T-CLL. DOCK10 neither was expressed at significant levels nor induced by IL4 in vitro in these patients, except for a weak induction in a common B-ALL case, suggesting that expression of DOCK10, and its induction with IL4, may be restricted to certain stages of B cell differentiation, and/or certain B cell malignancies. DOCK10 was distributed both in cytosolic and nuclear extracts of CLL cells, and IL4 increased its expression in both compartments. K562 clones stably transfected with DOCK10 using the inducible tet-off expression system showed significantly higher levels of DOCK10 in cytoplasm than in nucleus. Immunofluoresce analysis of HA-tagged DOCK10 K562 clones showed preferent staining of the cytoplasm, and dotted structures were frequently observed. GST-pulldown assays showed that DOCK10 bound to nucleotide-free (nf) Cdc42, but not to GTP- or GDP-loaded Cdc42. In addition, DOCK10 bound to nf Rac1, albeit with less affinity than to Cdc42. DOCK10 did not bind to RhoA. These results suggest that, like DOCK9 and DOCK11, DOCK10 may act as a novel Cdc42 guanine-nucleotide exchange factor (GEF) and, in addition, as a Rac1 GEF.

Down-regulation of neutrophil functions by the ELR+CXC chemokine platelet basic protein

Blood ◽  
2000 ◽  
Vol 96 (9) ◽  
pp. 2965-2972 ◽  
Author(s):  
Jan E. Ehlert ◽  
Andreas Ludwig ◽  
Tobias A. Grimm ◽  
Buko Lindner ◽  
Hans-Dieter Flad ◽  
...  
Keyword(s):  
Amino Acids ◽  
Binding Sites ◽  
Chemokine Receptors ◽  
Cell Activation ◽  
Basic Protein ◽  
Cathepsin G ◽  
Terminal Part ◽  
Down Regulation ◽  
Cxc Chemokine ◽  
Continuous Accumulation

Abstract The platelet-derived neutrophil-activating peptide 2 (NAP-2, 70 amino acids) belongs to the ELR+ CXC subfamily of chemokines. Similar to other members of this group, such as IL-8, NAP-2 activates chemotaxis and degranulation in neutrophils (polymorphonuclear [PMN]) through chemokine receptors CXCR-1 and CXCR-2. However, platelets do not secrete NAP-2 as an active chemokine but as the C-terminal part of several precursors that lack PMN-stimulating capacity. As we have previously shown, PMN themselves may liberate NAP-2 from the precursor connective tissue-activating peptide III (CTAP-III, 85 amino acids) by proteolysis. Instead of inducing cell activation, continuous accumulation of the chemokine in the surroundings of the processing cells results in the down-regulation of specific surface-expressed NAP-2 binding sites and in the desensitization of chemokine-induced PMN degranulation. Thus, NAP-2 precursors may be regarded as indirect mediators of functional desensitization in neutrophils. In the current study we investigated the biologic impact of another major NAP-2 precursor, the platelet basic protein (PBP, 94 amino acids). We show that PBP is considerably more potent than CTAP-III to desensitize degranulation and chemotaxis in neutrophils. We present data suggesting that the high desensitizing capacity of PBP is based on its enhanced proteolytic cleavage into NAP-2 by neutrophil-expressed cathepsin G and that it involves efficient down-regulation of surface-expressed CXCR-2 while CXCR-1 is hardly affected. Correspondingly, we found PBP and, less potently, CTAP-III to inhibit CXCR-2– but not CXCR-1– dependent chemotaxis of neutrophils toward NAP-2. Altogether our findings demonstrate that the anti-inflammatory capacity of NAP-2 is governed by the species of its precursors.

GMP-Grade Generation of B-Lymphocytes for Adoptive Immunotherapy in Patients After Allogeneic Stem Cell Transplantation

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4352-4352
Author(s):  
Julia Winkler ◽  
Michael Mach ◽  
Juergen Zingsem ◽  
Volker Weisbach ◽  
Andreas Mackensen ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cells ◽  
T Lymphocytes ◽  
B Cell ◽  
B Lymphocytes ◽  
Clinical Setting ◽  
One Step ◽  
Enrichment Protocol ◽  
The One

Abstract Abstract 4352 Background and objectives: We have recently shown that memory B-lymphocytes from murine CMV immune donor animals adoptively transferred into immunodeficient mice were highly effective in protecting from a viral infection indicating a therapeutic potential of virus specific memory B-cells. These preclinical data provided evidence that a cell-based strategy supporting the humoral immune response might be effective in a clinical setting of post-HSCT immunodeficiency (Klenovsek et al., 2007, Blood 110: 3472–9). As adoptive transfer of B-cells has not been used before in a clinical setting, it is necessary to establish a technology for the generation of GMP-grade B-cell products. Methods: Starting from the leukapheresis of healthy donors, B-cells were purified by two different separation strategies using GMP-grade microbeads and the CliniMACS∧TM device. A one-step protocol was used for positive enrichment of B-lymphocytes with anti-CD19 microbeads. In a two-step enrichment protocol, first T-lymphocytes were depleted by anti-CD3 microbeads and the remaining fraction was positively selected by anti-CD19 microbeads. Results: The leukapheresis contained a mean of 9.0×10∧8 CD19-positive B-cells (4.5–12.4 ×10∧8). After the one-step positive purification strategy a mean purity of CD20∧+ B-lymphocytes of 78.1% with a recovery of 32–41% was obtained. With the two-step T-cell depletion/B-cell enrichment protocol we achieved a mean purity of 96.4 % (93.4–97.8%) with a slightly lower recovery of 14–37%. The absolute B-cell numbers obtained in the product were 1.3 to 4.0 ×10∧8 and 1.7 to 2.6 ×10∧8 for the one-step positive enrichment and the two-step protocol, respectively. Importantly, the absolute number of T-cells was lower in cell products after the two-step protocol (0.1 to 0.9 ×10∧6 T-cells) as compared to the one-step positive CD19-enrichment (1.6 to 3.4 ×10∧6 T-cells). Assuming a patient with 70 kg body weight, the B-cell products obtained after the combined CD3-depletion and CD19-enrichment contained less then 4×10∧4 T-lymphocytes/kg bodyweight, which is a critical threshold number of T-cells in haploidentical HSCT. The B-cell products showed antibody production after in vitro stimulation in a limiting dilution assay and showed excellent viability after cryopreservation. Conclusions: A GMP-grade B-cell product can be obtained with high purity and very low T-cell contamination using the two-step enrichment protocol based on CliniMACS∧TM technology. (Supported by BayImmuNet) Disclosures: No relevant conflicts of interest to declare.

Down-regulation of neutrophil functions by the ELR+CXC chemokine platelet basic protein

Blood ◽  
2000 ◽  
Vol 96 (9) ◽  
pp. 2965-2972 ◽  
Author(s):  
Jan E. Ehlert ◽  
Andreas Ludwig ◽  
Tobias A. Grimm ◽  
Buko Lindner ◽  
Hans-Dieter Flad ◽  
...  
Keyword(s):  
Amino Acids ◽  
Binding Sites ◽  
Chemokine Receptors ◽  
Cell Activation ◽  
Basic Protein ◽  
Cathepsin G ◽  
Terminal Part ◽  
Down Regulation ◽  
Cxc Chemokine ◽  
Continuous Accumulation

The platelet-derived neutrophil-activating peptide 2 (NAP-2, 70 amino acids) belongs to the ELR+ CXC subfamily of chemokines. Similar to other members of this group, such as IL-8, NAP-2 activates chemotaxis and degranulation in neutrophils (polymorphonuclear [PMN]) through chemokine receptors CXCR-1 and CXCR-2. However, platelets do not secrete NAP-2 as an active chemokine but as the C-terminal part of several precursors that lack PMN-stimulating capacity. As we have previously shown, PMN themselves may liberate NAP-2 from the precursor connective tissue-activating peptide III (CTAP-III, 85 amino acids) by proteolysis. Instead of inducing cell activation, continuous accumulation of the chemokine in the surroundings of the processing cells results in the down-regulation of specific surface-expressed NAP-2 binding sites and in the desensitization of chemokine-induced PMN degranulation. Thus, NAP-2 precursors may be regarded as indirect mediators of functional desensitization in neutrophils. In the current study we investigated the biologic impact of another major NAP-2 precursor, the platelet basic protein (PBP, 94 amino acids). We show that PBP is considerably more potent than CTAP-III to desensitize degranulation and chemotaxis in neutrophils. We present data suggesting that the high desensitizing capacity of PBP is based on its enhanced proteolytic cleavage into NAP-2 by neutrophil-expressed cathepsin G and that it involves efficient down-regulation of surface-expressed CXCR-2 while CXCR-1 is hardly affected. Correspondingly, we found PBP and, less potently, CTAP-III to inhibit CXCR-2– but not CXCR-1– dependent chemotaxis of neutrophils toward NAP-2. Altogether our findings demonstrate that the anti-inflammatory capacity of NAP-2 is governed by the species of its precursors.

scholarly journals B Lymphocyte Chemotaxis Regulated in Association with Microanatomic Localization, Differentiation State, and B Cell Receptor Engagement

1998 ◽  
Vol 187 (5) ◽  
pp. 753-762 ◽  
Author(s):  
Conrad C. Bleul ◽  
Joachim L. Schultze ◽  
Timothy A. Springer
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Messenger Rna ◽  
B Lymphocyte ◽  
Somatic Hypermutation ◽  
Cell Receptor ◽  
B Cell Receptor ◽  
Cxc Chemokine

Migration of mature B lymphocytes within secondary lymphoid organs and recirculation between these sites are thought to allow B cells to obtain T cell help, to undergo somatic hypermutation, to differentiate into effector cells, and to home to sites of antibody production. The mechanisms that direct migration of B lymphocytes are unknown, but there is evidence that G protein–coupled receptors, and possibly chemokine receptors, may be involved. Stromal cell– derived factor (SDF)-1α is a CXC chemokine previously characterized as an efficacious chemoattractant for T lymphocytes and monocytes in peripheral blood. Here we show with purified tonsillar B cells that SDF-1α also attracts naive and memory, but not germinal center (GC) B lymphocytes. Furthermore, GC B cells could be converted to respond to SDF-1α by in vitro differentiation into memory B lymphocytes. Conversely, the migratory response in naive and memory B cells was significantly reduced after B cell receptor engagement and CD40 signaling. The receptor for SDF-1, CXC chemokine receptor 4 (CXCR4), was found to be expressed on responsive as well as unresponsive B cell subsets, but was more rapidly downregulated on responsive cells by ligand. Finally, messenger RNA for SDF-1 was detected by in situ hybridization in a layer of cells surrounding the GC. These findings show that responsiveness to the chemoattractant SDF-1α is regulated during B lymphocyte activation, and correlates with positioning of B lymphocytes within a secondary lymphoid organ.

scholarly journals Chemokine receptor specific for IP10 and mig: structure, function, and expression in activated T-lymphocytes.

1996 ◽  
Vol 184 (3) ◽  
pp. 963-969 ◽  
Author(s):  
M Loetscher ◽  
B Gerber ◽  
P Loetscher ◽  
S A Jones ◽  
L Piali ◽  
...  
Keyword(s):  
Amino Acids ◽  
T Lymphocytes ◽  
Chemokine Receptors ◽  
Reading Frame ◽  
Transmembrane Segments ◽  
Activated T Lymphocytes ◽  
Cc Chemokines ◽  
Receptor Cdna ◽  
G Protein Coupled ◽  
Human Receptor

A human receptor that is selective for the CXC chemokines IP10 and Mig was cloned and characterized. The receptor cDNA has an open reading frame of 1104-bp encoding a protein of 368 amino acids with a molecular mass of 40,659 dalton. The sequence includes seven putative transmembrane segments characteristic of G-protein coupled receptors. It shares 40.9 and 40.3% identical amino acids with the two IL-8 receptors, and 34.2-36.9% identity with the five known CC chemokine receptors. The IP10/Mig receptor is highly expressed in IL-2-activated T lymphocytes, but is not detectable in resting T lymphocytes. B lymphocytes, monocytes and granulocytes. It mediates Ca2+ mobilization and chemotaxis in response to IP10 and Mig, but does not recognize the CXC-chemokines IL-8, GRO alpha, NAP-2, GCP-2. ENA78, PF4, the CC-chemokines MCP-1, MCP-2, MCP-3, MCP-4, MIP-1 alpha, MIP-1 beta. RANTES, 1309, eotaxin, nor lymphotactin. The exclusive expression in activated T-lymphocytes is of high interest since the receptors for chemokines which have been shown so far to attract lymphocytes, e.g., MCP-1, MCP-2, MCP-3, MIP-1 alpha, MIP-1 beta, and RANTES, are also found in monocytes and granulocytes. The present observations suggest that the IP10/Mig receptor is involved in the selective recruitment of effector T cells.

scholarly journals The sequence of the mu transmembrane segment determines the tissue specificity of the transport of immunoglobulin M to the cell surface.

1990 ◽  
Vol 171 (3) ◽  
pp. 947-952 ◽  
Author(s):  
G T Williams ◽  
A R Venkitaraman ◽  
D J Gilmore ◽  
M S Neuberger
Keyword(s):  
Amino Acids ◽  
B Cells ◽  
Cell Surface ◽  
B Cell ◽  
B Lymphocytes ◽  
Tissue Specificity ◽  
Immunoglobulin M ◽  
Transmembrane Segment ◽  
Cos Cells ◽  
Associated Proteins

Membrane IgM is expressed on the surface of B lymphocytes. It is not transported to the surface of transfected plasmacytoma or COS cells. Here, we show that mutation of four hydrophilic amino acids in the microm transmembrane is sufficient to overcome the intracellular retention of membrane IgM in non-B cells. This suggests that the B cell-specific IgM-associated proteins that have been postulated to assist the transport of membrane IgM to the cell surface (3) act either by forming a hydrophobic sheath that surrounds the microm transmembrane segment or by displacing an interaction with this segment that would otherwise cause retention. Experiments with a CD8/mu hybrid H chain indicate that the proteins that assist the transport of membrane IgM to the B cell surface at most need the mu CH4 and transmembrane/cytoplasmic portion for interaction.

scholarly journals Autoantibody-associated cross-reactive idiotype-bearing human B lymphocytes: distribution and characterization, including Ig VH gene and CD5 antigen expression

Blood ◽  
1991 ◽  
Vol 78 (6) ◽  
pp. 1503-1515 ◽  
Author(s):  
G Inghirami ◽  
DR Foitl ◽  
A Sabichi ◽  
BY Zhu ◽  
DM Knowles
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Cell Subset ◽  
Antigen Expression ◽  
Lymphocytic Leukemia ◽  
Cell Subpopulation ◽  
Lymphoid Tissues ◽  
Vh Gene

Abstract Monoclonal antibodies (MoAbs) specific for autoantibody associated cross-reactive idiotypes (CRIs) frequently recognize the Igs of neoplastic B cells in patients with chronic lymphocytic leukemia (CLL) and/or Waldenstrom's macroglobulinemia. Very little is known regarding the normal B cells expressing CRIs (CRI-positive B cells). Using a variety of MoAbs against CRIs we investigated the distribution and topographic localization of CRI-positive B cells in normal adult human lymphoid tissues. We found that CRI-positive B cells represent a significant B-cell subpopulation expressing surface IgM (greater than 90%), IgG (approximately 5%), or IgA (approximately 2%). CRI-positive B cells are homogeneously distributed throughout all lymphoid tissues, accounting for 10% to 15% of all B lymphocytes, with the exception of the thymus, in which they represent the predominant B cell population. Immunophenotypic studies showed (1) that a small subpopulation (3.7% +/- 0.8%) of CRI-positive B cells are activated in vivo, based on CD25 and CD38 antigen expression; and (2) that approximately 50% of CRI-positive B cells express the 67-Kd pan-T-lymphocyte CD5 antigen, suggesting that the CRI-positive B-cell subset and the recently described CD5-positive B-cell subset are closely related. This hypothesis is supported by the fact that CRI-positive B cells produce oligo or polyreactive Igs, which are a characteristic feature of CD5-positive B cells, and also by the fact that both B-cell subpopulations appear to use similar and restricted Ig VH gene family members.

scholarly journals Current Perspectives on B Lymphocytes in the Immunobiology of Hepatocellular Carcinoma

2021 ◽  
Vol 11 ◽  
Author(s):  
Miaomiao Qin ◽  
Danping Wang ◽  
Yijiao Fang ◽  
Zhiying Zheng ◽  
Xinyang Liu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Tumor Microenvironment ◽  
T Lymphocytes ◽  
Cell Therapy ◽  
B Cell ◽  
B Lymphocytes ◽  
Immune Cells ◽  
Cell Biology ◽  
Therapy Strategy ◽  
The Way

Immune cells infiltrating tumors are capable of significantly impacting carcinogenesis through cancer promotion and anticancer responses. There are many aspects of hepatocellular carcinoma (HCC) related T lymphocytes that are undergoing extensive studies, whereas the effect exerted by B lymphocytes remains a less researched area. In this study, the latest research on the effect of B lymphocytes as they infiltrate tumors in relation to HCC is presented. Their prognosis-related importance is analyzed, along with their function in the tumor microenvironment (TME), as well as the way that B cell biology can be employed to help create a B cell therapy strategy for HCC.

Sign in / Sign up

Export Citation Format

Share Document