scholarly journals Establishment and Expression of Cytokines in a Theileria annulata-Infected Bovine B Cell Line

Genes ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 329 ◽  
Author(s):  
Rashid ◽  
Guan ◽  
Luo ◽  
Zhao ◽  
Wang ◽  
...  
Keyword(s):  
B Cells ◽  
Cell Surface ◽  
B Cell ◽  
Cell Line ◽  
Cytokine Production ◽  
Theileria Annulata ◽  
Transformed Cells ◽  
Specific Cell ◽  
Surface Markers ◽  
Cell Surface Markers

This study aimed to establish a pure single-cell Theileria annulata-infected B cell line for the assessment of cytokine production in transformed and lipopolysaccharide (LPS)-stimulated cells. Several studies have aimed to identify cell surface markers in T. annulata-transformed cells; however, no information on cytokine production in these cells is available. To investigate the potential of the transformed cells to produce cytokines and their potential responses to antigen-stimulation, we purified mature B cells (CD21) from the whole blood of cattle experimentally infected with the T. annulata Kashi strain by magnetic separation. The purity and specificity of the established cell line was assessed by the identification of specific cell surface markers (CD21, IgM, and WC4) by flow cytometry analysis. The transcript levels of the cytokines IL1A, IL1B, IL2, IL4, IL6, IL8, IL10, IL16, LTA, TGFB1, TNFA, IFNA, and IFNB in transformed, buparvaquone (BW720c)-treated cells, and antigen-stimulated cells were analyzed by quantitative polymerase chain reaction (qPCR) using cDNA from these cells. A T. annulata-infected bovine B cell line was successfully established with a purity of ~98.8% (CD21). IL4 and IL12A were significantly (p < 0.01) upregulated in the transformed cells. In BW720c-treated transformed cells, IL12B, TGFB1, and IFNB were significantly (p < 0.01) upregulated. Notably, no significant (p > 0.05) upregulation of cytokines was observed in LPS-stimulated transformed cells. Moreover, IL1A, IL1B, IL8, and IL16 were significantly (p < 0.01) upregulated in LPS-stimulated B cells. Our data signify the potential use of this cell line for cytokine production, observance of immunoglobulins, and production of an attenuated vaccine against tropical theileriosis.

Trogocytosis of multiple B-cell surface markers by CD22 targeting with epratuzumab

Blood ◽  
2013 ◽  
Vol 122 (17) ◽  
pp. 3020-3029 ◽  
Author(s):  
Edmund A. Rossi ◽  
David M. Goldenberg ◽  
Rosana Michel ◽  
Diane L. Rossi ◽  
Daniel J. Wallace ◽  
...  
Keyword(s):  
B Cells ◽  
Autoimmune Disease ◽  
Cell Surface ◽  
B Cell ◽  
Regulatory Proteins ◽  
Surface Markers ◽  
B Cell Antigen Receptor ◽  
Cell Surface Markers ◽  
Effector Cells ◽  
Key Points

Key Points Epratuzumab induces the reduction of multiple B-cell antigen receptor–modulating proteins on the surface of B cells via their trogocytosis to effector cells. Modulation of B cells by trogocytosis of key regulatory proteins may be an important mechanism of immunotherapy of autoimmune disease.

Characterization of a newly established feline lymphoma-derived cell line (BKD) lacking T and B cell surface markers

1986 ◽  
Vol 22 (5) ◽  
pp. 273-279 ◽  
Author(s):  
Robert W. Engelman ◽  
Katsuhiko Machida ◽  
Ross E. Longley ◽  
Wing T. Liu ◽  
Liem Q. Trang ◽  
...  
Keyword(s):  
Cell Surface ◽  
B Cell ◽  
Cell Line ◽  
Surface Markers ◽  
Cell Surface Markers

Towards an indirect screening technique facilitating detection of cellular populations bearing specific cell surface markers

2010 ◽  
Vol 26 (6) ◽  
pp. 1544-1550
Author(s):  
Sandeep K. Arora ◽  
Rohit Sharma ◽  
Gagandeep Kaur ◽  
Preeti Bhoria ◽  
Maryada Sharma ◽  
...  
Keyword(s):  
Cell Surface ◽  
Specific Cell ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Screening Technique ◽  
Specific Cell Surface

Expression of v-rel induces mature B-cell lines that reflect the diversity of avian immunoglobulin heavy- and light-chain rearrangements

1988 ◽  
Vol 8 (12) ◽  
pp. 5358-5368
Author(s):  
C F Barth ◽  
E H Humphries
Keyword(s):  
Cell Surface ◽  
B Cell ◽  
Cell Lines ◽  
Light Chain ◽  
Lymphoid Cells ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Chain Locus ◽  
Light Chain Locus

The infection of newly hatched chickens with reticuloendotheliosis virus strain T (REV-T) and a nonimmunosuppressive helper virus, chicken syncytial virus, induces rapidly metastatic B-cell lymphomas. In vivo analysis of these tumors with monoclonal antibodies detected the expression of the B-cell surface markers immunoglobulin M (IgM), CIa, Bu2, and CLA-1, but not IgG, Bu1, or a T-cell surface marker, CT-1. Cell lines derived from tumors exhibited the same pattern of staining, suggesting that expression of cell surface markers does not change during in vitro cell line development. All cell lines examined synthesized IgM in varying amounts. Northern (RNA blot) analysis confirmed abundant expression of v-rel mRNA, and Southern analysis revealed rearrangement of both heavy- and light-chain immunoglobulin loci. Analysis of the light-chain locus demonstrated that 20 of 22 lines contained a single rearranged allele. With respect to specific restriction enzyme sites within the V lambda 1 gene, the active allele in any given clone was either diversified or nondiversified. In contrast, examination of the heavy-chain loci within these lines demonstrated that 16 of the 22 had both alleles rearranged. Further diversification of the V lambda 1 locus did not occur after prolonged in vitro passage of the cell lines. We propose that v-rel expression arrests diversification of the light-chain locus in these lymphoid cells, allowing the production of stable, clonal B-cell populations. The development of these and similar cell lines will make it possible to identify specific stages of avian lymphoid ontogeny and to study the mechanism of rearrangement and diversification in the avian B lymphocyte.

Abstract 4744: CD22-targeting epratuzumab mediates trogocytosis of multiple cell-surface markers on normal, malignant, and lupus B cells.

2013 ◽  
Author(s):  
Edmund A. Rossi ◽  
David M. Goldenberg ◽  
Rosana Michel ◽  
Diane L. Rossi ◽  
Daniel J. Wallace ◽  
...  
Keyword(s):  
B Cells ◽  
Cell Surface ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Multiple Cell

CD81-Dependent Trafficking of CD19: Restoration of CD19 Surface Expression in Human B Cells Harboring A CD81 Mutation

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1049-1049
Author(s):  
Shoshana Levy ◽  
Chiung-Chi Kuo ◽  
Yael Sagi ◽  
Homer Chen ◽  
Neta Kela-Madar ◽  
...  
Keyword(s):  
B Cells ◽  
Cell Surface ◽  
B Cell ◽  
Cell Line ◽  
Conflicts Of Interest ◽  
Surface Expression ◽  
Antibody Deficiency ◽  
Cell Surface Expression ◽  
Dominant Negative Effect ◽  
Proximity Ligation Assay

Abstract Abstract 1049 Introduction: A 6-year-old girl, who was diagnosed with a primary antibody deficiency, had B cells lacking surface CD19. However, both her CD19 alleles were normal and the impairment was actually caused by a homozygous exon splice site mutation in CD81 (1). The patient's B cells also lacked surface CD81 and produced an immature glycosylated CD19 protein that was retained intracellularly. Interestingly, this human deficiency differed from that of CD81 knockout mice as the latter still express a low level of CD19 on their B cells. Methods: We used an EBV-transformed B cell line from this patient to better understand i) the difference between the human and mouse CD81 deficiencies and ii) how CD81 controls the trafficking of CD19 to the cell surface. We reasoned that the truncated human CD81 mutant (CD81mut) protein might be expressed intracellularly. Indeed, whereas most anti-CD81 mAbs did not recognize CD81mut, we identified one that bound the mutated form and used it in this study. We also expressed the human CD81mut in a CD81-deficient mouse B cell line to determine if it could negatively regulate CD19 surface expression. Results: We show that the CD81mut protein is indeed expressed intracellularly in the patient's EBV-transformed B cells. We then used a proximity ligation assay to demonstrate that the truncated CD81mut protein interacts intracellularly with CD19. However, this interaction with the CD81mut protein abrogated carbohydrate maturation and the trafficking of CD19 to cell surface. We therefore expressed the CD81mut in CD81KO mouse B cells, which still express low levels of surface CD19, and found that it did not exert a dominant negative effect on CD19 surface expression. Finally, we used this reconstitution system to identify specific CD81 domains that restored carbohydrate maturation and cell surface expression of the CD19 molecule in the patient's B cells. Conclusion: This specific case of antibody deficiency was manifested because of lack of surface expression of CD19, an important B cell signaling molecule. However, the maturation of CD19 and its trafficking to the cell surface require the presence of specific domains of the tetraspanin CD81 molecule. Disclosures: No relevant conflicts of interest to declare.

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