Human B Cell Differentiation Induced by Microbial Superantigens: Unselected Peripheral Blood Lymphocytes Secrete Polyclonal Immunoglobulin in Response toMycopusma ArthrztidzsMitogen

J chain expression was examined as a function of the stage in differentiation along the B cell axis in humans. Intracellular distribution of J and mu chains in leukemic HLA-DR+ null and pre-B cells, and in normal B cells stimulated with pokeweed mitogen (PWM) was determined by immunoelectron microscopy and radioimmunoassay (RIA). J chain was detected in leukemic null and pre-B cells on free and membrane-bound ribosomes in the cytoplasm, or on perinuclear cisternae. Mu chain was found on free ribosomes and ribosomal clusters in leukemic pre-B cells but was absent in the leukemic null cells. In pre-B cell lines, mu chain was seen within rough endoplasmic reticulum (RER) and the Golgi apparatus whereas J chain was not detected in these organelles. However, both mu and J chain were detected in RER and the Golgi apparatus of immature and mature plasma cells induced by PWM stimulation of normal peripheral blood lymphocytes. Low levels of J chain were also detected by RIA in lysates of leukemic null and pre-B cells. Most of the intracellular J chain became detectable after reduction and alkylation of cell lysates, and free J chain was not found in the culture supernatants. The amount of intracellular and secreted immunoglobulin-bound J chain increased dramatically after PWM stimulation of peripheral blood lymphocytes. The majority of J chain-positive cells seen over an 8 d culture interval were lymphocytes and lymphoblasts, while mu chain was found primarily in plasma cells. These results suggest that J chain expression precedes mu chain synthesis during B cell differentiation and that a combination of the two chains for secretion is not initiated until the onset of plasma cells maturation.

Download Full-text

Delineation of Human B Cell Differentiation: Immunological and Molecular Characterization of Human B Cell Differentiation Factor (BSF-2)

Mechanisms of Lymphocyte Activation and Immune Regulation - Advances in Experimental Medicine and Biology ◽

10.1007/978-1-4684-5323-2_17 ◽

1987 ◽

pp. 177-188 ◽

Cited By ~ 4

Author(s):

Tadamitsu Kishimoto ◽

Toshio Hirano ◽

Hitoshi Kikutani ◽

Atsushi Muraguchi

Keyword(s):

Cell Differentiation ◽

B Cell ◽

Molecular Characterization ◽

B Cell Differentiation ◽

Differentiation Factor ◽

Cell Differentiation Factor ◽

Human B Cell

Download Full-text

Induction of human B cell differentiation by Fc region activators

Clinical Immunology and Immunopathology ◽

10.1016/0090-1229(89)90133-5 ◽

1989 ◽

Vol 50 (2) ◽

pp. 251-263 ◽

Cited By ~ 4

Author(s):

Monte V. Hobbs ◽

Richard A. Houghten ◽

Jodee A. Janda ◽

William O. Weigle ◽

Edward L. Morgan

Keyword(s):

Cell Differentiation ◽

B Cell ◽

B Cell Differentiation ◽

Human B Cell

Download Full-text

Molecular structure and immunological function of human B cell differentiation factor (BSF2)

Molecular Basis of Lymphokine Action ◽

10.1007/978-1-4612-4598-8_12 ◽

1987 ◽

pp. 123-136 ◽

Cited By ~ 1

Author(s):

Tadamitsu Kishimoto ◽

Tetsuya Taga ◽

Kiyoshi Yasukawa ◽

Yasuo Watanabe ◽

Tadashi Matsuda ◽

...

Keyword(s):

Molecular Structure ◽

Cell Differentiation ◽

B Cell ◽

B Cell Differentiation ◽

Immunological Function ◽

Differentiation Factor ◽

Cell Differentiation Factor ◽

Human B Cell

Download Full-text

Multiple myeloma: circulating lymphocytes that express plasma cell antigens

Blood ◽

10.1182/blood.v64.2.352.bloodjournal642352 ◽

1984 ◽

Vol 64 (2) ◽

pp. 352-356

Author(s):

GJ Ruiz-Arguelles ◽

JA Katzmann ◽

PR Greipp ◽

NJ Gonchoroff ◽

JP Garton ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Plasma Cell ◽

Peripheral Blood ◽

Plasma Cells ◽

Peripheral Blood Lymphocytes ◽

Tumor Burden ◽

B Cell Differentiation ◽

Blood Lymphocytes ◽

Bone Marrow Plasma

The bone marrow and peripheral blood of 14 patients with multiple myeloma were studied with murine monoclonal antibodies that identify antigens on plasma cells (R1–3 and OKT10). Peripheral blood lymphocytes expressing plasma cell antigens were found in six cases. Five of these cases expressed the same antigens that were present on the plasma cells in the bone marrow. Patients that showed such peripheral blood involvement were found to have a larger tumor burden and higher bone marrow plasma cell proliferative activity. In some patients, antigens normally found at earlier stages of B cell differentiation (B1, B2, and J5) were expressed by peripheral blood lymphocytes and/or bone marrow plasma cells.

Download Full-text

Optimized Protocols for in Vitro T Cell-Dependent and T Cell-Independent Activation for B Cell Differentiation Studies Using Limited Cells

10.20944/preprints202111.0365.v1 ◽

2021 ◽

Author(s):

Casper Marsman ◽

Dorit Verhoeven

Keyword(s):

Cell Differentiation ◽

T Cell ◽

B Cells ◽

B Cell ◽

Plasma Cells ◽

Cell Formation ◽

B Cell Differentiation ◽

Differentiation Potential ◽

Human B Cell

Background/methods: For mechanistic studies, in vitro human B cell differentiation and generation of plasma cells are invaluable techniques. However, the heterogeneity of both T cell-dependent (TD) and T cell-independent (TI) stimuli and the disparity of culture conditions used in existing protocols makes interpretation of results challenging. The aim of the present study was to achieve the most optimal B cell differentiation conditions using isolated CD19+ B cells and PBMC cultures. We addressed multiple seeding densities, different durations of culturing and various combinations of TD stimuli and TI stimuli including B cell receptor (BCR) triggering. B cell expansion, proliferation and differentiation was analyzed after 6 and 9 days by measuring B cell proliferation and expansion, plasmablast and plasma cell formation and immunoglobulin (Ig) secretion. In addition, these conditions were extrapolated using cryopreserved cells and differentiation potential was compared. Results: This study demonstrates improved differentiation efficiency after 9 days of culturing for both B cell and PBMC cultures using CD40L and IL-21 as TD stimuli and 6 days for CpG and IL-2 as TI stimuli. We arrived at optimized protocols requiring 2500 and 25.000 B cells per culture well for TD and TI assays, respectively. The results of the PBMC cultures were highly comparable to the B cell cultures, which allows dismissal of additional B cell isolation steps prior to culturing. In these optimized TD conditions, the addition of anti-BCR showed little effect on phenotypic B cell differentiation, however it interferes with Ig secretion measurements. Addition of IL-4 to the TD stimuli showed significantly lower Ig secretion. The addition of BAFF to optimized TI conditions showed enhanced B cell differentiation and Ig secretion in B cell but not in PBMC cultures. With this approach, efficient B cell differentiation and Ig secretion was accomplished when starting from fresh or cryopreserved samples. Conclusion: Our methodology demonstrates optimized TD and TI stimulation protocols for more indepth analysis of B cell differentiation in primary human B cell and PBMC cultures while requiring low amounts of B cells, making them ideally suited for future clinical and research studies on B cell differentiation of patient samples from different cohorts of B cell-mediated diseases.

Download Full-text