Antigen presentation by the BCL1 murine B cell line: in vitro stimulation by LPS.

Harris, Mitchel; Kindle, Cathy S.; Abruzzini, Anthony F.; Pierce, C W; Cullen, Susan E.

doi:10.4049/jimmunol.133.3.1202

Cited by 7 publications

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“…Cell lines expressing I-A d (e.g., A20, K46R, 2PK3, BCL 1 B cell line, or PU5, WEHI-265 monocytic cell line) can be employed as antigen-presenting cells (APC) for activating the CDC35 or D1.6 T H cells (Walker et al, 1982;Harris et al, 1984;Zuckerman et al, 1988;Spencer et al, 1993). Use of cell lines for the APC source provides two advantages: first, mice do not have to be sacrificed, and, second, homogeneous preparations of B cells versus monocytic lines can be compared.…”

Section: Supplement 24mentioning

confidence: 99%

In Vitro Model for Modulation of Helper T Cell Differentiation and Activation

Heo

2005

CP Toxicology

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Influence of immunotoxicants on helper T cell reactivities can be investigated by assessing acquisition of IL-4- or IFN-γ-producing ability in vitro with antigen-primed naive or precursor helper T cells from the spleens of DO11.10 ovalbumin-specific transgenic mice. The in vitro differentiation model is believed to be close to the in vivo environmental conditions in which differentiation occurs. The effect of immunotoxicants on helper T cell activity can also be evaluated by antigen-specific activation of cloned type-1 or type-2 helper T cells in vitro, since the two subsets of helper T cells can be distinguished by patterns of cytokine secretion. This in vitro model will help investigators to examine the ability of toxicants to modulate helper T cell-mediated cellular or humoral immunity.

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Section: Supplement 24mentioning

confidence: 99%

In Vitro Model for Modulation of Helper T Cell Differentiation and Activation

Heo

2005

CP Toxicology

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Pulse profile analyses of endocytosis in capped B lymphocytes and BCL₁ cells

et al. 1988

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The effect of temperature on the kinetics of endocytosis by B lymphocytes and BCLl cells was examined by using flow cytometry. Mouse B cells were stained with FITC-RaMIg and induced to undergo cap formation in the temperature-controlled sample compartment of the flow cytometer. Capping and subsequent endocytosis were measured by continually monitoring the pulse profile descriptors (width and area) of the electronic signal curves generated during flow cytometric analyses. Decreases in area values which immediately followed cap formation were shown to result from a pH-dependent quenching of fluorescence emission as the internalized FITC-RaMIg entered acidic subcellular compartments. Similar results were obtained with BCLl cells, but, in addition to cap formation and acidification, cytoplasmic diffusion of the fluorescent ligand could also be discerned by flow cytometry. With either cell type the rates of cap formation and endocytosis were shown to be temperature dependent with temperature coefficient (&lo) values of 2.0-2.7. Based upon Arrhenius plots of width and area changes, activation energies for capping and endocytosis ranged from -12-18 kcallmol. Key terms: Flow cytometry, fluorescence, immunoglobulinFlow cytometry has been used to follow endocytosis in various cell types. In some studies the susceptibility of a ligand to labeling with either fluorescent antibodies (4,10,11) or liposomes (20) was used to follow internalization. However, in other studies workers capitalized on the pH sensitivity of fluorescein fluorescence emission to assay entry of fluorescein-conjugated ligands into acidic intracellular compartments (5,(12)(13)(14)(15)(16)18,21). Because similar studies might provide additional information regarding the endocytic pathway functioning in B lymphocytes and in leukemic B cells which have been employed in antigen processing and presentation studies (2,7), we have used flow cytometry to assess the kinetics of endocytosis following cap formation in B cells and in the BCLl cell line. For these studies we have made use of a previously described flow cytometric technique which involves the continuous monitoring of pulse area and pulse width values of the electronic signal curves generated by passage of the cells through the laser beam of the flow cytometer (3). As a result of these analyses, we were able to determine the rates of four consecutive steps in the endocytic pathway: capping, internalization, intracellular movement and entry of the fluorescent ligand into acidic subcellular compartments. In addition, by assaying the cells at various temperatures, we have been able to calculate Qlo and activation energy values for the four sequential steps. MATERIALS AND METHODSAnimals BALBk mice were obtained from a colony maintained in the Department of Microbiology at the University of Mississippi Medical Center. Isolation of B LymphocytesThe procedures used to obtain highly purified B lymphocytes have been described previously (1,3,23). Briefly, BALBic mice (2-3 months old) were killed by cervical disl...

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Internalization and processing of antibodies to surface antigens on human B cells. Monoclonal anti‐IgM antibodies are processed differently than monoclonal antibodies towards non‐Ig surface receptors

et al. 1986

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The internalization and intracellular processing of monoclonal antibody to immunoglobulin mu heavy chain (Mamu) have been investigated in two human Burkitt lymphoma cell lines (Ramos and Raji), in a human B cell lymphoma and in normal human peripheral blood B cells. In addition to the degradation of 125I-labeled Mamu to trichloroacetic acid (TCA) soluble material, a distinct pattern of larger 125I-Mamu fragments was detected in all sources of B cells tested. The particular fragmentation pattern, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis, involved the cleavage of both peptide bonds and disulfide bridges. This type of antibody fragmentation appeared to be a selective mechanism associated with sIgM, as no other degradation than that leading to TCA-soluble material could be detected after the internalization and degradation of radiolabeled monoclonal antibodies towards a variety of non-Ig B cell surface receptors. Three fragments of 125I-Mamu degradation were also detected in the supernatant of Ramos cells, implying that the recycling and exocytosis of certain 125I-Mamu fragments also took place.

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Antigen presentation by the BCL1 murine B cell line: in vitro stimulation by LPS.

Cited by 7 publications

References 0 publications

In Vitro Model for Modulation of Helper T Cell Differentiation and Activation

In Vitro Model for Modulation of Helper T Cell Differentiation and Activation

Pulse profile analyses of endocytosis in capped B lymphocytes and BCL₁ cells

Internalization and processing of antibodies to surface antigens on human B cells. Monoclonal anti‐IgM antibodies are processed differently than monoclonal antibodies towards non‐Ig surface receptors

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Antigen presentation by the BCL1 murine B cell line: in vitro stimulation by LPS.

Cited by 7 publications

References 0 publications

In Vitro Model for Modulation of Helper T Cell Differentiation and Activation

In Vitro Model for Modulation of Helper T Cell Differentiation and Activation

Pulse profile analyses of endocytosis in capped B lymphocytes and BCL1 cells

Internalization and processing of antibodies to surface antigens on human B cells. Monoclonal anti‐IgM antibodies are processed differently than monoclonal antibodies towards non‐Ig surface receptors

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Pulse profile analyses of endocytosis in capped B lymphocytes and BCL₁ cells