The relationship between storage and secretion of specific antibody by immune lymphoid cells: ultrastructural localization of anti‐peroxidase anti bodies in plaque‐forming cells of the rabbit popliteal lymph node

Zagury, Daniel; Bernard, Jacky; Lemieux, Suzanne; Mazié, J. C.; Avraméas, Stratis; Bussard, A

doi:10.1002/eji.1830060310

Cited by 16 publications

(6 citation statements)

References 18 publications

(4 reference statements)

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“…It is also unclear why many AFC exhibit such dilated RER in our cultures. It has been suggested that extensive RER dilatation may be associated with decreased antibody secretion (14), and we wonder whether dilated RER results in the strongest staining for cytoplasmic Ig in immunocytochemical assays. It is possible that the maturation into typical plasma cells functions to limit the amount of Ig secreted in an immune response.…”

Section: Discussionmentioning

confidence: 95%

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Mouse spleen lymphoblasts generated in vitro. Their replication and differentiation in vitro

Steinman¹,

Machtinger²,

Fried³

et al. 1978

The Journal of Experimental Medicine

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For many years it has been known that mouse spleen B cells can be stimulated to proliferate and secrete antibody when cultured in the presence of such substances as lipopolysaccharide (LPS)' and fetal calf serum (FCS; 1, 2). This stimulation is said to be "nonspecific" or "polyclonar' because large numbers of cells are activated, and individual antibody-forming cells (AFC) exhibit many different, probably noncross-reactive, specificities. The phenomenon of polyclonal B-cell activation or B-cell mitogenesis is thought to be a useful model for studying the normal immune response even though smaller proportions of specific lymphoid cells are activated.Most studies of nonspocific B-lymphocyte activation have simply documented the behavior of whole cultures rather than individual cells. For example, we know little about the extent to which stimulated B cells divide in vitro; how many of the blasts mature into typical plasma cells as opposed to other cell types; or the life-span of the various cell types. We also have little information comparing the behavior of B cells in response to antigen-specific and nonspecific stimulation.In this study, we take advantage of the availability of enriched populations of presumptive B lymphoblasts, obtained by floatation on dense bovine plasma albumin (BPA) columns as described in the accompanying paper (3). These populations consist almost entirely of blasts. They can be recovered in high yield in their first cycle of cell division in response to either LPS or FCS stimulation. By using these enriched populations, often in combination with a nontoxic [SH]thymidine radiolabeling technique, we show that the majority of mitogen-stimulated blasts mature into typical plasma cells. Maturation occurs after just two cell divisions, after which the plasma cell appears to withdraw from proliferative activity. The mature plasma cells are similar to those produced in response to antigen in situ in their cytologic features, content of abundant cytoplasmic immunoglobulin, diminished proliferative activity, and short life-span.

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Section: Discussionmentioning

confidence: 95%

“…Some cells ( Fig. 4 D) exhibited large dilated cisternae corresponding to Mott cell droplets (13,14). A sample differential of a typical 3-day culture of LPS blasts is presented in Table V.…”

Section: Differentiation Of the Cultured Lymphoblastmentioning

confidence: 99%

Mouse spleen lymphoblasts generated in vitro. Their replication and differentiation in vitro

Steinman¹,

Machtinger²,

Fried³

et al. 1978

The Journal of Experimental Medicine

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“…In contrast to anti-PO Ab-secreting cells from immunized animals which show heterogenous ultrastructural characteristics and are identified as either lymphocytes, proplasmocytes or plasmocytes, all the PO 772 C 2 cells showed the same ultrastructural pattern and were identified as proplasmocytes. Moreover, taking advantage of the PO penetration into fixed cells and the PO staining after enzymatic reaction [16], we studied the presence of Ab inside the cells and at the cell surface. EM observations showed Ab within the cisternae of the RER.…”

Section: Discussionmentioning

confidence: 99%

“…They were then processed for the detection of intracellular anti-PO Ab and finally for electron microscopic observation as described in detail elsewhere [16].…”

Section: Cytological Treatmentmentioning

confidence: 99%

Anti‐peroxidase antibody‐secreting hybrid lines. I. Identification, cloning and cell characterization

et al. 1979

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Anti-peroxidase antibody (Ab)-secreting hybrids have been produced by fusion of peroxidase (PO)-immunized mouse lymph node cells and immunoglobulin (Ig)-secreting P3-X63-Ag8 (X63) myeloma cells. Identification of Ab-secreting hybrids can be performed as early as day 5 after cell fusion by the hemolytic plaque assay. Immediately after identification, hybrids were directly isolated, by means of a micropipette, into Terasaki microchambers containing nutrient medium and a thymocyte filler layer. The yield of secreting hybrids is improved by using this procedure. All the cells of the PO 772 C2 clone show the same ultrastructural pattern and immunocytological properties; they are proplasmocytes, as are the parental X63 cells; they present intracisternae Ab and show no Ig or Fc receptors at the cell surface. Over 90% of viable PO 772 C2 cells form specific plaques. Isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis show that the cells of this clone secrete Ab; the secreted Ig are formed with chi and gamma 1 chains from the parental X63 cells and specific L and H chains from the lymphoid parent. These biological investigations demonstrate the relative stability of the PO 772 C2 clone secreting anti-peroxidase antibody.

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“…After antigenic challenge, proliferation of lymphocytes from the B lineage ultimately leads to the development of specific antibody-forming cells (Nieuwenhuis, 1981). Although plasma cells are generally considered to be the main antibody-producing cells, the population of cells responsible for antibody synthesis is morphologically heterogeneous: besides plasma cells, considerable numbers of antibody-forming lymphocytes, antibodyforming plasmablasts, and antibody-forming immature plasma cells were observed (Zagury et al, 1976;Geldof et al, 1984b). It has been observed that more mature antibody-forming cells tended to localize nearer the medulla and more immature cells nearer the cortex, in both the TD immune response against TNP-KLH (Van Rees et al, 1986) and the TI immune response against TNP-LPS (Van Rees et al, 1987).…”

Section: Morphology Of Specific Antibody-forming Cells In Lymph Nodesmentioning

confidence: 99%

The “in situ” immune response in lymph nodes: A review

Rooijen

1987

Anat. Rec.

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For many years data on the development of specific antibody-forming cells in lymph nodes were incomplete, fragmentary, and even contradictory. A number of recent studies have been performed, concerning 1) their overall architecture; 2) migration of B-lymphocytes; 3) localization of accessory cells and T-lymphocytes which are believed to be involved in humoral immune responses; and 4) localization patterns of specific antibody-forming cells developing during thymus dependent and thymus independent immune responses. Comparison of these new results with those of earlier studies suggests a single route of migration followed by all cells which will differentiate into antibody-forming cells. During their differentiation into antibody-forming plasma cells, antigen reactive B-cells migrate along the required accessory cells and/or T-lymphocytes.

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The relationship between storage and secretion of specific antibody by immune lymphoid cells: ultrastructural localization of anti‐peroxidase anti bodies in plaque‐forming cells of the rabbit popliteal lymph node

Cited by 16 publications

References 18 publications

Mouse spleen lymphoblasts generated in vitro. Their replication and differentiation in vitro

Mouse spleen lymphoblasts generated in vitro. Their replication and differentiation in vitro

Anti‐peroxidase antibody‐secreting hybrid lines. I. Identification, cloning and cell characterization

The “in situ” immune response in lymph nodes: A review

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