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A Rubartelli

Publications and source records attributed to A Rubartelli.

46 records · Page 3Linked to original sources
PubMed1988

The control of membrane and secreted heavy chain biosynthesis varies in different immunoglobulin isotypes produced by a monoclonal B cell lymphoma.

The control of production of the membrane (m) vs secreted (s) forms of immunoglobulin heavy chains was investigated in a panel of cell lines expressing different heavy chain classes but identical light chains (lambda) and variable regions. These cell lines could be induced towards Ig secretion by mitogen treatment. During this process a shift from m to s heavy chain production takes place. Here we show that, similarly to IgA- and IgE-producing B cells, in IgG2a-producing I.29 cells the gamma m-gamma s shift was accompanied by a shift in the corresponding mRNAs, with a decrease of gamma m mRNA and an increase of the gamma s mRNA in LPS-stimulated cells. By contrast, the micron mRNA was increased in LPS-stimulated IgM-producing cells, albeit these cells synthesized reduced amounts of micron polypeptides. The utilization of the translational level in the early steps of B lymphocyte maturation thus distinguishes the mode of regulation of mu chains from those of the other isotypes. In addition, in B cells a post-translational event blocks the secretion of IgM but not of IgG or IgE.

Animals↗
PubMed1987

Functional characterization of an antigen involved in an early step of T-cell activation.

An activation antigen, identified by the monoclonal antibody MLR3, is described that is present on activated T lymphocytes and thymocytes but not on resting T lymphocytes. Immunoprecipitation of radiolabeled membranes from an activated T-cell line showed that the MLR3-binding molecule has a molecular size of 28-34 kDa. Immunofluorescence analysis showed that the appearance of the MLR3 antigen is an early event and precedes that of the interleukin 2 receptor both in T lymphocytes and thymocytes. The proliferative response of resting T cells to OKT3-Sepharose and interleukin 1 or accessory cells, but not the interleukin 2-dependent proliferation, was inhibited by the addition of MLR3 monoclonal antibodies. Similar results wer also obtained in an interleukin 1-dependent human thymocyte proliferation assay. In addition when MLR3-positive cells were cultured with purified interleukin 1, MLR3 surface antigen expression was not observed. Thus MLR3 monoclonal antibody appears to recognize an antigen involved in an early step of T-cell activation related to interleukin 1-dependent functions and on both T lymphocytes and thymocytes.

Antibodies, Monoclonal↗
PubMed1986

In vitro production of interleukin 1 by normal and malignant human B lymphocytes.

In this study, the capacity of normal and neoplastic B lymphocytes to release interleukin 1 (IL 1) has been investigated. Peripheral blood B cells from normal donors were isolated by depletion of E rosetting cells and by positive selection of cells expressing surface immunoglobulin (sIg) or the B1 marker. Peripheral blood B cells from patients with B cell chronic lymphocytic leukemia (B-CLL) were purified by removal of E rosetting cells followed by complement-mediated cytotoxicity with selected monoclonal antibodies. All of the normal B cell suspensions and the large majority of the B-CLL cells produced in culture high amounts of IL 1 in the absence of any apparent stimulus. Control experiments ruled out that small numbers of monocytes in the B cell suspensions could represent the source of IL 1. These data support the contention that B cells participate to the immune response as accessory cells for T cell activation not only by physically presenting antigen, but also by releasing IL 1.

Animals↗
PubMed1985

Differentiation in the murine B cell lymphoma I.29: inductive capacities of lipopolysaccharide and Mycoplasma fermentans products.

Cells from the murine B lymphoma I.29, expressing IgM or IgA of identical idiotype, were found inducible by lipopolysaccharide to differentiate into plasma cells. Within 3 days, differentiating cells lost membrane-bound immunoglobulin (Ig) and accumulated large quantities of intracytoplasmic Ig. At day 6 of culture, IgA secretion increased 50-100-fold, as determined by enzyme-linked immunoassay. Proliferation increased for the first days of culture but decreased thereafter; by day 10 very few viable cells were present in lipopolysaccharide-stimulated cultures. Similar results were obtained by culturing I.29 cells in the presence of supernatants of certain B cell lines (e.g. BFO.3). The finding of a strict correlation between the inductive activity and presence of contaminating Mycoplasma fermentans suggested that factor(s) released by mycoplasma were responsible for the mitogenic activities. This was further indicated by the findings that: the supernatants of BFO.3 that were rendered free of mycoplasma were not inductive, and a nonactive cell line could be made active by infection with supernatants of BFO.3 cells containing viable microorganisms. Thus, supernatants of mycoplasma-infected cell lines may act as potent polyclonal activators on both normal and malignant B lymphocytes. The ability to induce membrane Ig on 70Z/3 cells indicates that mycoplasma-related mitogens are also active on pre-B cells. The possibility of mycoplasma contamination should thus be carefully excluded when presumptive factors of cloned cell lines are being evaluated.

Animals↗
PubMed1985

Maturation of chronic lymphocytic leukemia B cells: correlation between the capacity of responding to T-cell factors in vitro and the stage of maturation reached in vivo.

Circulating malignant cells from 15 patients with B-cell chronic lymphocytic leukemia (B-CLL) were examined by the electron microscope (EM) and tested for their capacity to produce immunoglobulin (Ig) molecules using endogenous labeling techniques in vitro. In agreement with previous observations, B-CLL clones from various patients could be subdivided in three distinct groups: immature (type 1) clones, that comprised mainly small resting lymphocytes which synthesized, but degraded Ig of the secretory type intracellularly; mature (type 3) clones consisting mainly of cells with an extended Golgi apparatus and numerous strands of rough endoplasmic reticulum (RER) that secreted Ig molecules; and clones (type 2) at an intermediate maturational stage. When stimulated with T-cell supernatants that contained T-cell replacing factor(s) (TRF), type 3 and, to a lesser extent, type 2 clones differentiated further and secreted increased amounts of Ig. This maturation was not observed when type 1 clones were stimulated with the same supernatants. However, these cells were not incapable of further maturation since they could differentiate in response to phorbol ester (TPA). The present data reinforce the concept that different CLL clones undergo a process of maturation in vivo that may be arrested at different levels and demonstrate that the various clones respond differently to physiological stimuli depending upon the level of maturation already reached in vivo.

B-Lymphocytes↗
PubMed1985

The regulation of membrane-bound and secreted alpha-chain biosynthesis during the differentiation of the B cell lymphoma I.29.

The regulation of the synthesis of membrane-bound and secreted IgA was investigated in the murine B lymphoma I.29 during the differentiation from IgA-bearing lymphocytes to IgA-secreting cells, as caused by treatment with lipopolysaccharide (LPS). LPS induced a threefold to fivefold increase in the amount of IgA synthesized, and induced a shift from the synthesis of the membrane form of alpha-chain (alpha m) to the synthesis of the secreted form of alpha-chain (alpha s), resulting in a 60-fold increase in the amount of IgA secreted. In vitro translation of sucrose gradient-fractionated RNA indicated that two mRNA molecules, 3.1 and 2.1 kilobase pairs (kb), encode alpha m-chains, whereas a smaller RNA molecule, 1.7 kb, encodes alpha s. Analyses by RNA blotting showed that the relative amounts of the three alpha mRNA changed rapidly during LPS-induced differentiation. The amount of the 3.1 and 2.1 kb alpha mRNA decreased, and the amount of the 1.7 kb alpha s mRNA increased in LPS-stimulated cells as compared with controls. These observations suggest that the regulation of alpha m/alpha s synthesis is controlled mostly at the pretranslational level.

Animals↗
PubMed1984

The role of glycosylation in secretion and membrane expression of immunoglobulins M and A.

The role of glycosylation in membrane expression and secretion of IgM and IgA was investigated in murine lymphoma and hybridoma cell lines, derived from I.29 tumor, which synthesize IgM or IgA with identical variable regions. Tunicamycin, a selective inhibitor of N-linked glycosylation, prevented the membrane expression of both isotypes, as demonstrated by immunofluorescence, radioiodination and endogenous labeling experiments. Selective immunoprecipitation and immunochemical analysis of membrane, intracellular and secreted molecules permitted us to determine the amount of membrane heavy chain externalized in the presence or absence of tunicamycin. Id 150 and Id 43, two I.29-derived hybridomas secreting IgA and IgM respectively, were differently affected by tunicamycin. While secretion of IgM was inhibited to greater than 95%, no inhibition of secretion of non-glycosylated IgA could be detected in Id 150 cells. These results indicate that different requirements for glycosylation exist in the biosynthetic pathways of immunoglobulin isotypes, and suggest that distinct intracellular transport systems may operate for membrane and secreted alpha-chains.

Animals↗
PubMed1983

Differentiation of chronic lymphocytic leukemia cells: correlation between the synthesis and secretion of immunoglobulins and the ultrastructure of the malignant cells.

The capacity of synthesizing and secreting Ig molecules was studied in 11 patients with B-cell chronic lymphocytic leukemia (B-CLL) whose cells expressed surface IgM, in 3 patients with surface IgG-bearing cells, and in 2 IgM prolymphocytic leukemias (IgM-PLL). Three types of mu chains were detected by SDS-polyacrylamide gel electrophoresis analysis of the endogenously labeled molecules isolated by specific immunoprecipitation. Two of them were isolated from the cell lysates and were identified as the membrane mu chain and the precursor of the secreted molecules, respectively. The latter also possibly contained precursors of the membrane molecules. The third type of molecule was detected only in the culture medium and was identified as secretory mu chain. Not all of the malignant clones possessed the three types of mu chains. Only 7/13 of the IgM-bearing malignant cell clones were capable of secretion, whereas the remaining synthesized the secretory mu chains but degraded them intracellularly. Two types of molecules (membrane and secreted) were found in the IgG-bearing CLL cells from three patients. In all of them, secretion was detected. Ultrastructural analysis demonstrated that cells from the secreting clones had the features of more mature lymphocytes than the cells from nonsecreting clones. These features were represented by a developed Golgi apparatus, various types of vesicles (smooth and coated), and strands of the rough endoplasmic reticulum. A certain heterogeneity of the degree of maturation of the cells was observed within these clones. The data are consistent with the hypothesis that CLL clones are heterogeneous and can be distinguished through the different degrees of maturation of their cell components.

Antibodies, Anti-Idiotypic↗
PubMed1982

Membrane-bound and secreted IgA contain structurally different alpha-chains.

Three different forms of alpha-chains are synthesized by BF0.3 and 615.2, two cloned cell lines derived from the murine B lymphoma 1.29. The three forms of alpha-chains differ in size, pI, cellular location, and rate of turnover. They were identified by means of lactoperoxidase-catalyzed radioiodination, internal 14C or 35S labeling, and immunofluorescence techniques as membrane-bound(alpha m), secreted (alpha s), and intracellular (alpha ic) proteins. Comparison of immunoglobulin products of the two lymphoma lines with those of a hybridoma cell line, Id 150, which secretes IgA of the 1.29 idiotype but lacks membrane IgA, confirmed the assignments of alpha m, alpha s, and alpha ic. Results of biosynthetic labeling of BF0.3, 615.2, and Id 150 in the presence and absence of tunicamycin suggest that the difference in m.w. and charge observed between alpha m and alpha s can be attributed to differences in primary amino acid structure rather than different degrees of glycosylation.

Animals↗