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G Tertian

Publications and source records attributed to G Tertian.

43 records · Page 3Linked to original sources

Long-term in vitro culture of murine mast cells. II. Purification of a mast cell growth factor and its dissociation from TCGF.

Procedures are described for the isolation of a mast cell growth factor (MCGF) from medium conditioned by mitogen-activated splenic leukocytes (CM). Although optimal conditions for the production of MCGF in CM are identical to those for the production of T cell growth factor (TCGF), MCGF can be dissociated from TCGF after the first stage of purification on a DEAE-cellulose column. MCGF elutes from the column in the breakthrough fraction, whereas TCGF binds avidly to DEAE and is eluted only at high salt concentration. MCGF also differs from TCGF with respect to m.w. (as estimated by Sephadex G-150 chromatography) and sensitivity to trypsin. In addition, MCGF is produced by the murine myelomonocytic leukemia WEHI-3 and the radiation induced thymic lymphoma LBRM-33 cells, whereas TCGF is produced only by the latter in the presence of a mitogen. Another hemopoietically active factor, granulocyte colony-stimulating factor (G-CSF) present in media conditioned by WEHI-3 and LBRM-33 cells, however, shares a number of properties with MCGF. Although studies with purified or partially purified MCGF have thus far failed to reveal a correlation between MCGF and G-CSF, further biochemical analyses are necessary to dissociate MCGF from G-CSF.

Animals↗

Long-term in vitro culture of murine mast cells. I. Description of a growth factor-dependent culture technique.

A feeder layer independent long-term in vitro culture system for murine mast cells is described. Concanavalin A-activated murine splenic leukocyte-conditioned medium, prepared under conditions optimal for T cell growth factor production, has been found also to contain a growth-promotion activity for murine mast cells identified by their morphology, characteristic ultrastructure of the granules, positive reactions with toluidine blue and alcian blue, presence of receptors for IgG and IgE, as well as presence of histamine, serotonin, L-Dopa, 5-hydroxytryptophan, and sulfated products within the cytoplasm. After 2 to 3 wk of culture in the presence of the conditioned medium, mast cell lines were established from various sources initially devoid of matured mast cells. Such sources included spleen and bone marrow of athymic nude mice, long-term cultured marrow cells as well as T cell-depleted normal marrow. Cultured mast cells are absolutely dependent upon the conditioned medium-derived growth factor(s) for growth and viability. Death ensues within 24 hr in the absence of the factor(s). Established mast cell lines have been maintained in exponential growth for over 1 yr by passaging in the conditioned medium every 3 to 7 days.

Animals↗

[Production of a mastocyte growth factor in lymphocyte cultures stimulated by a mitogen or by allogenic cells].

Supernatants from mouse spleen leukocytes stimulated by concanavalin A or from a secondary MLC between donor and recipient of a skin allograft can induce in bone marrow cells an exponential growth of mononuclear cells with cytoplasmic basophilic granules. These cells can be maintained in liquid suspension culture for up to 6 months. They lack the properties of T or b lymphocytes, monocytes or macrophages. They have the morphological features of mast cells or basophils; the cytoplasmic granules are metachromatic for toluidine blue, alcian blue-positive at pH 0.3 and contain histamine. In addition, these cells bind monoclonal mouse IgE. They may be considered as cells of the basophil-mast cell lineage.

Animals↗

Induction and long-term maintenance of Thy-1 positive T lymphocytes: derivation from continuous bone marrow cultures.

In the present study we investigated the presence of T-lymphocyte progenitors in the long-term murine bone marrow culture system described by Dexter: mature Thy-1 antigen-bearing T lymphocytes are lost in these cultures after a few days. By culturing nonadherent cells from such cultures in the presence of a supernatant of concanavalin A-stimulated spleen cells, a source of T-cell growth factor, we found that Thy-1 positive blast cells proliferated together with a second population of Thy-1 negative cells. These two populations of cells have been maintained in long-term in vitro cultures by passaging the cells in fresh conditioned medium at regular intervals. Moreover, we have been able to establish pure cultures of the Thy-1 bearing blast cells after separating them from the non-T cells using their adherence property to plastic surfaces. Long-term cultures of T lymphocytes can thus be established from long-term marrow cultures as well as from the spleen, thymus or fresh bone marrow.

Animals↗

IL-6 mRNA expression in CD25 positive malignant lymphomas.

We recently demonstrated that IL-2 is produced by reactive T cells in CD25-positive malignant lymphomas (ML). Using in situ hybridization, we investigated IL-6 mRNA expression in these CD25-positive ML. The ML tested included 9 anaplastic large cell lymphomas and 3 B-diffuse large cell lymphomas. Five CD25-negative ML were studied as controls. We show that IL-6 producing cells are present in all these ML. The density of positive cells was heterogeneous from case to case. However 3 cases of CD25-positive ML showed a dramatically higher density of IL-6 producing cells (70, 50, 43 producing cells per 10,000 cells, respectively) as compared to the other 9 cases of CD25-positive ML (mean 6.03 +/- 2.1 per 10,000). Morphological and topographical data suggested that several types of cells including fibroblasts, lymphocytes, macrophages and endothelial cells may synthesize IL-6. A combination of immunohistochemistry and in situ hybridization showed that reactive T cells and endothelial cells express the IL-6 gene whereas CD30-positive ML cells do not express this gene. Previous studies showed that IL-6 was capable to induce IL-2 receptor expression as well as production of IL-2 and stimulation of lymphomatous cells growth. Our present results indicate that the paracrine production of this cytokine may play a role in the proliferation of malignant lymphomas.

Adolescent↗