Preparation of lymphocyte-activating factor from continuous murine macrophage cell lines

Lachman, Lawrence B.; Hacker, Miles P.; Blyden, Gershwin T.; Handschumacher, Robert E.

doi:10.1016/0008-8749(77)90263-5

Cited by 113 publications

(44 citation statements)

References 14 publications

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“…1 × 10 T cells/ml were stimulated with phytohemagglutinin-M at a concentration of 0.75-1.0%. Thymocytes, at 5 × 106 cells/ml were stimulated with eoneanavalin A at 5/~g/ml (23). At the times indicated in the text, the cells were labeled as shown in table legends.…”

Section: Preparation Of Mt~-conditioned Mediamentioning

confidence: 99%

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Effect of interleukin 1 on human thymocytes and purified human T cells.

Maizel¹,

Mehta²,

Ford³

et al. 1981

The Journal of Experimental Medicine

100

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Human Interleukin 1 (IL-1) purified by molecular weight fractionation, isoelectric focusing, and gel electrophoresis has been tested on human thymocytes and highly purified human T cells. IL-1 prepared in this manner could not support the long-term growth of T cells yet would augment lectin-stimulated mitogenesis. The IL-1 preparations were shown to possess the lectin-augmenting activity at dilutions containing less than 1 ng of the measurable protein. These data are in agreement with the model that IL-1 stimulates production of IL-2 from lectin-stimulated lymphocytes.

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Section: Preparation Of Mt~-conditioned Mediamentioning

confidence: 99%

“…The conditioned media were utilized as 20% of the final volume in the mitogenic assay as preliminary data indicated this concentration to yield optimum mitogenesis. § The Mdg-derived conditioned media were initially tested on murme thymocytes (23).…”

Section: Preparation Of Mt~-conditioned Mediamentioning

confidence: 99%

Effect of interleukin 1 on human thymocytes and purified human T cells.

Maizel¹,

Mehta²,

Ford³

et al. 1981

The Journal of Experimental Medicine

100

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“…IL-1 standards included that derived from the J774A. 1 cell line, as described by Lachman et al (23), and purified IL-1 (Genzyme, Boston, MA).…”

Section: Methodsmentioning

confidence: 99%

Macrophage-derived growth factor for osteoblast-like cells and chondrocytes.

Rifas¹,

Shen²,

Mitchell³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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Rat resident peritoneal macrophages in primary culture were found to elaborate a mitogenic factor (or factors) for rat osteoblast-like cells and chondrocytes but not for skin fibroblasts. Growth-promoting activity appeared in the incubation medium within the first 20 hr of macrophage culture and was released in amounts that paralleled the number of macrophages per culture. After their proliferative response, as judged by increases in DNA synthesis and cell number, the osteoblast-like cells became enriched in alkaline phosphatase, an index of osteoblast specialization. The macrophage-derived activity was nondialyzable and heat-stable, and it was eliminated by exposure to trypsin. Inhibition of prostaglandin cyclooxygenase failed to modify its generation. Partial purification (Amicon filter concentration, gel filtration) disclosed principal peaks of activity corresponding to Mr of 43,000 and 10,000. The crude conditioned medium and the Mr 43,000-peak, but not the low-molecular-weight peak, exhibited interleukin 1 activity, as judged by the ability to stimulate the proliferation of mouse thymic lymphocytes. The macrophagederived growth factor described herein may participate in bone remodeling and repair and in primary bone and cartilage growth.The mammalian skeleton is a dynamic tissue, undergoing virtually continuous turnover after its initial development (1). Skeletal turnover stems from a complex remodeling process of microscopic dimensions in which osteoclasts resorb bone mineral and organic matrix and osteoblasts proceed to repair the resorption cavity (2, 3). Resorptive and formative events at each remodeling locus are congruent in space, sequentially coupled in time, and balanced in quantity, so that the mass of bone does not change, at least not in the short term. This spatial, temporal, and quantitative coordination points to the participation of local signals in the control of remodeling, and it has prompted a search for locally elaborated regulatory factors. Cells of the monocyte macrophage series may well generate such factors. Not only are they thought to be the precursors of mature osteoclasts (4-12), they are known to release agents that promote bone resorption both directly and indirectly (13). Furthermore, macrophage-like cells occupy resorption cavities during the time interval between resorption and formation, so that they are appropriately positioned to influence formative events (14,15). We have examined the possibility that macrophages elaborate a factor (or factors) that promote(s) the proliferation of osteoblasts, thereby acting to couple formation and resorption. We describe herein a heat-stable peptide activity in macrophage-conditioned medium (MCM), which promotes the growth of osteoblasts and chondroblasts in vitro. Abbreviations: CDM, chemically defined medium; PDS, plateletpoor plasma-derived serum; IL-1, interleukin 1 (lymphocyte-activating factor); MDGF, macrophage-derived growth factor; MCM, macrophage-conditioned medium. *To whom reprint requests should be addressed. MATER...

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“…Since the demonstration in 1976 that a soluble factor in culture supernatants could support the continued growth of T cells in culture (1), the central role of interleukin 2 (IL 2) in the proliferation of human and murine T cells has been established by many investigators. The result of these studies has been several models describing the role of soluble mediators in T-lymphocyte activation by mitogens (4,7,10). A central tenet of one of these models is that monocytes are required for lectin mitogen stimulation because of a product (interleukin 1; IL 1) that they elaborate.…”

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confidence: 99%

“…The importance of soluble mediators in the activation of T lymphocytes has become increasingly clear in the past several years (1)(2)(3)(4)(5)(6)(7)(8)(9)(10). Since the demonstration in 1976 that a soluble factor in culture supernatants could support the continued growth of T cells in culture (1), the central role of interleukin 2 (IL 2) in the proliferation of human and murine T cells has been established by many investigators.…”

mentioning

confidence: 99%

Evidence for an interleukin-independent pathway for human lymphocyte activation.

Koretzky¹,

Daniele²,

Greene³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

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Though lectin mitogen stimulation of T-cell proliferation is an interleukin 1-(IL 1), interleukin 2-(IL 2) dependent process, the calcium ionophore A23187 may be able to initiate T-lymphocyte proliferation by an additional pathway. That the action of A23187 is IL 1 independent was demonstrated by its ability to stimulate monocyte-depleted cells without the addition of exogenous IL 1. The IL 2 independence of A23187 was indicated by (i) the inability of exogenous IL 2 to augment A23187-induced proliferation and (ii) the inability of the monoclonal antibody anti-Tac (with specificity for the human IL 2 receptor) to inhibit proliferation mediated by A23187. By contrast, in cultures stimulated with the lectin mitogen phytohemagglutinin, additional IL 2 considerably increased proliferation, whereas anti-Tac routinely caused 60-90% inhibition. Although the ionophore caused some IL 2 production and resulted in IL 2 receptor expression, quantitative studies showed that our results could not be explained by excessive amounts of endogenous IL 2 interfering with the blockdng action of the antibody. Therefore, these data suggest that the action of A23187 as a human lymphocyte mitogen may be the result of at least two pathways-one dependent on the interleuidn-cell interactions and the other independent of these mediators.The importance of soluble mediators in the activation of T lymphocytes has become increasingly clear in the past several years (1-10). Since the demonstration in 1976 that a soluble factor in culture supernatants could support the continued growth of T cells in culture (1), the central role of interleukin 2 (IL 2) in the proliferation of human and murine T cells has been established by many investigators. The result of these studies has been several models describing the role of soluble mediators in T-lymphocyte activation by mitogens (4, 7, 10). A central tenet of one of these models is that monocytes are required for lectin mitogen stimulation because of a product (interleukin 1; IL 1) that they elaborate. Mitogen and IL 1 then together interact with T cells, resulting in the production of IL 2 (7, 10) and the acquisition of IL 2 receptors (11). However, it remains unclear whether this is the only means whereby proliferation of T lymphocytes can occur.To approach the possibility of an alternative pathway for Tcell activation that might be independent of IL 1 or IL 2, or both, we used the calcium ionophore A23187, which is mitogenic for human lymphocytes (12, 13). We present evidence that the ionophore is capable of stimulating monocyte-depleted human lymphocytes (MDC) to proliferate without added replacing factors [e.g., IL 1, 12-0-tetradecanoylphorbol 13-acetate (TPA)], thus suggesting that the action of A23187 is IL 1 independent. We also present evidence that the monoclonal antibody anti-Tac, which appears to identify the human IL 2 receptor (14-17) does not inhibit cultures stimulated by A23187, although this antibody is a potent inhibitor of activation induced by other mitogens or antigens. I...

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Preparation of lymphocyte-activating factor from continuous murine macrophage cell lines

Cited by 113 publications

References 14 publications

Effect of interleukin 1 on human thymocytes and purified human T cells.

Effect of interleukin 1 on human thymocytes and purified human T cells.

Macrophage-derived growth factor for osteoblast-like cells and chondrocytes.

Evidence for an interleukin-independent pathway for human lymphocyte activation.

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