Growth of B-lymphocyte colonies in vitro.

Metcalf, Donald; Nossal, G. J. V.; Warner, Noel L.; Miller, J. F. A. P.; Mandel, T. E.; Layton, Judith E.; Gutman, George A.

doi:10.1084/jem.142.6.1534

Cited by 151 publications

(40 citation statements)

References 12 publications

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“…Colony formation is mitogen dependent (17) and the addition of the B-cell mitogen LPS, or SRBC enhances colony formation, particularly when few B cells are cultured (20). We therefore compared the dose response of lymph node cells cultured with or without these agents.…”

Section: Comparison Of Macrophage-containing Underlayers With Other Pmentioning

confidence: 99%

Regulation of B-lymphocyte clonal proliferation by stimulatory and inhibitory macrophage-derived factors

Kurland¹,

Kincade²,

Moore³

1977

The Journal of Experimental Medicine

126

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A functional subpopulation of murine B lymphocytes proliferate in semisolid agar culture in the presence of 2-mercaptoethanol to form colonies. The effects of diffusible macrophage-derived factors on this focal proliferation was investigated using a two-layer culture system which prevented macrophage-lymphocyte contact and permitted B-cell activation to be critically assessed under conditions of extremely low cell densities. Adherent peritoneal macrophages incorporated within underlayers of spleen or lymph node cell cultures potentiated both the number and size of developing B-cell colonies. These effects were most striking when low numbers of spleen or lymph node cells, or macrophage- depleted lymphoid cell suspensions were used. Thus, macrophage-depleted lymph node ceils gave rise to virtually no colonies, but colony-forming ability was restored by the presence of an optimal number of macrophages. When the number of macrophages exceeded that required for optimal stimulation, colony formation was suppressed; an effect which was largely prevented by indomethacin, an inhibitor of prostaglandin synthesis. Under these conditions, stimulation and inhibition of B-cell activation by macrophages could be dissociated, indicating that each signal is selectively controlled by individual molecules elaborated by the macrophage. With an appropriate number of macrophages required for B-cell activation, and sufficient indomethacin to inhibit the accumulation of macrophage-derived prostaglandin, B-lymphocyte clonal proliferation was a linear function of the number of B cells placed in culture. In the absence of macrophages, B-cell colony formation was potentiated by both lipopolysaccharide and intact sheep erythrocytes through a mechanism different from that of the macrophage-derived stimulatory factor. In addition to their direct stimulatory effect on B-cell proliferation, lipopolysaccharide and sheep erythrocytes were each capable of modulating the production and/or release of B-cell stimulatory and inhibitory factors by the macrophage. Parallel studies of conventional mitogen- stimulated lymphocyte cultures did not show a requirement for macrophages and confirm that the semisolid assay is uniquely suited to studies on the regulatory role of the macrophage in B-cell activation.

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Section: Comparison Of Macrophage-containing Underlayers With Other Pmentioning

confidence: 99%

Regulation of B-lymphocyte clonal proliferation by stimulatory and inhibitory macrophage-derived factors

Kurland¹,

Kincade²,

Moore³

1977

The Journal of Experimental Medicine

126

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“…Thus, in 10 experiments in which control cultures contained 0X1 ml normal saline, the mean number of colonies developing was 10 + 5. In cultures containing 0a1 ml of 10% fresh sheep red cells, colony formation is potentiated and becomes linear with respect to the number of cells cultured (Metcalf et al, 1975b). Thus, in the above experiments, a mean of 207 ± 93 colonies was observed in cultures containing 0-1 ml of 10% sheep red cells.…”

Section: General Effects Of Human Serummentioning

confidence: 72%

“…Because of the deliberate use of low numbers of cultured cells to obtain suboptimal culture conditions (Metcalf et al, 1975b) some fluctuation occurred between different experiments in the level of Blymphocyte colony formation obtained in the control cultures. Preliminary studies in 10 control experiments showed that cultures of 25,000 C57BL spleen cells usually produced 10 ± 5 colonies with the addition of 041 ml of normal saline, 150-250 colonies with the addition of 041 ml of 10% sheep red cells, and 10-20 colonies with 041 ml of the 3 control human sera (usually at a dilution between 1 : 4 and 1 : 16).…”

Section: Methodsmentioning

confidence: 99%

“…Morphological examination of colony cells showed that they were mononuclear cells, many of which were identifiable as early plasma cells. Analysis of membrane immunoglobulin showed that the majority of colony cells reacted positively with a fluorescein-conjugated anti-it serum, which is again typical of B-lymphocyte colony cells (Metcalf et al, 1975b). so Spleen populations contain some granulocytic and macrophage progenitor cells, and many of the human sera tested contained high levels of GM-CSF.…”

Section: General Effects Of Human Serummentioning

confidence: 99%

“…However, this unique behaviour may be more apparent than real. The cultures exhibit a marked non-linearity between the number of cells cultured and the number of colonies developing (Metcalf et al, 1975b). Furthermore, some evidence has been produced that 2-mercaptoethanol may induce the formation in the culture dish itself of material with stimulating or potentiating activity on B-lymphocyte colony proliferation (Metcalf, 1976).…”

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confidence: 99%

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Stimulation of B-lymphocyte colony formation in vitro by sera from patients with leukaemia or lymphoma

Metcalf¹,

Kolber²

1976

Br J Cancer

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Studies were made on the effects of 665 sera, from normal donors or patients with various diseases, on B-lymphocyte colony formation in agar by mouse spleen cells. Undiluted serum from most normal donors inhibited colony formation, but 43-53% of sera from patients with histiocytic lymphoma, lymphocytic lymphoma or Hodgkin's disease stimulated colony formation, serum activity correlating with the stage of the disease. Moderate colony-stimulating activity was observed with serum taken from patients with acute lymphoid or myeloid leukaemia following, but not prior to, chemotherapy. Colony stimulating activity was not correlated with the blood group of serum donors and could not be ascribed to the presence of endotoxin, red cells or mouse red cell haemagglutinins in the active sera. Elevated colony stimulating activity was not observed in sera from patients with non-neoplastic disorders ot haemopoiesis or with diseases of other organ systems.

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In vitro growth of lymphoma colonies from children with non-hodgkin's lymphoma

Smith

Wood

Fried

et al. 1981

Cancer

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Growth in vitro of malignant lymphoid cells has been previously reported, but those studies involved direct establishment of cell lines from the patients' cells. In the present study, an in vitro system for both primary growth and passage of malignant lymphoblastic colony-forming cells (ML-CFC) was established, and for the first time, colony-forming cells from non-Hodgkin's lymphoma patients were cultured, and cell lines were established from individual colonies. Malignant lymphoid colonies grew from 9 of 16 (56%) bone marrow aspirates, which were histologically involved with lymphoma and from 5 of 29 (17%) morphologically normal bone marrow samples. ML-CFCs grew concomitantly with myeloid and monocytic CFCs and were distinguishable by their morphologic characteristics on agar, cytochemical staining characteristics, and cell surface markers. Successful passage of ML-CFCs was accomplished in five patients, and ten distinct cell lines (six null cell, three B cell, and one T cell) were established. This agar assay system provides an opportunity to study malignant lymphoma cell growth both as a primary colony and a cell line established from a primary colony.

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Growth of B-lymphocyte colonies in vitro.

Cited by 151 publications

References 12 publications

Regulation of B-lymphocyte clonal proliferation by stimulatory and inhibitory macrophage-derived factors

Regulation of B-lymphocyte clonal proliferation by stimulatory and inhibitory macrophage-derived factors

Stimulation of B-lymphocyte colony formation in vitro by sera from patients with leukaemia or lymphoma

In vitro growth of lymphoma colonies from children with non-hodgkin's lymphoma

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