Effect of hydrocortisone and bacterial lipopolysaccharide on colony-stimulating activity production from mouse marrow adherent cells, spleen cells and peritoneal macrophages in vitro

The C3H/HeJ mouse strain, previously shown to be a nonresponder to bacterial lipopolysaccharide (LPS)-induced mitogenesis in vitro, was demonstrated by the present studies to be competent to respond mitogenically to LPS, but only to LPS preparations obtained by selected extraction methods. These preparations appear to be confined to LPS isolated by mild extraction techniques, such as TCA or butanol. In contrast, those obtained by techniques utilizing phenol were only weakly stimulatory or completely nonstimulatory for spleen cells from the C3H/HeJ. All LPS preparations tested, on the other hand, were highly stimulatory for cells from another mouse strain, namely the C3H/St. The critical importance of the method of extraction of LPS on its mitogenic activity for C3H/HeJ cells was stressed by experiments in which LPS was prepared from Escherichia coli K235 using either of two procedures. In these experiments, phenol-extracted LPS, although mitogenic in the C3H/St, was completely nonstimulatory in the C3H/HeJ; whereas, butanol-extracted LPS was highly stimulatory in both strains of mice. This striking difference was attributed to a destructive effect of phenol on LPS, as demonstrated by the fact that treatment of butanol LPS with phenol resulted in a total loss of its mitogenic activity in the C3H/HeJ, but in only a partial loss in the C3H/St. In general, the mitogenic response observed with selected LPS preparations in the C3H/HeJ was quantitatively lower and more transient than that seen with the C3H/St, although qualitatively these responses appeared to be similar. This was evidenced by the observation that in both mouse strains LPS was a specific mitogen for B cells, a property which was also attributed in both strains to the same distinct structural region of the LPS molecule, that is lipid A. A preparation of LPS that failed to stimulate B cells from the C3H/HeJ nonetheless had the capacity to block activation of these B cells by a stimulatory preparation of LPS. These results strongly suggest that mitogenic stimulation of B cells by LPS is a function of the structural integrity of both the LPS molecule and putative B-cell receptors for LPS.

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Soluble factors from rabbit spleen cells kill and lyse Treponema pallidum in vitro

Canadian Journal of Microbiology ◽

10.1139/m90-120 ◽

1990 ◽

Vol 36 (10) ◽

pp. 711-717 ◽

Cited By ~ 4

Author(s):

Thomas J. Fitzgerald ◽

Barbara J. Elmquist

Keyword(s):

Cytochalasin B ◽

Treponema Pallidum ◽

Adherent Cells ◽

Spleen Cells ◽

Soluble Factors ◽

Macrophage Phagocytosis ◽

Immune Mediated ◽

Nonadherent Cells ◽

Immune Rabbit

Antibody and complement immobilize (kill) Treponema pallidum in vitro. Recent evidence also documents immobilization by soluble factors released by activated macrophages and lymphocytes. Immune-mediated lysis of treponemes, however, has not been reported. The findings in this paper focus on apparent treponemal lysis by rabbit splenic cell preparations. Using cells from animals infected testicularly for 9 to 12 days, unfractionated splenic preparations, as well as adherent and nonadherent preparations, killed and lysed T. pallidum. Phagocytosis alone could not explain the detrimental effects of adherent cells. When cytochalasin B was used to block phagocytosis, decreases in treponemal numbers were still detected. In related studies, immune rabbit sera did not enhance treponemicidal activity of the adherent cells. To assess the specificity of these reactions, T. pallidum was incubated with two monocyte-like cell lines (human U937 and mouse P388D1). Neither cell line was detrimental, and treponemal numbers were not lowered. The soluble nature of the treponemicidal factors from adherent and nonadherent preparations was shown by physically separating these cells from the organisms and demonstrating treponemal killing and lysis. In summary, clearance of T. pallidum from infected tissues is probably at least partially attributed to macrophage phagocytosis. Our findings suggest another mechanism involving lytic factors secreted by activated adherent and nonadherent cells. Key words: syphilis, splenic treponemicidal activity, Treponema pallidum.

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Induction of cystine transport activity in mouse peritoneal macrophages by bacterial lipopolysaccharide

Biochemical Journal ◽

10.1042/bj3100547 ◽

1995 ◽

Vol 310 (2) ◽

pp. 547-551 ◽

Cited By ~ 81

Author(s):

H Sato ◽

K Fujiwara ◽

J Sagara ◽

S Bannai

Keyword(s):

Interleukin 1 ◽

Peritoneal Macrophages ◽

Bacterial Lipopolysaccharide ◽

Tnf Alpha ◽

Transport Activity ◽

Necrosis Factor Alpha ◽

Ifn Gamma ◽

Factor Alpha ◽

Mouse Peritoneal Macrophages

The transport of cystine has been investigated in mouse peritoneal macrophages cultured in vitro. The transport activity for cystine was very low in freshly isolated macrophages but was potently induced during culture in the presence of bacterial lipopolysaccharide (LPS) at concentrations as low as 0.1 ng/ml. The transport activity for cystine was enhanced when the cells were incubated with tumour necrosis factor-alpha (TNF-alpha), but not with interferon-gamma (IFN-gamma) or interleukin-1. IFN-gamma was rather repressive in the induction of the activity by LPS or TNF-alpha. The transport activity for cystine induced by LPS has been characterized. Cystine was transported mainly by Na(+)-independent system and the uptake of cystine was inhibited by extracellular glutamate and homocysteate, but not by aspartate, indicating that the transport of cystine in macrophages treated with LPS is mediated by System xc-. Glutathione content of the macrophages increased when they were exposed to LPS, and this increase was, at least in part, attributable to the induced activity of the cystine transport.

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THE EFFECT OF 2-MERCAPTOETHANOL ON MURINE MIXED LYMPHOCYTE CULTURES

Journal of Experimental Medicine ◽

10.1084/jem.139.4.1025 ◽

1974 ◽

Vol 139 (4) ◽

pp. 1025-1030 ◽

Cited By ~ 85

Author(s):

Michael J. Bevan ◽

Ruth Epstein ◽

Melvin Cohn

Keyword(s):

Fetal Calf Serum ◽

Calf Serum ◽

Mouse Spleen ◽

Adherent Cells ◽

Spleen Cells ◽

Cytotoxic Cells ◽

Cytotoxic Response ◽

Lymphocyte Cultures ◽

Mixed Lymphocyte Cultures

Mouse spleen cells which have been depleted of adherent cells do not respond to allogeneic lymphocytes in vitro. Their cytotoxic response can be restored by inclusion of mercaptoethanol in the medium. Mercaptoethanol is shown to have a stimulatory effect also on the response of normal (unseparated) spleen cells to alloantigens. The enhancement of the DNA-synthetic and cytotoxic response is similar, varying from 3.5–15-fold. Cytotoxic cells also appear in unmixed lymphocyte cultures in the presence of mercaptoethanol and fetal calf serum. The specificity of these background cytotoxic cells is not known.

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Effects of lichen heteroglycans on proliferation and IL-10 secretion by rat spleen cells and IL-10 and TNF- secretion by rat peritoneal macrophages in vitro

Phytomedicine ◽

10.1016/j.phymed.2004.03.012 ◽

2005 ◽

Vol 12 (6-7) ◽

pp. 461-467 ◽

Cited By ~ 14

Author(s):

S. Omarsdottir ◽

J. Freysdottir ◽

H. Barsett ◽

B. Smestad Paulsen ◽

E.S. Olafsdottir

Keyword(s):

Peritoneal Macrophages ◽

Spleen Cells ◽

Tnf Secretion

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Antigen recognition. IV. Discrimination by antigen-binding immunocompetent B cells between immunity and tolerance is determined by adherent cells.

Journal of Experimental Medicine ◽

10.1084/jem.143.4.805 ◽

1976 ◽

Vol 143 (4) ◽

pp. 805-821 ◽

Cited By ~ 33

Author(s):

E Diener ◽

N Kraft ◽

K C Lee ◽

C Shiozawa

Keyword(s):

B Cell ◽

Short Pulse ◽

Tolerance Induction ◽

Mouse Spleen ◽

Adherent Cells ◽

Antigen Receptors ◽

Spleen Cells ◽

A Cells ◽

Tolerant State

Mouse spleen cells capable of specifically binding intrinsically tritium-labeled polymerized flagellin (POL) (labeling by biosynthesis of flagellar protein) via IgM receptors were found to comprise a distinct population of about 20-50 cells per 10(6) lymphocytes. Evidence is presented that the majority of mouse spleen cells binding tritium-labeled POL undergoes blastogenesis after antigen capping, antigen shedding, and receptor reformation. Under conditions of tolerance induction in vitro, however, loss of antigen from the cell surface was inhibited. Such inhibition of antigen redistribution and shedding was reversed by a short pulse of colchicine and new antigen receptors were formed. In spite of this, colchicine had no effect on the tolerant state. However, tolerance could be broken, regardless of presence or absence of the alkaloid, with radioresistant theta-negative accessory (A) cells (adherent cells) from normal but not from tolerant spleen cell populations. "Tolerant" A cells, although they were incapable of cooperating in a response to POL, were capable of participating in a response to a second unrelated antigen. It is concluded that tolerance to POL in vitro is induced by mechanisms other than the physical blocking of bone marrow-derived (B) cell receptors by antigen. Most likely, the discrimination by the B cell between a tolerogenic and immunogenic signal is mediated by A cells.

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THE EFFECTS OF MERCAPTOETHANOL AND OF PERITONEAL MACROPHAGES ON THE ANTIBODY-FORMING CAPACITY OF NONADHERENT MOUSE SPLEEN CELLS IN VITRO

Journal of Experimental Medicine ◽

10.1084/jem.136.3.604 ◽

1972 ◽

Vol 136 (3) ◽

pp. 604-617 ◽

Cited By ~ 194

Author(s):

Chang Chen ◽

James G. Hirsch

Keyword(s):

Antibody Formation ◽

Peritoneal Macrophages ◽

Lymphoid Cells ◽

Red Cells ◽

Mouse Spleen ◽

Spleen Cells ◽

Sheep Red Cells ◽

Low Concentrations ◽

Nonadherent Cells

Nonadherent mouse spleen cells exhibited poor viability and little or no capacity to form antibodies to sheep red cells in the Mishell-Dutton culture system. Viability and antibody-forming capacity could be restored by addition to these cultures of low concentrations of mercaptoethanol (10–4–10–5 M), or by addition of appropriate numbers of mouse peritoneal macrophages. Macrophage concentrations lower than optimal resulted in lower lymphoid cell viability and correspondingly fewer plaque-forming cells, whereas excess macrophages resulted in marked inhibition of antibody formation despite good viability of the lymphocytes. Restoration of the nonadherent cells with mercaptoethanol was thus much simpler and more reproducible than it was with macrophages; furthermore, the number of plaque-forming cells developed in cultures restored with mercaptoethanol was approximately fourfold higher than it was in cultures restored with optimal numbers of macrophages. In the presence of mercaptoethanol, the plaque-forming capacity of the nonadherent spleen cells was not increased when small numbers of macrophages were added to the system, nor was it decreased when the few macrophages present in the nonadherent cells were further reduced or eliminated. Excess macrophages inhibited antibody formation in the cultures containing mercaptoethanol as they did in control cultures. Optimal restoration of plaque-forming capacity to the nonadherent spleen cells with mercaptoethanol required the reducing agent to be present throughout the 4 or 5 day culture period. Addition of mercaptoethanol 1 or more days after initiation of culture, or transfer of the cells to a medium free of mercaptoethanol before completion of the culture resulted in a reduction in the numbers of plaque-forming cells. The results suggest that mouse lymphoid cells do not require macrophages in order to form antibodies to sheep red cells in vitro, provided mercaptoethanol is present in the culture medium. The mechanism of action of mercaptoethanol under these conditions is not completely clear, but one of its effects is to promote the viability of lymphoid cells in the cultures.

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