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

1990 ◽  
Vol 36 (10) ◽  
pp. 711-717 ◽  
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.

Dual effect of meningococcal antigens on a T cell dependent immune response

1983 ◽  
Vol 29 (12) ◽  
pp. 1619-1625 ◽  
Author(s):  
Brian G. Sparkes
Keyword(s):  
Immune Response ◽  
Inhibitory Activity ◽  
Sheep Erythrocyte ◽  
Subsequent Development ◽  
Plaque Formation ◽  
Adherent Cells ◽  
Spleen Cells ◽  
Dual Effect ◽  
Adjuvant Activity ◽  
Nonadherent Cells

Meningococcal antigens (MA) showed adjuvant activity when administered to mice at the same time as antigen (sheep erythrocyte (SE)), by increasing the splenocyte plaque-forming response in a dose-related manner. However, when SE were given 1 day after MA administration, the subsequent plaque formation was diminished from normal in proportion to the dose of MA injected. Splenocytes taken from mice up to 5 days after MA injection actively inhibited plaque formation when mixed with splenocytes immunized with SE 4 days earlier. Two days after MA injection the nonspecific inhibition of plaque formation was mainly due to adherent spleen cells, while at 5 days nonadherent cells had acquired the inhibitory activity. It appears that it is the degree of activation of adherent cells resulting from the timing and dosage of MA which modulates the subsequent development and secretion of antibody-forming cells.

The mitogenic activity of lipopolysaccharide for spleen cells from germfree, conventional, and gnotobiotic rats

1979 ◽  
Vol 25 (9) ◽  
pp. 1087-1093 ◽  
Author(s):  
Carol Wells ◽  
Edward Balish
Keyword(s):  
Spleen Cell ◽  
Lipid A ◽  
Mitogenic Activity ◽  
Adherent Cells ◽  
Spleen Cells ◽  
E Coli ◽  
Blastogenic Response ◽  
Pseudomonas Species ◽  
Nonadherent Cells ◽  
Germfree Rats

Spleen cells from germfree rats, conventionally reared rats, and gnotobiotic rats associated with two Pseudomonas species gave no positive blastogenic response when incubated with each of four lipopolysaccharide (LPS) preparations from Escherichia coli, with glycolipid extracted from Salmonella minnesota R595 or with S. minnesota R595 lipid A. However, spleen cell preparations from athymic mice demonstrated a positive blastogenic response when incubated with E. coli LPS. Removal of adherent cells from germfree and conventional-flora rat spleen cells did not increase the mitogenic activity of LPS for nonadherent cells (< 0.5% esterase-positive cells). All rat spleen cell preparations gave positive blastogenic responses to phytohemagglutinin and concanavalin A. This study indicates that LPS may not be a mitogenic agent for rat spleen cells.

scholarly journals THE EFFECT OF 2-MERCAPTOETHANOL ON MURINE MIXED LYMPHOCYTE CULTURES

1974 ◽  
Vol 139 (4) ◽  
pp. 1025-1030 ◽  
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.

scholarly journals POTENTIATION OF THE T-LYMPHOCYTE RESPONSE TO MITOGENS

1972 ◽  
Vol 136 (1) ◽  
pp. 143-155 ◽  
Author(s):  
Igal Gery ◽  
Byron H. Waksman
Keyword(s):  
T Cells ◽  
Bone Marrow Cells ◽  
Mouse Spleen ◽  
Adherent Cells ◽  
Spleen Cells ◽  
Con A ◽  
Cell Responses ◽  
Donor Cells ◽  
Peripheral Leukocytes ◽  
Nonadherent Cells

Effective supernatants (SUP), which potentiate mouse T-cell responses to phytohemagglutin (PHA), are obtained from cells of several species (human, rabbit, rat, mouse) and indeed from syngeneic spleen, thymus, or bone marrow cells. Unstimulated cells release some SUP activity but more is produced after stimulation. Lipopolysaccharide (LPS) produced very active SUP in all cultures tested. PHA was similarly active on human leukocytes only, whereas concanavalin A (Con A) gave highly efficient SUP only with mouse spleen cells. SUP production is not correlated with a mitotic response of the donor cells and is observed in cultures unable to respond mitotically to the stimulant. Adherent mouse spleen cell populations, consisting largely or entirely of macrophages, produce active SUP, while nonadherent cells do not. Similarly, purification of human peripheral leukocytes on nylon columns, with removal of macrophages and other adherent cells, destroys their ability to produce SUP. The importance of indirect effects in stimulating mitotic responses of T cells is emphasized by the fact that the mitotic response of mouse thymocytes to LPS and its ability to potentiate the response of these cells to PHA disappears with removal of adherent cells from the thymocyte population. Conversely the production of SUP from spleen cells stimulated by Con A requires the presence of T cells.

scholarly journals Establishment in culture and characterization of a strain with mast cell and monocytic properties from the bone marrow of a child with diffuse cutaneous mastocytosis

Blood ◽  
1991 ◽  
Vol 78 (2) ◽  
pp. 290-303 ◽  
Author(s):  
SA Krilis ◽  
SG Warneford ◽  
J Macpherson ◽  
S Kyradji ◽  
L Dalla-Pozza ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
Mast Cell ◽  
Mast Cells ◽  
Platelet Activating Factor ◽  
Prostaglandin D2 ◽  
Adherent Cells ◽  
Phenotypic Features ◽  
Nonadherent Cells

Abstract Bone marrow was isolated from a child with congenital mastocytosis. Upon prolonged in vitro culture, initially in the presence of interleukin-3 (IL-3), a population of relatively large fusiform, strongly adherent cells grew out plus a subpopulation of smaller nonadherent cells. The morphology of the adherent cells was not typical of fibroblasts, epithelial cells, nor of standard hematopoietic cell types, whereas the morphology of the nonadherent cells resembled mast cells. Neither cell type required the presence of IL-3 nor a feeder layer of fibroblasts for continued growth. Attempts to isolate the two populations were unsuccessful. This cell strain comprised of both cell populations has been termed human bone marrow-derived mastocytosis cells (HBM-M). These cells were found to possess some of the cytochemical, ultrastructural, and surface phenotypic features of degranulated mast cells. They reacted with the mast cell marker, monoclonal antibody YB5.B8, but not with the basophil specific monoclonal antibody Bsp-1 and released the inflammatory mediators histamine, leukotriene C4, prostaglandin D2, and platelet-activating factor constitutively. This release was not potentiated by immunologic- or nonimmunologic-activating stimuli. In addition, they exhibited cytochemical and surface phenotypic features of monocytes. Our results indicate that a population of abnormal proliferative cells exist in the marrow of this patient; that these cells may be responsible for the patient's pronounced systemic proliferation of mast cells and the associated symptoms; and that the cell's mast cell, monocyte properties may be indicative of a common bone marrow-derived mast cell/monocyte precursor.

scholarly journals Evidence that Lyb-2 is critical to specific activation of B cells before they become responsive to T cell and other signals.

1982 ◽  
Vol 155 (5) ◽  
pp. 1309-1316 ◽  
Author(s):  
H Yakura ◽  
F W Shen ◽  
E Bourcet ◽  
E A Boyse
Keyword(s):  
B Cells ◽  
Sheep Erythrocyte ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Cell Surface Molecules ◽  
Subsequent Generation ◽  
Spleen Cells ◽  
Soluble Factors ◽  
Surface Molecules

The generation of plaque-forming cells (PFC) to T-dependent antigen, but not to T-independent antigen, is reduced in vitro by Lyb-2 antibody. Monoclonal Lyb-2 antibody, added to Mishell-Dutton cultures within the first 2 d, but not later, greatly reduces the generation of alpha-sheep erythrocyte (SRBC) PFC from T-depleted spleen cells whether help is provided in the form of intact T cells or as soluble factors contained in mixed lymphocyte culture (MLC) supernatants. Generation of alpha-SRBC PFC from purified B cells, assisted by soluble factors in MLC and macrophage (P388D.1 cell) supernatants, is similarly reduced by Lyb-2 antibody. The initial 2-d period, during which cultures are diminishingly sensitive to reduction of PFC generation by Lyb-2 antibody, is not affected by the time at which such soluble factors are added. Thus, Lyb-2 cell surface molecules evidently do not function as receptors for these differentiative signals. Reduction of PFC generation by Lyb-2 antibody is antigen dependent in the sense that reduction of the PFC response to one antigen (SRBC) does not affect subsequent generation of PFC to a second antigen (horse erythrocytes) from the same cell population. These findings accord with the view that the Lyb-2 molecule participates in a B cell differentiative phase, probably proliferative, which begins with binding of antigen and precedes the phase in which B cells become fully receptive to signals from T and other cells.

scholarly journals Establishment in culture and characterization of a strain with mast cell and monocytic properties from the bone marrow of a child with diffuse cutaneous mastocytosis

Blood ◽  
1991 ◽  
Vol 78 (2) ◽  
pp. 290-303
Author(s):  
SA Krilis ◽  
SG Warneford ◽  
J Macpherson ◽  
S Kyradji ◽  
L Dalla-Pozza ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
Mast Cell ◽  
Mast Cells ◽  
Platelet Activating Factor ◽  
Prostaglandin D2 ◽  
Adherent Cells ◽  
Phenotypic Features ◽  
Nonadherent Cells

Bone marrow was isolated from a child with congenital mastocytosis. Upon prolonged in vitro culture, initially in the presence of interleukin-3 (IL-3), a population of relatively large fusiform, strongly adherent cells grew out plus a subpopulation of smaller nonadherent cells. The morphology of the adherent cells was not typical of fibroblasts, epithelial cells, nor of standard hematopoietic cell types, whereas the morphology of the nonadherent cells resembled mast cells. Neither cell type required the presence of IL-3 nor a feeder layer of fibroblasts for continued growth. Attempts to isolate the two populations were unsuccessful. This cell strain comprised of both cell populations has been termed human bone marrow-derived mastocytosis cells (HBM-M). These cells were found to possess some of the cytochemical, ultrastructural, and surface phenotypic features of degranulated mast cells. They reacted with the mast cell marker, monoclonal antibody YB5.B8, but not with the basophil specific monoclonal antibody Bsp-1 and released the inflammatory mediators histamine, leukotriene C4, prostaglandin D2, and platelet-activating factor constitutively. This release was not potentiated by immunologic- or nonimmunologic-activating stimuli. In addition, they exhibited cytochemical and surface phenotypic features of monocytes. Our results indicate that a population of abnormal proliferative cells exist in the marrow of this patient; that these cells may be responsible for the patient's pronounced systemic proliferation of mast cells and the associated symptoms; and that the cell's mast cell, monocyte properties may be indicative of a common bone marrow-derived mast cell/monocyte precursor.

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