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At least 19 recordsLinked to original sources
PubMed1979

In vitro proliferation of haemopoietic cells in the presence of adherent cell layers. II. Differential effect of adherent cell layers derived from various organs.

Mouse bone marrow-derived adherent cell populations promoted proliferation of haemopoietic cells in vitro in a liquid culture system for at least 4 weeks. Adherent cell layers derived from other haemopoietic organs (foetal liver, adult spleen) and fibroblasts from embryonic tissues did not maintain haemopoietic cells in this system. Medium, conditioned by adherent cells derived from foetal liver, spleen and embryonic tissues displayed a considerable colony stimulating activity (CSA). Marrow-derived adherent cells produced no detectable CSA. A possible relationship between the in vitro expression of a growth-promoting activity of an adherent cell population on haemopoietic cells, and its endogenous CSA production, is discussed.

Animals↗
PubMed1976

The role of adherent cells in the immune response. Fibroblasts and products released by fibroblasts and peritoneal cells can substitute for adherent cells.

The primary immune response to sheep erythrocytes in adherent cell-depleted cultures was restored by adding a critical number of peritoneal cells. Complete substitution was achieved also with supernatants from allogeneic and syngeneic peritoneal cells. Both living fibroblasts and supernatants from fibroblast cultures were found to be highly efficient substitutes for adherent cells in both syngeneic and allogeneic systems. Supernatants from non-antigen-reated peritoneal cells and fibroblasts caused increased DNA synthesis and induction of polyclonal antibody synthesis in normal spleen cells. Thus, adherent cells need not function in the immune response by presenting antigen to the B cells via 'IgT' or by releasing signal-2 activity, which acts on lymphocytes that have already received signal 1.

Animals↗
PubMed1976

A radioimmunoassay for antibodies against surface membrane antigens using adhering cells.

A radioimmunoassay using cells adhering to plastic is described. In this assay, A-10 mammary carcinoma attached to the surface of plastic in microtiter plates were permitted to bind antibody and the bound antibody was detected with purified rabbit 125I-anti-mouse-Fab. The bound radioactive material was eluted with glycine-HCI buffer (pH 2.5), and the acid eluates were counted in a gamma counter. This assay can be used to detect cytolytic or noncyctolytic antibody to cell surface antigens in studies with any tumor or normal cell that will adhere to a solid surface.

Adenocarcinoma↗
PubMed1979

In vitro proliferation of haemopoietic cells in the presence of adherent cell layers. I. Culture conditions and strain dependence.

The culture system, in which a marrow-derived adherent cell population, established in vitro, exerts a long-term promoting influence on proliferation of haemopoietic cells, is reproduced. Essential parameters of the system are investigated; it is confirmed that the system is critically dependent on horse serum, and on the in vitro age of the adherent cell layer. The growth-promoting effect on haemopoietic cells seems to be independent of the number of marrow cells per culture flask initially inoculated into the cultures to establish the adherent cell layer. In vitro established marrow-derived adherent cell layers from RFM (H-2f) and BALB/c (H-2d) mice can promote the long-term proliferation of syngeneic and allogenic haemopoietic cells; haemopoietic marrow cells from C3H (H-2k) cannot be maintained on syngeneic or allogeneic (BALB/c, H-2d) adherent cell layers; adherent cell layers of C3H (H-2k) can maintain haemopoietic cells of the H-2d (BALB/c) genotype. This culture system does not reproduce the in vivo phenomenon of allogeneic resistance. The relevance of these findings to the suggestion that the growth-promoting activity of adherent marrow cells on haemopoietic stem cells in vitro duplicates aspects of the in vivo haemopoietic microenvironment is discussed.

Animals↗
PubMed1977

Stimulation of human haemopoietic cells by colony stimulating factors: adherent cell dependent colony stimulating activity in human serum.

The number of granulocyte-macrophage clones formed in agar culture of bone marrow is dependent on levels of colony stimulating activity (CSA) a proposed in vivo haemopoietic regulator. A dose-response relationship for stimulation of human haemopoietic cells by CSA is demonstrated, which could be explained by thresholds of stimulation to cell division following a normal distribution. A simple method for the comparison of activities of test and control sources of CSA is presented. The apparent potentiating effect of the addition of two sources of CSA is explained by this dose-response relationhip. Haemopoietic cells from patients with chronic granulocytic or acute myeloid leukaemia showed the same dose-response relationship. CSA levels in normal human sera were greatly reduced by assay in the absence of adequate numbers of bone marrow "adherent cells" (cells adherent to nylon or plastic) or peripheral blood leucocytes, suggesting the presence of two kinds of CSA in human serum, one dependent on the presence of bone marrow adherent cells and one effective in their absence. Reduction of numbers of nonspecific esterase positive cells in the bone marrow sample correlated with reduction in the stimulating effect of serum. In all sera tested, adherent cell dependent CSA was the major component.

Cell Adhesion↗
PubMed1992

Characterization of murine CD10, an endopeptidase expressed on bone marrow adherent cells.

The CD10/neutral endopeptidase (NEP) gene was identified in murine genomic DNA using a human CD10 cDNA probe. It is transcribed most abundantly in kidneys resulting in RNA transcripts of 3.4, 6.0, and 6.2 kb. The activity of the murine CD10/NEP shows identical kinetic parameters, Km and Ki, to those observed for this enzyme in other species. Two mAbs raised against rabbit NEP detect a 100 kDa protein by Western blot analysis; the antigen immunoprecipitated from extracts of lung shows specific NEP activity. CD10/NEP, as analyzed by Western blot and enzymatic activity, is expressed at high levels in kidney and lung, and at lower levels in liver, brain, thymus, spleen, and bone marrow. Analysis of bone marrow subpopulations indicate that the majority of CD10/NEP is associated with cells adherent to plastic and with subpopulations that do not express the surface markers AA4.1, B220, Mac-1, and Gr-1. These results suggest that CD10 is primarily associated with the stromal elements in murine bone marrow. A bone marrow stromal line, BMS 2.2, also expresses high levels of CD10/NEP. This peptidase activity on the surface of stromal cells may influence lymphopoiesis or other hematopoietic processes through the hydrolysis of regulatory peptides in the microenvironment.

Animals↗
PubMed1977

Role of T and adherent cells in in vitro response of nude mouse spleen cells to bacterial lipopolysaccharide.

Thy 1.2-positive cells and adherent cells were depleted from nude mouse spleen cell suspensions by treatment with anti-Thy 1.2 antisera plus complement and by passage through Sephadex G-10 column, respectively. The results of 3H-TdR uptake and appearance of IgM containing cells induced in vitro by lipopolysaccharide (LPS) were similar in both untreated and depleted spleen cells. Our findings confirm T and adherent cell independence of LPS response in nude mice.

Animals↗
PubMed1976

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

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.

Animals↗
PubMed1979

The requirement for adherent cells in the Fc fragment-induced proliferative response of murine spleen cells.

The proliferative response of mouse B lymphocytes induced by Fc fragments was found to be dependent upon an adherent cell population. The adherent cell is esterase positive, irradiation resistant, and not susceptible to lysis by anti-thymus serum and complement. The mechanism(s) by which Fc fragments induce B-cell proliferation could be the result of the interaction of Fc with both B cells and adherent cells or with adherent cells which then release factors that trigger the B cells to proliferate. Spleen cells from the C3H/HeJ mouse were shown to be unable to respond to Fc fragments. The addition of adherent cells from either C3H/St or C3H/HeN mice to adherent cell depleted C3H/HeJ cells enabled them to respond to Fc, indicating the defect was in the adherent cell population.

Animals↗
PubMed1992

Erythropoiesis in long term cultures of foetal liver cells is transiently obtained on adult but not on foetal adherent cell layers.

We have previously reported long term erythroid differentiation of adult bone marrow cells seeded onto adherent cells derived from adult bone marrow. In this paper, we show that the adherent cells obtained from foetal liver do not support the erythroid differentiation of either adult bone marrow cells or foetal liver cells. Adherent layers derived from bone marrow of adult W/Wv mice supported differentiation of adult bone marrow precursors, but foetal liver progenitors only produced erythrocytes for a few weeks and the foetal origin of these red cells was confirmed by haemoglobin typing. The duration and extent of erythropoiesis was generally inversely proportional to the cell dose. Foetal progenitors were as sensitive to erythropoietin as adult cells, but were optimally stimulated at a lower plateau concentration. These results suggest that inhibitory cells present in foetal liver may block erythropoiesis and their growing importance with age may provide an explanation for the arrest of erythropoiesis in the liver at late developmental stages.

Anemia↗
PubMed1977

Stimulation of human peripheral blood lymphocytes by periodate, galactose oxidase, soybean agglutinin, and peanut agglutinin: differential effects of adherent cells.

Blastogenic responses of normal human peripheral lymphocytes to three distinct groups of mitogens were studied: Group I--phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM); Group II--soybean agglutinin (SBA) and peanut agglutinin (PNA); and Group III--galactose oxidase (GO) and sodium periodate (IO4-). SBA was mitogenic for human cells, and this effect was enhanced by treating the cells with neuraminidase (NA). PNA was mitogenic only after cells had been treated with NA. GO was effective before and activity was increased after lymphocytes were treated with NA. Responses to Group II and III mitogens were more variable than were those to Group I mitogens. Studies with purified T and B cells indicated that SBA and PNA were T cell mitogens, whereas IO4- and GO failed to stimulate either T or B cells. Adding macrophages back to this system indicated that they were both T cell mitogens with strict macrophage requirements. T cell responses to SBA and PNA were enhanced over responses to unfractionated cells to a degree that could not be explained simply by enrichment of the cultures with T cells. Removal of adherent cells from unfractionated cell suspensions again revealed a marked enhancement of responses to SBA and PNA, a consistent decrease in responses to IO4-, and a variable decrease in responses to GO. Similar results were found with 14C-leucine and 3H-uridine incorporation, as well as 3H-thymidine for the assessment of bastogenic response. Mechanisms responsible for these differential effects of macrophage depletion on lymphocyte responses to different groups of mitogens are yet to be determined. Either different mitogens require different lymphocyte to macrophage ratios for optimal stimulation, or some mitogens (i.e., SBA and PNA) form inhibitory complexees in the lymphocyte-macrophage mixture. In any case, variability in response to mitogenic agents in normal as well as pathologic states may be dependent on adherent cell populations, rather than on the lymphocytes themselves.

Alcohol Oxidoreductases↗
PubMed1978

Adherent cell function in murine T lymphocyte antigen recognition. II. Definition of genetically restricted and nonrestricted macrophage functions in T cell proliferation.

The mechanisms by which adherent cells, presumably of mononuclear phagocytic lineage, influence in vitro antigen-specific activation of murine T lymphocytes was examined. Two distinct functions for macrophages could be discerned. One macrophage function is dependent on a soluble factor produced by cultured adherent cells and is most easily studied with complex multideterminant antigens. This factor is neither antigen-specific nor MHC-restricted in its action in that PEC, regardless of haplotype, produce factor in the absence of antigen. A second function, antigen-specific T cell activation, is seen when antigens of more restricted heterogeneity are used, such as those under the control of Ir genes. This latter activity demands identity or partial identity between the antigen-presenting cell and the primed T cell, thus suggesting an additional specific, genetically restricted function for macrophages in in vitro antigen recognition. Whether these adherent cell functions are mediated by all or distinct subsets of cells was not established.

Animals↗
PubMed1979

T cell recognition in the mixed lymphocyte response. I. Non-T, radiation-resistant splenic adherent cells are the predominant stimulators in the murine mixed lymphocyte reaction.

The ability of subpopulations of murine spleen cells to stimulate a mixed lymphocyte response (MLR) was studied. It was found that T cells (nylon-nonadherent spleen cells) and B cells [G-10 passed and treated with rabbit anti-mouse brain serum (RAMB) and complement (C)] were poor stimulators of an MLR. In contrast, whole spleen cells or B cells plus adherent cells (RAMB +C-treated spleen cells) produced good stimulation. However, a non-T, radiation-resistant splenic adherent cell (SAC) population was up to 20 to 50 times more efficient as a stimulator of an MLR on a per cell basis than an unseparated spleen population. These SAC were shown to express Ia determinants encoded by genes in I-A and I-E/C. These results suggest that Ia+ SAC may be the predominant stimulating cells in spleen cell populations, and the preferential target for T cell recognition in cell interaction events.

Animals↗
PubMed1977

Enhancement of phytohemagglutinin and concanavalin A responses of splenic lymphocytes by adherent cell removal.

Removal of glass wool-adherent cells resulted in an enhancing effect on DNA synthesis as measured by increased [3H]-thymidine uptake by normal swine spleen cells stimulated with phytohemagglutinin or concanavalin A (Con A). Depletion of nylon-adherent cells caused decreased DNA synthesis in response to Con A, except at high doses of Con A, whereas similar cultures stimulated with phytohemagglutinin showed enhanced DNA synthesis throughout the dose range employed. Titration experiments revealed that enhancing effects of adherent cell removal are most pronounced at higher cell and mitogen concentrations. Glass wool was more efficient than nylon in the enhancement of mitogen-induced DNA synthesis.

Animals↗
PubMed1979

Quantitative development of adherent cell colonies in bone marrow cell culture in vitro.

Quantification of the formation of adherent cell colonies in bone marrow cell culture was attempted. By secondary transfer of the bone marrow cells as a single cell suspension after 4 days' culture of fine marrow fragments, a linear relationship was obtained between the number of adherent cell colonies developing and the number of cells secondarily inoculated into the culture bottle. This suggests that 4 days' culture of the bone marrow cells with close intercellular interactions is sufficient for the 'conditioning' of the cells to develop adherent cell colonies. Activity of such colonies to support haemopoietic stem cell proliferation was also shown.

Animals↗
PubMed1992

Correlation between cell-adherent activity and surface structure in Porphyromonas gingivalis.

The cell-adherent ability of 6 strains of Porphyromonas gingivalis (381, ATCC 33277, SU63, KD1, W50 and W83) was compared by using radiolabeled bacterial cells and human gingival fibroblasts (Gin 1), human periodontal ligament fibroblasts (HPLF) and human epithelial cells (Ca9-22) that had been grown on collagen beads. The cell-adherent activity of these organisms varied among strains; P. gingivalis strains 381, ATCC 33277 and SU63 bound to the target cells at a range of 14% to 72%, but the other 3 strains (KD1, W50 and W83) were scarcely bound (0.6% to 3.5%). On the other hand, whole bacterial cells and culture supernatants of all strains showed distinct hemagglutinating activity. The 3 strains showing high cell-adherent activity were hydrophobic and the other strains showing less activity were relatively hydrophilic. Furthermore, a number of peritrichous fimbriae were found on the surface of P. gingivalis strains 381, ATCC 33277 and SU63, which showed high adherent activity, whereas, fimbriae on the other 3 strains showing low adherent ability were barely apparent. Therefore, it was assumed that the cell-adherent activity of P. gingivalis was related to the hydrophobicity of the cell surface, which was related to the number of fimbriae.

Bacterial Adhesion↗
PubMed1976

The role of adherent cells in the secondary cell-mediated response in vitro to a natural poxvirus pathogen.

The role of adherent cells in an in vitro secondary response to ectromelia virus infection was investigated. Spleen cells from ectromelia-primed mice ("responder" cells) depleted of adherent cells by either carbonyl iron treatment, adherence to plastic or passage through cotton wool columns had a markedly decreased capacity to produce a secondary response, as indicated by decreased T cell-mediated cytotoxicity against virus-infected target cells, when cultured with virus-infected "stimulator" cells. The secondary response was restored by the addition of peritoneal cells from either normal or ectromelia-immune mice. Small numbers of peritoneal cells completely reconstituted the response within a certain dose range but larger numbers produced a marked inhibition of the response. Spleen cells were less effective in restoring the response. The peritoneal cells were not merely acting as additional, infected "stimulator" or antigen-presenting cells, since they could be added as late as 3 days after culture. Reconstituting activity was not affected by pretreatment with anti-theta serum and complement and cell separation studies showed that the activity was associated mainly with Ig-negative cells and that the active cell probably bears Ia antigens on its surface. These results indicate that the adherent cells involved are probably macrophages and that they act non-specifically to produce optimum conditions for the specific response of T cells.

Animals↗