Polyanions and lipopolysaccharide acts on different subpopulations of B cells

Diamantstein, Tibor; Blitstein-Willinger, E; Schulz, G.

doi:10.1038/250596a0

Cited by 54 publications

(21 citation statements)

References 2 publications

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“…These data are in agreement with those reported by Gronowicz and Coutinho (13) who showed that DxS stimulates a less mature population of LPS-sensitive cell subsets which can also be found in bone marrow (14).…”

Section: Discussionsupporting

confidence: 83%

“…In mice, Melchers and Andersson (12) concluded that PPD and lipepelysaccharide (LPS) I stimulate the same cell, whereas Gronowicz and Coutinho (13) obtained an additive effect after double stimulation with PPD and LPS. Diamanstein et al (14) have also obtained an additive effect ai~r double stimulation with dextran sulfate (DxS) and LPS. Finally, it was recently suggested that LPS-reactive B cells differ from those B cells stimulated by lipoprotein (15,16) or peptidoglycan (17) prepared from Escherichia coli.…”

mentioning

confidence: 98%

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Mitogenic analysis of murine B-cell heterogeneity.

Bona¹,

Yano²,

Dimitriu³

et al. 1978

The Journal of Experimental Medicine

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Murine B lymphocytes appear to be heterogeneous from a functional point of view. Thus, those B cells that give rise to antibody-producing cells can be divided into a subset that requires cooperation with T-derived lymphocytes and another subset that is T independent (1-6). Recently, it has been shown that the B-cell population also contains suppressor cells that regulate various functions of T cells (7)(8)(9)(10)(11) and that these suppressor cells seem to be different from antibody-producing cells (10, 11).There is contradictory evidence as to whether the response to different mitogens defines different B-cell subpopulations. In mice, Melchers and Andersson (12) concluded that PPD and lipepelysaccharide (LPS) I stimulate the same cell, whereas Gronowicz and Coutinho (13) The aim of this work was to test directly various alternative hypotheses that could explain the differences in the responsiveness of routine B lymphocytes with regard to various mitogens: (a) One cell possesses the receptors for various mitogens and can be stimulated by all known murine B-cell mitogens separately; (b) the ability to respond to various mitogens depends on the stage of maturation of the B cell; (c) there are different subsets of lymphocytes for different mitogens.We have chosen three well-known B-cell mitogens for this study; LPS (12), Nocardia water-soluble mitogen (NWSM) (18) and DxS (13). We have used the 5-bromo-deoxyuridine (BUdR) suicide technique of Zoschke and Bach (20). This should be a more critical method for ascertaining whether different cells respond to different mitogens by eliminating or inactivating those cells that respond to an initial mitogen.The specific killing by rabbit anti-mouse immunoglobulin serum and by antiIa alloantisera was used to investigate the presence of Ig and Ia on the surface of cells reactive to the above mitogens.1 Abbreviations used in this paper: BUdR, 5-bromo-deoxyuridine; CNRS, Centre National de la Recherche Scientifique; Con A, concanavalin A; DxS, dextran sulfate; LPS, lipopolysaccharide; NWSM, Nocardia water-soluble mitogen. 136J. ExP. MED.

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Section: Discussionsupporting

confidence: 83%

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confidence: 98%

Mitogenic analysis of murine B-cell heterogeneity.

Bona¹,

Yano²,

Dimitriu³

et al. 1978

The Journal of Experimental Medicine

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“…Additive effects of two mitogens indicate differences in the mitogens and possibly also in the responding cell populations, whereas, greater than additive colony stimulation suggests that some cells may require multiple mitogen encounters for proliferation. Both phenomena have previously been described for liquid culture systems (6)(7)(8). Resistance to alkali treatment, absolute dependence on the presence of mercaptoethanol, differential sensitivity of the stimulated cells to anti-/~ antibodies, and different patterns of Ig class expression by proliferating cells further distinguishes agar mitogen from LPS.1 Agar has been used by others as a supporting matrix for studying cell-cell interactions in mitogen responses (9), to clone mitogen-activated cells (10)(11)(12), and in culture systems where agar was assumed to be inert (13).…”

Section: Discussionmentioning

confidence: 99%

Growth of B-lymphocytes clones in semisolid culture is mitogen dependent.

Kincade¹,

Ralph²,

Moore³

1976

The Journal of Experimental Medicine

104

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In vivo cloning techniques have made it possible to estimate the frequency, triggering requirements, and proliferative potential of individual B lymphocytes, as well as the diversity of major class and specificity of their end products (1-3). Recently, an in vitro technique for cloning murine B lymphocytes in agar has been described (4). In this report, we present evidence that a polysaccharide B-cell mitogen/polyclonal activator is present in unrefined agar and it or another appropriate B-cell mitogen is essential to B-cell proliferation in semisolid cultures. Semisolid Cultures. Single spleen cell suspensions were prepared, freed of coarse debris by settling and washed once with RPMI 1640 medium containing 10% fetal calf serum. Single strength McCoy's modified 5a medium plus 15% fetal calf serum, 2 mM L-glutamine, 16 ~g/ml Lasparagine, 8 ~g/ml L-serine, and 5 x 10 -3 M 2-mercaptoethanol (2-ME) was warmed to 37°C and mixed with a 1/10 volume of boiled 3% Bacto agar in H20. Spleen cells (2 x 104) were added and 1-ml aliquots were allowed to gel in 35-mm tissue culture dishes. Alternatively, agarose at 0.2% final concentration or 1% methyl cellulose were used instead of agar. Granulocyte-macrophage progenitors (CFU-c) were stimulated with an optimal concentration of medium conditioned by murine myelomonocytic leukemia (WEHI 3) cells in cultures of 7.5 x 104 bone marrow cells using the same medium without mercaptoethanol. Cultures were held at 37°C in a humidified atmosphere of 10% CO2-90% air for 5-7 days before examination with a dissecting microscope. Materials and MethodsLiquid Cultures. Duplicate cultures of spleen cells (2 x 10~/ml) were prepared in RPMI 1640 medium containing 5% fetal calf serum and 5 x 10 -5 M 2-ME in 0.2-ml volumes plus 0.01 ml of mitogen or phosphate-buffered saline (PBSL After 3 days incubation at 37°C and 10% CO2, the cultures were labeled by the addition of 0.2 ~Ci/well of '2~I-iododeoxyuridine ('25IUdR) (200 Ci/ mmol; New England Nuclear, Boston, Mass.). After an additional 3 h at 37°C, cultures were harvested on filter paper, washed with TCA using a semiautomatic cell harvester, and counted in a gamma counter. Lipopolysaccharide from Salmonella thyphosa WO901 (Difco Laboratories, Detroit, Mich.) and dextran sulfate (500,000 mol wt; Sigma Chemical Co., St. Louis, Mo.) were used as mitogens.Extraction and Chemical Characterization of Agar Mitogen. Bacto agar Clot no. 504388 and 551975; Difco Laboratories) was extracted with 37°C H20 for 1 h with occasional shaking, filtered through Whatman no. 1 paper on a scintered glass funnel, and lyophilized. Up to 80 mg was recovered/g of agar. Before use the material was dissolved in 37°C H20 and subjected to one of the *

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“…Al though there is a possibility that (-/-bearing lymphocytes were found in this site [16] and responded to LPS, it provides strong cir cumstantial evidence for nonthymic origin of the blasts that in the control (nu/ + ) the dividing cells were present in the follicular area. Appearance of dividing blasts is shown in the periarteriolar sheath of nu/nu mice following intracutaneous injection of Shigella dysenteriae antigen, the other thy mus-independent antigen [4], It has been shown that B cells can be di vided into several subsets with regard to the different responsiveness to B cell mitogens and that one specific subset responds prefer entially to LPS [5,7,12]. In nu/+ mice, this LPS-responsive subset appears to popu late in the follicular area where the blasts were observed mainly after LPS stimula tion.…”

Section: Discussionmentioning

confidence: 99%

<i>In vivo</i> Maturation of B Cells in the Spleen of Nude Mice Following Administration of Bacterial Lipopolysaccharide

Takigawa¹,

Hanaoka²

1978

Int Arch Allergy Immunol

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Morphological changes of the splenic white pulp in athymic nude mice (nu/nu) and their normal littermates (nu/+) following intraperitoneal injection of bacterial lipopolysaccharide were studied by light and electron microscopy, immunofluorescence and autoradiography. Early blast formation and subsequent appearance of IgM-containing cells were observed by 72 h and at 120 h, respectively, in the periarteriolar sheath of nu/nu mice and in the follicular area of nu/+ mice. Ultrastructural details of blasts and the time course of their development were similar in both nu/nu and nu/+ mice. Lymphoblasts showed a large nucleus with a prominent nucleolus, many polysomes and poorly developed smooth endoplasmic reticulum. Plasmablasts had a nucleus with coarse heterochromatin and copious cytoplasm filled with dilated rough endoplasmic reticulum. Generally, lymphocytes proliferated and differentiated through lymphoblasts to plasmablasts by 72 h and finally to plasma cells at 120 h. However, this development was asynchronous since lymphoblasts, plasmablasts and plasma cells were observed simultaneously at 72h. It was suggested that a B cell subset responsive to bacterial lipopolysaccharide matures to antibody-forming cells in the thymus-dependent area in nu/nu mice and in the thymus-independent area in nu/+ mice.

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Polyanions and lipopolysaccharide acts on different subpopulations of B cells

Cited by 54 publications

References 2 publications

Mitogenic analysis of murine B-cell heterogeneity.

Mitogenic analysis of murine B-cell heterogeneity.

Growth of B-lymphocytes clones in semisolid culture is mitogen dependent.

<i>In vivo</i> Maturation of B Cells in the Spleen of Nude Mice Following Administration of Bacterial Lipopolysaccharide

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