Suppressive effect of immature erythroid cells on the B-cell proliferation

Самарин, Д. М.; Селедцова, Г. В.; Селедцов, В. И.; Taraban, Vadim Y.; Козлов, В. А.

doi:10.1007/bf02764380

Cited by 8 publications

(10 citation statements)

References 8 publications

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“…Furthermore, stimulation of T suppressor cells with prostaglandin E 2 [11] can be the cause of weak immune response in mice. Erythroid cells can also produce suppressive effects: immature cells inhibit B lymphocyte proliferation, while mature cells affect APC formation [4,13].…”

Section: Resultsmentioning

confidence: 99%

Peculiarities of primary humoral immune response in mice with cytostatic disease

Masnaya¹,

Ratner²

2000

Bull Exp Biol Med

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Single intraperitoneal injection of cyclophosphamide in a maximum permissible dose (250 mg/kg) followed by immunization wih thymus-dependent antigen (sheep erythrocytes) markedly suppressed the formation of antibody-producing cells in the spleen, especially at early stages of cytostatic disease, and delayed the synthesis of specific antibodies in male CBA/CaLac mice. Platidiam (cisplatin) injected in comparable doses 4 days before immunization practically did not suppress the formation of antibody-producing cells and their functional activity, but being injected 30 days before immunization reduced the number of antibody-producing cells.

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Section: Resultsmentioning

confidence: 99%

Peculiarities of primary humoral immune response in mice with cytostatic disease

Masnaya¹,

Ratner²

2000

Bull Exp Biol Med

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“…Immunofluorescence analysis of cells [3] showed that the purity of isolated T and B lymphocytes was close to 100%.…”

Section: Methodsmentioning

confidence: 99%

“…Splenocytes from normal BDF1 mice were activated with concanavalin A (Pharmacia, 5 ktg/ml) or lipopolysaccharide (E. coli 055 :B5:, Sigma, 20 ~tg/ml) in 25 cm2-plastic flasks (Linbro) (10 ml) tor 24 h. T and B lymphocytes activated with concanavalin A and lipopolysaccharide, respectively, were isolated by the positive penning method [3]. Immunofluorescence analysis of cells [3] showed that the purity of isolated T and B lymphocytes was close to 100%.…”

Section: Methodsmentioning

confidence: 99%

Activated lymphocytes produce mediators potentiating the antitumor cytostatic activity of bone marrow cells

et al. 1999

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Mitogen-activated murine T and B Iymphocytes produce soluble mediators that potentiate the inhibitory effect of bone marrow cells on the growth of leukemic cells in vitro. 7-Interferon is a T-cell product increasing the cytostatic activity of bone marrow cells. An increase in the cytostatic activity under the effect of T-cell soluble mediators was characteristic of bone marrow cells from nude mice. Key Words: T and B lymphocytes; bone marrow cell; tumor growth inhibitionBone marrow cells (BMC) phenotypically similar to bone marrow natural suppressor cells inhibit the growth of leukemic cells in vitro [1,2,[8][9][10]. Tumor growth suppression by BMC is not associated with tumor cell destruction and is mediated, at least partially, by solutile cytostatic products [9].We investigated the ability of activated T and B lymphocytes to regulate the cytostatic activity (CSA) of BMC by producing soluble factors. Cells were cultured in RPMI-1640 with 10 mM HEPES, 2 mM L-glutamine, 5x10 -5 M 2-mercaptoInstitute oi" Clinical Immunology, Siberian Division, Russian Academy of Medical Sciences, Novosibirsk ethanol, antibiotics, and 7% fetal calf serum (all reagents from Sigma) in a humidified atmosphere with 5% CO 2. MATERIALS AND METHODS(Splenocytes from normal BDF1 mice were activated with concanavalin A (Pharmacia, 5 ktg/ml) or lipopolysaccharide (E. coli 055 :B5:, Sigma, 20 ~tg/ml) in 25 cm2-plastic flasks (Linbro) (10 ml) tor 24 h. T and B lymphocytes activated with concanavalin A and lipopolysaccharide, respectively, were isolated by the positive penning method [3]. Immunofluorescence analysis of cells [3] showed that the purity of isolated T and B lymphocytes was close to 100%.For preparing culture supernatants, activated lymphocytes (3-4x106 cells/ml) were cultured in a 24-well plate (Linbro) for 24 h. The supernatant was obtained by centrifugation and stored at -20~ until use.In the cytostatic test, BMC (3xl05/well) were cultured with (25%) or without the supernatant from activated T or B lymphocytes in round-bottom 96-well plates (BDSL) tot 20 h. After the medium replacement, P815 or L1210 ceils were added to BMC into wells (104 cells/well) and cultured for 24 h. In the control tests, tumor cells were cultured without adding any cells or with thymocytes incapable of suppressing leukemic cell growth in vitro [1,8]. The level of proliferative activity was measured routinely by-incorporation of 3H-thymidin (Izotop) added in a dose of 0.75 lxCi to all wells 5 h belore the end of culturing.

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“…At least 60% of suspended cells of erythropoietic spleen were NEC [3,10]. NEC were separated from other cells as described elsewhere [1]: blast cells were isolated by centrifugation in a Percoll density gradient (1.075 g/ liter), treated with anti-Thy 1.2 monoclonal antiInstitute of Clinical Immunology, Siberian Division of Russian Academy of Medical Sciences, Novosibirsk bodies (clone 5a-8, IgG2b, ascites dilution 1:500, Cedaflane), macrophages, and T-and B-lymphocytes were removed by adhesion to plastic coated with anti-immunoglobulin antibodies, and the cells that failed to adhere to the plastic were treated with leucyl-methyl ether (Fluka) toxic for large granular lymphocytes and macrophages [14]. Cell viability was assessed by the trypan blue exclusion test.…”

Section: Methodsmentioning

confidence: 99%

Role of transforming growth factor-β in erythroid cell-mediated suppression of B-cell blastogenesis

et al. 1997

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Soluble products spontaneously secreted by murine nuclear erythroid cells directly suppress the proliferation of activated B-lymphocytes induced by bacterial lipopolysaccharides. Blocking of the transforming growth factor-13 synthesis in nuclear erythroid cells by antisense oligonucleotides binding mRNA and blocking of this factor's functional activity by neutralizing antibodies were associated with a marked decrease in suppressor activity of the medium conditioned by nuclear erythroid cells. Key Words: erythroblast; natural immunosuppression; transforming growth factor-f~Nuclei-containing erythroid cells (NEC) suppress humoral immune response induced by both thymusdependent and thymus-independent antigens [2][3][4]. This immunoregulatory effect of NEC is mediated, at least partially, by secretion of soluble suppressor factors [10]. We tried to prove that transforming growth factor-13 (TGF-~) is one of such factors. MATERIALS AND METHODS(CBA• F 1 (CBF1, H-2VH-2 b) mice aged 4-8 months from Breeding Center of Siberian Division of the Russian Academy of Medical Sciences were used.Splenic erythropoiesis was induced by intraperitoneal injection of phenylhydrazine (1.2 mg/ml, Sigma) as described previously [2,3,10]. At least 60% of suspended cells of erythropoietic spleen were NEC [3,10]. NEC were separated from other cells as described elsewhere [1]: blast cells were isolated by centrifugation in a Percoll density gradient (1.075 g/ liter), treated with anti-Thy 1.2 monoclonal antiInstitute of Clinical Immunology, Siberian Division of Russian Academy of Medical Sciences, Novosibirsk bodies (clone 5a-8, IgG2b, ascites dilution 1:500, Cedaflane), macrophages, and T-and B-lymphocytes were removed by adhesion to plastic coated with anti-immunoglobulin antibodies, and the cells that failed to adhere to the plastic were treated with leucyl-methyl ether (Fluka) toxic for large granular lymphocytes and macrophages [14]. Cell viability was assessed by the trypan blue exclusion test. Analysis of cell smears stained by the Nocht--Maksimov method showed that at least 95% of the resultant cell population were basophilic NEC.For preparing the supernatant, NEC were cultured in serum-free RPMI-1640 medium with 20 mM HEPES, 2 g/iitcr NaHCO3, 4 mM L-glutamine, and antibiotics (all reagents from Sigma) in a humidified atmosphere with 5% CO 2. After 24 h, the culture medium was collected by centrifugation, sterilized by fdtration through 0.22 ~t fdter, and stored at -20~ Lymphocytes were cultured in RPMI-1640 with 2 g/liter NaHCO3, 20 mM HEPES, 2 mM L-glutamine, 5x 10 -s M 2-mercaptoethanol, antibiotics, and 7% fetal calf serum in humidified atmosphere with 5% CO 2.For preparing activated B-lymphocytes, 10 ml of splenocyte suspension (5 x 106 cells/ml) was cultured

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Suppressive effect of immature erythroid cells on the B-cell proliferation

Cited by 8 publications

References 8 publications

Peculiarities of primary humoral immune response in mice with cytostatic disease

Peculiarities of primary humoral immune response in mice with cytostatic disease

Activated lymphocytes produce mediators potentiating the antitumor cytostatic activity of bone marrow cells

Role of transforming growth factor-β in erythroid cell-mediated suppression of B-cell blastogenesis

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