共查询到20条相似文献，搜索用时 15 毫秒
1.
Virus-induced interferon (IFN) production in canine lymphoid cells was studied, using Newcastle disease virus as principal inducer. It was found that spleen cells at a concentration of 5 X 10(6) cells/ml with Newcastle disease virus at a multiplicity of infection of 1, incubated at 37 C in 5% CO2 for 24 hours, produced highest titers of IFN. Among the lymphoid cells from different tissues, IFN production was in the order of spleen = bone marrow greater than thymus greater than mesenteric lymph node greater than or equal to peripheral blood lymphocytes. Macrophages did not produce IFN, and virus-induced IFN production in spleen cells did not depend on the presence of phagocytic mononuclear cells. These optimal conditions and macrophage independence are different from those of mitogen-induced IFN production in canine lymphoid cells. 相似文献
2.
Optimum conditions for turkey lymphocyte transformation tests were determined. Thrice-washed turkey buffy-coat cells obtained after slow centrifugation (40 x g, 10 minutes) responded well to mitogenic stimulation. Turkey lymphocytes isolated on Ficoll-containing separation media largely lost their ability to respond to mitogens. Maximum responses were obtained with 2 x 10(7) lymphoid cells/ml. Responses to the mitogens were greatest when bovine fetal serum was used at a 2.5% concentration or pooled turkey serum and autologous plasma were used at a 1.25% concentration. Higher concentrations of turkey serum or plasma decreased the responses when sub-optimum doses of concanavalin-A (Con A) or phytohemagglutinin-P (PHA-P) were used. Serum-free cultures gave higher stimulation indices than cultures with serum only when sub-optimum doses of Con A or PHA-P were used. Optimum mitogen concentrations varied with individual birds, timing of the culture, temperature of incubation, and serum concentration in the cultures. Responses were usually greatest with final concentrations of 5 micrograms Con A/ml, 10 micrograms PHA-P/ml, and 20 micrograms pokeweed mitogen (PWM)/ml and when the cultures were incubated in 96-well microplates at 40 C in humidified air with 5% CO2 for 40-42 hours with pulsing with 3H-thymidine during the final 16 hours of incubation. 相似文献
3.
试验旨在探索使较高比例的淋巴细胞富集在有丝分裂G2/M期的最佳条件。运用植物血凝素(PHA)和刀豆蛋白A(ConA)对淋巴细胞进行刺激使其增殖,培养一定时间后加秋水仙素对淋巴细胞进行同步化处理,用流式细胞仪检测G2/M期的细胞数量进行比对,观察试剂的最佳作用浓度和加秋水仙素的最佳时间。结果表明,采用PHA和ConA刺激淋巴细胞增殖的最佳作用浓度是60和5 μg/mL,且淋巴细胞经ConA刺激培养45 h再加秋水仙素处理5 h G2期的比例最高为58.38%。结果提示,就绒山羊的淋巴细胞来说,ConA刺激绒山羊淋巴细胞增殖的效果比PHA好,且摸索出细胞G2/M期的时间点至关重要。 相似文献
4.
Y Sanada H Noda H Nagahata 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(3):455-460
A fluorometric assay was applied to evaluate blastogenesis of equine lymphocytes. Optimal culture conditions were as follows; concentrations of phytohaemagglutinin-P (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM) were 1 microgram/ml, 40 micrograms/ml and 10 micrograms/ml, respectively, when 5 X 10(5) lymphocytes were incubated with culture medium containing 20% pooled horse serum (PHS) for 120 hours. The relative mean stimulation index of healthy non-pregnant mares were 5.107 +/- 0.323 (M +/- SE) with PHA, 4.019 +/- 0.183 with Con A and 3.610 +/- 0.131 with PWM. Sequentially the blastogenic responses of lymphocytes from twenty mares were observed during various stages of the perinatal period. Response decreased gradually before parturition was lowest at the time of parturition (PHA: 1.923 +/- 0.174, Con A: 1.698 +/- 0.206 and PWM: 1.706 +/- 0.177), and then increased gradually after parturition towards non-pregnant levels. 相似文献
5.
Modulation of sheep lymphocyte responses by Fasciola hepatica excretory-secretory products 总被引：2，自引：0，他引：2
The effect of Fasciola hepatica excretory-secretory products (FhESPs) on mitogen-induced proliferation of sheep peripheral blood mononuclear cells (PBMCs) and PBMC subsets (CD2(+), CD4(+), CD8(+), gammadeltaTCR(+) or CD21(+) cells) were studied. PBMCs were incubated with Concanavalin A (ConA) or phytohemagglutinin (PHA) at optimal (1 microg per well) or suboptimal (0.25 microg per well) doses and with FhESPs at several doses (1.25-20 microg per well). PBMC subsets were incubated with ConA at a suboptimal dose and with FhESPs at 5 microg per well. These cells were incubated with or without monocytes (CD14(+) cell). FhESPs slightly increased the proliferation of PBMCs stimulated with optimal doses of PHA. FhESPs (10 and 20 microg per well) inhibited the PBMCs stimulated with optimal doses of ConA. FhESP dose-dependent inhibition was observed on PBMCs stimulated with suboptimal doses of ConA. CD21(+) lymphocytes (B lymphocytes), CD14(+) cells (monocytes) and gammadeltaTCR(+) cells were not stimulated by ConA. T lymphocyte subsets (CD2(+), CD4(+) or CD8(+) cells) proliferation was decreased by FhESPs at 5 microg per well. FhESPs inhibits the ConA-induced stimulation of sheep PBMCs and sheep T lymphocyte subsets. Further studies should be done to investigate the mechanism of this FhESP immunomodulatory effect. 相似文献
6.
Optimum conditions for the chicken lymphocyte transformation test. 总被引：16，自引：0，他引：16
Optimum conditions for chicken (Gallus gallus) lymphocyte transformation tests were determined. Thrice-washed chicken buffy-coat cells obtained after slow centrifugation (40 x g for 10 minutes) responded substantially better to mitogenic stimulation than lymphocytes isolated on separation media containing Ficoll. Maximum responses were obtained with 2 x 10(7) lymphoid cells/ml. Responses to the mitogens were greatest when fetal bovine serum was used at a 5% concentration or pooled chicken serum and autologous plasma were used at a 1.25% concentration. Optimum mitogen concentrations varied with individual birds, timing of the culture, temperature of incubation, and serum concentration in the cultures. When 1.25% chicken serum was used in the cultures, responses were usually greatest with final concentrations of 30-50 micrograms/ml of concanavalin A (Con A) and 30-50 micrograms/ml of phytohemagglutinin-P (PHA-P). The optimum concentration of pokeweed mitogen (PWM) varied from 1 to 40 micrograms/ml among the birds and was practically impossible to establish in general. The incubation in humidified air with 5% CO2 was significantly better at 40 C than at 37 C. The total culture time of 40 hours including pulsing with 3H-thymidine during the final 16 hours of incubation was the best for Con A- and PHA-P-stimulated cells, whereas a longer incubation of 64 hours gave the highest results with PWM stimulations. 相似文献
7.
Kok-Mun Tham Colin R. Wilks Michael J. Studdert 《Veterinary immunology and immunopathology》1982,3(5):485-498
A microculture technique was developed for the in vitro blastogenesis of feline lymphocytes. Blastogenesis of ficoll-diatriazoate gradient separated mononuclear cell, washed blood and whole blood were compared. In general the whole blood cultures yielded higher stimulation indices (SI) than the washed blood or separated mononuclear cell cultures.The effect of several variables on the stimulation of lymphocyte cultures was examined. A cell concentration of 3 × 105 cells/well and a 1:20 dilution of washed and unwashed whole blood gave optimal stimulation with concanavalin A (Con A). Phytohaemagglutinin-P (PHA-P) did not give significant levels of stimulation. Inactivated fetal calf serum (FCS) at levels of 2.5% (for washed blood) and 5% (for separated mononuclear cell and whole blood) gave highest SI. Supplementation with FCS was preferable to autologous, homologous or horse sera for all cultures. Optimal SI was obtained in all cultures incubated for 3 days and labelled with 1 μCi tritiated thymidine (3H-TdR) for the last 16 hours. The highest SI were in the range of 70 to 105 (18,764 to 42,681 counts per minute (CPM) for separated mononuclear cell culture, 100 to 165 (28,403 to 45,334 CPM) for washed blood culture and 105 to 186 (41,076 to 69,999 CPM) for whole blood culture. 相似文献
8.
Several experimental parameters were examined to determine optimal conditions for proliferative responses of mammary mononuclear cells (MMC) obtained from six nonlactating dairy cows. These parameters were: pre-incubation of cells in medium prior to assay, mitogen concentration, assay incubation time, and type of culture medium. Response variables included viability of cells and the rate of proliferation as assessed by tritiated thymidine incorporation. Pre-incubation of cells in medium had no effect on the proliferative response of MMC. Whereas Concanavalin A (ConA; 3.3 or 6.6 micrograms/ml) and phytohemagglutinin (PHA; 1, 5, 10 micrograms/ml) did stimulate proliferation of MMC, the higher doses did not stimulate greater proliferation than the lower doses of mitogens. The greatest mitogenic response was obtained on days 2 and 3 of incubation. Proliferative responses were significantly higher at all mitogen levels tested in a 50-50 mixture of Rosewell Park Memorial Institute medium 1640 and Liebovitz-15 medium (RPMI/L-15) than in RPMI alone. Viability of MMC was also significantly higher in the RPMI/L-15 medium. To test whether the significant effect of media on blastogenesis was specific for mononuclear cells from the bovine mammary gland, peripheral blood lymphocytes (PBL) from four dairy cows were cultured with ConA and PHA in a mitogen assay in both RPMI and RPMI/L-15. Viability was measured on day of collection and on all culture days. PBL were stimulated equally in both media. PBL viability decreased significantly on day 1 in both RPMI and RPMI/L-15. These results suggest that the optimal culture conditions for blastogenic responses of mammary mononuclear cells and peripheral blood lymphocytes may differ. 相似文献
9.
试验旨在研究不同浓度的天门冬多糖(ASP),在ConA或LPS的协同刺激下对猪脾淋巴细胞体外增殖的影响。猪脾淋巴细胞体外培养体系中加入不同浓度的天门冬多糖使其终浓度为400、200、100、50、25、12.5μg/ml,在ConA(5μg/ml)或者LPS(10μg/ml)的协同刺激作用下,细胞培养24、48、72 h时,观察猪脾淋巴细胞增殖情况。结果表明,天门冬多糖及其协同ConA或LPS能显著或极显著的促进猪脾淋巴细胞体外增殖(P<0.05或P<0.01)。 相似文献
10.
为了探讨毒害艾美耳球虫(E.necatrix)感染雏鸡后,其免疫器官白细胞介素(IL-2)和T、B淋巴细胞增殖能力的变化,本研究应用组织匀浆涂片和酸性α-醋酸萘酯酶(ANAE)染色及细胞培养技术和四甲基偶氮唑盐(MTT)测定法对E.necatrix感染雏鸡免疫器官的T淋巴细胞百分数、IL-2诱生活性、T和B淋巴细胞对C... 相似文献
11.
鬼臼多糖的分离纯化及药理学活性 总被引：2，自引：0，他引：2
目的：分离纯化鬼臼多糖（PEP）,测定PEP的单糖组成,观察PEP对小鼠脾脏、胸腺指数、溶菌酶含量及脾脏淋巴细胞增殖的影响.方法：用热水抽提及醇沉法纯化鬼臼多糖,采用纸层析法和红外光谱对PEP的组成进行研究,用比浊法测溶菌酶含量,用MTT法研究PEP对脾脏淋巴细胞增殖的影响.结果：PEP的得率为60%,PEP含有葡萄糖、果糖、甘露糖、和阿拉伯糖,红外光谱扫描表现出一般多糖类物质的特征吸收峰.PEP能够提高脾脏指数,但对胸腺指数和血清中的溶菌酶含量的影响不明显.MTT结果显示,PEP能够显著刺激小鼠脾脏淋巴细胞增殖,且与ConA有协同作用. 相似文献
12.
The optimum conditions for the culture of cells from dissociated spleens were determined. Routinely, 10(7) cells were seeded per ml of RPMI 1640 medium supplemented with 20% pre-tested foetal calf serum. For the assay of the immune response, cultures were supplemented with 30 muMolar mercaptoethanol. The immune responses to sheep erythrocyte and bluetongue virus antigens were determined by the haemolytic plaque-forming cell assays described by Oellermann (1974) and Oellermann, Carter & Marx (1976a). The optimum sheep erythrocyte antigen concentration was 2 X 10(6) erythrocytes per 10(7) spleen cells and maximum IgM plaque-forming cells were detected after 4 days in culture. Successful stimulation of the immune response to bluetongue virus was achieved in spleen cell cultures from mice previously primed with bluetongue virus. The optimum antigen concentration was 30-40 ng bluetongue virus per 10(7) spleen cells and the maximum plaque-forming cell response was observed after 4 days in culture. 相似文献
13.
E.W. Liewes R.H. Van Dam M.G. Vos-Maas R. Bootsma 《Veterinary immunology and immunopathology》1982,3(3):325-343
The optimization of a leukocyte stimulation microassay with carp (Cyprinus carpio L.) leukocytes is described. Leukocytes were isolated from the thymus, anterior kidney, spleen, mid-kidney and peripheral blood.Leukocyte cultures were stimulated with PHA-P, LPS (Escherichia coli 055: B5) PWM, ConA and PPD from Mycobacterium fortuitum. The optimum incubation temperature for leukocyte cultures differed depending on the mitogen used. The optimum incubation period was 3.5 days for leukocyte cultures derived from lymphoid organs and 4.5 days for peripheral blood lymphocyte cultures. Leukocytes from various organ sources showed similar reactivity patterns to stimulation in vitro by different mitogens. The results of these mitogen stimulations did not present sufficient arguments in favour of compartmentation. 相似文献
14.
Balb/c mice, injected i.p. with extracellular products (ECP) of Aeromonas salmonicida subsp. achromogenes (Asa), displayed symptoms similar to toxic shock syndrome. The LD(50) observed was between 1.5 and 2.0 microg g(-1) and the mice died within 19 h. Four inflammatory cytokines were measured in mice receiving sublethal ECP doses. TNF-alpha and IL-6 showed a sharp peak in the serum while IL-1 beta and IL-2 were not detected. When peritoneal macrophages were cultivated in the presence of ECP, AsaP1 (a toxic caseinolytic metallo-protease purified from ECP) or LPS, all cultures produced TNF-alpha, IL-6 and IL-1 beta. The same antigens were mitogenic in spleen cell cultures. Furthermore, IL-2 production, which is a normal T-cell response to ConA stimulation, was downregulated in spleen cell cultures from mice injected with ECP. 相似文献
15.
The lymphocyte transformation (LT) test was performed using duck blood lymphocytes stimulated with phytohaemagglutinin (PHA), concanavalin A (Con A), lentil lectin (LC), Roman snail lectin (HP), peanut agglutinin (PNA), Bandeiraea simplicifolia seed lectin (BSS), wheat germ agglutinin (WGA), horseshoe crab lectin (HSC), pokeweed mitogen (PWM) and E. coli lipopolysaccharide (LPS). Cells were cultured in microtitre trays, at 41.6 degrees C, 8 x 10(5) cells in 200 microliters medium (= 4 x 10(6) cells/ml) supplemented with 10% pooled duck serum. Mitogens were added at final concentrations of 0.1-100 micrograms/ml and triplicate cultures at each concentration were harvested daily for scintillation counting 6 hr after addition of 1 microCi [3H]thymidine. Three patterns of response were observed. The responses to Con A, LC, HP and HSC were greatest at high mitogen concentrations (40-100 micrograms/ml) throughout the 7 days of culture. With PHA, PNA, WGA and LPS maximum stimulation was obtained at 3-5 days, at which time the cells were responding to lower concentrations of mitogen than were required at other times during the experiment. The response to BSS and PWM showed increasing sensitivity to lower concentrations of mitogen during the first 3 days of culture and then stimulated most strongly at 2-10 micrograms/ml in cultures harvested after 4-7 days. Cells from two ducks were cultured for 3 and 5 days with selected concentrations of these mitogens; the results confirmed the variation in response to different mitogens. It is possible that these patterns of response are the outcome of stimulating different populations of duck lymphocytes. 相似文献
16.
Lymphocytes from healthy adult cats were separated into T and B cell-enriched subfractions by centrifuging rosetted cells on sodium metrizoate/Ficoll gradients. The responsiveness of unseparated lymphocytes (T + B), and T and B cell-enriched subfractions to stimulation with mitogens (phytohemagglutinin-P (PHA-P), concanavalin A (ConA), and pokeweed mitogen (PWM) was tested. Cultures of unseparated lymphocytes and those enriched in T cells showed similar responsiveness of PHA, ConA, and PWM stimulation; however, only a weak response to ConA and PWM was observed in B cell-enriched cultures. The mitogenic effects of PHA-P, ConA, and PWM on feline lymphocytes appeared to be due primarily to T-cell activation. 相似文献
17.
感染鸡贫血病毒雏鸡接种新城疫疫苗后免疫功能和免疫调节的变化 总被引：1，自引：1，他引：0
雏鸡1日龄感染鸡贫血病毒，8日龄接种Lasota疫苗，以未感染免疫雏鸡为对照，于免疫后7、14、28d检测血清免疫球蛋白IgG、IgM、IgA,在凝抑制抗体（HI）滴度；胸腺、法氏囊、脾脏T细胞、IgG^ 、IgM^ 、IgA^ ,抗体生成细胞数量及T、B细胞增殖反应；胸腺、脾脏细胞因子IL－2、IFN活性的变化。结果发现，感染CAV雏鸡Lasota疫苗免疫后，其血清IgG、IgM、IgA免疫球蛋白含量明显减少，HI抗体滴度降低；胸腺、法氏囊、脾脏T细胞、抗体生成细胞数量降低及T、B细胞增殖反应减弱，胸腺、脾脏IL－2及TNF诱生活性降低，表明其细胞免疫和体流免疫功能以及细胞因子免疫调节作用均未感染免疫雏鸡明显减弱。 相似文献
18.
J M Kaneene F C Okino R K Anderson C C Muscoplat D W Johnson 《Veterinary immunology and immunopathology》1981,2(1):75-85
Incubation of Brucella abortus (field strain) infected and strain 19 vaccinated bovine peripheral blood lymphocytes with B. abortus antigen and levamisole caused a consistently significant increase in [3H] thymidine uptake when compared to cultures without levamisole. Levamisole did not potentiate B. abortus-induced blastogenic response of lymphocytes from non-exposed cattle. A dose response study showed that 10 micrograms/culture induced maximum potentiation of B. abortus-induced lymphocyte stimulation. Using the 10 micrograms/well concentration of levamisole, further studies were conducted to determine the net potentiation of the blastogenic responses in lymphocytes from B. abortus (field strain) infected cattle. B. abortus strain 19 vaccinated but nonresponsive and non-exposed cattle. Levamisole significantly potentiated the B. abortus-induced lymphocyte blastogenesis in lymphocytes from unresponsive cattle. 相似文献
19.
R C Manak 《Journal of animal science》1986,62(3):759-771
The ability of lymphocytes from newborn calves to undergo blastogenic responses to the mitogens Concanavalin A (Con A), phytohemagglutinin (PHA), or Pokeweed Mitogen (PWM), and immunomodulation of these responses by neonatal calf serum was assessed as a function of age. Lymphocytes were obtained from thymus, spleen, and mesenteric lymph nodes of 1-, 2- to 3-, 5- to 7-, and 9- to 10-d-old calves, aliquoted and incubated (+/- mitogens) in sera from 1-, 2-, 3-, or 7- to 10-d-old calves. Lymph-node lymphocytes responded least when cultured in sera from 1-d-old calves, regardless of mitogen or age of cell donor; the response increased as age of serum donor increased (P less than .05). Splenic lymphocytes responded similarly (P less than .005). However, when cultured in sera from older calves, splenic lymphocytes from older calves responded greater to PWM than did those from younger calves. Thymic lymphocytes responded minimally to PWM and PHA. Their response to Con A increased (P less than .005) with age of serum donor calf, but the effect was greatest on lymphocytes from 5- to 7-d-old calves. Mixing experiments with varying ratios of 1-d-old calf serum: 10-d-old calf serum suggested that serum from 1-d-old calves contained suppressive activity. Serum cortisol level (measured by radioimmunoassay) was 30 +/- 4.6 ng/ml in calves at 6 h of age and declined to 5.5 +/- 1.1 ng/ml by 10 d. Charcoal treatment to remove steroids did not enhance blastogenesis. Addition of cortisol (50 ng/ml) to charcoal-treated sera resulted in inhibition of response to PHA, but no change in response to Con A or PWM. Further investigation is indicated to characterize this immunosuppressive activity and to establish its relationship to disease susceptibility. 相似文献
20.
A microculture system used in conjunction with a semiautomatic sample harvester is described to determine the in vitro properties of turkey peripheral blood lymphocytes. By this new procedure, multiple tests were done rapidly, relatively few cells were used, and results were highly reproducible. Analysis indicated that the conditions for optimal concanavalin A (con A) stimulation, as measured by incorporation of 3H-thymidine (3H-TdR) included (a) 2 times 10(6) cells per culture in RPMI 1640 medium in the absence of any serum; (b) 0.4 mug of con A per culture, incubated in flat-bottom microtitration wells for 72 hours at 37 C; and (c) 1 muCi of 3H-TdR per culture added 12 to 24 hours before termination. Conditions for optimal stimulation with pokeweed mitogen were similar to those used for con A. The exceptions were that 1 times 10(6) cells per culture were incubated in round-bottom microtitration wells. The pokeweed mitogen gave the highest degree of stimulation when used at a concentration of 80 mug per culture. 相似文献