紫锥菊多糖对鸡淋巴细胞体外增殖影响

为了评价紫锥菊多糖免疫增强效果,将4种浓度的紫锥菊多糖分别加入体外培养的鸡脾脏淋巴细胞中,培养44 h后,用MTT法测定淋巴细胞增殖变化。结果表明,4种浓度的紫锥菊多糖对鸡淋巴细胞均有促进增殖及协同的作用,其中在50-100μg/mL浓度范围内效果最为显著。     

Age-related changes in mitogen-induced lymphocyte proliferation and polymorphonuclear neutrophil function in the piglet

Changes in concentration of plasma cortisol, mitogen-induced lymphocyte proliferation and the phagocytic and killing abilities of polymorphonuclear neutrophils (PMN) were assessed in 24 crossbred piglets at .5, 1, 3 and 6 wk of age. Concentrations of blood cortisol were high at birth but decreased (P less than .001) thereafter. Spontaneous lymphocyte proliferation was approximately 10-fold higher in the newborn than in pigs 6 wk old. The methods by which data on lymphocyte proliferation were expressed greatly influenced the results of the age-related changes in lymphocyte response. When lymphocyte proliferation was expressed as a stimulation index, there was an increase in mitogen-induced lymphocyte proliferation with advancing age. However, when lymphocyte proliferation was expressed as total counts per minute, mitogen-induced lymphocyte proliferation decreased with increasing age. In general, no significant age-related changes were observed in PMN function except for a transient decrease in PMN phagocytic ability at wk 3. Piglets were born with high concentrations of blood cortisol; this was associated with age-related changes in lymphocyte proliferation.     

Effect of recombinant human interferon-alpha in vitro and in vivo on mitogen-induced lymphocyte blastogenesis in cats

The effect of recombinant human interferon-alpha (rHuIFN-alpha) in vitro and in vivo on mitogen-induced lymphocyte blastogenesis was evaluated in specific-pathogen-free cats. Pre-incubation of isolated feline peripheral blood lymphocytes (PBL) in vitro with either 10(4) or 10(3) International Units (U) of rHuIFN-alpha for 24 h significantly suppressed (P less than 0.001 and 0.01, respectively) blastogenic responses to the phytomitogens concanavalin A (Con A) and pokeweed mitogen (PWM). Lower doses of IFN (range, 10-10(-3) U/ml) neither suppressed nor enhanced mitogenesis. In the absence of phytomitogens, incubation of PBL with 10(4) - 10(2) U (P less than 0.001) or 10 U (P less than 0.05) of rHuIFN-alpha/ml resulted in a significant decrease in incorporation of [methyl-3H] thymidine into newly synthesized cellular DNA. Cultures of PBL exposed continuously for 4 days to rHuIFN-alpha doses of 10(4) U/ml or less did not demonstrate specific reductions in cell viability, indicating that the observed antiproliferative actions of IFN apparently were independent of any direct cytotoxic effects. To investigate the dose-response effects of rHuIFN-alpha in vivo on lymphocyte blastogenesis, individual groups of cats were evaluated on 3 consecutive days before and then 24 h after each cat was inoculated intramuscularly with either a high dose (10(6) U/kg), moderate dose (10(4) U/kg), or a relatively low dose (10(2) U/kg) of rHuIFN-alpha. Cats inoculated with 10(6) U of rHuIFN-alpha/kg had significantly reduced (P = 0.037) blastogenic responses to Con a at 24 h postinoculation compared to preinoculation values; mean PWM responses were also decreased, but this effect was not statistically significant. In contrast, inoculation of cats with either 10(4) or 10(2) U of rHuIFN-alpha/kg significantly enhanced (P = 0.05 or 0.008, respectively) Con A-induced blastogenesis and had no discernible effect on PWM responses. These findings suggest that very high doses of rHuIFN-alpha given parenterally may be associated with suppression of certain T-cell responses in cats; conversely, much lower doses may be immunoenhancing.     

Detection of mitogen-induced lymphocyte proliferation by bromodeoxyuridine (BrdU) incorporation in the chicken

3H-thymidine (3H-TdR) incorporation assay has been generally used to measure lymphocyte proliferation in the chicken. Disadvantages of this assay are that radioisotope is biological hazard to the person and environment and that it can not measure which subset of lymphocytes proliferates. In this study, bromodeoxyuridine (BrdU) incorporation assay by flow cytometry was compared with 3H-TdR incorporation assay. As a result, BrdU incorporation assay showed a strong correlation with 3H-TdR incorporation assay, and it could be applied simultaneously to detect BrdU incorporation and expression of cell surface marker antigens. These results suggest that the BrdU incorporation assay by flow cytometry is useful to analyze lymphocyte proliferation in detail.     

The effect of cimetidine on lymphocyte subpopulations in vivo in experimental mice.

The effects of cimetidine, an H2 receptor antagonist, on subpopulation of splenic lymphocytes were studied in mice. Cimetidine (50 mg/kg) was given to groups of mice by intraperitoneal injection. The splenic mononuclear cells from treated and control animals were evaluated for relative number of lymphocyte subpopulations (i.e. LYT1, LYT2, L3T4 and GAM cells) by flow cytometry. The percentages of LYT2 cells (suppressor equivalent) were significantly lower (5.5 vs 12.0%) in mice who were treated with cimetidine vs control animals. The percentages of L3T4 (helper equivalent) were not statistically different in any of the experimental groups.     

The effect of Rhodiola quadrifida extracts on cellular immunity in mice and rats

Rhodiola quadrifida (Rq) roots and rhizomes are traditionally used in Asia as a tonic, adaptogen, antidepressant and anti-inflammatory drug. The aim of this work was to study the in vivo effect of aqueous and 50% hydro-alcoholic extracts of Rq rhizomes on some parameters of cellular immunity in mice and rats. The metabolic activity of blood phagocyting cells was determined based on the measurement of intracellular respiratory burst after stimulation by PMA in RBA test. Potential bactericidal activity of phagocyting cells was determined in isolated blood leukocytes stimulated with killed microorganisms, according to the PKA test. Proliferative response of lymphocytes stimulated by mitogen concanavaline A (ConA) or lipopolysaccharide (LPS) were determined by MTT assay. Both extracts stimulated granulocytes activity in vitro and increased lymphocyte response to mitogens. The ability of parental strain mice lymphocytes to induce local cutaneous graft-versus-host reaction (GVH) in F1 hybrids was stimulated by 50% hydro-alcoholic extract only.     

Effect of hydrocortisone on circulating lymphocyte numbers and their mitogen-induced blastogenesis in lambs

The effect of hydrocortisone on the number of circulating lymphocytes and their blastogenic response was studied in 20 feedlot lambs given combinations of 3 treatments: hydrocortisone (25 mg/kg of body weight, 4 times a day, IM), feed changes (100% roughage to 90% concentrate over a 6-day period), and oral inoculation of Pasteurella haemolytica biotype T (10(9) to 10(11) bacteria/day via stomach tube) to develop a model for reproduction of septicemic pasteurellosis. Hydrocortisone caused lymphopenia and inhibited the blastogenic response of peripheral blood lymphocytes to phytohemagglutinin and concanavalin A mitogens. A synergistic effect was observed between hydrocortisone injections and feed changes resulting in higher than expected serum hydrocortisone concentrations and lower circulating lymphocyte counts. Seemingly, stress-induced increases in serum hydrocortisone concentrations cause suppression of the immune response of feedlot lambs. The combined effect of feed changes and stress on the immune response of lambs may explain the role of these 2 factors in the pathogenesis of septicemic pasteurellosis.     

五加芪粉对小鼠淋巴细胞增殖功能和抗体形成细胞的影响

为了考察五加芪粉对小鼠淋巴细胞增殖功能和抗体形成细胞的影响,为临床应用提供理论依据,设不同剂量的(60、120、240 mg·kg·d~(-1))五加芪粉组,同时设黄芪多糖粉对照组和空白对照组。连续灌胃7 d后,无菌取小鼠脾脏,制备脾淋巴细胞,采用MTT法检测小鼠淋巴细胞增殖功能,定量溶血分光光度(QHS)法测定小鼠抗体形成细胞。结果显示:五加芪粉中、高剂量组可显著(0.01P≤0.05)或极显著(P≤0.01)的提高小鼠淋巴细胞的吸光度值,具有提高机体细胞免疫力的作用;五加芪粉各给药组均显著和极显著的提高了小鼠抗体形成细胞水平(0.01P≤0.05,P≤0.01),具有提升动物体液免疫功能的作用。表明五加芪粉可同时提高机体的细胞免疫和体液免疫功能,增强机体免疫力。     

Phytotherapeutic effects of Echinacea purpurea in gamma-irradiated mice

Echinacea (E.) purpurea herb is commonly known as the purple coneflower, red sunflower and rudbeckia. In this paper, we report the curative efficacy of an Echinacea extract in γ-irradiated mice. E. purpurea was given to male mice that were divided into five groups (control, treated, irradiated, treated before irradiation & treated after irradiation) at a dose of 30 mg/kg body weight for 2 weeks before and after irradiation with 3 Gy of γ-rays. The results reflected the detrimental reduction effects of γ-rays on peripheral blood hemoglobin and the levels of red blood cells, differential white blood cells, and bone marrow cells. The thiobarbituric acid-reactive substances (TBARs) level, Superoxide dismutase (SOD) and glutathione peroxidase (GSPx) activities and DNA fragmentation were also investigated. FT-Raman spectroscopy was used to explore the structural changes in liver tissues. Significant changes were observed in the microenvironment of the major constituents, including tyrosine and protein secondary structures. E. purpurea administration significantly ameliorated all estimated parameters. The radio-protection effectiveness was similar to the radio-recovery curativeness in comparison to the control group in most of the tested parameters. The radio-protection efficiency was greater than the radio-recovery in hemoglobin level during the first two weeks, in lymphoid cell count and TBARs level at the fourth week and in SOD activity during the first two weeks, as compared to the levels of these parameters in the control group.     

Inhibitory effect of African swine fever virus on lectin-dependent swine lymphocyte proliferation

The incubation of swine peripheral blood mononuclear cells (PBMC) with African swine fever (ASF) virus preparations strongly inhibited the proliferative response of lymphocytes to PHA and other lectins. The inhibition, which persisted after inactivation of the virus by UV radiation, was dependent upon the dose and the time that virus preparations were present in cultures. When virus preparations were fractionated by ultracentrifugation, the inhibitory activity resulted to be soluble, whereas no activity was found in the sedimented viral fraction. However, the preincubation during 4 days of this sedimented fraction with swine PBMC, before the addition of the mitogen, restored the inhibitory activity. The results obtained suggest that the inhibition is mediated by one or more soluble factors released by swine PBMC after coincubation with ASF virus in a time dependent process. These factors show a molecular weight between 40 and 80 kDa by gel filtration chromatography. The inhibitory activity described in the present paper is an indication of inhibition of lymphocyte function produced by ASF virus which can help to understand how this virus escapes from the host immune system.     

交感神经对小鼠脾脏中淋巴细胞转化率的影响

神经、内分泌、免疫诸系统间存在着相互作用,这是近30年来新兴的神经免疫学中主要的研究成果。交感神经及其递质去甲肾上腺素(NE)在体内和体外条件下对免疫细胞功能的调节作用是神经免疫学中研究最早、报道最多的内容之一。脾脏中T淋巴细胞和B淋巴细胞转化率分别代表了机体的细胞免疫和体液免疫,但是,对于外周交感神经损毁后,脾脏中淋巴细胞转化率的影响还未见报道。因此,本研究利用6-OHDA损毁小鼠交感神经模型,检测了脾脏中不同浓度ConA诱导的T淋巴细胞转化率,不同浓度LPS诱导的B淋巴细胞转化率,以探讨交感神经对机体免疫状态的影响…     

Influence of different tumour types on natural cytotoxicity (NK cell activity) and mitogen-induced lymphocyte proliferation in isolated blood lymphocytes from 110 dogs with tumours

The cell-mediated immune response of blood lymphocytes from 110 untreated dogs with different tumours was evaluated. The influence of different tumour types on the cellular immune system was examined by assessing the percentage of isolated large granular lymphocytes (LGL), in vitro natural cytotoxicity and mitogen-induced lymphocyte proliferation. Although the overall natural cytotoxicity of dogs with different tumours was decreased, the overall difference from control values was not statistically significant. However, mitogen-induced lymphocyte proliferation was significantly depressed in dogs with tumours in comparison with the controls. Dogs with mammary carcinomas showed significantly lower natural cytotoxicity than controls and dogs with myeloid neoplasms showed significantly lower mitogen-induced lymphocyte proliferation. Abnormalities exist not only in natural cytotoxicity but also in mitogen-induced lymphocyte proliferation. For the dog, this is the first study to assess the influence of different tumours using a combined evaluation of natural cytotoxicity and mitogen-induced lymphocyte proliferation in such a large number of animals.     

The effect of different isolation procedures on canine leucocyte populations and on lectin-induced lymphocyte proliferation

Proliferation assays performed on peripheral blood mononuclear cells (PBMC) are commonly used in experimental and clinical immunology. A prerequisite for an in vitro assay is the ability to obtain relatively pure populations of mononuclear cells from whole blood, as contaminating polymorphonuclear cells may affect the proliferation of lymphocytes. Purification of canine leucocytes from whole blood is associated with difficulties in obtaining pure lymphocytes in high yields. The aim of this study was to optimize the lymphocyte purification from canine whole blood in terms of total cell recovery and purity, while not influencing the proliferation capacity of the isolated cells. To acquire optimal isolation of canine lymphocytes several density gradient media of different densities and osmolalities were examined. For optimal phagocyte removal, pre-treatment of whole blood with carbonyl iron/arabic gum and/or adherence to fibrinogen pre-coated polystyrene tissue flasks were examined. Lectin-induced proliferation was used as measurement of cell activity of the obtained cell fractions after the different separation procedures. Canine blood pre-treated with carbonyl iron/arabic gum followed by density gradient centrifugation with medium 'G' (density: 1.079 g/cm(3), osmolality: 256 mOsm) and adherence to pre-coated polystyrene tissue flask obtained the best PBMC cultures with a median lymphocyte purity of 88% and a median yield of recovered lymphocytes of 54%. This culture also resulted in the highest proliferation and subsequently the highest stimulation index upon lectin stimulation.     

金线莲多糖对免疫抑制小鼠脾淋巴细胞增殖及免疫器官的影响

目的：探讨金线莲多糖对免疫抑制小鼠脾淋巴细胞增殖及免疫器官的影响。方法：按大、中、小剂量给小鼠灌胃金线莲多糖,腹腔注射环磷酰胺（CY）建立免疫抑制模型;测定小鼠体重及胸腺、脾脏重量,计算胸腺、脾指数;采用MTT法检测脾淋巴细胞的增殖。结果：3个金线莲多糖剂量组小鼠的体重及免疫器官指数均低于空白组而高于模型组,脾淋巴细胞增殖均显著（P〈O．05）高于模型组。结论：金线莲多糖能提高免疫抑制小鼠体重及免疫器官指数,促进脾淋巴细胞增殖。     

Effects of racing on lymphocyte proliferation in horses

OBJECTIVE: To measure the lymphocyte proliferation response in horses 12 to 16 hours after completion of a race. ANIMALS: 8 Thoroughbreds that competed in 14 races and 3 control Thoroughbreds that did not race. PROCEDURE: Horses participated in races during the late afternoon or evening. Venous blood samples were collected on a morning before a race (1 or 2 days before the race or on the day of the race), on the afternoon of a race (40 to 60 minutes after the race), and on the morning of the day after a race (12 to 16 hours after the race). Lymphocyte proliferation responses and WBC count were measured in samples obtained in the mornings. Plasma cortisol was measured in all samples. RESULTS: Lymphocyte proliferation responses were significantly reduced and WBC counts significantly increased 12 to 16 hours after a race. Plasma cortisol concentrations were significantly increased 40 to 60 minutes after a race. In samples from the control horses, lymphocyte proliferation responses, WBC counts, or plasma cortisol concentrations did not differ significantly among time periods. CONCLUSIONS AND CLINICAL RELEVANCE: A decrease in proliferative responses of circulating lymphocytes can be found as late as 12 to 16 hours after a horse participates in a race. Although the clinical consequences of these exercise-related alterations of the immune response are not yet known, managers of horses should take into account that the immune system of a horse may be affected by racing.     

Comparative studies on in vitro mitogen-induced proliferation of peripheral blood lymphocytes in dog and breeding fox.

In vitro blastogenesis of dog and fox lymphocytes was compared by a microculture technique. The highest 3H-thymidine incorporation in cultures of dog lymphocytes was observed at day 3, while in those of fox at day 2, incubated either at 37 degrees C or at 39 degrees C. Lymphocytes cultured at 39 degrees C incorporated more tritiated thymidine than did cells cultured at 37 degrees C. The stimulation index (SI) of dog peripheral blood lymphocytes to both mitogens concanavalin A (Con A) and leucoagglutinin (LA) was in a similar range, while pokeweed mitogen (PWM) showed a weaker but significant stimulatory action. The blastogenesis of fox lymphocytes was the greatest in Con A stimulated cultures. The mitogenic potency of LA and PWM was about half of that of Con A, with no essential difference between them. Maximum lymphocyte proliferation of dog and fox was observed when culture media were supplemented with 10% fetal calf serum (FCS).     

硫酸化人参总皂苷的制备及其对鸡外周血淋巴细胞增殖的影响

用氯磺酸-吡啶试剂制备了硫酸化人参总皂苷衍生物，采用红外光谱仪检测到制备的硫酸化人参总皂苷衍生物中在1230cm^-1和810cm^-1有硫酸酯键的特征吸收峰。为了初步探讨硫酸化人参总皂苷的免疫学活性．以MTT法测定了人参总皂苷及其衍生物对鸡外周血淋巴细胞增殖的影响。结果表明，150μg／m1人参总皂苷和衍生物均能显著抑制鸡外周血淋巴细胞的增殖（P〈0．05），10μg／ml的衍生物能极显著促进淋巴细胞增殖，与同浓度人参总皂苷相比活性极显著增强（P〈0．01）。实验结果提示，对人参总皂苷进行硫酸化修饰是可行的，经硫酸化后人参总皂苷衍生物的免疫活性明显增强。     

Optimal conditions for in vitro mitogen-induced proliferation of peripheral blood lymphocytes in breeding foxes

The proliferative response of fox peripheral blood lymphocytes to nonspecific mitogens: leucoagglutinin (LA), concanavalin A (Con A) and pokeweed mitogen (PWM) was studied. Microcultures were kept at 39 degrees C in a humidified atmosphere containing 5% CO2. The highest 3H-thymidine incorporation was observed, when Con A was used, while LA and PWM showed weaker but significant stimulatory action. Optimal doses of mitogens were: 5 micrograms/ml for Con A, 5 micrograms/ml for LA and a dilution of 1:100 for PWM. The maximal stimulation index for Con A was about 240 and up to 100 for LA or PWM. The maximal lymphocyte proliferation was observed when culture media were supplemented with 10% serum. When proliferation kinetics were studied, the peak response was observed on Day 2.     

黄芪多糖对肉鸡脾淋巴细胞转化及信息分子的影响

本实验研究了黄芪多糖对肉鸡脾淋巴细胞转化率及细胞内信息分子 -一氧化氮 (NO)、Ca2 + 及 c AMP、c GMP的影响。发现黄芪多糖能剂量依赖性促进脾淋巴细胞转化率及细胞内 NO、Ca2 + 浓度 ;当作用时间为 2 0 min时 ,4 0 μg/ ml黄芪多糖能增加 c AMP浓度 ,降低 c GMP浓度 (P<0 .0 1) ;而当作用时间为 6 0 m in,对 c AMP和 c GMP浓度无显著影响。以上结果提示 ,黄芪多糖能显著提高脾淋巴细胞免疫功能 ,其作用是通过改变细胞内信息传导实现的。