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1.
Equine recurrent airway obstruction (RAO) is thought to result from an aberrant immune response to inhaled antigens, modulated by T lymphocytes via the secretion of pro-inflammatory cytokines. However data relating to the phenotypes of the T lymphocytes present in peripheral blood and bronchoalveolar lavage fluid of RAO horses and their cytokine profiles are contradictory. The aim of this study was to further investigate the cytokine (IL-4, IL-5, IL-13 and INF-gamma) mRNA expression profile in peripheral blood lymphocytes and bronchoalveolar lavage lymphocytes from RAO and control horses, before and at 48 h after horses were exposed to hay/straw. In contrast to previous studies, cytokine expression was quantified in populations of CD4 and CD8 T lymphocytes which were purified using magnetic bead antibody cell separation. Hay/straw exposure induced clinical airway obstruction, airway neutrophilia and airway lymphocytosis in RAO horses, and, induced a mild, but significant, airway neutrophilia in controls. However, hay/straw exposure had no significant effect on peripheral blood lymphocyte or bronchoalveolar lavage lymphocyte cytokine expression in either group. In conclusion, RAO was not associated with alterations in lymphocyte cytokine expression that are consistent with Th1 or Th2 responses, but rather with a general down-regulation in expression of the measured cytokines in peripheral blood lymphocytes and bronchoalveolar lavage lymphocytes.  相似文献   

2.
通过检测奶牛C D3+、CD4+和CD8+淋巴细胞亚群的动态及细胞因子IL-2、IL-4在分娩前后21d内的基因转录水平的变化,以探讨此阶段奶牛免疫机能受损的机理.应用流式细胞术检测CD3+、CD4+、CD8+淋巴细胞亚群的动态变化及实时荧光定量检测细胞因子IL-2、IL-4 mRNA的表达量.结果表明:血液中CD3+淋巴细胞的比例在产前21 d内占(50.8±4.61)%,产后14 d内降低到(39.21±4.98)%,两者差异极显著(P<0.01) ;CD4+淋巴细胞在产前21 d内保持较高水平,达(21.38±0.65)%,产后21 d内维持在(6.89±0.32)%的较低水平,且与产前差异显著(P<0.05) ;CD8+淋巴细胞的比例在产犊前逐渐升高,到产犊时达到最高水平(16.90±2.02)%,差异显著(P<0.05) ;CD4+/CD8+淋巴细胞的比率在产前>1,产后<1.IL-2 mRNA的表达量在产后0~14 d内下调,之后上升.IL-4 mRNA的表达量分娩时期达到最高,随后下降,至第7天降到最低.奶牛在围产期阶段尤其是分娩前后14 d内机体处于免疫抑制状态,可能是由于自身的CD3+、CD4+和CD8+T淋巴细胞的亚群的数量和比例发生改变及细胞因子IL-2、IL-4 mRNA表达量下调引起的.  相似文献   

3.
Expression of CD25 (interleukin-2 receptor alpha chain) was used to monitor antigen-specific activation of T lymphocyte subsets (CD4+, CD8+, and gamma delta T cells) from cattle immunized with modified-live virus (MLV) bovine viral diarrhea virus (BVDV) vaccines. Two groups of 15 animals each were vaccinated with one dose of either BVDV genotype 1 (BVDV-1) or BVDV-1 and BVDV genotype 2 (BVDV-1/2). Six animals negative for both BVDV antibody and BVDV virus were used as negative controls. Three animals vaccinated 7 and 5 weeks before the start of the experiment with MLV BVDV-1 vaccine served as positive controls. Blood samples were taken from the negative control group, the positive control group, and the BVDV-1/2 group 0, 21, 35, 60, and 90 days after vaccination. Blood samples were taken from the BVDV-1 group 0, 21, and 90 days after vaccination. Isolated peripheral blood lymphocytes from immunized and control animals were incubated for 5 days with and without BVDV-1 or BVDV-2. Compared with nonvaccinated animals, a significant (P <.05) increase in expression of CD25 by CD4+ (60 days), CD8+, and gammadelta T (35 to 90 days) lymphocytes from the group given BVDV-1/2 was detected following in vitro exposure to BVDV-1 or BVDV-2 after vaccination. The CD8+ and gammadelta T cells from the group vaccinated with BVDV-1 had significantly (P <.05) increased expression of CD25 compared with nonvaccinates following postvaccination exposure to in vitro BVDV-1 but not to BVDV-2. There was no significant difference between the two vaccinated groups in CD25 expression on any of the T cell subsets in response to BVDV-1 or BVDV-2 exposure. A single administration of MLV BVDV vaccine may be more effective at stimulating CD8+ and gammadelta T cell-specific immune responses to the homologous genotype than to the heterologous genotype.  相似文献   

4.
Fifteen Holstein dairy cattle were monitored for lymphocyte subsets and expression of adhesion molecules on cells in milk and blood at parturition and at intervals up to 21 days post-partum. Using flow cytometry, we determined percentages of T cells (CD4+, CD8+, gammadelta) and B cells from milk and blood of these cows. We also measured expression of adhesion molecules (CD62L, LFA-1, LPAM-1, and CD44) on lymphocytes in milk and blood. Significantly higher percentages of CD8+ cells were found in milk than in blood at all time points while significantly higher percentages of B cells were found in blood than in milk at all time points. There were minimal to no significant differences in percentages of CD4+ or gammadelta+ cells between milk and blood. Expression of adhesion molecules was consistently higher on all subsets of milk lymphocytes compared with blood lymphocytes. These differences were most pronounced and statistically significant at calving and in the first week following calving. CD62L, LPAM-1 and CD44 were expressed on a significantly higher percentage of lymphocytes in milk at calving than in milk at subsequent sampling times, while LFA-1 expression on lymphocytes in milk was significantly lower at calving than at subsequent times.  相似文献   

5.
Lymphocyte subsets in peripheral blood of dogs--a flow cytometric study   总被引:4,自引:0,他引:4  
Slight differences in the results of papers describing lymphocyte subsets distribution in the peripheral blood of healthy dogs may be explained by differences in monoclonal antibody clones and sources, breed and age of animals examined, methods of sample treatment, or methods of result analysis. In this paper, we described the effect of sample processing and of sample storage as well as the effect of age, breed, and gender of dogs on lymphocyte subset distribution. No significant differences were found between samples processed following a whole-blood lysis method and samples processed after density gradient separation. Furthermore, no significant differences were found between samples processed within 2h after collection and those stored at 4 degrees C for 12-16 h before processing. Age-related changes were evident in lymphocyte subset distribution in the peripheral blood of 38 Beagles divided according to their age into the six groups: (1) 5-6 days; (2) 2 months; (3) 6 months; (4) 1-2 years; (5) 3-5 years; and (6) >5 years. The percentage of B-lymphocytes (CD21-like positive cells) in the peripheral blood of newborn pups was 39.5+/-5.7 and decreased with advancing age. The percentage of CD8+ lymphocytes was 7.7+/-3.4 after birth and increased with advancing age. No age-related changes were observed in the percentages of CD4+ lymphocytes. The CD4+:CD8+ ratio decreased with advancing age. No significant age-related change was observed for lymphocytes bearing the gammadelta-TCR. Some breed differences were evident. Adult (1-5-year-old) Beagles, German Shepherds, Dalmatians, and Dachshunds were examined. The percentages of lymphocytes were higher in Beagles and Dachshunds than in Dalmatians and German Shepherds. The highest and the lowest absolute lymphocyte counts were found in Beagles and German Shepherds, respectively. As a consequence, German Shepherds showed the lowest absolute counts of the individual lymphocyte subpopulations and the widest neutrophil:lymphocyte ratio. Dalmatians showed the lowest percentage of CD3+ cells, the highest percentage of CD21+ cells, and the lowest CD4+:CD8+ ratio. German Shepherds showed the lowest percentage of CD21+ cells and the highest CD4+:CD8+ ratio. Females in Beagles and Dachshuns had nonsignificantly higher percentages of total lymphocytes, CD3+, CD4+, and nonsignificantly lower percentages of CD21+ lymphocytes. We concluded that there are age-, breed-, and perhaps also gender-related differences in lymphocyte subset distribution in the peripheral blood of dogs. Therefore, there is need to use appropriate control group in the experimental protocols. Among-breed differences could explain, at least partly, breed predisposition for some diseases.  相似文献   

6.
Twelve dairy cows infected with Mycobacterium avium subsp. paratuberculosis were monitored for lymphocyte subsets and expression of adhesion molecules on cells in blood and milk at parturition and at intervals up to 21 days post-partum. Using fluorescent antibody labeling of cells and analysis by flow cytometry, we determined percentages of T cell subsets (CD4+, CD8+, gammadelta+) and expression of adhesion molecules (CD62L, LFA-1, LPAM-1, and CD44) on cells from blood and milk of these cows. Significantly higher percentages of CD8+ cells were found in milk than in blood at all time points; there were no significant differences in percentages of CD4+ or gammadelta+ cells. CD62L, LFA-1, and LPAM-1 were expressed on a significantly higher percentage of all T cell subsets in milk than in blood at various times after parturition. No differences were seen in expression of CD44. Increased percentages of T lymphocytes expressing adhesion molecules in milk compared to blood suggest that a migratory population of cells is being selectively recruited to the mammary gland from the circulation.  相似文献   

7.
为了研究荷斯坦奶牛受到双翅目昆虫侵袭时奶牛个体所表现的护身行为、外周血液中淋巴细胞亚群的变化规律,试验期间每日跟踪观察奶牛护身行为1 h,记录每5 min内侵袭奶牛的双翅目昆虫数目和奶牛的护身行为出现次数,定时采血用流式细胞仪检测外周血液中CD3+、CD4+、CD8+ T淋巴细胞和CD21+B淋巴细胞等的变化规律.结果表明:奶牛护身行为随飞来昆虫的数量而变化;外周血液中的CD3+、CD4+、CD21+水平随双翅目昆虫数量的增多而降低;CD8+水平随双翅目昆虫数量的增多而升高.  相似文献   

8.
Lymphocyte subsets in canine umbilical cord blood were flow cytometrically analyzed and compared with those of the dams' peripheral blood. The proportion of CD3+ T lymphocytes, CD21+CD3- B lymphocytes, and CD3-CD21- non-T non-B lymphocytes in umbilical cord blood was 52.9%, 30.4%, and 16.7%, respectively. T lymphocyte/B lymphocyte ratio was significantly lower in the umbilical cord blood than in the dams' peripheral blood (2.1 +/- 1.4 versus 11.0 +/- 8.1, P < 0.001). In contrast, CD4+ lymphocyte/CD8+ lymphocyte ratio was significantly higher in the umbilical cord blood than in the dams' peripheral blood (7.6 +/- 2.2 versus 1.8 +/- 0.6, P<0.001). These findings clarified the phenotypic characters of canine umbilical cord blood lymphocytes.  相似文献   

9.
The objective of the study was to analyze changes in peripheral blood leukocyte subsets in cases of naturally occurring exudative epidermitis (EE) in pigs. Five of ten piglets developed the chronic clinical form of EE 2-5 days after weaning (PW). Blood samples were obtained at 7, 14 and 21 days from both normal and clinically affected piglets for routine haematology and for the determination of CD45, CD21, CD4, CD8 and gammadeltaTCR cell markers by flow cytometry. When compared with clinically normal piglets EE affected pigs showed significantly decreased values of monocytes at 14 and 21 days PW, and increased numbers of neutrophils and leukocytes at 21 days PW. The EE affected pigs also had an early significant CD4(+) and CD8(high+) T lymphocyte proliferative response at 7 days PW. However affected pigs had a significantly reduced number of B (CD21(+)) and gammadeltaTCR(+) T lymphocytes in blood at 21 days PW. Although all values remained within the normal range, the significant differences in some peripheral blood leukocyte subsets between the two groups of piglets suggest that the generalised cutaneous infection with Staphylococcus hyicus is severe enough to induce a systemic inflammatory and immune responses.  相似文献   

10.
The present study was undertaken to establish reference values for the composition of blood leucocyte populations in neonatal calves by differential leucocyte counts and immunophenotyping. Neonatal calves 1 h post partum (p.p.) were found to have a very high absolute number of granulocytes while the number of peripheral blood mononuclear cells was lower than in calves aged 3-9 weeks. The relative numbers of T cell subpopulations were similar in newborn and older calves, but newborn calves had lower percentages of B cells and MHC class II positive cells. Within the first 4 h of life the relative numbers of CD2+, CD6+, and CD8+ T cells declined in colostrum-fed as well as in colostrum-deprived calves. In contrast, the percentage of MHC class II positive cells and monocytes increased from 1 h to 4 h p.p. particularly in colostrum-fed calves. Although there is some evidence for immaturity of lymphocytes in neonatal calves, the immune system of these animals seems to be fully present at birth.  相似文献   

11.
12.
Gnotobiotic calves given intramuscular injections of dexamethasone (DM, 0.5 mg kg-1 day-1) showed marked changes in haematological parameters including a neutrophilia and a lymphopaenia. Not only was there a reduction in the numbers of circulating mononuclear cells, but there was also a significant (P less than 0.01) decrease in the in vitro responsiveness of the remaining circulating peripheral blood lymphocytes to the mitogens, phytohaemagglutinin (PHA), concanavalin A (ConA) and pokeweed mitogen (PWM). Responses to all three mitogens were suppressed to a similar degree. Analysis of the circulating mononuclear cell sub-populations before and during DM treatment demonstrated a selective depletion of B cells; the T lymphocyte sub-population that expresses the gamma/delta form of T cell receptor, are CD2-, CD5+, CD8-, CD4- and constitute a major population in peripheral blood of calves. In vitro studies in gnotobiotic and conventional calves confirmed that DM was highly inhibitory for PHA responses but, in contrast to the in vivo findings, showed little effect of DM on ConA responses. Expression of surface antigens after 72 h in vitro culture in the presence of DM were little affected with the exception of BoCD8 and MHC II, which showed increased and decreased expression, respectively. These observations would suggest that distinct mechanisms are involved in glucocorticosteroid suppression of the responses to these two mitogens.  相似文献   

13.
Bestatin, a low-molecular weight dipeptide, is a potent inhibitor of aminopeptidase N which has been demonstrated to have antitumor and immunomodulatory effects. The effects of bestatin (10, 1 and 0.1 mg/kg) administered intraperitoneally once, five or ten times to mice on the total number of lymphocytes in the thymus, spleen and mesenteric lymph nodes and the percentage and the absolute number of T cell subsets (CD4+CD8+, CD4-CD8-, CD4+, CD8+) in the thymus and T (CD3+, CD4+, CD8+) and B (CD19+) lymphocytes in the spleen and mesenteric lymph nodes were studied. It has been found that bestatin administered ten times at doses of 10, 1 and 0.1 mg/kg increased the total number of thymocytes, splenocytes and lymphocytes of mesenteric lymph nodes. Bestatin also changed the percentage and the absolute number of T cell subsets in the thymus and T and B lymphocytes in the peripheral lymphatic organs. Five and ten exposures to bestatin (10, 1 and 0.1 mg/kg) increased the absolute count of both immature CD4+CD8+ and CD4-CD8- thymic cells. Moreover, both a single and multiple administration of bestatin (1 and 0.1 mg/kg) decreased the percentage and absolute count of CD3+ splenocytes and mesenteric lymph node cells with corresponding decreases in the percentage and absolute count of CD4+ and CD8+ cells. Both a single and multiple administration of bestatin at all the doses under investigation augmented the percentage and the absolute count of CD19+ (B lymphocytes) in the peripheral lymphatic organs. The results of the study show that there is a relationship between the effect induced by bestatin and the dose of the drug as well as the number of doses applied. The strongest effect on the T and B lymphocyte subsets was noted after five injections of bestatin at doses of 1 and 0.1 mg/kg.  相似文献   

14.
Leucocyte counts and subpopulations were studied in peripheral blood from calves experimentally infected in the respiratory tract with either bovine virus diarrhoea virus (BVDV) or Mannheimia haemolytica (Mh), or with a combination of both agents (BVDV/Mh). A non-inoculated control group was included. Peripheral blood samples were obtained for total leucocyte counts, and for neutrophil, lymphocyte and monocyte counts. The numbers of blood lymphocytes expressing the surface antigens CD4, CD8, WC1, B and IL-2R were analysed using flow cytometry. The results showed that BVDV inoculation induced a significant decrease in total leucocyte counts and in neutrophil and lymphocyte numbers, while Mh inoculation induced significant increases in total leucocyte counts and neutrophils, while the lymphocyte count decreased. In the BVDV/Mh group, the total leucocyte count and the lymphocyte numbers decreased significantly. In this group, the lymphocyte numbers remained on a very low level throughout the rest of the study. The numbers of CD4+, CD8+ and WC1+ lymphocytes decreased significantly compared with before inoculations mainly in the BVDV and BVDV/Mh groups. The drops were most pronounced in the BVDV/Mh group. The numbers of B+ lymphocytes and IL-2R+ cells did not change significantly.  相似文献   

15.
OBJECTIVE: To characterize changes in lymphocyte subsets over time in foals from birth to 18 weeks of age, accounting for differences among individuals, and to determine the effect of overnight storage of blood samples on foal lymphocyte subset concentrations. ANIMALS: 8 healthy Quarter Horse foals from birth to 18 weeks of age. PROCEDURE: Blood samples were collected longitudinally from birth to 18 weeks of age and a CBC performed on each sample. The samples were stained for lymphocyte markers, either immediately or after overnight storage and analyzed by flow cytometry. RESULTS: Total leukocytes, total lymphocytes, and the absolute concentrations of all lymphocyte subsets increased significantly with age. The proportions of B29A+, CD21+, and-equine major histocompatability complex class-II molecule+ lymphocytes increased significantly with age. The proportion of equine (Eq) CD5+, EqCD8+, and EqWC4+ lymphocytes decreased significantly with age. Significant differences among foals were found with respect to initial concentrations with respect to initial concentrations, but not with respect to the rate of increase of the various subsets tested. Significant differences were not found in subset values when comparing blood samples stained on the day of collection or after overnight storage at room temperature (approx 21 C) or under refrigeration. CONCLUSIONS AND CLINICAL RELEVANCE: These results are consistent with an increase in subset numbers and proportions over time, but with individual differences among foals. The observation of individual differences in subsets among foals suggests that there may be individual differences in susceptibility to infectious disease during the perinatal period. The absence of an effect of overnight storage makes field studies of lymphocyte subset concentrations more feasible.  相似文献   

16.
采用流式细胞仪检测1、3、5、7、14、21、28、35、42、49日龄AA肉鸡血液中的CD3、CD4、CD8阳性T细胞比例。研究结果表明:1~5日龄T细胞逐渐进入血液参与细胞免疫,7、21日龄注射疫苗起免疫应答作用,28日龄后基本形成稳固的细胞免疫水平。  相似文献   

17.
A longitudinal study of peripheral blood lymphocyte subsets from 7 to 15 months of age was performed in Beagle dogs employing a multiparametric flow cytometry. The data were compared with data obtained from adult Beagle dogs that were housed in the same animal facilities and that were subjected to the same controls during the 34 weeks of the study. Absolute counts of total lymphocytes and CD3+ T, CD3+CD4+ Th and CD21+ B lymphocytes decreased during the entire 34 weeks period of the study in the young dogs group. The same was observed with regard to the percentage of CD3+CD4+ Th lymphocytes and the CD4/CD8 ratio, while the percentage of CD3+CD8+ Tc lymphocytes increased from 7 to 15 months of age. These age-related changes found in lymphocyte subsets distribution of young dogs led to level the absolute and relative values of adult dog lymphocytes. The observations of this longitudinal study illustrate the changes related to maturation of lymphocyte subsets that occur during early life in Beagle dogs.  相似文献   

18.
The aim of the study was to determine whether treatment with recommended doses of meloxicam or flunixin had an effect on the apoptosis of peripheral blood T lymphocytes in calves. The study was carried out on 4-5 months old calves (n = 24, 8 per group). Experimental animals were injected subcutaneously with a single dose of 0.5 mg x kg(-1) of meloxicam or intravenously with 3 doses of 2.2 mg x kg(-1) day(-1) of flunixin. The non-treatment animals served as control. Blood samples were taken at day 0 and at days 1, 2, 3, 5, 7 and 14 after the first NSAIDs injection. Apoptosis was determined by flow cytometry using Annexin V-PE/7-AAD staining. The kinetic analysis of apoptosis in the total lymphocyte population, as well as in the CD4+ and CD8+ subsets did not reveal significant differences in the frequency of early apoptotic cells between control and experimental groups throughout the period studied. Although, 24 h after administration of the first dose of NSAIDs, late-stage apoptosis/necrosis was significantly increased in the total lymphocyte population (the meloxicam group), as well as in the CD4+ (the meloxicam group and the flunixin group) and CD8+ (the flunixin group) subsets of T cells. However, this disturbance was transient, relatively poorly expressed and, thus, unlikely to be of clinical significance. Our results indicate that the use of meloxicam or flunixin in accordance with the recommended dosage regimen in cattle do not have a clinically significant influence on apoptosis of peripheral blood T cells.  相似文献   

19.
OBJECTIVE: To assess the effects of inhalation of feed flour dust and dustborne endotoxin on respiratory tracts of pigs. ANIMALS: 29 healthy Belgian Landrace pigs. PROCEDURE: Pigs housed in an environmental chamber were exposed for 6 days to feed flour dust (1 to 15 mg/m3) and dustborne endotoxins (50 to 2,500 ng/m3). Effects were evaluated by measuring albumin concentration, lactate dehydrogenase (LDH) activity, cell composition of nasal lavage (NL) and bronchoalveolar lavage (BAL) fluids and blood, and percentages of CD4+ and CD8+ T lymphocytes in blood and lavage fluids. Dustborne endotoxin was obtained by mixing endotoxins from Escherichia coli (serotype O127:B8) with feed flour before spraying the flour in the environmental chamber. RESULTS: Exposure did not affect cell composition of NL fluid or blood. Total cell counts of BAL fluids were increased in all groups exposed to dust. Macrophage counts were increased in pigs exposed to inhalable dust concentrations as low as 4.4 mg/m3, and lymphocyte counts were increased in groups exposed to high dust concentrations. Percentages of CD4+ and CD8+ T lymphocytes in blood and lavage fluids were unchanged. In all dust-exposed groups, albumin content of BAL fluid was increased, whereas LDH activity was unaffected. Macrophage and lymphocyte infiltration and edema in the bronchi were identified by light microscopy. Effects attributable to E. coli endotoxin exposure were not identified. CONCLUSIONS: Inhalation of feed flour dust did not affect nasal mucosa but did induce bronchial airway inflammation. Dustborne endotoxins did not have effects attributable to endotoxin alone.  相似文献   

20.
为探讨治疗奶牛乳房炎的复方中药秦公散对小鼠脾脏和外周血中T淋巴细胞和B淋巴细胞亚群的影响,采用流式细胞仪测定各处理组小鼠脾脏和外周血中用CD3+、CD4+ 、CD8+标记的T淋巴细胞百分率及CD4+/CD8+的比值和CD19+标记的B淋巴细胞百分率。结果表明,与蒸馏水组相比,秦公散高剂量组小鼠脾脏和外周血中CD3+、CD4+细胞百分率和CD4+/CD8+的比值均显著提高,对环孢菌素(CsA)抑制小鼠有显著恢复作用(P<0.05);秦公散高剂量(20 g/kg体重)可显著提高健康小鼠脾脏和外周血中CD19+细胞百分率(P<0.05),对环孢菌素抑制小鼠的CD19+细胞百分率有显著恢复作用(P<0.05)。结果表明,治疗奶牛乳房炎的复方中药秦公散通过提高机体中CD3+细胞、CD19+细胞百分率和调节CD4+/CD8+之间的平衡来显著提高小鼠机体的细胞免疫功能和恢复由环孢菌素(CsA)抑制的小鼠机体免疫力。  相似文献   

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