Alpha-naphthyl acetate esterase activity is not a specific marker for ovine T lymphocytes

Alpha-naphthyl acetate esterase (ANAE) activity was demonstrated in ovine lymphocytes harvested from blood on Ficoll-metrizoate gradients. The enzyme's specificity for T lymphocytes, identified by immunofluorescent staining of T cell-specific antigens, was assessed. Correlation analysis of the results obtained using unfractionated lymphocytes from 12 sheep showed no correlation between ANAE activity and the expression of T cell antigens (r = 0.22). When lymphocytes from 4 sheep were fractionated on nylon wool columns a mean of only 43.2% of the cells in the non-adherent population were ANAE-positive whereas 94.7% of these cells were identified as T lymphocytes. Blood lymphocytes from 5 animals were separated into 3 fractions using Percoll discontinuous density gradients. No significant relationship was seen between ANAE activity and T cells in Fractions 1 and 3 (r = 0.41 and 0.21). Fraction 2 cells, however, did show a significant positive relationship (r = 0.91) between these two features but the biological significance of this relationship is unknown. It was concluded that ANAE activity is not a specific marker for ovine T lymphocytes.     

Use of alpha-naphthyl acetate esterase staining to identify T lymphocytes in cattle

Peripheral blood lymphocytes (PBL), lymph nodes, and/or spleens from clinically normal cattle were examined for cytochemical staining of alpha-naphthyl acetate esterase (ANAE). Two types of positive-staining patterns in ANAE staining resulted. By a combination of ANAE staining and latex-ingesting test, diffuse ANAE-positive cells were considered as mononuclear phagocytic cells. Using erythrocyte rosettes, erythrocyte antibody complement rosettes, nylon-wool column technique, surface immunoglobulin (SIg) staining, and the ANAE staining technique, granular ANAE-positive lymphocytes were shown to be T lymphocytes. The frequency of T and B lymphocytes in PBL, spleens, and lymph nodes of clinically normal cattle was measured, using ANAE staining and SIg staining. In PBL, 47.7% were ANAE-positive and 26.9% were SIg-positive; in spleens, 22.4% were ANAE-positive and 53.7% were SIg-positive; and in lymph nodes, 38.5% were ANAE-positive and 28.3% were SIg-positive. The frequencies of T and B lymphocytes in PBL, spleens, and/or lymph nodes from cattle with enzootic bovine leukosis (EBL) and cattle with persistent lymphocytosis and in tumor cells from cattle with EBL were measured. When compared with those of clinically normal cattle, PBL, spleens, and lymph nodes of cattle with EBL and the PBL of cattle with persistent lymphocytosis contained numerous SIg-positive cells and few ANAE-positive cells. Tumor cells from cattle with EBL contained 7.3% ANAE-positive and 78.0% SIg-positive cells.     

Light and electron microscopic studies on liver histology in chicks fed aflatoxin

This study was carried out under both light and electron microscopy to investigate the effects on liver carbohydrate and lipid metabolism caused by aflatoxin (AF) fed to chicks. Twenty broiler chicks were used. The birds were housed in electrically heated battery cages and exposed to light for 24 h. Feed and water were provided ad libitum. The animals were allocated to two groups each made up of 10 broilers. Total aflatoxin levels of zero (0) and 5 mg of AF/kg feed (81.05% AFB1, 8.79% AFG1, 6.06% AFB2, and 4.10% AFG2) added to a commercial diet, were fed to chicks from hatching up to 3 weeks of age, when the experiment was terminated. The chicks were executed by cervical dislocation and liver samples were obtained for light and electron microscopy. Oil red 'O', Sudan Black B, periodic acid Schiff (PAS) and Best's carmine stains were used to reveal fat and glycogen in the liver. Histological changes in hepatocytes included increased lipid droplets, high glycogen content, and mild mononuclear cell infiltration in the portal areas. Ultrastructural findings were destruction of rough endoplasmic reticulum (rER), reduction in mitochondrial size, enlargement of bile canaliculi, and cisternal dilatation of the smooth endoplasmic reticulum (sER).     

高铜对雏鸡外周血T-淋巴细胞ANAE阳性率的影响

选用1日龄艾维菌健康肉鸡420只,随机分为7组,分别喂以对照日粮(Cu 11 mg/kg)和高铜日粮(Cu 100mg/kg),高铜Ⅰ组,Cu 200 mg/kg,高铜Ⅱ组;Cu 300 mg/kg,高铜Ⅲ组;Cu 400 mg/kg,高铜Ⅳ组,Cu 500 mg/kg,高铜Ⅴ组;Cu 600 mg/kg,高铜Ⅵ组)6周,观察日粮铜添加水平对雏鸡外周血T-淋巴细胞ANAE阳性率的影响.与对照组比较,高铜Ⅰ、Ⅱ组雏鸡外周血T-淋巴细胞ANAE阳性率增加,但差异不显著;高铜Ⅲ、Ⅳ、Ⅴ、Ⅵ组雏鸡外周血T-淋巴细胞ANAE阳性率降低,且与日粮含铜量呈负相关,同时胸腺淋巴细胞减少,凋亡细胞比例增高.结果表明,与对照组比较,日粮铜含量100～200 mg/kg对雏鸡外周血中T-淋巴细胞ANAE阳性率有促进作用,但差异不显著;日粮铜含量300～600 mg/kg程度不同地降低了T-淋巴细胞ANAE阳性率,细胞免疫功能受损.     

Ultrastructural cytochemical characterization of alpha-naphthyl acetate esterase in chicken monocytic cell lines.

In order to study the differentiation level of two chicken monocytic cell lines, IN24 and LSCC-NP1 cells, alpha-naphthyl acetate esterase (ANAE) was examined by ultrastructural cytochemical technique. Morphologically and functionally, IN24 cells showed immature character compared to LSCC-NP1 cells. In IN24 and LSCC-NP1 cells, ANAE activity was localized on the external face of the plasma membrane and coated vesicles, but not in lysosomes and phagocytic vacuoles. ANAE reactivity showed a dense linear reaction product on the entire cell surface in IN24 cells and granular reaction product in LSCC-NP1 cells. On treatment with interferon-gamma (IFN-gamma), ANAE reactivity of IN24 cells became similar to that of LSCC-NP1 cells. IN24 cells treated with lipopolysaccharide (LPS) had less amounts of ANAE surface product, and reactivity was partially dispersed on the cell surface.     

Light and scanning electron microscopic study on the blood vascular system of the donkey placenta

The donkey placenta is diffuse and epitheliochorial with numerous microplacentomes consisting of a fetal microcotyledonary and a maternal microcaruncular part. The microplacentomal vasculature during the last third of pregnancy has been investigated by light microscopy in comparison to scanning electron microscopy of the materno-fetal contact surface and corrosion casts of blood vessels after plastic instillation from either the microcotyledonary or the microcaruncular side, and, for the first time in a perissodactyle, from both sides. Morphological data were semiquantitatively evaluated. The supplying parts of both, the microcotyledonary and the microcaruncular vascular system are strictly proximo-distally oriented, thus reaching the capillary systems or working parts in the shortest way possible. The straight course of the vasculature, particularly on the fetal side, suggests the occurrence of venulo-arteriolar back diffusion. The fetal capillary system consists of convolutes confronting the maternal septal capillary complexes in a countercurrent way. This materno-fetal blood flow interrelationship is highly efficient in terms of placental exchange, which is further supported (1) by dilations and increasing coiling of the fetal venular capillary limbs in particular and (2) by a decrease in the interhaemal distance from 12.5 to 7.2 microm between the two capillary systems. Besides the countercurrent blood flow interrelationship, some maternal branch arterioles reach the septal capillary system from the maternally oriented pole of the microplacentome or microcaruncle, respectively, resulting in the less efficient crosscurrent blood flow. Hence, in the donkey placenta fetal and maternal blood vessels meet in a mix of countercurrent and crosscurrent flow patterns.     

左旋咪唑对犬外周血T淋巴细胞转化的影响

左族咪唑又名左咪唑、左旋四咪唑，是人工合成的广谱驱虫药。本品具有旋光性，为噻咪唑左旋异构体，常用其盐酸盐(盐酸左旋咪唑)，为白色或微带黄色晶粉，性质稳定，易溶于水(25C：62．7％)，酸性水溶液中稳定。本品对体内多种寄生虫如蛔虫、肺丝虫、钩虫等具有强大的驱杀作用，并且给药方便，可皮下     

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).     

Blastocyst implantation in the domestic cat. Light and electron microscopic study]

Blastocyst implantation in the domestic cat A light and electronmicroscopic investigation Implantation in the cat begins 121/2 days post coitum and lasts to the 14th day. Several hours before implantation begins the extraembryonic trophoblast, in what could be considered a precontact or sensitization phase, shows fine structural evidence of secretory activity. The uterine epithelium at the same times gives evidence of being at first resorptive and, after a differentiation, of being secretory. Secretion also in the gland epithelium can be observed. The attachment of the embryo to the endometrium begins with an appositional phase. The trophoblast is now distinctly resorptive and forms with the uterine epithelium microvilli-free areas of contact lying between the openings of the uterine glands. Then follows an adhesion phase during which additional junctional complexes are formed between an actively proliferating trophoblast and the uterine epithelium which shows now symplasmic differentiation. Finally, in the intrusion phase the uterine epithelium completely disintegrates. In this manner the syncytiotrophoblast which differentiates from the cytotrophoblast invades the maternal tissues until it reaches the endothelium of the uterine blood vessels.     

Phagocytic activity in blood and proliferation of peripheral blood lymphocytes during the perinatal period in primiparous sows

The objective of this study was to determine whether phagocytic activity in blood and proliferation of peripheral blood lymphocytes are impaired during perinatal period. The study comprised 18 primiparous sows (Landras × Large White) free from clinical signs of diseases. During the experiment blood samples were collected three times from each sow. Sampling was performed on three different dates, always from all sows at once. At the first date of sampling sows were 21 ± 3 days before parturition, at the second date ± 1 day around parturition time and at the third date 21 ± 3 days after parturition. Phagocytic activity of monocytes and granulocytes was assessed in heparinized whole blood with addition of fluorescein isothiocyanate (FITC)-labelled opsonized bacteria Escherichia coli and the percentage of phagocytes which have ingested bacteria was measured as fluorescence activity by flow cytometry. The percentage of phagocyting monocytes and granulocytes was lowest at parturition (72.6 ± 16.37, 52.4 ± 20.59) and significantly increased within the next 21 ± 3 days (86.5 ± 6.16, 69.89 ± 5.80). Similarly, the phytohemagglutinin (PHA) (10 μg/ml) stimulated in vitro lymphocyte response was suppressed by parturition in primiparous sows (p < 0.001).     

Histological and electron microscopic studies of endemic ethmoidal carcinomas in cattle.

In five cases of endemic ethmoidal carcinoma in cattle from the Dominican Republic three tumor types could be classified: undifferentiated carcinoma (3), adenocarcinoma (1), and squamous cell carcinoma (1). Electron microscopy showed that the tumor cells in undifferentiated carcinomas closely resembled the cells of the normal olfactory mucosa. This was especially true for the dark cells of Bowman's gland. Ultrastructurally, the lymphoid cells of the undifferentiated bovine carcinoma resembled the lymphoid cells of human nasopharyngeal carcinoma being closely associated with Epstein-Barr Virus. This and epidemiological observations suggested a viral cause of endemic ethmoidal carcinoma.     

CD56 is expressed exclusively on CD3+ T lymphocytes in canine peripheral blood

CD56+ cells in canine blood leukocytes were characterized by flow-cytometric analysis of peripheral blood of 30 healthy adult beagle-dogs (15 males and 15 non-pregnant females). In 19 of the 30 dogs, anti human CD56 antibody, Leu-19, reacted with 8.8-21.7% of peripheral blood lymphocytes. All CD56+ cells simultaneously expressed CD3 molecules on their surface. Further phenotypic analysis revealed that 50.6+/-13.1% of the CD56+ cells showed CD4-CD8+ phenotype and 43.7+/-10.1% showed CD4+CD8- phenotype. Expression intensity of CD56 on the CD4-CD8+CD56+ cells was significantly higher than that on CD4+CD8-CD56+ cells (P<0.001). These findings indicate that CD56, which is a neural cell adhesion molecule, is uniquely expressed on subsets of T lymphocytes in canine peripheral blood.     

Flow cytometric analysis of ovine peripheral blood lymphocytes.

Leukocyte suspensions were prepared from the peripheral blood of 12 sheep three times at two month intervals beginning at 12 months of age. Monoclonal antibodies and flow cytometric analysis were used to characterize the cells. There were no significant differences over time therefore the data from the three time intervals were pooled. The mean percentages and ranges (minimum to maximum) of the major lymphocyte subtypes were: B-cells (29.6%, 11-50), gamma delta T-cells (36.6%, 22-68), CD4+ T-cells (14.1%, 8-22) and CD8+ T-cells (12.0%, 4-22). Lymphocyte subtype percentages appeared less variable within than between individuals. Two populations of B-cells were noted; one population had more cytoplasm and light scatter characteristics similar to monocytes while a second population of B-cells was more typical of small lymphocytes. The nature of the large B-cells requires further study.     

PD-L1+ HeLa细胞对人外周血活化T细胞的免疫调节作用

目的 探讨人宫颈癌细胞表达PD-L1对人外周血T细胞的免疫调节作用.方法 采用PD-L1质粒转染人宫颈癌HeLa细胞株,将PD-L1+ HeLa细胞与PHA刺激活化的人外周血T淋巴细胞混合培养,MTT法检测T细胞的增殖水平,流式细胞术分析T细胞凋亡率和CD8+/CD4+T细胞比例;ELISA检测PD-L1+ HeLa细胞与PHA刺激活化的人外周血T淋巴细胞混合培养上清液IFN-γ的含量.结果 MTT检测显示与PD-L1+ HeLa混合培养组的T细胞增殖光密度值为(0.31±0.02),明显低于与PD-L1 - HeLa混合培养组的T细胞和对照组T细胞,分别为[(0.62±0.02)和(0.59±0.03),P＜0.01];T细胞凋亡率明显高于与PD-L1 - HeLa混合培养组和对照组T细胞,分别为(32.7%、17.9%和18.3%);CD8 +/CD4+T细胞比值稍低于PD-L1 - HeLa混合培养组和对照组T细胞(分别为0.86、0.91和0.89);与PD-L1+ HeLa混合的T细胞培养上清液IFN-γ的含量低于与PD-L1 - HeLa混合培养组和对照组T细胞,分别为[(801.1±20.5) pg/ml、(1 006.2±30.2 )pg/ml和(1 070.2±113.3) pg/ml,P＜0.01].结论 PD-L1明显抑制人外周血活化T细胞增殖促进其凋亡并下调其免疫功能.     

Peripheral neuropathy associated with dietary riboflavin deficiency in the chicken. I. Light microscopic study

Chickens fed a riboflavin-deficient diet from hatching had leg weakness and paralysis as early as 12 days of age. Signs worsened through day 16; after 35 days, recovery was evident. Sciatic nerves from affected chickens were enlarged. Significant microscopic lesions were confined to peripheral nerves and included tissue separation (suggesting interstitial edema), Schwann cell swelling, perivascular leukocytic infiltration, and segmental demyelination accompanied by accumulation of osmiophilic debris in Schwann cell cytoplasm. Axon degeneration was present, but was not a primary lesion. Acid phosphatase enzyme activity of Schwann cells was increased in affected nerves. These results demonstrate that dietary riboflavin deficiency causes a demyelinating peripheral neuropathy in young, rapidly growing chickens.