Polymorphonuclear leukocyte function in cattle with left displaced abomasum with or without concurrent infections

Cattle submitted to the University of Minnesota for surgical correction of left displaced abomasum (LDA) were examined for the in vitro phagocytic and bactericidal activities of their polymorphonuclear leukocytes (PMN). The PMN from cattle with LDA with or without concurrent infection had depressed phagocytic function when compared with PMN from healthy animals (controls). Those with concurrent infection had phagocytic activities lower than those in the group of cattle with LDA without any concurrent infection, and the former group was also observed to have depressed intracellular killing. Cattle with LDA complicated by infection were the only group in which phagocytic function was altered during surgical correction of LDA (and recovery). Treatment of PMN from both groups of affected cattle with levamisole in vitro enhanced intracellular killing, but had no effect on phagocytosis.     

Polymorphonuclear neutrophil leukocyte function in clinical bovine patients and in cows with or withoutStaphylococcus aureus mastitis

A fluorochrome microassay was used to investigate peripheral blood polymorphonuclear leukocyte (PMNL) function in cattle. Glass-adherent PMNL were reacted withStaphylococcus aureus princubated in 20% bovine serum for 30, 60 and 90 min. Coverslips were stained with acridine organge (AO) followed by crystal violet to quench extracellular bacterial fluorescence. PMNL function was evaluated by counting the number of dead (stained red with AO) and live (stained green with AO)S. aureus contained within 100 PMNL. A phagocytic index was calculated as the average number of bacteria contained within PMNL. The percentage killing ofS. aureus was calculated from the average proportion ofS. aureus within PMNL that were dead.Six clinically normal Holstein calves, 3–4 months of age, were sampled on 6 consecutive days. PMNL phagocytosis and killing did not vary significantly (p>0.05) among repeated samplings per calf. PMNL function increased with increasing time of incubation of PMNL withS. aureus. Means (± SD) for percentage killing were 46.7±13.1, 57.4±11.6, and 62.1±9.8% for 30, 60 and 90 min of reaction, respectively. Means (± SD) for the phagocytic index were 2.9±0.8, 3.6±1.0, and 4.2±1.1 bacteria/PMNL for 30, 60 and 90 min of reaction, respectively. PMNL function was determined in 30 normal cattle of various breeds, age and sex, and these values were pooled to provide normal values for PMNL function.When values for bovine clinical patients (n=25) with various diagnoses were compared with normal values (defined by the mean ± 2SD for the 30 normal cattle) for PMNL function, only one patient was observed to exhibit PMNL hypofunction. A cow with disseminated intravascular coagulation in association with peracute coliform mastitis exhibited decreased PMNL killing capacity. Abnormal PMNL function was uncommon in the hospital population studied.Peripheral blood PMNL function was evaluated in lactating Holstein cows with (n=15) or without (n=15) chronic subclinicalS. aureus mastitis. There was no significant (p>0.05) difference in PMNL function among these cows.     

Evaluation of bovine polymorphonuclear leukocyte function

Bovine polymorphonuclear leukocytes (PMNs) were isolated from the peripheral blood of cattle. Five in vitro procedures were utilized to evaluate PMN function: 1) Random migration under agarose, 2) Ingestion of ¹²⁵I-iododeoxyuridine labeled $\text{Staphylococcus aureus}$, 3) Quantitative nitroblue tetrazolium reduction, 4) Chemiluminescence and 5) Iodination. Normal values for bovine PMNs are reported and interpretation of results is discussed. The PMN function tests were designed so that all 5 procedures may be performed in a short period of time on the same cell preparation. This allows for the detection and partial characterization of a potential PMN dysfunction.     

Bovine leukocyte adhesion deficiency: in vitro assessment of neutrophil function and leukocyte integrin expression.

Bovine leukocyte adhesion deficiency (BLAD) was identified in a two-month-old Holstein heifer calf using DNA-polymerase chain reaction analysis of the affected calf and other clinical parameters. Neutrophil integrin expression (CD18, CD11a, CD11c), aggregation, and transendothelial migration were studied in vitro. Neutrophils were isolated from the affected calf and from normal, healthy, age-matched control Holstein calves. Neutrophils isolated from the affected BLAD calf had decreased expression of leukocyte integrins on their cell surface, decreased ability to aggregate in response to chemotactic stimuli, and decreased ability to migrate across bovine endothelial cell monolayers in vitro. Transendothelial migration of neutrophils from normal calves was reduced to levels comparable to the BLAD neutrophils by treatment with an anti-CD18 monoclonal antibody (MAb 60.3). Peripheral-blood lymphocytes from the BLAD calf also expressed negligible levels of leukocyte integrins, similar to their neutrophil counterparts. Our experimental findings in vitro correlate well with the clinical observations of decreased leukocyte trafficking and diminished host defense in leukocyte adhesion-deficient animals. The syndrome of BLAD may be a suitable model for one of the human leukocyte adhesion deficiency disorders.     

Effect of ovariohysterectomy on canine postsurgical leukocyte function

The effect of surgery on phagocytic activity of blood leukocytes and mitogen-induced blastogenesis of lymphocytes was studied in fourteen dogs. Simple ovariohysterectomy with anaesthesia induced by ketamine and xylazine or by ketamine, xylazine and halothane caused a short nonsignificant depression of phagocytic activity that persisted for four hours after surgery. Ingestion capacity of leukocytes decreased significantly immediately after surgery. Mitogen-induced blastogenesis of lymphocytes was depressed significantly in the first 48 hours and despite partial recovery this parameter did not reach the value of the control groups until the end of observation (7 days). A more conspicuous decrease of blastogenic response of blood lymphocytes to mitogens was found after the use of ketamine and xylazine in a dose maintaining anaesthesia. Anaesthesia with ketamine and xylazine in the lower dose and maintained with halothane resulted in a later improvement of the blastogenic response of lymphocytes.     

Characterization of peripheral blood and pulmonary leukocyte function in healthy foals

Studies in infants and foals indicate an age-dependent maturation of peripheral lymphocyte subsets. The age-dependent relationship for maturation of cellular immune responses, such as phagocytosis and lymphocyte responses of the peripheral and pulmonary-derived leukocytes, has not been characterized in foals. Lymphocyte subpopulations, mitogen stimulation response of lymphocytes, lymphokine-activated killing cell activity, phagocytosis and oxidative burst activity, and serum immunoglobulin (Ig) classes G and M concentrations were determined in developing foals. This study illustrates age-dependent changes in immunoglobulin class concentrations, lymphocyte subsets, and EqMHC Class II expression in cells of the peripheral blood and lungs of developing neonatal-to-weanling foals. The increase in peripheral blood and BAL B-lymphocytes and serum immunoglobulins in developing foals suggests expansion of immune cell populations during a time in which environmental pathogen exposure is great. General immune function, mitogenic responses, LAK cell activity, opsonized phagocytosis, and oxidative burst activity of newborns was similar to the adult horse. Total immune-cell numbers, rather than function, seemed to be the limiting factor in the development of the equine neonatal immune system. There was an age-related percent increase in the appearance of pulmonary lymphocytes, but a percent decrease in macrophages. Although development of the respiratory immune system follows changes in the peripheral blood, cellular expansion, activation, and migration may occur at a slower pace, making the respiratory environment susceptible to pathogens prior to optimal immune system maturity.     

Evaluation of orally administered valacyclovir in experimentally EHV1-infected ponies

The purpose of the current study was to investigate the therapeutic efficacy of valacyclovir against EHV1 in a controlled study. Eight na?ve Shetland ponies were inoculated with 10(6.5) TCID(50) of the neuropathogenic strain 03P37. Four ponies were treated with valacyclovir at a dosage of 40mg/kg bodyweight, 3 times daily, for 5 (n=2) or 7 (n=2) consecutive days, while the other four ponies served as untreated controls. The treatment regimen started 1h before inoculation. Ponies were monitored daily for clinical signs. At 0, 1, 2, 3, 4, 5, 7, 9, 11, 14, 17 and 21 days post inoculation (d pi), a nasopharyngeal mucus sample was taken to determine viral shedding. At the same time points, blood was collected and peripheral blood mononuclear cells (PBMC) were isolated to determine viremia. During the treatment, blood samples were collected 6 times daily, i.e. just before valacyclovir administration and 1h later, to determine the concentration of acyclovir in plasma. Also a nasopharyngeal swab was taken to measure the acyclovir concentration in nasal secretion. No differences could be noticed between valacyclovir-treated and untreated ponies. The clinical signs, the viral shedding and the viremia were similar in both the groups. Plasma acyclovir concentration could be maintained above the EC(50)-value of EHV1 during 50% of the entire treatment period in valacyclovir-treated ponies. Acyclovir could be detected in nasal swabs at concentrations varying from 50% to 100% of the corresponding plasma concentration. Although sufficiently high acyclovir levels could be reached in plasma and nasal mucus, no effect was seen of the treatment with valacyclovir on clinical signs, viral shedding and viremia of EHV1-infected ponies.     

Effect of Rhodococcus equi on equine polymorphonuclear leukocyte function

A procedure was developed for isolating large numbers of purified polymorphonuclear leukocytes (PMNs) from the peripheral blood of horses. Equine PMN function was evaluated by three procedures: 1) Staphylococcus aureus ingestion, 2) nitroblue tetrazolium reduction, and 3) iodination. Four preparations of R. equi were added to polymorphonuclear leukocytes (PMNs) in each test system. Live bacteria, heat-killed bacteria, the washed pellet from heat-killed bacteria, and the supernatant fluid from heat-killed bacteria were evaluated for effects on equine PMN function. None of the R. equi preparations had an effect on S. aureus ingestion by equine PMNs. Nitroblue tetrazolium reduction by PMNs, a measure of oxidative metabolism, was suppressed by pellet and supernatant fractions. Values for the iodination reaction were depressed by all R. equi preparations, indicating decreased activity of the myeloperoxidase-H2O2-halide system of the PMN. Further evaluation of the supernatant from heat-killed R. equi showed that it retained its inhibitory effect on iodination following autoclaving and/or passage through a 10,000 MW filter. R. equi fractions did not alter the enzymatic conversion of 125I to a protein-bound form in a PMN-free assay developed to evaluate this reaction. The presence of a surface component capable of inhibiting bactericidal mechanisms of the PMN may play an important role in intracellular survival of R. equi.     

Polymorphonuclear leucocyte function: relationship between induced migration into the bovine mammary gland and in vitro cell activity

Low doses of 10(-7) mg Escherichia coli endotoxin applied as intramammary infusion into single bovine quarters induced a rise in milk cell count without other inflammatory signs. Significantly fewer quarters responded in early lactation than in mid lactation. Maximum cell count was also somewhat later and less pronounced in early lactation. The rise in milk cell count after infusion of E. coli endotoxin was related to in vitro chemotactic activity of blood polymorphonuclear leucocytes (PMN). PMN isolated from cows which did not respond with a rise in milk cell count upon endotoxin infusion showed a diminished chemotactic activity in vitro as compared to PMN isolated from animals which did respond to an intramammary endotoxin infusion with a rise in milk cell count. No differences in phagocytic and metabolic activity were observed in vitro between the PMN isolated from the two groups of animals.     

Depression of polymorphonuclear leukocyte function associated with experimentally induced Escherichia coli mastitis in sows

In a study of susceptibilities of sows from 2 herds to experimentally induced Escherichia coli mastitis, a marked difference was seen. The "susceptible" sows were from a conventional herd and "resistant" sows were from a specific-pathogen-free herd. The purpose of the study was to determine whether deficient neutrophil function was associated with increased susceptibility to E coli-induced mastitis. Four in vitro procedures were used to evaluate polymorphonuclear leukocyte (PMN) function: (i) random migration under agarose, (ii) ingestion of 125I-iododeoxyuridine-labeled Staphylococcus aureus, (iii) quantitative nitroblue tetrazolium reduction, and (iv) iodination. After parturition and intramammary inoculation with E coli, sows from the susceptible herd were neutropenic and the neutrophils which were present in the peripheral blood had reduced function. Specifically, there were depressed random migration under agarose, S aureus ingestion, and iodination when compared with PMN function in resistant sows. These data indicate that susceptibility to E coli mastitis was associated with deficiencies in PMN numbers and function. Potential causes of the neutrophil dysfunction are discussed and include possible systemic hormonal aberrations or the presence of an inapparent viral or bacterial infection.     

Effects of bovine respiratory disease viruses and isoprinosine on bovine leukocyte function in vitro

Peripheral blood mononuclear cells obtained from 4- to 6-month-old-calves were inoculated in vitro with bovine herpesvirus-1, parainfluenza-3, or bovine virus diarrhea viruses. No increase in infectious virus progeny was observed; however, the viruses were detected in the cells for at least 96 h post-infection without any significant reduction in cell viability. The three viruses, either alone or in combination, suppressed phytohemagglutinin-induced proliferation of the mononuclear cells. The greatest suppression was observed in cultures inoculated with bovine virus diarrhea virus. Addition of isoprinosine partially restored this viral-induced suppression of proliferative response, and the efficiency of reversal was greater in bovine virus diarrhea virus-infected cells. Interleukin-2 activity was higher in cultures of virus-infected mononuclear cells than in cultures of non-infected cells.     

Effects of ACTH administration on bovine polymorphonuclear leukocyte function and lymphocyte blastogenesis

Yearling steers were treated with ACTH to determine the effect of increased plasma cortisol concentration on bovine lymphocyte and polymorphonuclear leukocyte (PMN) function. The administration of ACTH caused a significant (P less than 0.01) increase in serum cortisol concentration and depression of lymphocyte blastogenesis in response to phytohemagglutinin and concanavalin A. The response to pokeweed mitogen was also depressed, but not significantly. Random migration by PMN was significantly enhanced by ACTH treatment, but there was no effect on ingestion of Staphylococcus aureus, nitroblue tetrazolium reduction, or antibody-dependent cell-mediated cytotoxicity by PMN. The iodination reaction, which evaluates the activity of the myeloperoxidase-hydrogen peroxide-halide antibacterial system of the PMN, was significantly impaired after ACTH treatment. These data indicate that specific parameters of lymphocyte and neutrophil function were impaired directly or indirectly by elevated in vivo concentrations of plasma cortisol.     

The Origin of Amniotic Polymorphonuclear Leucocytes in the Mare

The objective of this study was to investigate the presence and origin of polymorphonuclear leucocytes (PMNLs) in the amniotic fluid of mares giving birth to healthy foals. Material from 25 mares was included. Amniotic fluid was collected during parturition before breakage of the amniotic vesicle. Manual microscopic cytologic evaluation was made on cytospin preparations after staining. PMNLs were found in all amniotic samples examined. The genomic DNA was extracted from 12 of the amniotic fluid samples and was genotyped. The results indicate that the PMNLs originate from the foetus.     

科学家发现了与HIV-1感染传播有关的蛋白

某国际研究小组已证实发现了与HIV-1入侵有关的宿主细胞的一个蛋白家族，因此对该病毒的传播有了更本质的认识。对这些蛋白及其之相关蛋白的研究，将带领人类找到治疗HIV-1和有同样入侵机制的其它病毒疾病的更好方法。和其它囊膜病毒一样，HIV-1首先得侵入宿主细胞，然后将感染传播给其它细胞。HIV的箝制蛋白包含一个特殊的氨基酸序列，该蛋白能启动人肿瘤基因101(TSG101)。然后，HIV病毒利用TSGIOI完成对蛋白排列和囊泡形成机制的控制，并达到自己的入侵目的。     

穿心莲对肉鸡淋巴细胞活性及白细胞吞噬功能影响的研究

本试验研究了不同剂型穿心莲对肉鸡细胞免疫的影响.将1日龄肉鸡随机分为七个试验组,处理如下:A组为1%超微粉;B组为2%超微粉;C组为0.2%浓缩颗粒(1g浓缩颗粒相当于5g生药);D组为0.4%浓缩颗粒;E组为1%浓缩颗粒;F组为2%细粉;G组为不加中药的空白对照.在试验的第14、21,28、35、42 d分别采血,进行淋巴细胞转化率、白细胞吞噬率及吞噬指数测定.试验结果表明,穿心莲对肉鸡淋巴细胞活性及白细胞活性(吞噬率和吞噬指数)有显著的提高作用(P＜0.05).同时,穿心莲超微粉和浓缩颗粒提高细胞免疫的效果优于细粉;相同剂量下,超微粉的效果有优于浓缩颗粒的趋势.     

Phagocytosis and Related Phenomena in Polymorphonuclear Leukocytes from Cow's Milk

There were fewer efficient phagocytes among leukocytes collected from artificially irritated mammary glands than among the leukocytes from blood of the same animals. The milk polymorphonuclear (PMN) leukocytes adhered poorly to a column of siliconised glass beads when compared with the blood cells. However, investigations of the O₂ uptake and CO₂ production of the milk PMN leukocytes revealed that these cells appeared to utilize metabolic pathways similar to those used by human and guinea pig PMN leukocytes during phagocytosis. These pathways are associated with degranulation and the production of H₂O₂ following particle ingestion. It is therefore suggested that the milk PMN leukocytes appear not to have lost the ability to produce this bactericidal substance.