Simultaneous Flow Cytometric Analysis of Phagocytosis and Oxidative Burst Activity in Equine Leukocytes

This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst, for the detection of impaired mechanisms of microbial destruction. Furthermore, the technique allows evaluation of opsonization activity in this context.     

IL-17细胞因子家族调节黏膜免疫的研究进展

皮肤和黏膜上皮细胞既是机体的物理屏障,又是机体防御微生物的第一道防线。上皮细胞与白细胞、树突细胞(DCs)等之间的相互作用,是机体产生适应性免疫反应的重要因素。IL-17细胞因子家族是最近新发现的具有强大的促炎症作用的细胞因子,它们在机体的固有和适应性免疫中发挥着重要作用,IL-17A、IL-17C和IL-17F能够直接作用于组织的上皮细胞,诱导各种免疫反应来对抗病原体,且能够促进组织的修复。IL-17E最基本的作用是作用于白细胞和诱导Ⅱ型免疫,这在其对抗寄生虫的作用中是非常关键的,此外,IL-17E还可以反向调节白细胞对IL-17A和IL-17F的产生;而对于IL-17B和IL-17D的研究则相对较少一些。     

IN VITRO EVALUATION OF FELINE LEUKOCYTES RADIOLABELED IN WHOLE BLOOD WITH 99MTC STANNOUS COLLOID

Technetium-99m stannous colloid (^99mTcSnC) has been used to radiolabel human leukocytes to investigate various inflammatory disorders. We investigated the in vitro behavior of feline leukocytes labeled in whole blood with ^99mTcSnC. Heparinized blood samples were collected from healthy cats and divided into control and test aliquots. The latter were labeled with ^99mTcSnC using a standard procedure. Leukocyte viability was determined for each sample using a trypan blue exclusion test. Labeling efficiency was determined for test aliquots. Test aliquots were layered onto Histopaque-1077^® and centrifuged before measurement of radioactivity of the blood components. Leukocytes from radiolabeled and control samples were washed and incubated with opsonized zymosan particles to allow assessment of phagocytic function. Aliquots were taken from radiolabeled feline leukocyte samples at 1, 3, 4, and 7 h postlabelling. After centrifugation of each aliquot, radioactivity of the supernatant and pellet was measured and the labeling retention determined. Leukocyte viability in both radiolabeled and control samples was >98%. The labeling efficiency was 95.2±0.14%. The distribution of radioactivity in feline blood was found to be 3.4±0.18% in plasma, 39.0±0.37% in erythrocytes, and 57.6±0.38% in leukocytes. Labeled feline leukocytes had phagocytic activity of 90.9±0.18% (control 91.3±0.15%). The radiolabeled leukocytes retained 93.4±0.19% of the radioactivity up to 7 h postlabeling. ^99mTcSnC efficiently labeled feline leukocytes with no effect on viability and minimal effect on phagocytic function. The percentage retention of radioactivity by the leukocytes was still high at 7 h postlabeling.     

A Preliminary Comparison of Plasma Fibrinogen Concentrations,Leukocyte Numbers and Erythrocyte Sedimentation Rate as Non-Specific Indicators of Inflammatory Conditions in Buffalo (Bubalis Bubalis)

Zargham Khan, M., Muhammad, G., Umar, A. and Ali Khan, S., 1997. A preliminary comparison of plasma fibrinogen concentrations, leukocyte numbers and erythrocyte sedimentation rate as non-specific indicators of inflammatory conditions in buffalo (Bubalis bubalis). Veterinary Research Communications, 21 (4), 265-271The plasma fibrinogen concentration (Fib), total leukocyte count (TLC), neutrophil, lymphocyte and monocyte numbers, and the erythrocyte sedimentation rate (ESR) were determined in 153 buffaloes suffering from different clinical conditions. Fib increased significantly (p < 0.05) in chronic mastitis, pyrexia, pyometra, cutaneous abscesses, tail gangrene and acute indigestion, whereas in most of the other conditions studied it varied non-significantly. TLC increased significantly in chronic mastitis, pyrexia, endometritis, cutaneous abscesses and infected skin wounds. An increase in neutrophils was associated with an increased TLC. Numbers of lymphocytes varied non-significantly in most of the conditions. Monocytes decreased significantly in most of the acute conditions. ESR was significantly elevated in all clinical conditions. Significantly increased mean Fib values in the different conditions varied from 703 ± 119 to 725 ± 140 mg/dl, while the maximum individual value was 1510 mg/dl in a case of cutaneous myiasis. The significantly increased mean TLC ranged from 9.48 ± 2.91 to 11.1 ± 3.5 × 103/µl, while it was 21.7 × 103/µl in a case of meningitis. ESR values in sick buffaloes varied from 57 to 111 mm in the first hour.     

Leukocyte numbers and intestinal mucosal morphometrics in horses with no clinical intestinal disease

Healthy horses and other animals have large numbers of resident leukocytes in the intestinal wall, but there is scant information regarding which and how many leukocytes are normally present in the equine intestinal wall. Our aim was to provide a reference range of leukocytes in the intestinal mucosal and submucosal propria of normal horses. We included in our study intestinal tissues from 22 Thoroughbred racehorses with no clinical intestinal disease, which had been euthanized because of catastrophic musculoskeletal injuries. Neutrophils, lymphocytes, eosinophils, macrophages, and plasma cells were counted in 5 random 17,600-µm² areas of villus lamina propria of the duodenum, jejunum, and ileum, and deep lamina propria of the duodenum, jejunum, ileum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, and small colon. Other features investigated in the same intestinal segments included villus height and width (small intestine), presence of ciliated protozoa, Paneth cells number, subcryptal leukocyte layers (number of leukocyte layers between the bottom of the crypts and the muscularis mucosae), and submucosal leukocytes. Lymphocytes were the most numerous cells in all segments analyzed, followed by plasma cells, eosinophils, macrophages, and neutrophils. Eosinophil numbers were significantly higher in both lamina propria and submucosa of the large intestine than in the small intestine. The duodenum had shorter and thinner villi than either jejunum or ileum. The data provided from our study will be useful for diagnosticians examining inflammatory processes in the intestinal tract of horses.     

Channel catfish, Ictalurus punctatus, leukocytes secrete immunoreactive adrenal corticotropin hormone (ACTH)

The interrelationships between the neuroendocrine and immune systems are becoming more understood, at least in mammalian systems. The most characterized of these relationships is that of hormonal signaling within the hypothalamo-pituitary-adrenal (HPA) axis. CNS-perceived signals stimulate the release of corticotropin releasing hormone (CRH) which in turn stimulates the release of pituitary corticotropin (ACTH) and ultimately the release of adrenal-cortex-derived corticosteroids. We demonstrate that channel catfish peripheral blood mononuclear cells, a channel catfish B-cell line (1G8) and a T-cell line (28S.1), constitutively and in response to CRF, secrete a molecule that is reactive with a mammalian RIA for ACTH (irACTH). The T-cell line was the most responsive to CRH and may provide a valuable model for understanding the interrelationships between the neuroendocrine and immune systems in lower vertebrates. Lymphoid derived ACTH, or ACTH-like products, in fish, as well as higher vertebrates, may represent a paracrine or autocrine control on lymphocyte function and immune regulation.     

Characterization of blood cells in the Leopard Cat (Prionailurus bengalensis)

Background: The Leopard Cat (Prionailurus bengalensis) is the most frequently encountered wild cat in most of Southeast Asia. Limited hematologic investigation exists for this species. Objectives: The objectives of this study were to assess routine hematologic measurements and parameters and characterize the morphology, cytochemical staining, and ultrastructural features of blood cells in Leopard Cats. Methods: Blood samples were collected from 12 adult healthy captive Leopard Cats (7 males and 5 females). Complete blood counts were performed using an automated hematology analyzer and manual differential counts. Cytochemical staining (Sudan black B [SBB], peroxidase [PO], periodic acid‐Schiff [PAS], α‐naphthyl acetate esterase [ANAE], and β‐glucuronidase [BG]) and scanning and transmission electron microscopy were performed using standard methods. Results: Median (range) hematologic results were as follows: PCV 0.46 L/L (0.30–0.55 L/L), hemoglobin 136.5 g/L (100–183 g/L), WBC 9.0 × 10⁹/L (6.9–15.2 × 10⁹/L), band neutrophils 0.07 × 10⁹/L (0–0.30 × 10⁹/L), segmented neutrophils 2.9 × 10⁹/L (1.2–6.34 × 10⁹/L), lymphocytes 5.3 × 10⁹/L (2.7–8.1 × 10⁹/L), eosinophils 0.14 × 10⁹/L (0–0.73 × 10⁹/L), basophils 0/L (0–0.22 × 10⁹/L), and monocytes 0.08 × 10⁹/L (0–0.30 × 10⁹/L). Neutrophils stained strongly positive for SBB, PO, and PAS; lymphocytes had fine granular positivity for ANAE and BG; monocytes were weakly positive for ANAE and BG; and basophils were strongly positive for BG. Ultrastructurally, eosinophils contained many large rod‐shaped granules with prominent crystalloid core structures, ribosomes, and mitochondria. Basophils contained many round to oval specific granules with homogeneous contents. Low number of basophils contained a few small vacuoles that usually were not detected by light microscopy. Conclusion: These findings will facilitate interpretation of hematologic results for future investigative and diagnostic studies of this species.     

Morphologic evaluation of acute endometritis in mares with differing resistance to uterine infections

The study was designed to determine differences between normal mares and mares with endometrial pathology in the inflammatory response after bacterial challenge. Six normal mares (biopsy category I) and 4 mares with pathological endometrial changes (biopsy category II) were given an intrauterine infusion of β-hemolytic streptococci on the second day of estrus. All mares had a similar kind of inflammatory response after the bacterial inoculation as assessed by rectal and vaginal examinations. There were no significant differences in the amount of discharge, uterine tone, uterine size and cervical relaxation between the groups. Leukocytic response, as determined by endometrial smears and biopsies, was of the same magnitude in both groups. Two mares from the pathological group were not able to eliminate the infection, but had vaginal discharge and bacteriologically positive uterine swabs until the end of the experiment. It is concluded that the inability of some mares to clear uterine infections cannot be explained by a deficient inflammatory response.     

Diurnal variation of canine blood leukocyte counts

A diurnal variation was detected in the numbers of total leukocytes, neutrophils, lymphocytes and eosinophils in the blood of 19 dogs (10 beagles and nine German shepherds). Blood samples were collected every fourth hour for 24 hours and once a day for 7 days. The neutrophil count increased during the day and reached its maximum in the late afternoon. The lymphocyte count had its maximum values in the late evening and its minimum values in the early morning. The eosinophil numbers were low around noon, increased during the afternoon and reached their maximum numbers in the late evening. Leukocyte numbers had statistically significant diurnal variation, so in designing research protocols with repeated blood sampling and closely controlled factors it may be important to take blood samples at the same time every day. The mild normal leukocyte changes during the day are not likely to confuse interpretation of clinical cases where patient results are compared with wide reference ranges.     

Studies on Bovine Leukosis

Summary

An enzyme linked immunosorbent assay (ELISA) and the agar gel immunodiffusion test with bovine leukosis virus glycoprotein as antigen (AGIDT‐BLV gp) were further used to test 633 bovine sera for antibodies to BL V. Both tests detected the same number of sera positive (149) or negative (464) for antibodies. Nine sera were negative in the ELISA but found to be weakly positive (2 sera) or bending the control line (7) in the AGIDT‐BLV gp. On the other hand 11 sera were scored negative in the AGIDT‐BLV gp but were weakly positive (9 sera), positive (1), and strongly positive (I) in the ELISA. Both tests are used routinely in this Institute as they complement each other, specially if sera with low antibody titers are under investigation. It is concluded that ELISA can fully replace radioimmunoassays in the serodiagnosis of enzootic bovine leukosis.