Percentage Of Phagocytic Cells Research Articles

The Effect of Orally Administered Multi-Strain Probiotic Formulation (Lactobacillus, Bifidobacterium) on the Phagocytic Activity and Oxidative Metabolism of Peripheral Blood Granulocytes and Monocytes in Lambs.

Probiotic feed additives have attracted considerable research interest in recent years because the effectiveness of probiotics can differ across microbial strains and the supplemented macroorganisms. The present study was conducted on 16 lambs divided equally into two groups (C-control and E-experimental). The examined lambs were aged 11 days at the beginning of the experiment and 40 days at the end of the experiment. The diet of group E lambs was supplemented with a multi-strain probiotic formulation (Lactobacillus plantarum AMT14, Lactobacillus plantarum AMT4, Lactobacillus rhamnosus AMT15, and Bifidobacterium animalis AMT30), whereas group C lambs did not receive the probiotic additive. At the beginning of the experiment (day 0) and on experimental days 15 and 30, blood was sampled from the jugular vein to determine and compare: phagocytic activity (Phagotest) and oxidative metabolism (Phagoburst) of peripheral blood granulocytes and monocytes by flow cytometry. An analysis of the phagocytic activity of granulocytes and monocytes revealed significantly higher levels of phagocytic activity (expressed as the percentage of phagocytic cells and mean fluorescence intensity) in lambs that were administered the multi-strain probiotic formulation compared with lambs in the control group. The probiotic feed additive also exerted a positive effect on the oxidative metabolism of both granulocytes and monocytes (expressed as the percentage of oxidative metabolism and mean fluorescence intensity) after stimulation with Escherichia coli bacteria and with PMA (4-phorbol-12-β-myristate-13-acetate). These findings suggest that the tested probiotic formulation may have a positive effect on the immune status of lambs.

Mesenchymal Stem/Stromal Cells Derived from Cervical Cancer Promote M2 Macrophage Polarization

Macrophages with the M2 phenotype promote tumor development through the immunosuppression of antitumor immunity. We previously demonstrated the presence of mesenchymal stem/stromal cells (MSCs) in cervical cancer (CeCa-MSCs), suggesting an immune protective capacity in tumors, but to date, their effect in modulating macrophage polarization remains unknown. In this study, we compared the capacities of MSCs from normal cervix (NCx) and CeCa to promote M2 macrophage polarization in a coculture system. Our results demonstrated that CeCa-MSCs, in contrast to NCx-MSCs, significantly decreased M1 macrophage cell surface marker expression (HLA-DR, CD80, CD86) and increased M2 macrophage expression (CD14, CD163, CD206, Arg1) in cytokine-induced CD14+ monocytes toward M1- or M2-polarized macrophages. Interestingly, compared with NCx-MSCs, in M2 macrophages generated from CeCa-MSC cocultures, we observed an increase in the percentage of phagocytic cells, in the intracellular production of IL-10 and IDO, the capacity to decrease T cell proliferation and for the generation of CD4+CD25+FoxP3+ Tregs. Importantly, this capacity to promote M2 macrophage polarization was correlated with the intracellular expression of macrophage colony-stimulating factor (M-CSF) and upregulation of IL-10 in CeCa-MSCs. Furthermore, the presence of M2 macrophages was correlated with the increased production of IL-10 and IL-1RA anti-inflammatory molecules. Our in vitro results indicate that CeCa-MSCs, in contrast to NCx-MSCs, display an increased M2-macrophage polarization potential and suggest a role of CeCa-MSCs in antitumor immunity.

Ageing-Induced Decline in Primary Myeloid Cell Phagocytosis Is Unaffected by Optineurin Insufficiency

Optineurin is a ubiquitin-binding adaptor protein involved in multiple cellular processes, including innate inflammatory signalling. Mutations in optineurin were found in amyotrophic lateral sclerosis, an adult-onset fatal neurodegenerative disease that targets motor neurons. Neurodegeneration results in generation of neuronal debris, which is primarily cleared by myeloid cells. To assess the role of optineurin in phagocytosis, we performed a flow cytometry-based phagocytic assay of apoptotic neuronal debris and E. coli bioparticles in bone marrow-derived macrophages (BMDMs), and primary neonatal microglia from wild-type (WT) and optineurin-insufficient (Optn470T) mice. We found no difference in phagocytosis efficiency and the accompanying cytokine secretion in WT and Optn470T BMDMs and microglia. This was true at both steady state and upon proinflammatory polarization with lipopolysaccharide. When we analysed the effect of ageing as a major risk factor for neurodegeneration, we found a substantial decrease in the percentage of phagocytic cells and proinflammatory cytokine secretion in BMDMs from 2-year-old mice. However, this ageing-induced phagocytic decline was unaffected by optineurin insufficiency. All together, these results indicate that ageing is the factor that perturbs normal phagocytosis and proinflammatory cytokine secretion, but that optineurin is dispensable for these processes.

Modern approaches to the application of photoimmunomodulating technologies in the treatment of HPV-associated cervicitis

We aimed to investigate the effects of the combined use of polychromatic visible and infrared polarized light with topical application of Imiquimod 5% cream on the macrophage link of immunity in patients with chronic human papillomavirus (HPV)-associated cervicitis.
 Materials and methods. The study included 90 patients aged 2035 years, who had a history of HPV-associated cervicitis for at least 1 year. Patients were distributed into three groups by random sampling. The main group included 30 patients who were exposed to blue monochromatic polarized incoherent light on the projection of the carotid arteries and polychromatic visible and infrared polarized light on the uterine cervix in combination with topical application of Imiquimod 5% cream (complex one). The comparison group included 30 patients who were exposed to polychromatic visible and infrared polarized light on the uterine cervix in combination with Imiquimod 5% cream (complex 2), and the control group included 30 patients who received local exposure to the uterine cervix with Imiquimod 5% cream. Examination data of 20 healthy women of the same age were taken as normal values. The macrophage link activity was assessed using the nitroblue tetrazolium test.
 Results. Under the influence of the developed complexes 1 and 2, the percentage of phagocytic cells of the macrophage link increased significantly to normal values (by 36% and 31%, respectively; p 0.01), the phagocytic number and index of phagocytosis completeness increased to near-normal values (p 0.001). In addition, the percentage of spontaneous (excessive) redox processes of neutrophils decreased from 21.1% 1.3% to 11.8 1.01% in the main group (p 0.001) and to 14.4 1.5% in the comparison group (p 0.01). Less significant results were obtained in the control group where the patients received topical application of Imiquimod 5% cream on the uterine cervix, and the studied parameters were 26%51.7% lower than that in the main group.
 Conclusion. The complex application of blue monochromatic polarized incoherent light on the projection of the carotid arteries in combination with polychromatic visible and infrared polarized light and external application of Imiquimod 5% cream on the cervix of patients with chronic HPV-associated cervicitis has a pronounced immunocorrecting effect on the immunity macrophage link. This is confirmed by an improvement in the indicators of the functional activity of neutrophils, in the form of a decrease in the percentage of excessive redox reactions and an increase in the index of phagocytosis completeness, and indicates an increase in the productivity of the macrophage link functioning.

Вплив реабілітаційно-профілактичного комплексу на стан імунної системи в дітей з рекурентною респіраторною патологією на тлі гастроезофагеальної рефлюксної хвороби

Вплив реабілітаційно-профілактичного комплексу на стан імунної системи в дітей з рекурентною респіраторною патологією на тлі гастроезофагеальної рефлюксної хвороби

Coralline Skeleton Biomaterial Reduces Phagocytosis in Mouse Blood in vitro

AbstractInflammatory and immunogenic response to foreign bodies presents a challenge in the use of biomaterials as implants for tissue restoration. Therefore, there is a need to understand the interactions between such implants and the blood. One such material, currently in clinical use for bone replacement in humans, is the skeleton of corals, in the form of crystalline aragonite. This biomaterial has been shown to impart a protective and supportive influence on several types of cells ex vivo. The carbonate skeleton activates secretion in phagocytes in vitro, however its effects on these cells in the blood, and on the process of phagocytosis itself, remain unknown. Using 1–500 μm particles of coral skeleton, we show that these particles bind blood proteins and alter the leukocyte population, reducing the proportion of granulocytes by more than 3‐fold with no effect on the proportion of monocytes. In addition, the presence of coral skeleton in the blood causes a reduction in phagocytosis. Specifically, we observed a decrease in the percentage of phagocytic cells by 27 % in the granulocytes and by 73 % in monocyte family, as well as a 41.6 % reduction in the MFI of granulocytes, but with no such effect on monocytes. Taken together, the results suggest that the coral skeleton biomaterial may act as a strong, promotive scaffold for tissue regeneration due to its ability to reduce its rejection by inflammatory reactions such as phagocytosis.

The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats.

Simple SummaryThe main focus of industrial livestock production is to maximise production output without compromising the well-being of animals, which is why animal diets are supplemented with various feed additives. Feed additives boost immunity and protect animals against pathogens. The list of potential feed additives includes β-hydroxy-β-methylbutyrate (HMB) which occurs naturally in small quantities in citrus fruit, avocado, asparagus, cauliflower, selected fish species, red wine, milk, and alfalfa. However, its mechanism of action and effects on immune system cells have not been thoroughly investigated in animals, including goats. In the present study, the experimental goats whose diets were supplemented with HMB over a period of 60 days were characterised by higher levels of chemotactic and phagocytic activity and a higher rate of oxidative metabolism of peripheral blood granulocytes and monocytes than control group animals whose diets were not supplemented. Granulocytes and monocytes constitute the first line of defence against pathogens and protect animals against disease. They play a particularly important role in young animals which are more susceptible to viral and bacterial infections. Feed additives can deliver numerous benefits by boosting immunity and preventing the spread of infectious diseases in goats.The objective of this study was to determine the effect of β-hydroxy-β-methylbutyrate (HMB) on the chemotactic activity, phagocytic activity, and oxidative metabolism of peripheral blood granulocytes and monocytes in goats. Goat kids aged 30 ± 3 days were divided into two groups of 12 animals each: I—control, and II—experimental. Experimental group animals were fed a diet supplemented with HMB in the amount of 50 mg/Kg BW; whereas the diets of control goats were not supplemented. At the beginning of the experiment (day 0) and on experimental days 15, 30, and 60, blood was sampled from the jugular vein to determine and compare chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit), and oxidative metabolism (BURSTTEST® kit) of peripheral blood granulocytes and monocytes by flow cytometry. The analyses of the chemotactic and phagocytic activity of granulocytes and monocytes revealed statistically higher levels of phagocytic activity in the experimental group than in the control group, as expressed by the percentage of phagocytic cells and mean fluorescence intensity. HMB also enhanced the oxidative metabolism of both granulocytes and monocytes, expressed by the rate of oxidative metabolism and mean fluorescence intensity after stimulation with Escherichia coli bacteria and PMA (4-phorbol-12-β-myristate-13-acetate).

The Phagocytosis of Blood Leukocytes from Cystic Fibrosis Patients is not Impaired in General.

Impaired phagocytosis of Pseudomonas aeruginosa was found in isolated monocytes of peripheral blood of cystic fibrosis patients, but not in their neutrophils, as reported some years ago. In the present study, we analysed the phagocytic capacity of peripheral blood neutrophils and monocytes of cystic fibrosis patients and of healthy controls. Phagocytosis was determined using a commercial phagocytosis "in whole blood" assay on the basis of fluorescence-labelled opsonized Escherichia coli bacteria and flow cytometry. Venous blood of cystic fibrosis patients and of healthy controls was collected and the phagocytosis assay was performed. No differences in the percentage of phagocytic cells or in the overall phagocytic capacity were found between samples of cystic fibrosis patients and healthy controls either in monocytes or in neutrophils. Thus, our results did not support the hypothesis of a generally reduced phagocytic ability in the peripheral blood immune cells of cystic fibrosis patients.

Effect of vitamin C on innate immune responses of rainbow trout (Oncorhynchus mykiss) leukocytes

Vitamin C, also known as ascorbic acid, is an essential micronutrient that influences a wide variety of physiological processes, including immunological functions. Although the positive effects of vitamin C supplementation on the immunological status of fish has been established in different species, the bases for these positive effects are still unknown. Hence, the aim of our study was to evaluate the in vitro effect of vitamin C on several innate immune functions of rainbow trout (Oncorhynchus mykiss) leukocyte populations. For this, we assessed the effects exerted on the established rainbow trout monocyte-macrophage cell line RTS11, and compared them to those observed in trout head kidney leukocytes. Our results demonstrate that vitamin C increases the production of reactive oxygen species and the percentage of phagocytic cells in both cell populations. On the other hand, vitamin C had no effect on the surface MHC II levels and only in the case of RTS11 cells increased the capacity of these cells to migrate towards the CK9 chemokine. Finally, vitamin C also increased the transcription of several pro-inflammatory and antimicrobial genes elicited by Escherichia coli, with some differences depending on the cell population studied. Our results contribute to further understand how vitamin C supplementation regulates the fish immune system.

Culture medium associated changes in the core proteome of macrophages and in their responses to copper oxide nanoparticles.

The physiology of cells cultured in vitro depends obviously on the external conditions, including the nutrients present in the culture medium. In order to test the influence of this parameter, J774 macrophages grown either in RPMI or in DMEM were compared by a combination of targeted analyses and a proteomic approach. The two media differ in their glucose, amino acids, and vitamins concentrations, but there were no significant differences in the cell cycle or in the percentage of phagocytic cells in both media, although the phagocytic capacity (i.e. the number of phagocytized particles) was higher in DMEM. Conversely, we found that J774 cells grown in RPMI produced more nitric oxide in response to lipopolysaccharide. The proteomic study highlighted differences affecting the central metabolism and nucleotide metabolism, cytoskeleton, protein degradation, and cell signaling. Furthermore, proteomics showed that J774 cells grown in RPMI or in DMEM and exposed to copper oxide nanoparticles respond rather differently, with only a few proteins similarly modulated between cells grown in both media. Taken together, our results show that the basal state of cells grown in two different media is different, and this may affect the way they respond to an external stimulus or stress.

Effect of a plant preparation Citrosept on selected immunity indices in blood of slaughter turkey hens.

The objective of this study was to determine the effect of per os administration of 3 various dosages of a Citrosept preparation (a grapefruit extract)to growing turkey hens on changes in their selected haematological and immunological blood indices. An attempt was also undertaken to select the most efficient dose of the preparation with respect to the mentioned indices in turkey hens. The experiment was conducted on 180 turkey hens allocated at random to 4 groups, 45 birds in each group. Samples of their full blood were analyzed for haematological indices, such as red blood cell count (RBS), haemoglobin content (Hb), haematocrit value (Ht), and white blood cell count (WBC). Samples of blood plasma were assayed to determine the activity of lysozyme (chamber-diffusive method) and heterophils capability to reduce nitro blue tetrazolium (stimulated and spontaneous NBT test). Phagocytic activity of leucocytes against Staphylococcus aureus 209P strain was assessed and expressed as the percentage of phagocytic cells (% PC) and phagocytic index (PI). The administration of the grapefruit extract to turkey hens with drinking water caused a significant increase in haemoglobin content in blood, as well as an increase in non-specific humoral immunity marker (activity of lysozyme) and non-specific cellular immunity marker (percentage of phagocytic cells; P ≤ 0.05). The results obtained enabled the positive evaluation of the advisability of applying the Citrosept preparation in the feeding of turkey hens at the age of 6-9 weeks. Among the doses examined, the most efficient with respect to the stimulation of the non-specific humoral and cellular immunity was the dose of 0.021 ml/kg of body weight.

Exposure of single-walled carbon nanotubes impairs the functions of primarily cultured murine peritoneal macrophages

It is increasingly important to understand the single-walled carbon nanotubes' (SWCNTs) immune response as their increasingly biomedical researches and applications. Macrophages and T cells play important roles in scavenging foreign materials and pathogens and regulating immune response. In this work, primarily cultured murine peritoneal macrophages and purified splenic T cells were utilised to determine the toxic effects of SWCNTs and acid-functionalised SWCNTs (AF-SWCNTs) on the immune system, especially on macrophage functions. Macrophages were exposed to 0–50 μg/ml of CNTs for 24 h and no significant cytotoxicity was found by live/dead and annexin-V-FITC/PI analyses. The TEM images revealed that AF-SWCNTs were engulfed mostly through phagocytosis and located in lysosomes of macrophages. Measurement of mitochondrial membrane potential and proteasome subunit gene expression demonstrated that 10 and 50 μg/ml AF-SWCNTs could damage mitochondrial function and proteasome formation in a concentration-dependent manner. Functional analyses revealed that the percentage of phagocytic cells were affected significantly by 20 μg/ml CNTs, and 5 μg/ml AF-SWCNTs inhibited the phagocytic efficiency of latex beads in macrophages. The accessory cell function was affected by both AF-SWCNTs and SWCNTs at concentrations of 10 and 50 μg/ml, respectively. Furthermore, AF-SWCNT biased naïve T-cell differentiation to Th1 type by inducing the production of IFN-γ and TNF, implying the potential risk of Th1-associated diseases (e.g. autoimmune diseases and inflammation) on AF-SWCNT exposure.

Effect of Oral Administration of Zinc Sulphate with Simultaneous Use of Nonspecific Immunostimulation on the Course of Trichophytosis in Beef Cattle

Abstract The paper assesses the treatment of cutaneous fungal infection in young beef cattle caused by Trichophyton verrucosum using orally administered zinc sulphate for 28 d at 2 g per animal per day and subcutaneous application of 2.5 mg/kg levamisole administered three times every 7 d. The study was conducted on animals at the age of 110 to 122 d, which were divided into two experimental groups: group I (animals suffering from ringworm with low levels of zinc in serum) and group II (animals suffering from ringworm with normal levels of zinc in serum). The remaining animals (control group) were divided into three subgroups: K (clinically healthy animals, which had normal serum levels of zinc), KI (animals infected with ringworm and showing a reduced level of zinc), and KII (animals suffering from ringworm with normal serum zinc levels). During the study the following parameters were investigated: clinical signs of fungal infection, zinc content in serum, percentage of phagocytic cells, and phagocytic index. The symptoms of the disease were observed only in group II on day 56. In group I, an increase in the percentage of phagocytic cells was observed, as compared to the original values, on days 7, 21, and 56 and a fall in this parameter was noted on days 14 and 28. In group II, there was a decrease on day 14 and an increase in comparison to the original values, followed by a gradual fall till day 56 of the experiment. In group I, phagocytic index remained on a stable level till day 56 and in group II until day 28, and subsequently began to increase gradually. It was found that oral supplementation of zinc sulphate combined with nonspecific immunostimulation may be applied in fighting skin fungal infection in cattle, and that zinc and levamisole may exhibit synergism affecting the evolution of nonspecific immunity.

Modulation of the immune parameters and expression of genes of gilthead seabream (Sparus aurata L.) by dietary administration of oxytetracycline

The oxytetracycline (OTC) is an antibiotic widely used in the aquaculture, principally in the treatment of infections caused by the pathogenic Gram-negative bacteria. In the present study, the consequences of the oral administration of OTC on the main humoral (natural haemolytic complement activity, IgM levels and seric peroxidase level) and cellular (respiratory burst, intracellular peroxidase level and phagocytosis) immune parameters of gilthead seabream (Sparus aurata, L) were evaluated after 7, 14 and 21days of treatment. Furthermore, the expression levels of immune-associated genes (MHC-IIα, C3, IL-1β, COX-2 and β-defensin) were analysed by real-time polymerase chain reaction in head-kidney and gut. Three experimental groups were established: control group (fed with non-supplemented OTC food) and experimental groups (4 and 8mg OTC g−1 feed, respectively) (N=30). The results reveal a decrease of the natural haemolytic complement activity to the higher dose of OTC administered. By contrast, after 7days of treatment, fish fed the lowest concentration of OTC were observed to have a statistically significant increase in the percentage of phagocytic cells, while the phagocytic capacity and respiratory burst increased after 14days of treatment for both concentrations of OTC. No significant effects were observed after 21days of treatment for both concentrations of OTC with regard to the values obtained in the control group. In head-kidney and gut, the dietary administration of OTC not caused statistically significant variation, only in C3 gene expression at 7days in gut. The present results suggest that after dietary administration of OTC, the humoral and cellular innate immune parameters of gilthead seabream are depressed and stimulated respectively, and induce up- or down-regulation of the immune-relevant genes analysed depending on the concentration of OTC supplied in diet.

Enhanced Cellular Immunity in Shrimp (Litopenaeus vannamei) after ‘Vaccination’

It has long been viewed that invertebrates rely exclusively upon a wide variety of innate mechanisms for protection from disease and parasite invasion and lack any specific acquired immune mechanisms comparable to those of vertebrates. Recent findings, however, suggest certain invertebrates may be able to mount some form of specific immunity, termed ‘specific immune priming’, although the mechanism of this is not fully understood (see Textbox S1). In our initial experiments, either formalin-inactivated Vibrio harveyi or sterile saline were injected into the main body cavity (haemocoel) of juvenile shrimp (Litopenaeus vannamei). Haemocytes (blood cells) from V. harveyi-injected shrimp were collected 7 days later and incubated with a 1∶1 mix of V. harveyi and an unrelated Gram positive bacterium, Bacillus subtilis. Haemocytes from ‘vaccinated’ shrimp showed elevated levels of phagocytosis of V. harveyi, but not B. subtilis, compared with those from saline-injected (non-immunised) animals. The increased phagocytic activity was characterised by a significant increase in the percentage of phagocytic cells. When shrimp were injected with B. subtilis rather than vibrio, there was no significant increase in the phagocytic activity of haemocytes from these animals in comparison to the non-immunised (saline injected) controls. Whole haemolymph (blood) from either ‘immunised’ or non-immunised’ shrimp was shown to display innate humoral antibacterial activity against V. harveyi that was absent against B. subtilis. However, there was no difference in the potency of antibacterial activity between V. harveyi-injected shrimp and control (saline injected) animals showing that ‘vaccination’ has no effect on this component of the shrimp's immune system. These results imply that the cellular immune system of shrimp, particularly phagocytosis, is capable of a degree of specificity and shows the phenomenon of ‘immune priming’ reported by other workers. However, in agreement with other studies, this phenomenon is not universal to all potential pathogens.

Evaluation of peripheral blood neutrophil function in tumor‐bearing dogs

Peripheral blood neutrophils of untreated human cancer patients have been shown to have normal, increased, and decreased phagocytic activity, killing capacity, and/or oxidative burst activities. The objectives of this study were to evaluate oxidative burst and phagocytic activities of peripheral blood neutrophils from tumor-bearing dogs before therapy and compare them with neutrophil function of healthy control dogs. Heparinized whole blood was obtained from dogs with high-grade lymphoma (n=23), sarcoma (n=13), or carcinoma (n=11), and healthy control dogs (n=11) for flow cytometric evaluation of oxidative burst and phagocytic activities. Percentage of bursting cells and amount of oxidative burst activity were determined after stimulation with phorbol 12-myristate 13-acetate (PMA) or Escherichia coli. Percentage of phagocytic cells and amount of phagocytic activity were determined after incubation with fluorescent E. coli. Compared with control dogs, dogs with sarcoma (P=.004) and carcinoma (P=.05) had a lower percentage of neutrophils exhibiting oxidative burst activity after stimulation with PMA. Phagocytic activity was significantly lower in dogs with sarcomas compared with control dogs (P<.0001) and dogs with lymphoma (P=.01). Untreated carcinomas and sarcomas in dogs may suppress the percentage of neutrophils capable of oxidative burst when stimulated by PMA. Furthermore, sarcomas also may suppress the amount of phagocytic activity per neutrophil. Until further studies can be performed, the clinical significance of these findings is unknown.

Human Brucellosis Is Characterized by an Intense Th1 Profile Associated with a Defective Monocyte Function

In animal models, a defective Th1 response appears to be critical in the pathogenesis of brucellosis, but the Th1 response in human brucellosis patients remains partially undefined. Peripheral blood from 24 brucellosis patients was studied before and 45 days after antibiotherapy. Twenty-four sex- and age-matched healthy donors were analyzed in parallel. Significantly increased levels of interleukin 1beta (IL-1beta), IL-2, IL-4, IL-6, IL-12p40, gamma interferon (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha), but not of IL-10, in serum and/or significantly increased percentages of samples with detectable levels of these cytokines, measured by enzyme-linked immunosorbent assays (ELISA), were found for untreated brucellosis patients, but these levels were reduced and/or normalized after treatment. Flow cytometry studies showed that the intracytoplasmic expression of IFN-gamma, IL-2, and TNF-alpha, but not that of IL-4, by phorbol myristate-activated CD4(+) CD3(+) and CD8(+) CD3(+) T lymphocytes was significantly increased in untreated brucellosis patients and was also partially normalized after antibiotherapy. The percentage of phagocytic cells, the mean phagocytic activity per cell, and the phagocytic indices for monocytes at baseline were defective and had only partially reverted at follow-up. T lymphocytes from untreated brucellosis patients are activated in vivo and show Th1 cytokine production polarization, with strikingly high serum IFN-gamma levels. In spite of this Th1 environment, we found deficient effector phagocytic activity in peripheral blood monocytes.

Nitric Oxide and Superoxide Anion Generation in Hemocytes ofCrassostrea ariakensisStimulated with Rickettsia-Like Organisms and Zymosan

ABSTRACT To investigate the in vitro generation of nitric oxide (NO) and superoxide anion (O2 -) by Crassostrea ariakensis hemocytes stimulated with rickettsialike organisms (RLOs), RLOs were purified from infected C. ariakensis. BALB/c mice were infected with the purified RLOs and the infectious vitality of RLOs for the mice was detected using the smears and histological sections stained with Machiavello and transmission electron microscope (TEM) technology. These experiments proved that the purified RLOs retained considerable vitality. The study of the influence of temperature on phagocytosis activity revealed that the percentage of phagocytic cells was lower at 10°C than at 20°C and 30°C. The in vitro generation of the cytotoxic molecules such as NO and O2 - by C. ariakensis hemocytes was detected when incubated at 25°C with the purified RLOs and zymosan. The generation of O2 - and NO indicated a possible formation of the highly bactericidal oxidant peroxynitrite by the hemocyte of C. ariakensis. The g...

Assessment of oxidative stress in leukocytes and granulocyte function following oral administration of a silibinin-phosphatidylcholine complex in cats

To determine the effect of oral administration of a silibinin-phosphatidylcholine complex (SPC) on oxidative stress in leukocytes and granulocyte function in healthy cats. 10 purpose-bred adult cats. Cats were administered SPC (10 mg/kg/d) orally for 5 days; blood samples were collected prior to and immediately after the 5-day treatment period. Leukocytes were incubated with monochlorobimane for detection of reduced glutathione (GSH) via flow cytometry. Leukocytes were also incubated with dihydrorhodamine 123 and mixed with Escherichia coli conjugated to a fluorescent marker to measure E coli phagocytosis and the subsequent oxidative burst via flow cytometry. Activities of the antioxidant enzymes superoxide dismutase and glutathione peroxidase, along with the reduced glutathione-to-oxidized glutathione (GSH:GSSG) ratio and a measure of lipid peroxidation (malondialdehyde concentration [micromol/L of blood]), were measured spectrophotometrically. The mean fluorescence intensity (MFI), representing GSH content, increased significantly in feline lymphocytes and granulocytes following 5 days of oral administration of SPC. Mean +/- SD lymphocyte MFI significantly increased from 27.8 +/- 9.0 to 39.6 +/- 6.7, and the granulocyte MFI increased from 508.6 +/- 135.6 to 612.1 +/- 122.9. Following 5 days of SPC administration, the percentage of phagocytic cells that were responding optimally significantly increased (from 37 +/- 11.8% to 45 +/- 17.5%). Other measures of oxidative stress did not change significantly. In cats, oral administration of supplemental SPC appears to increase granulocyte GSH content and phagocytic function, both of which would be potentially beneficial in cats with diseases associated with oxidative stress.

Immune responses of the scallop Chlamys farreri after air exposure to different temperatures

The present study examined the influence of air exposure at different temperatures: a common perturbation associated with aquaculture handling practices, on immune responses in zhikong scallop Chlamys farreri. Scallops were exposed to air for 2 h, 6 h, 12 h and 24 h at 5 °C, 17 °C and 25 °C respectively. Thereafter, a recovery period of 24 h at 17 °C was applied. Haemocyte mortality, phagocytosis and reactive oxygen species (ROS) production of haemocytes, acid phosphatase (ACP) and superoxide dismutase (SOD) activity in haemocyte lysates were chosen as immunomarkers of anoxic stress. The results showed that an increase of haemocyte mortality and a decrease of phagocytosis and ACP activity were observed after 2 h of air exposure for all temperatures tested. Moreover, a significant increase of ROS production occurred following 2 h of air exposure at 25 °C and 24 h of air exposure at 17 °C. Significant differences were also observed in haemocyte mortality, percentage of phagocytic cells and ACP and SOD activity depending on the temperature of air exposure. Finally, after 24 h of recovery at 17 °C, percentage of phagocytic haemocytes and ACP activity did not return to initial values. ROS production was significantly higher than before the recovery period and initial values for scallops subjected to air exposure at 5 °C. In our study, scallops showed a relative low anoxia tolerance under a high temperature. All the scallops air exposed to 25 °C died after the 6 h sampling. In conclusion, air exposure associated to aquaculture practices was demonstrated to strongly affect functional immune activities of scallop haemocytes, and high temperature air exposure caused reduced survival of scallops.