Decreased phagocytic activity of Kupffer cells in a rat nonalcoholic steatohepatitis model

AIM: To investigate Kupffer cell dynamics and phagocytic activity,using a rat nonalcoholic steatohepatitis (NASH) model. METHODS: Male F344 rats were fed either a control diet or a choline-deficient L-amino acid-defined (CDAA) diet,followed by contrast enhanced ultrasonography (CEUS) using Levovist. The uptake of latex beads by the Kupffer cells was determined by fluorescent microscopy. The status of the Kupffer cells was compared between the two groups,using the immunohistochemical staining technique. RESULTS: After 4 or more wk of the CDAA diet,CEUS examination revealed a decrease in the signal intensity,20 min after intravenous Levovist. Fluorescent microscopic examination showed that the uptake of latex beads by the Kupffer cells was reduced at week 1 and 2 in the study group,compared with the controls,with no further reduction after 3 wk. Immunohistochemical staining revealed no significant difference in the Kupffer cell counts between the control group and the CDAA group. CONCLUSION: CEUS examination using Levovist demonstrated reduced contrast effect and phagocytic activity in the liver parenchymal phase,although the Kupffer cell numbers were unchanged,indicating reduced phagocytic function of the Kupffer cells in the rat NASH model. We believe that CEUS examination using Levovist is a useful screening modality,which can detect NASH in fatty liver patients.

Assessment of platelet activation and phagocytic activity in gastric cancer patients

AIM:To assess the activation of platelets and their phagocytic activity in the course of gastric cancer.METHODS:Forty-three gastric cancer patients were recruited to the study.The patients were divided into 3 groups depending on tumor stage.Group E included 6 patients with early gastric cancer;group A 18 patients with locally advanced cancer;and group M-19 with metastatic cancer.The investigations were performed twice,prior to surgery and 12-14 d afterwards.RESULTS:The platelet count and the level of soluble platelet selectin(sP-selectin) were found to increase with the disease progression.The level of sP-selectin was lowest in early cancer and was observed to increase after surgery in all the study patients.Irrespective of tumor stage,a statistically significant decrease was noted in the percentage of phagocytizing platelets and in the phagocytic index in gastric cancer patients as compared to healthy subjects.Despite increasedplatelet count and stimulation of thrombocytopoiesis,the phagocytic functions of blood platelets were markedly impaired.Tumor development seems to impair metabolic processes.CONCLUSION:A decreasing phagocytic activity can promote both inflammatory processes and cancer growth.

Effects of Traditional Chinese Herbal Medicine on Immune Organ Indexes and Macrophages Phagocytic Indexes in Chickens Infected by Infectious Bronchitis Virus

[ Objective] To study the therapeutic effects of traditional Chinese medicine prescriptions on infectious bronchitis （IB） and find a novel avenue for prevention and treatment of viral diseases in poultry. [Method] A total of 160 cockerels at the age of 15 d were divided into four groups randomly, including traditional Chinese medicine group, moroxydine control group, challenge control group and healthy control group. Except the healthy control group, other groups were challenged with infectious bronchitis virus （IBV） on Day 15. After 48 h post challenge, the traditional Chinese medicine groupand moroxydine control group were respectively administrated with Chinese herbal medicine prescription and moroxydine, continuously for 5 d. The immune organ indexes and macrephage phagocytic indexes were detected on Day 18, 24 and 30, respectively. [ Result] The immune organ indexes and macrophage phagocytic indexes were not significantly different between traditional Chinese medicine group and moroxydine control group on Day 18. But all the indexes of the traditional Chinese medicine groups were increased significantly （ P 〈 0.05） on Day 24 and 30, and showed extremely significant difference （ P 〈 0.01 ） with those of challenge control group on Day 30. [ Conclusion] The traditional Chinese herbal medicine can enhance macrophage phagocytic indexes and immune organs indexes of chickens infected by IBV.

Assessment of platelet activation and phagocytic activity in gastric cancer patients

AIM: To assess the activation of platelets and their phagocytic activity in the course of gastric cancer. METHODS: Forty-three gastric cancer patients were recruited to the study. The patients were divided into 3 groups depending on tumor stage. Group E included 6 patients with early gastric cancer; group A included 18 patients with locally advanced cancer; and group M included 19 patients with metastatic cancer. The investigations were performed twice, prior to surgery and 12-14 d afterwards. RESULTS: The platelet count and the level of sP-selectin were found to increase with the disease progression. The level of sP-selectin was lowest in early cancer and was observed to increase after surgery in all the study patients. Irrespective of tumor stage, a statistically significant decrease was noted in the percentage of phagocytizing platelets and in the phagocytic index in gastric cancer patients as compared to healthy subjects. Despite increased platelet count and stimulation of thrombocytopoiesis, the phagocytic functions of blood platelets were markedly impaired. Tumor development seems to impair metabolic processes. CONCLUSION: A decreasing phagocytic activity can promote both inflammatory processes and cancer growth.

Determination of phagocytic functions of macrophages, vascular density and NPY-nerve within autotransplanted splenic tissue in adult rats

Objective: To estimate directly phagocytic functions of the macrophages because of the importance in innate immunity, determine blood vessel density and re-innervation density which are basis of function. Methods: Eighty adult Wistar rats were randonjy divided into experimental and control groups.The formers underwent splenotomy and a half splenie slice was transplanted into greater omentum. The latter only moved. After 6 months, examination was made as follows: ① After injection of 0.4% carbon particles by vein, spleme tissues were taken out at different times for estimating phagocytosis by light microscope. ② When splenic tissues had been intubated into left ventricle under total anesthesia, animals were perfused by formalin and India ink mixture suspension. Splenic tissues were taken out for making sections for measurement of area density of blood vessels.③ Immunohistochemical procedure was used for detecting neuropeptide Y(NPY). Results: Phagocytie functions had no difference between two groups, hut the area density of blood vessels and NPY-positive fibers re-dued (P<0.01) in experimental group. Conclusion:Autotransplanted splenic tissues show good innate immunity though regeneration of blood vessels and nerves do not reach normal level.

Antimicrobial roles of phagocytosis in teleost fish:Phagocytic B cells vs professional phagocytes

The defense system of teleost fish organized on innate and adaptive immunity protects them against a wide variety of pathogenic microorganisms in the aquatic environment.Phagocytosis is one of the most effective defense strategies against microbial challenge mainly performed by classical‘professional’phagocytes(including monocytes,macrophages and granulocytes).They contain,kill and process the internalized pathogens for antigen presentation by providing antigenic ligands to initiate activation and clonal expansion of T and B cells,which bridge the innate and adaptive immunity.The discovery of phagocytic B cells in teleost fish has broken the paradigm that primary vertebrate B cells are lack of phagocytosis of particulates,as well as led to the investigation of phagocytic activity of mammalian B-1 B cells.The active phagocytic,microbicidal capabilities and antigen presentation in teleost phagocytic B cell have demonstrated to be similar as professional phagocytes,providing a potential impact on development of new vaccination strategies to prevent and control infectious diseases.In this review,we aim to address current progress on the antimicrobial role of phagocytic B cells in teleost fish by comparing it with other professional phagocytes and mammalian B-1 B cells,and provide the application prospect of phagocytic B cells in developing vaccines as well as the prevention of fish diseases.

Ruxolitinib treatment for bronchiolitis obliterans syndrome following hematopoietic stem cell transplant in a patient with primary hemophagocytic lymphohistiocytosis

To the Editor:A 20-year-old man who developed hemophagocytic lymphohistiocytosis(HLH)was treated with methylprednisolone combined with etoposide(VP-16)for 24 weeks.One month after treatment completion,HLH recurred.The patient visited the Hematology Department of the Beijing Friendship Hospital and was diagnosed with primary HLH with PRF1 fusion gene mutation.

Successful resolution of hemophagocytic lymphohistiocytosis during the chemotherapy course of acute myelocytic leukemia with ruxolitinib

To the Editor:Hemophagocytic lymphohistiocytosis(HLH)is a severe or even fatal inflammatory condition.Malignant HLH(M-HLH)is one of the most common types of secondary HLH(sHLH),especially in adults.HLH can occur either as the initial manifestation of malignant disease(here referred to as“malignancy-triggered HLH”)or in the setting of iatrogenic immuno-suppression from chemotherapy(“HLH during chemotherapy”or Ch-HLH).

Studies on Antiviral and Immuno-Regulation Activity of Low Molecular Weight Fucoidan from Laminaria japonica

The antiviral activity in vitro and in vivo and the effect of the immune system of two fucoidan fractions with low molecular weight and different sulfate content from Laminaria japonica(LMW fucoidans) were investigated in order to examine the possible mechanism. In vitro, I-type influenza virus, adenovirus and Parainfluenza virus I were used to infect Hep-2, Hela and MDCK cells, respectively. And 50% tissue culture infective dose was calculated to detect the antiviral activity of two LMW fucoidans. The results indicated that compared with the control group, 2 kinds of LMW fucoidans had remarkable antiviral activity in vitro in middle and high doses, while at low doses, the antiviral activity of 2 kinds of LMW fucoidans was not statistically different from that in the blank control group. And there was no statistically difference between two LMW fucoidans in antiviral activity. In vivo, LMW fucoidans could prolong the survival time of virus-infected mice, and could improve the lung index of virus-infected mice significantly, which have statistical differences with the control group significantly(p < 0.01). However, the survival time of the two LMW fucoidans was not statistically significant(p > 0.05). In this study, it was shown that both of two LMW fucoidans(LF1, LF2) could increase the thymus index, spleen index, phagocytic index, phagocytosis coefficient and half hemolysin value in middle and high doses, which suggested that LMW fucoidans could play an antiviral role by improving the quality of immune organs, improving immune cell phagocytosis and humoral immunity.

Immunological Effect of PM_(2.5) on Cytokine Production in Female Wistar Rats

Objective To investigate the immunological effect of PM2.5 on cytokine production in female Wistar rats. Methods Female Wistar rats were given 0.3 mg, 0.75 mg, 2 mg, 5 mg of PM2.5 per 0.5 mL saline, respectively. Saline was used as the negative control. TNF-α and IL-6 levels in the branchoalveolar lavage were measured by ELISA, and mRNA expression levels in lung tissue were detected by RT-PCR. Alveolar macrophages were collected for testing phogacytic function. Results Exposure to PM2.5 stimulated TNF-α production in a dose-dependent manner （P〈0.05）, However, no statistically significant difference was found. No time-dependent change in TNF-α and IL-6 production was found. TNF-α and IL-6 mRNA expressions were induced by PM2.5-exposure. The phagocytic rate （PR） was significantly decreased by PM2.5 treatment. Conclusion PM2.5 exposure increases inflammation response of the lung in a dose-dependent manner. Moreover, tissue injury induced by PM2.5 may be related to altered production of cytokines. PM2.5 may impair the phagocytic activity of alveolar macrophages.

Preparation and in vitro Studies of Stealth PEGylated PLGA Nanoparticles as Carriers for Arsenic Trioxide

The aim of this study was to prepare arsenic trioxide （ATO）-loaded stealth PEGylated PLGA nanoparticles （PEG-PLGA-NPs） and to assess the merits of PEG-PLGA-NPs as drug carriers for ATO delivery. PEG-PLGA copolymer was synthesized with methoxypolyethyleneglycol （Mw=5000）, D, L-lactide, and glycolide by the ring-opening polymerization method. Amorphous ATO was transformed into cubic crystal form to increase its solu-bility in the organic solvent. ATO-loaded PEG-PLGA-NPs were prepared by the modified spontaneous emulsification solvent diffusion （SESD） method, and the main experimental factors influencing the characteristics of nanopar- ticles were investigated, to optimize the preparation. To confirm the escape of PEG-PLGA-NPs from phagocytosis by phagocytes, PEG-PLGA-NPs labeled rhodamine B uptake by murine peritoneal macrophages （MPM） were analyzed by flow cytometry. The results showed that the physicochemical characteristics of PEG-PLGA-NPs were affected by the type and concentration of the emulsifiers, polymer concentration, and drug concentration. ATO-loaded PEG-PLGA-NPs, with particle size of 120.8nm, zeta potential of-10.73mV, encapsulation efficiency of 73.6%, and drug loading of 1.36%, were prepared under optimal conditions. The images of transmission electron micros-copy （TEM） indicated that the optimized nanoparticles were near spherical and without aggregation or adhesion. The release experiments in vitro showed the ATO release from PEG-PLGA-NPs exhibited consequently sustained release for more than 26d, which was in accordance with Higuchi equation. The uptake of PEG-PLGA-NPs by MPM was found to decrease markedly compared to PLGA-NPs. The experimental results showed that PEG-PLGA-NPs were potential nano drug delivery carriers for ATO.

Impairment of innate immune responses in cirrhotic patients and treatment by branched-chain amino acids

It has been reported that host defense responses, such as phagocytic function of neutrophils and natural killer (NK) cell activity of lymphocytes, are impaired in cirrhotic patients. This review will concentrate on the impairment of innate immune responses in decompensated cirrhotic patients and the effect of the treatment by branched-chain amino acids (BCAA) on innate immune responses. We already reported that phagocytic function of neutrophils was significantly improved by 3-mo BCAA supplementation. In addition, the changes of NK activity were also significant at 3 mo of supplementation compared with before supplementation. Also, Fisher&#x02019;s ratios were reported to be significantly increased at 3 mo of BCAA supplementation compared with those before oral supplementation. Therefore, administration of BCAA could reduce the risk of bacterial and viral infection in patients with decompensated cirrhosis by restoring impaired innate immune responses of the host. In addition, it was also revealed that BCAA oral supplementation could reduce the risk of development of hepatocellular carcinoma in cirrhotic patients. The mechanisms of the effects will also be discussed in this review article.

Immune stimulatory activity of BRP-4,an acidic polysaccharide from an edible plant,Basella rubra L.

Objective:To evaluated the immunomodulatory effect of BRP-4,an acidic polysaccharide from Basella rubra(B.rubra) L on the macrophage activity.Methods:Phagocytic activity was determined by the ingestion of Latex Beads-Rabbit IgC-FITC using the fluorescent microscopy and flow cytometry analysis and nitric oxide production was measured using Griess reaction assay.Results:An enhanced production of NO was observed at 10 and 100 μg/mL of BRP-4.The phagocytic activity of macrophage was enhanced in BRP-4 treated RAW264.7 cells.BRP-4combined with concanavalin A(Con A) provided obvious promotion and strengthening of the proliferation of the splenocytes.Conclusions:BRP-4,polysaccharide isolated from B.rubra,is suggested to activate macrophage function and stimulate splenocyte proliferation.The strong immunomodulatory activity of BRP-4 confirmed its good potential as an immunotherapeutic adjuvant.

Immunomodulatory activity of butanol fraction of Gentiana olivieri Griseb. on Balb/C mice

Objective:To explore the immunomodulatory properties of 80% ethanol extract and butanol fraction of Gentiana olivieri(G. olivieri) Griseb on Balb/C mice.Methods:The study was performed with basic models of immunomodulation such as the humoral antibody response(hemoglutination antibody titres), cell mediated immune response(delayed type hypersensitivity and in vivocarbon clearance or phagocytosis). Ethanol(80%) extract of flowering aerial parts of G.olivieriand its butanol fraction were administered p.o.(orally) to the mice. Levamisole, 2.5 mg/kg was used as standard drug.Results:There was a potentiation of immune response to sheep red blood cells by cellular and humoral mediated mechanisms comparable to levamisole(2.5 mg/kg) by both 80% ethanol extract and the butanol fraction at doses of 50-200 mg/kg in male Balb/C mice. Both significantly(P<0.01) potentiated the humoral immune response in cyclophosphamide(250 mg/kg)immunosupressed mice at 100 and 200 mg/kg of each extract and fraction as compared to control.The potentiation of delayed type hypersensitivity response was statistically significant(P<0.01) at200 mg/kg of ethanol extract and 100, 200 mg/kg of butanol fraction as compared to control. The phagocytosis was significant at 200 mg/kg with butanol fraction ofG. olivieri.Conclusions:The results reveal the immunostimulant effects of plantG. olivieriin mice by acting through cellular and humoral immunity in experimental models of immunity in mice. Butanol fraction is the most effective at a dose level of 200 mg/kg.

Functional Activity of Circulating Phagocytes as Potential Pretreatment Marker of Tumor Drug Resistance

The aim of this work was a study of the functional activity of neutrophils and peripheral blood monocytes in rats with the transplanted Walker carcinosarcoma as potential predictors of the sensitivity of this tumor to doxorubicin treatment. This study provides an evidence that such indices of the functional activity of circulating phagocytes of the tumor-bearing rats as the quantity and the phagocytosis intensity of monocytes, as well as the intensity of ROS production by monocytes and neutrophils, may reflect the degree of sensitivity of the tumor to doxorubicin. So it was shown that the growth of the resistant tumor caused a significant increase of the number of circulating phagocytic cells and the intensity of phagocytosis by more than 100% (p < 0.001) compared with the corresponding indices of intact rats and rats with the parental variant of the tumor. The ability of blood mono-cytes and neutrophils of rats with a resistant tumor to produce ROS was also significantly different from that in intact rats and animals with the parental carcinosarcoma variant. The predictive value of these indices is especially important in the dynamic monitoring of the development of tumor drug resistance during long-term cancer chemotherapy. Considering the standard 2 - 3 week interval between the courses of cancer therapy and the short lifetime of circulating phagocytes, an assessment of the indices of their functional activity before each subsequent course can be considered as a pretreatment assessment. Meanwhile, further studies are needed to determine the spectrum of malignant neoplasms for which the degree of tumor drug resistance correlates with the functional activity of circulating phagocytes.

Effect of Hot Water Extract from Agaricus Blazei Murill on Chemotaxis of Neutrophils

Hot water extract from the edible Brazilian mushroom, Agaricus Blazei Murill (ABM), is used for both traditional and alternative medicine. ABM is reported to stimulate anti-tumor, anti-infection, and immune activity. However, there are few reports of how ABM affects neutrophils. Therefore, in this study, we examined the effect of hot water ABM extract on neutrophil migration, phagocytosis, and reactive oxygen species production using neutrophils from guinea pig. Migratory direction and velocity as indicators of chemotactic activity of neutrophils were significantly (p 0.001) increased at concentration of 50 and 100 mg/ml in ABM extract compared with control. Phagocytic activity of neutrophil was significantly (p 0.01) increased at concentration of 5 mg/ml in ABM extract compared with control. Production of reactive oxygen species (ROS: H2O2 or ) by neutrophils was significantly (p 0.01) increased at concentration of 5 mg/ml in ABM extract compared with control. These results suggest that enhancement in neutrophil chemotactic activity, phagocytic activity and ROS production are mechanisms by which ABM extract inhibits bacterial infection in the skin and dermatitis.

Coxiella burnetii:living inside the host cell

Coxiella burnetii is an obligate intracellular pathogen and the causative agent of Q fever.In this brief review,we describe how recently described mechanisms help our understanding of C.burnetii invasion and its survival in the host cell by the formation of a replicative niche:the Coxiella-containing vacuole.We describe the actin-associated proteins involved in the internalization of C.burnetii,and we discuss the contribution of diverse degradation pathways of the cell during the formation and stabilization of the Coxiella-containing vacuole.

The immunity-promoting activity of porcine placenta in mice as an immunomodulator for functional foods

This study was to explore the immunity-promoting activity of porcine placenta as a potential raw material for functional foods.Porcine placenta was subjected to the analysis for its bioactive substances,and their immunity-promoting activity was determined in mice supplemented with porcine placenta extract(PPE)and freeze-dried porcine placenta powder at high(PPH)and low(PPL)dosage.Results showed that porcine placenta contained placental peptides and 15 free amino acids,and the amounts of estrogen and progesterone in products developed from porcine placenta were within the limit of national standard.Mice model experiment revealed that compared with the control,the PPH treatment significantly improved the spleen index(P<0.05)by increasing the phagocytic rate of macrophages from 20%to 60%and the conversion rate of T lymphocytes from 8%to 60%.The q PCR analysis disclosed that the porcine placenta powder enhanced mice immunity via promoting the expression of Th1 cytokines of interleukin-2(IL-2)and IFN-γ,especially the former,by almost 8 times in the spleens of male mice,while inhibited Th2 cytokines of IL-4 and IL-10.This investigation has provided a reference for the development of porcine placenta as a raw material applied in functional foods to improve human immunity.

Alveolar macrophage functions and DNA damage in cigarette smoke-exposed mice

Alveolar macrophages (AM) are known to play an essential role in lung defense through their ability to remove the foreign matters reaching the lung alveoli. Cigarette smoke (CS) is a critical risk factor for many lung diseases. CS is inhaled into the lung by respiretion and affects AM. It has been previously reported that CS induces inhibition of cytokine production, cell surface receptor expression and antigen presentation in AM. However, the relationship of immune suppression and DNA damage caused by CS in AM is still unclear. Therefore, in this study, we investigated AM immune function and DNA damage in CS-exposed mice. Mice were exposed to CS of 20 cigarettes/day during 10 days using a HambrugⅡsmoking machine. After exposure, AM were obtained by bronchoalveolar lavage. The number of AM was significantly increased in CS-exposed mice compared with non-CS-exposed mice. Phagocytic activity of AM was significantly inhibited by CS exposure. Percentage of CD11b-, CD14-, Toll-like receptor (TLR)2- or TLR4-positive cells was significantly decreased in CS-exposed mice compared with non-CS-exposed mice. Interleukin-1β mRNA expression in lipopolysaccharide-stimulated AM was significantly inhibited by CS exposure. Intracellular reactive oxygen species (ROS) (, H2O2) production of AM was significantly increased, and DNA damage was induced by CS exposure. These results suggest that impaired immune functions by CS exposure may be related to DNA damage via excessive ROS induced by CS. These alterations of AM caused by CS could be associated with infection and development of pulmonary diseases.

Physiological Studies on the Effect of Fenugreek on Productive Performance of White New-Zealand Rabbit Does

The objective of the current work is to investigate the possible beneficial effects of forced feeding of fenugreek to rabbit does on their milk composition, milk yield, anemia screening parameters, immunity and the growth performance of bunnies. Twenty white New Zealand female rabbits were randomly assigned into 5 groups: normal control and 4 treated groups (vehicle (molasses), fenugreek powdered seeds + molasses, sprouted fenugreek + molasses, and fenugreek oil + molasses). All treated groups took the given doses orally for two days before parturition and 13 days after parturition for 2 successive pregnancies. Blood samples were collected from each doe on the 13th day of lactation for determination of hematological, biochemical and immunological parameters. Milk samples were taken for evaluation of milk composition. All rabbit does and their bunnies were weighed for calculation of the milk yield. The fenugreek seeds and oil administered groups (G3 & G5) recorded the highest percent values of fat, protein, lactose, total solids and solid not fat contents of the rabbits’ milk in both 1st and 2nd lactation periods relative to other tested groups. At the beginning and mid of lactation periods, the milk yields recorded non-significant differences either between tested groups or between 1st and 2nd pregnancy in the same group. Whereas, at the end of lactation, germinated seeds (G4) and oil (G5) treated groups recorded significantly higher rates of milk yield relative to control and fenugreek seeds groups. Bunnies’ body weights suckling from dams of group III (G3) recorded the highest weights relative to other tested groups in both 1st and 2nd pregnancy. During the 1st lactation period, all experimental groups showed significant reduction of white blood cells, while, G3 and G4 recorded significant elevation during 2nd lactation period. Rabbits treated with fenugreek seeds powder (G3) showed a significant raise of phagocytic activity and phagocytic index. Fenugreek seeds group (G3) and germinated one (G4) showed significant red blood corpuscles and packed cell volume raise relative to other groups during both 1st and 2nd lactation periods.