Zinc supplementation ameliorates lung injury by reducing neutrophil recruitment and activity.

INTRODUCTION: Zinc is well known for its anti-inflammatory effects, including regulation of migration and activity of polymorphonuclear neutrophils (PMN). Zinc deficiency is associated with inflammatory diseases such as acute lung injury (ALI). As deregulated neutrophil recruitment and their hyper-activation are hallmarks of ALI, benefits of zinc supplementation on the development of lipopolysaccharides (LPS)-induced ALI were tested. METHODS: 64 C57Bl/6 mice, split into eight groups, were injected with 30 µg zinc 24 hours before exposure to aerosolised LPS for 4 hours. Zinc homoeostasis was characterised measuring serum and lung zinc concentrations as well as metallothionein-1 expression. Recruitment of neutrophils to alveolar, interstitial and intravascular space was assessed using flow cytometry. To determine the extent of lung damage, permeability and histological changes and the influx of protein into the bronchoalveolar lavage fluid were measured. Inflammatory status and PMN activity were evaluated via tumour necrosis factor α levels and formation of neutrophil extracellular traps. The effects of zinc supplementation prior to LPS stimulation on activation of primary human granulocytes and integrity of human lung cell monolayers were assessed as well. RESULTS: Injecting zinc 24 hours prior to LPS-induced ALI indeed significantly decreased the recruitment of neutrophils to the lungs and prevented their hyperactivity and thus lung damage was decreased. Results from in vitro investigations using human cells suggest the transferability of the finding to human disease, which remains to be tested in more detail. CONCLUSION: Zinc supplementation attenuated LPS-induced lung injury in a murine ALI model. Thus, the usage of zinc-based strategies should be considered to prevent detrimental consequences of respiratory infection and lung damage in risk groups.

Targeted Complement Inhibition at Synapses Prevents Microglial Synaptic Engulfment and Synapse Loss in Demyelinating Disease.

Multiple sclerosis (MS) is a demyelinating, autoimmune disease of the central nervous system. While work has focused on myelin and axon loss in MS, less is known about mechanisms underlying synaptic changes. Using postmortem human MS tissue, a preclinical nonhuman primate model of MS, and two rodent models of demyelinating disease, we investigated synapse changes in the visual system. Similar to other neurodegenerative diseases, microglial synaptic engulfment and profound synapse loss were observed. In mice, synapse loss occurred independently of local demyelination and neuronal degeneration but coincided with gliosis and increased complement component C3, but not C1q, at synapses. Viral overexpression of the complement inhibitor Crry at C3-bound synapses decreased microglial engulfment of synapses and protected visual function. These results indicate that microglia eliminate synapses through the alternative complement cascade in demyelinating disease and identify a strategy to prevent synapse loss that may be broadly applicable to other neurodegenerative diseases. VIDEO ABSTRACT.

Protection of salvianolate against atherosclerosis via regulating the inflammation in rats.

Inflammation plays an essential role in the pathophysiology of atherosclerosis. Our study was aimed to investigate whether salvianolate, a novel water-soluble phenolic compound of Danshen, alleviates atherosclerosis via regulating the inflammation in rats. High fat diet feeding plus vitamin D3 injection was used to induce atherosclerosis in rats. Salvianolate (60, 120 or 240 mg/kg) or placebo was given to atherosclerotic rats. The plasma lipids, interleukin 6 (IL-6) and C reactive protein (CRP) were measured by ELISA. CD4+CD25+Foxp3+ cells were determined by flow cytometry. Histological changes were examined by hematoxylin and eosin staining. The results showed that the levels of plasma IL-6 and CRP were elevated in the rats fed on high fat diet, and the histological analysis demonstrated the successful establishment of atherosclerosis models. Treatment with salvianolate alleviated the atherosclerotic process and decreased the levels of plasma IL-6 and CRP. Also the number of CD4+CD25+Foxp3+ cells was increased in salvianolate-treated rats. It was concluded that salvianolate could treat atherosclerosis via modulating the inflammation at cytokine and cell levels.

Vitamin D3 mediated effects on postprandial leukocyte activation and arterial stiffness in men and women.

Postprandial inflammation is considered to be pro-atherogenic. Vitamin D can reduce inflammation and arterial stiffness. We hypothesized that vitamin D3 improves postprandial arterial elasticity by the modulation of leukocyte activation. Healthy volunteers underwent two oral fat-loading tests (OFLTs). The augmentation index (AIx) and flow cytometric quantification of leukocyte activation markers were measured. After the first OFLT, 100 000 IU of vitamin D3 was administered and a second OFLT was carried out 7 days later. Six men and six women were included. A favorable reduction in AIx was found after vitamin D3 supplementation (P=0.042) in both genders. After vitamin D3, exclusively in women a reduction in the area under the postprandial curve for monocytes CD11b and CD35 by 10.5% (P=0.016) and 12.5% (P=0.04) and neutrophil CD11b by 17.0% (P=0.014) was observed. In conclusion, vitamin D3 probably increased postprandial arterial elasticity in men and women, but reduced postprandial leukocyte activation exclusively in women.

IgG antibodies in food allergy influence allergen-antibody complex formation and binding to B cells: a role for complement receptors.

Allergen-IgE complexes are more efficiently internalized and presented by B cells than allergens alone. It has been suggested that IgG Abs induced by immunotherapy inhibit these processes. Food-allergic patients have high allergen-specific IgG levels. However, the role of these Abs in complex formation and binding to B cells is unknown. To investigate this, we incubated sera of peanut- or cow's milk-allergic patients with their major allergens to form complexes and added them to EBV-transformed or peripheral blood B cells (PBBCs). Samples of birch pollen-allergic patients were used as control. Complex binding to B cells in presence or absence of blocking Abs to CD23, CD32, complement receptor 1 (CR1, CD35), and/or CR2 (CD21) was determined by flow cytometry. Furthermore, intact and IgG-depleted sera were compared. These experiments showed that allergen-Ab complexes formed in birch pollen, as well as food allergy, contained IgE, IgG1, and IgG4 Abs and bound to B cells. Binding of these complexes to EBV-transformed B cells was completely mediated by CD23, whereas binding to PBBCs was dependent on both CD23 and CR2. This reflected differential receptor expression. Upon IgG depletion, allergen-Ab complexes bound to PBBCs exclusively via CD23. These data indicated that IgG Abs are involved in complex formation. The presence of IgG in allergen-IgE complexes results in binding to B cells via CR2 in addition to CD23. The binding to both CR2 and CD23 may affect Ag processing and presentation, and (may) thereby influence the allergic response.

[Effect of Shenfu injection on the erythrocyte immune function of patients undergoing cardiopulmonary bypass].

OBJECTIVE: To observe the effect of Shenfu Injection (SFI) on erythrocyte immunity function of patients undergoing cardiopulmonary bypass (CPB). METHODS: Twenty patients scheduled for valve replacement were randomly assigned to two groups, i.e. , the SFI group and the control group, 10 in each. SFI 1 mL/kg was intravenously dripped before induction of anesthesia and SFI 1 mL/kg administered in priming solution in the SFI group, while only normal saline was given to those in the control group. Venous blood samples (5 mL) were collected before induction of anesthesia (T1), 30 min CPB (T2), immediate by the end of CPB (T3), and postoperative 24 h (T4) respectively in all groups. The levels of the rosette rate of RBC-C3b receptor (RBC-C3bRR), the rosette rate of RBC-immune complex (RBC-ICR), plasma malondialdehyde (MDA), free hemoglobin (FHB), and interleukin-6 (IL-6) were detected. RESULTS: There was no significant difference in the levels of RBC-C3bRR, RBC-ICR, plasma MDA, FHB, and IL-6 at T1 in both groups (P > 0.05). RBC-C3bRR at the rest time points was lower in the two groups than before induction of anesthesia. There was no statistical difference in FHB or IL-6 between T4 and T1 in the SFI group. The levels of RBC-ICR, MDA, FHB, and IL-6 increased in the two groups more than before induction of anesthesia at T2-4 ( P < 0.05). Besides, the RBC-C3b RR was lower, and levels of RBC-ICR, MDA, FHB, and IL-6 higher in the control group than in the SFI group, showing significant difference (P <0.05). CONCLUSION: SFI could decrease the generation of inflammatory mediators during CPB, improve the erythrocyte immune function of patients during CPB, and reduce the risk of postoperative infection.

[Construction expression and biologic activity of recombinant human sCR1 eucaryotic cell].

AIM: To express human soluble complement receptor type 1(sCR1)protein using ferment cell secreting type carrier and study the extraorgan biologic activity of recombinant human sCR1 fusion protein. METHODS: Total human RNA was extracted from peripheral blood. The full length cDNA of human sCR1 gene was obtained by RT-PCR and them, cloned into Pichia pastoris eukaryotic expression vector pPIC9k to construct the recombinant plasmid pPIC9k-sCR1 containing human sCR1.After identified by DNA sequencing, the recombinant plasmid pPIC9k-sCR1 was transformed into Pichia pastoris SMD1168. The ferment cell line of the recombinant sCR1 which was chosen by G418 resistance was identified by PCR, After methanol induction, the expressed protein products were verified by SDS-PAGE and Western blot, purified by Ni(2+)-NTA agarose affinity chromatography, and its biologic activity was identified. RESULTS: The obtained Pichia pastoris secretion type yeast carrier pPIC9k-sCR1 was chosen by G418 and identified by PCR to get a highly copied and integral recombinant ferment cell line. The recombinant human sCR1 fusion protein was expressed by yeast cells containing pPIC9k-sCR1 induced by methanol. It was a protein band about M(r) 31 000 in gel, which could be identified by CD35 of anti-sCR1 protein monoclonal antibody with Western blotting technique. The highly purified sCR1 fusion protein and its biologic activity were detected obtained by Ni(2+)-NTA agarose affinity chromatography. CONCLUSION: The recombinant human sCR1 fusion protein can be highly expressed in the Pichia pastoris expression system, which resembles the human natural protein's antigenicity and biologic activity.

[Study on the effects of two kinds of cactus polysaccharide on erythrocyte immune function of S180 mice].

OBJECTIVE: To study the effects of two kinds of cactus polysaccharide on erythrocyte immune function in S180 mice. METHOD: Classical pharmaceutical method and test kit. RESULT: The cactus polysaccharide increased the content of RBC-CaR, RFER, decreased the content of RFIR, raised the content of sialic acid. And the effect of median dose group of medical cactus polysaccharide and high dose group of edible cactus polysaccharide is very remarkable (P < 0.01) compared with model group. CONCLUSION: The cactus polysaccharide improved the erythrocyte function of tumor-mice, which may be one of anti-tumor mechanisms.

beta2-Integrin and lipid modifications indicate a non-antioxidant mechanism for the anti-atherogenic effect of dietary coenzyme Q10.

Dietary supplementation with coenzyme Q (CoQ) has been proposed to have anti-atherogenic effects by virtue of its antioxidant capacity. To investigate this question, the leukocyte status of 5 males and 5 females (52-68 years) was evaluated before and after supplementation with 200mg CoQ/day for 5 and 10 weeks. CoQ was selectively taken up by mononuclear cells and alpha-tocopherol increased in polynuclear and mononuclear cells. The expression of beta2-integrin CD11b and complement receptor CD35 on the plasma membrane of resting and stimulated monocytes was significantly decreased upon dietary CoQ. Fatty acid and aldehyde analysis revealed that there was a selective increase of arachidonic acid and plasmalogens in only mononuclear cells. These selective lipid changes are not consistent with a general improvement in antioxidant status and indicate that CoQ most likely inhibits a phospholipase A2. Thus, these results strongly suggest that the anti-atherogenic effects of CoQ be mediated by other mechanisms beside its antioxidant protection.

[Study on therapeutic effect and immunological mechanism of fuzhengfang herbs in mice with Lewis lung cancer].

OBJECTIVE: To observe the therapeutical effect of Fuzhengfang (FZF). METHODS: Therapeutic effect of FZF herbs on mice with Lewis lung cancer was observed, and its regulation on function of red blood cell immune system of the mice was also studied. RESULTS: FZF herbs could inhibit the growth of tumor, and increase the weight of mice with Lewis lung cancer. The quality of life of the mice was improved after treatment with FZF herbs. FZF herbs could increase the activity of red blood cell C3b receptors, reduce the quantity of red blood cell immune complexes, elevate the immune adherent function of red blood cells to tumor cells, raise activity of red blood cell immune adherent enhancing factors in serum, and lower the activity of red blood cell immune adherent inhibitory factors. Chemotherapy could inhibit the growth of tumor, but decrease the weight of mice with Lewis lung cancer. The quality of life of the mice was decreased after chemotherapy. Chemotherapy could reduce the activity of red blood cell C3b receptors, increase the quantity of red blood cell immune complexes, depress the immune adherent function of red blood cells to tumor cells, lower the activity of red blood cell immune adherent enhancing factors in serum, and raise the activity of red blood cell immune adherent inhibitory factors in serum. There was a significant difference between FZF herbs and chemotherapy. CONCLUSION: FZF herbs have a positive therapeutic effect in mice with Lewis lung cancer, and its mechanism might be associated with improving function of red blood cell immune system of mice with Lewis lung cancer.

[Effect of yishen jianpi drugs on T lymphocyte subsets, soluble interleukin-2 receptor and red blood cell immunity of senile deficiency syndrome patients].

UNLABELLED: To evaluate the effect of Yishen Jianpi drugs (YJD) on immunological function of the Senile Deficiency Syndrome (SDS) patients, T lymphocyte subsets, soluble interleukin-2 receptor (SIL-2 R), the red blood cell immunity, the rate of red blood cell C 3b receptor rosette (RBC-C 3 bRR) and red blood cell immune complex rosette (RBC-ICR) were dynamically investigated in 31 SDS patients as well as in normal controls. RESULTS: the percentage of CD3, CD4 and CD4/CD8 ratio and the rate of RBC-C 3 bRR in SDS were all significantly lower than that of control (P < 0.01). The further analysis also revealed that between the immunological indices of CD4/CD8 ratio and SIL-2 R in SDS patients were markedly negative correlated (r = -0.572, P < 0.01) while positive correlated (r = 0.435, P < 0.02) markedly between CD4/CD8 and RBC-C 3 bRR. After treatment with YJD, in comparing pretreatmentally, all the above-mentioned immunological indices, following the relief of clinical manifestation of SDS, they were corresponding negatively changed, and there were marked significance (P < 0.01) except red blood cell immunity. It suggested that YJD would markedly improve and/or regulate immunological function in SDS.

In vitro stimulation of polymorphonuclear cell adhesion by ribomunyl and antibiotic + ribomunyl combinations: effects on CD18, CD35 and CD16 expression.

Several functions of polymorphonuclear cells (PMNs) require adhesion to occur. Various membrane proteins' functions such as CD18 (beta 2 chain of integrin), CD35 (CR1) and CD16 (F c gamma Receptor III) participate in adhesion. In vivo treatment with Ribomunyl (R), an immunomodulating agent, was shown to enhance adhesion and migration of PMNs. To explore the direct effect of R on PMNs, cells from healthy subjects were treated in vitro with R. A significant increase of PMN adhesion and expression of CD18 and CD35 molecules were observed with 50 and 100 micrograms/ml of R after 2 h incubation. However, R-treatment decreased the PMN reactivity towards anti-CD16 (F c gamma RIII) monoclonal antibody. The effect of R on adhesion and membrane molecule expression was independent of the presence of serum and of polymixin B. Thus, this effect cannot be due to lipopolysaccharide (LPS) contaminants and does not require interactions with serum components. In previous studies, it was shown that in vitro amoxicillin increased some PMN functions whereas josamycin decreased them. The in vitro incubation of PMNs with R and amoxicillin (100 micrograms/ml) potentiated the positive effect of amoxicillin on adhesion and the antibiotic counterbalanced the negative effect of R on CD16 expression. In addition, R compensated the negative effect of josamycin (100 micrograms/ml) on PMN adhesion and on CD18 and CD35 expression. This study indicates: (1) the direct effect of R on PMN adhesion and on expression of molecules involved in adhesive-mediated functions, and (2) the beneficial effect of the association of R with antibiotics which can stimulate PMN activity.