This study aimed to examine the number of circulating Toll-like receptor 4 (TLR4) + CD14+ monocytes in patients with different stages of chronic kidney disease (CKD), their responses to lipopolysaccharide (LPS), and to explore the potential association of the number of TLR4+CD14+ monocytes with clinical laboratory measures. The numbers of TLR4+CD14+, LPS-stimulated TNF-α+CD14+ and interleukin (IL)-6+CD14+ monocytes were determined by flow cytometry in 9 patients with stage 3 CKD, 11 with stage 4 CKD, 16 with stage 5 CKD, and 19 healthy controls (HCs). Their laboratory tests were performed by routine methods and the potential association among these measures was analyzed by Pearson's correlation analysis. The numbers of CD14+, CD14+TLR4+, LPSstimulated TNF-α+CD14+ and IL-6+CD14+ monocytes in patients with CKD were significantly less than those of HCs (all P<0.05), and were negatively associated with patient disease severity. The number of CD14+TLR4+ monocytes was positively correlated with estimated glomerular filtration rate (eGFR, P<0.001) and the levels of hematocrit (P<0.01), but negatively correlated with the levels of blood urine nitrogen, serum creatinine, and C-reactive protein (P<0.001 for all), in the CKD patients. Our data indicate that significant reduction in the number of TLR4+ monocytes and their impaired responses to LPS may be associated with the progression of CKD in Chinese patients.
Cytokines ; metabolism ; Humans ; Kidney Failure, Chronic ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Lipopolysaccharides ; pharmacology ; Monocytes ; metabolism ; Toll-Like Receptor 4 ; metabolism

This study was aimed to explore the killing effect of PBMNC induced by IL-23 alone or combined with IL-2 on K562 cells and its mechanism. The PBMNC were induced in vitro by IL-23 (50 ng/ml) alone or IL-23 combined with IL-2 (100 U/ml) for 72 h, and then were co-cultured with leukemia cell line K562. The CCK-8 method was used to detect the effect of PBMNC induced at different times on K562 cells, the ELISA was performed for detecting IFN-γ level in culture supernatant, and the perforin and granzymes B were detected by RQ-PCR. The results showed that the killing effect of PBMNC induced by IL-23 alone or IL-23 combined with IL-2 on K562 cells was observed, and obviously enhanced with prolonging of time, moreover, there was statistical difference among different time points (P < 0.05). The IFN-γ level in supernatant of PBMNC cultured with cytokines significantly increased, and the IFN-γ levels in group of IL-23 combined with IL-2 were higher than that in other groups (P < 0.05). The mRNA expressions level of perforin and granzymes B of the expanded PBMNC in groups cultured with cytokines were higher than that in control group (P < 0.05), and the mRNA expressions of perforin and granzymes B in group of IL-23 combined with IL-2 were significantly higher than that in others (P < 0.05). It is concluded that IL-23 can promote the killing effect of PBMNC on K562 cells. The combination of IL-2 with IL-23 displays synergic effect and a time-dependent manner. IL-23 also enhances the expression of IFN-γ, perforin and granzyme B in PBMNC. Its combination with IL-2 displays synergistic effect, suggesting that the anti-leukemic activity of IL-23 may be realized through inducing PBMNC to express IFN-γ, perforin and granzyme B.
Granzymes ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-2 ; pharmacology ; Interleukin-23 ; pharmacology ; K562 Cells ; Monocytes ; drug effects ; metabolism ; Perforin ; metabolism

The immunocytes microglia in the central nervous system (CNS) were reported to play a crucial role in neurodegeneration. As a member of P2 receptors family, purinoceptor P2Y6 has attracted much attention recently. Previous studies showed that purinoceptor P2Y6 mainly contributed to microglia activation and their later phagocytosis in CNS, while in immune system, it participated in the secretion of interleukin (IL)-8 from monocytes and macrocytes. So there raises a question: whether purinoceptor P2Y6 also takes part in neuroinflammation? Thus, this review mainly concerns about the properties and roles of purinoceptor P2Y6, including (1) structure of purinoceptor P2Y6; (2) distribution and properties of purinoceptor P2Y6; (3) relationships between purinoceptor P2Y6 and microglia; (4) relationships between purinoceptor P2Y6 and immunoinflammation. Itos proposed that purinoceptor P2Y6 may play a role in neuroinflammation in CNS, although further research is still required.
Animals ; Humans ; Inflammation ; immunology ; metabolism ; Microglia ; drug effects ; metabolism ; Monocytes ; metabolism ; Phagocytosis ; physiology ; Receptors, Purinergic P2 ; chemistry ; genetics ; metabolism

Astragalus membranaceus ; chemistry ; Cell Line ; Cytokines ; metabolism ; Humans ; Inflammation ; Macrophages ; drug effects ; metabolism ; Monocytes ; drug effects ; metabolism ; Polysaccharides ; pharmacology

OBJECTIVETo investigate effects of serum HDL(1) on the formation of foam cells from human peripheral blood monocyte-derived macrophages.

METHODSSectie density polyacrylamide gel electrophoresis (sd-PAGE) was applied for isolation and preparation of HDL(1) simultaneously. Monocytes were isolated from human peripheral blood by Ficoll-Hypaque density gradient centrifugation and plastic adsorptive process. The isolated monocytes were stimulated by phorbol 12-myristate 13-acetate (PMA) at a concentration of 50 nmol/L for 48 h and transferred to macrophages. The monocyte-derived macrophages were then coincubated with 80 mg/L ox-LDL and HDL(1) (0, 0.1, 1.0 and 10.0 mg/L) for 6, 12 and 24 h, respectively. The formation of foam cells was identified by transmission electron microscope (TEM), total cholesterol (TC), free cholesterol (FC) and protein (Pro) in cultured cells were quantitatively analyzed by high performance chromatography (HPLC) and modified lowry protein assay, respectively.

RESULTSHDL(1) isolated from human serum by sd-PAGE could significantly decrease TC/Pro ratio in foam cells in a concentration-dependent (0 mg/L: 36.9 +/- 1.1, 10.0 mg/L: 6.2 +/- 0.4, P < 0.01) and time-dependent (10.0 mg/L HDL(1) 6 h: 16.9 +/- 0.9, 24 h: 6.4 +/- 0.6, P < 0.01) manner.

CONCLUSIONHDL(1) is capable of inhibiting and attenuating the formation of foam cells by decreasing cellular TC, therefore, might play an important role in attenuating atherosclerosis.

Atherosclerosis ; Cells, Cultured ; Cholesterol, LDL ; metabolism ; Foam Cells ; cytology ; metabolism ; Humans ; Lipoproteins, HDL ; blood ; Lipoproteins, LDL ; Monocytes ; cytology ; metabolism

BACKGROUND: Autoanalyzer ADVIA2120 uses intracellular peroxidase concentration to perform white blood cell (WBC) differential. Therefore, in specimens containing neutrophils with low peroxidase concentration, neutrophils can be miscounted as monocytes or large unstained cells resulting in pseudoneutropenia. Myeloperoxidase deficiency can be detected by the mean peroxidase index (MPXI). The aims of this study are to establish the reference interval of MPXI and define a cut off value for manual slide review to discriminate pseudoneutropenia. METHODS: We calculated reference intervals as mean+/-2SD according to the indirect method of CLSI C28-A3 guideline from MPXI data of 5,802 individuals who took routine health checkup from April 2010 to June 2012. We performed manual slide review on specimens with low MPXI and compared neutrophil differential count of manual method with that of autoanalyzer. When neutrophil differential in manual slide review was >20%P higher than autoanalyzer result, it was regarded as a pseudoneutropenia. We performed ROC analysis using the MPXI results of samples with and without pseudoneutropenia to define a cutoff to discriminate pseudoneutropenia. RESULTS: The reference intervals of MPXI in total population, male, and female were -4.9 to 7.5, -5.5 to 7.3, and -4.5 to 7.5, respectively. The mean value of MPXI was significantly higher in female than in male and there was no difference by age. Twenty-two pseudoneutropenia samples from 7 patients were identified. ROC analysis yielded cutoff value of -20.7 with 94.9% of sensitivity and 77.3% of specificity. CONCLUSIONS: MPXI may be used in the manual slide review guideline for detecting pseudoneutropenia.
Discrimination (Psychology) ; Female ; Humans ; Leukocytes ; Male ; Metabolism, Inborn Errors ; Monocytes ; Neutrophils ; Peroxidase ; ROC Curve

IL-32 acts as a pro-inflammatory cytokine by inducing the synthesis of inflammatory molecules as well as promoting the morphological changes involved in the transformation of monocytes into osteoclasts (OCs). Evaluation of the functions of IL-32 has mainly focused on its inflammatory properties, such as involvement in the pathogenesis of various autoimmune diseases. Recently, IL-32 was shown to be involved in bone metabolism, in which it promotes the differentiation and activation of OCs and plays a key role in bone resorption in inflammatory conditions. IL-32γ also regulates bone formation in conditions such as ankylosing spondylitis and osteoporosis. In this review, we summarize the results of recent studies on the role of IL-32γ in bone metabolism in inflammatory arthritis.
Arthritis* ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Bone Resorption ; Inflammation ; Metabolism* ; Monocytes ; Osteoblasts ; Osteoclasts ; Osteogenesis ; Osteoporosis ; Spondylitis, Ankylosing

Deficiencies in the clearance of peripheral amyloid β (Aβ) play a crucial role in the progression of Alzheimer's disease (AD). Previous studies have shown that the ability of blood monocytes to phagocytose Aβ is decreased in AD. However, the exact mechanism of Aβ clearance dysfunction in AD monocytes remains unclear. In the present study, we found that blood monocytes in AD mice exhibited decreases in energy metabolism, which was accompanied by cellular senescence, a senescence-associated secretory phenotype, and dysfunctional phagocytosis of Aβ. Improving energy metabolism rejuvenated monocytes and enhanced their ability to phagocytose Aβ in vivo and in vitro. Moreover, enhancing blood monocyte Aβ phagocytosis by improving energy metabolism alleviated brain Aβ deposition and neuroinflammation and eventually improved cognitive function in AD mice. This study reveals a new mechanism of impaired Aβ phagocytosis in monocytes and provides evidence that restoring their energy metabolism may be a novel therapeutic strategy for AD.
Animals ; Mice ; Alzheimer Disease ; Amyloid beta-Peptides ; Monocytes ; Cognition ; Energy Metabolism ; Phagocytosis

OBJECTIVETo investigate the changes in the expression of HLA-DR on CD14+ monocytes of burn patients with delayed resuscitation, and to analyze the relationship between it and sepsis.

METHODSTwenty-five patients with total burn surface area over 30% TBSA and delayed resuscitation were enrolled in the study, among which 7 were complicated by sepsis during hospitalization. Peripheral blood was collected on 1, 3, 7, 14 and 28 post-burn days (PBD), and the blood of the patients with sepsis were also collected on the 1 and 2 days after the occurrence of sepsis. Twenty healthy volunteers were enrolled as controls. Expression rate of HLA-DR on CD14+ monocytes was determined by flow cytometry. The level of TNF-alpha and IL-10 were measured by ELISA.

RESULTSExpression rate of HLA-DR antigen on CD14+ monocytes in burn patients without sepsis on 1, 3, 7, 14, 28 PBD were (15 +/- 6)%, (7 +/- 5)%, (26 +/- 17)%, (28 +/- 16)% and (47 +/- 16)%, respectively, which were obviously lower than that of healthy people [(92 +/- 10)%, P < 0.01], and it was also markedly lower on 1 and 2 days after the occurrence of sepsis than that of controls and those of patients without sepsis on 1, 7, 14, 28 PBD (P < 0.01). The positive rate and concentration of TNF-alpha in patients with sepsis were obviously higher than that of healthy people and patients without sepsis (P < 0.05 or P < 0.01). There was a negative correlation between the expression rate of HLA-DR on CD14+ monocytes and IL-10 levels, and it showed significant difference on 1, 7, and 28 PBD (r = -0.9963, -0.7459, -0.8474, respectively, P < 0.01).

CONCLUSIONImmune function is suppressed and proinflammatory mediators are excessively released in severely burn patients after delayed resuscitation, especially when complicated with sepsis. Expression of HLA-DR on CD14+ monocytes may be an useful parameter for monitoring the immune function of burn patients.

Burns ; immunology ; metabolism ; HLA-DR Antigens ; metabolism ; Humans ; Interleukin-10 ; blood ; Lipopolysaccharide Receptors ; metabolism ; Monocytes ; immunology ; metabolism ; Sepsis ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVE: To investigate the expression of M3 subtype of muscarinic receptors (M3R) during the incised wound healing of the skin in mice and the characteristics of its time-dependent. METHODS: The change of M3R in skin incised wound was detected by immunohistochemical staining and Western blot. RESULTS: M3R-positive cells were detected in epidermis, hair follicle, sebaceous glands, sweat glands, dermomuscular layer in normal mouse skin. Expression of M3R was mainly detectable in polymorphonuclear cells (PMNs) in the wound specimens aged from 6h to 12h after injury. Afterwards, the M3R-positive cells were mostly mononuclear cells (MNCs) and fibroblastic cells (FBCs) at 1 d to 3d post-injury, whereas the M3R-positive cells were mostly FBCs aged from 5 d to 14d. Morphometrically, the ratio of the M3R-positive cells increased aged from 6h to 12h after injury, with a peak at 12h. The ratios kept a high relatively level aged from 1 d to 5 d, but significantly that lowered as compared with aged 12h after injury. The ratio reached the peak at 7 d again after injury, and then decreased gradually. The M3R protein also revealed a time-dependent tendency with double peaks at 12h and 7 d after injury as detected by Western blotting. CONCLUSION M3R is time-dependently expression in PMNs, MNCs and FBCs suggesting that it may play roles during the skin incised wound healing, and M3R may be used as a marker for wound age determination.
Animals ; Blotting, Western ; Fibroblasts/metabolism* ; Immunohistochemistry ; Male ; Mice ; Monocytes/metabolism* ; Neutrophils/metabolism* ; Receptor, Muscarinic M3/metabolism* ; Skin/metabolism* ; Time Factors ; Wound Healing ; Wounds and Injuries/metabolism*