Macrophage-like Human Cells Research Articles

Lactoferrin, Quercetin, and Hydroxyapatite Act Synergistically against Pseudomonas fluorescens.

Pseudomonas fluorescens is an opportunistic, psychotropic pathogen that can live in different environments, such as plant, soil, or water surfaces, and it is associated with food spoilage. Bioactive compounds can be used as antimicrobials and can be added into packaging systems. Quercetin and lactoferrin are the best candidates for the development of a complex of the two molecules absorbed on bio combability structure as hydroxyapatite. The minimum inhibiting concentration (MIC) of single components and of the complex dropped down the single MIC value against Pseudomonas fluorescens. Characterization analysis of the complex was performed by means SEM and zeta-potential analysis. Then, the synergistic activity (Csyn) of single components and the complex was calculated. Finally, the synergistic activity was confirmed, testing in vitro its anti-inflammatory activity on U937 macrophage-like human cell line. In conclusion, the peculiarity of our study consists of optimizing the specific propriety of each component: the affinity of lactoferrin for LPS; that of quercetin for the bacterial membrane. These proprieties make the complex a good candidate in food industry as antimicrobial compounds, and as functional food.

Variation in Macrophage Phagocytosis of Streptococcus agalactiae Does Not Reflect Bacterial Capsular Serotype, Multilocus Sequence Type or Association with Invasive Infection

Background:Group B Streptococcus (GBS) is an encapsulated Gram-positive coccus that is an important cause of infections in adults with chronic medical conditions, pregnant women, and neonates. GBS causes a range of clinical syndromes, from asymptomatic colonization to deep-seated invasive and highly lethal infections. Macrophages are important sentinels of innate immunity, protecting host tissues from infection when bacteria advance beyond cutaneous or mucosal barriers. We hypothesized that the capacity for macrophages to phagocytose unopsonized GBS would vary across distinct clinical strains, and such differences would reflect serotype diversity.Methods:A high-throughput screen using the phorbol ester-differentiated THP-1 macrophage-like human cell line was used to quantify phagocytosis of a diverse group of 35 different human clinical isolates of GBS representing a wide variety of capsular serotypes. Validation studies were conducted using human primary phagocytes.Results:Phagocytosis of GBS differed widely across clinical isolates but this was not related to capsular serotype, genetic sequence type, pilus type, or clinical source of the GBS isolate (colonizing or invasive strain).Conclusions:Structural and/or biochemical differences among diverse GBS strains are reflected in a diverse capacity for macrophages to ingest them through non-opsonic phagocytosis. Mechanisms explaining these differences are not clear.

Microarray analysis of nicotine‐induced changes in gene expression in a macrophage‐like human cell line

Cigarette smoking has been suggested as a risk factor for periodontitis. Thousands of components are present in cigarette smoke, including nicotine, which may play an important role in the observed effects of smoking on cell metabolism. However, the mechanisms underlying these effects are unclear. Using DNA microarrays, we monitored differentially expressed genes, responsive to nicotine, in a macrophage-like human cell line. Human U937 cells were treated for 1 h, with or without 1.0 microg/ml of nicotine. For differentiation, cultures were incubated with 10 nm phorbol myristate acetate for 48 h. Analysis of gene expression was performed using a DNA microarray of 8500 genes. The expression of 4914 genes was detected. Screening was carried out on those genes whose expression in three separate experiments showed an average change of twofold or greater, and 118 up-regulated genes and 97 down-regulated genes were identified. Among these were genes related to inflammation and other immune responses, such as phospholipase A2 and interferon. Consistent with the array findings, we found similar changes in mRNA expression after analysis using the real-time polymerase chain reaction. The results suggest that nicotine causes excess inflammation and disturbs host defense mechanisms against pathogens.

Differential cytokine induction by two types of Porphyromonas gingivalis

The presence of Porphyromonas gingivalis with type II fimA is strongly associated with adult periodontitis. However, the importance of specific fimA types in the immune response is unknown. Two types of P. gingivalis (type I and type II) and Actinomyces naeslundii were assessed for their degree of cytokine induction in the macrophage-like human cell line U937. Real-time reverse transcriptase polymerase chain reaction was used to determine mRNA expression of 12 cytokines. Significant levels of interleukin (IL)-8 induction and a similar cytokine expression pattern were observed at 6 h postinfection for all three bacterial strains. However, type II P. gingivalis infection showed statistically higher levels of IL-1beta, IL-8, IL-12 and tumor necrosis factor-alpha mRNA induction than those of control at 24 h postinfection, whereas type I P. gingivalis and A. naeslundii showed no significant induction of these cytokines. These data suggest that compared with A. naeslundii and type I P. gingivalis, type II P. gingivalis prolongs the cytokine response. Although other factors may also be involved, the sustained cytokine response induced by type II P. gingivalis may play an important role in enhanced periodontal tissue inflammation and destruction.

Differential gene induction in macrophage-like human cells by two types of Porphyromonas gingivalis: a microarray study

Several studies have provided clinical evidence that FimA clonal variation may contribute to the periodontopathogenicity of Porphyromonas gingivalis (P.g.). We studied the gene expression profiling of the macrophage-like human cell line U937 after infection of two types of P.g. (fimA type I; Pg-I and fimA type II; Pg-II) using microarray. Of 1088 genes examined, 394 genes were detectable. Bioinformatics algorithms were used to analyze the detectable genes. Hierarchical clustering analysis showed that gene expression patterns of Pg-II and the control (no infection) were grouped together. K-means clustering grouped 79 genes into Pg-II dominance and 88 genes into Pg-I dominance. A large number of genes related to cell signaling, extracellular communication proteins, cell receptors (by ligands), protein turnover and cell adhesion receptors/proteins were grouped into clusters of Pg-I dominance. Our results indicate that compared with Pg-I, Pg-II induces a low host response as measured by its weak induction of gene expression.