Peripheral Blood Mononuclear cell (PBMC)

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Third-stage Gnathostoma spinigerum larva excretory secretory antigens modulate function of Fc gamma receptor I-mediated monocytes in peripheral blood mononuclear cell culture

Third-stage Gnathostoma spinigerum larva excretory secretory antigens modulate function of Fc gamma receptor I-mediated monocytes in peripheral blood mononuclear cell culture

In all, 3 × 10 6 PBMC (CD27 − ) were cultured in RPMI 1640 supplemented with 10 % FBS alone or plus IL-2 [9], TGF-β1 [18], or ESA (0.1 μg/ml). After incubation for 15, 30, 60, or 90 min [19], the cultured PBMC were harvested and immediately washed with PBS. Total RNA was extracted from the PBMC using the RNeasy total RNA isolation kit and a DNase treatment step (QIAGEN, Hilden, Germany). Then cDNA was synthesized using 1 μg of total RNA with a SuperScript III First-Strand Synthesis System for RT-PCR (Invitrogen, USA) according to the manufacturer’ s protocol. Each 20 μl of PCR reaction con- tained 10 μl of LightCycler 480 SYBR Green I Master mix (Roche Diagnostic, Mannheim, Germany) and was mixed with 100 ng of cDNA and a specific primer (1 μM) in a LightCycler 480 instrument. The FcγRI gene was amplified with the following primers: FcγRI 5′-GTGTCATGCGTG GAAGGATA-3′ (forward) and FcγRI 5′-GCACTGGAGC TGGAAATAGC-3′ (reverse) (212 base pair product) [20]. The PCR reactions were subjected to 1 cycle of 95 °C for 5 min, followed by 45 cycles of 95 °C for 30 s, 60 °C for 30 s, and 72 °C for 45 s. The beta actin gene [20] was used to normalize the relative amounts of mRNA expression of FcγRI gene for the same sample. The equation 2 − ( ΔΔ Ct)

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HIV Coreceptor Tropism in Paired Plasma, Peripheral Blood Mononuclear Cell, and Cerebrospinal Fluid Isolates from Antiretroviral Naïve Subjects

HIV Coreceptor Tropism in Paired Plasma, Peripheral Blood Mononuclear Cell, and Cerebrospinal Fluid Isolates from Antiretroviral Naïve Subjects

A survey of HIV coreceptor usage in cerebrospinal fluid (CSF) samples, peripheral blood mononuclear cells (PBMCs), and plasma samples from naïve seropositive patients was conducted. One hundred patients were enrolled in this study. Of the 100 patients, 36 had a primary or recent infection (P-RI), 31 had an early chronic infection (>350 CD4 cells) (ECI), and 33 had a late chronic infection (LCI). All 3 compartments were sampled in a subset of 33 participants, while the remaining 67 patients provided plasma samples and PBMCs only. Seventy-seven patients harbored the R5 virus in plasma samples and had a significantly higher median and percentage of CD4 ⴙ T cells than patients with X4 virus (437 and 281 cells/ ␮ l, respectively; P ⴝ 0.0086; 20.6% and 18.6%, respectively). The X4 strain was detected more frequently in patients with LCI than in patients with P-RI or ECI (39.3%, 19.4%, and 9.6%, respectively; P ⴝ 0.0063). PBMC and plasma tropism was concordant in 90 patients, and 73 had the R5 strain. Among patients with discordant results, 4 had the R5 virus in their plasma and the X4 virus in PBMCs; 6 showed the opposite profile. Plasma, PBMC, and CSF tropism determinations were concordant in 26/33 patients (21 patients had R5, and 5 had X4). The tropism was discordant in 5/33 patients, with the X4 virus in plasma and R5 in CSF; the HIV tropism in PBMCs was X4 in 3 patients. The remaining 2/33 patients had the R5 virus in plasma and PBMCs and the X4 virus in CSF; one of these patients had a P-RI. The discordant tropism in CSF and blood may have implications for chemokine (C-C motif) receptor 5 (CCR5) antagonist use in patients with limited response to antiretroviral therapy (ART) or in responding patients evaluated for simplification of treatment.

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Differences in peripheral blood mononuclear cell gene expression and triglyceride composition in lipoprotein subclasses in plasma triglyceride responders and non-responders to omega-3 supplementation

Differences in peripheral blood mononuclear cell gene expression and triglyceride composition in lipoprotein subclasses in plasma triglyceride responders and non-responders to omega-3 supplementation

In the present exploratory study, we found that intake of omega-3 fatty acids differentially altered PBMC gene ex- pression in TG responders and non-responders. Specific- ally, enriched pathways in responders compared to non-responders were related to development and apop- tosis, immune function, and LPA signaling. These results lend further support to the findings of Rudkowska et al. who investigated transcriptomic and metabolomics pro- files of TG responders and non-responders to omega-3 supplementation. They reported that 1020 transcripts were altered within the non-responder group, and 252 transcripts were altered within the responder group with only 10 transcripts overlapping between the groups [9]. In the current study, we also report that responders had higher baseline TG in M- and S-HDL subclasses and a greater reduction in TG levels in four of six VLDL and three of four HDL subclasses than non-responders.

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Identification and precursor frequency analysis of a common T cell epitope motif in mitochondrial autoantigens in primary biliary cirrhosis

Identification and precursor frequency analysis of a common T cell epitope motif in mitochondrial autoantigens in primary biliary cirrhosis

The immunodominant antimitochondrial antibody response in patients with primary biliary cirrhosis (PBC) is directed against the E2 component of the pyruvate dehydrogenase complex (PDC-E2). Based on our earlier observations re- garding peripheral blood mononuclear cell (PBMC) T cell epitopes, we reasoned that a comparative analysis of the precursor frequencies of PDC-E2 163-176–specific T cells isolated from PBMC, regional hepatic lymph nodes, and from the liver of PBC patients would provide insight re- garding the role of T cells in PBC. Results showed a disease- specific 100–150-fold increase in the precursor frequency of PDC-E2 163-176–specific T cells in the hilar lymph nodes and liver when compared with PBMC from PBC patients. Interestingly, autoreactive T cells and autoantibodies from PBC patients both recognize the same dominant epitope. In addition, we demonstrated cross-reactivity of PDC-E2 pep- tide 163-176–specific T cell clones with PDC-E2 peptide 36- 49 and OGDC-E2 peptide 100-113 thereby identifying a common T cell epitope “motif” ExETDK. The peptide 163- 176–specific T cell clones also reacted with purified native PDC-E2, suggesting that this epitope is not a cryptic deter- minant. These data provide evidence for a major role for PDC-E2 peptide 163-176 and/or peptides bearing a similar motif in the pathogenesis of PBC. ( J. Clin. Invest. 1998. 102: 1831–1840 . ) Key words: autoreactive T cells • primary biliary

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Type specific Real time PCR for detection of human herpes virus 6 in schizophrenia and bipolar patients: a case control study

Type specific Real time PCR for detection of human herpes virus 6 in schizophrenia and bipolar patients: a case control study

BD: Bipolar disorder; BHQ-1: Black hole quencher 1; CNS: Central nervous system; CSF: Cerebrospinal fluid; DNA: Deoxyribonucleic acid; DSM-IV: Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition; FAM: 6-carboxyfluorescein; JOE: 6-carboxy-4,5-dichloro-2,7-dimethyoxyflurorescein; HC: Healthy control; HHV-6: Human herpes virus-6; PBMC: Peripheral blood mononuclear cell; PCR: Polymerase chain reaction; SC: schizophrenia; SCID-I: Structured Clinical Interview for DSM-IV Axis I Disorders;

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Background: Cell proliferation occurs not only in normal but also in cancer cells. Most of cell proliferation inhibition can be done by inhibiting the DNA synthesis, notably by intervening the formation of purine or pyrimidine. In purine de novo synthesis, it was assumed that biotin plays a role as a coenzyme in carboxylation reaction, one of the pivotal steps in the purine de novo pathways. The aim of this study was to see the avidin potency to bind biotin and inhibit mitosis. Methods: Peripheral blood mononuclear cell (PBMC) was cultured in RPMI-1640 medium and stimulated by phytohemagglutinin (PHA) in the presence or absence of interleukin-2 (IL-2), with or without avidin. The effect of avidin addition was observed at 24, 48, and 72 hours for cell proliferation, viability, and cell cycle. Statistical analysis was done by one-way ANOVA.

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Highlights of the 31st annual meeting of the Society for Immunotherapy of Cancer (SITC), 2016

Highlights of the 31st annual meeting of the Society for Immunotherapy of Cancer (SITC), 2016

Following the introduction to tumor immunology, Kristin Kreamer, CRNP, MSN, AOCNP, APRN-BC (Fox Chase Cancer Center) delved into aspects of clinical manage- ment of immunotherapeutic agents, offering first a brief explanation of the CTLA-4 and PD-1/L1 pathways before providing an overview of immunotherapy agents currently approved for the treatment of melanoma, NSCLC, renal cell carcinoma, Hodgkin lymphoma, HNSCC, and bladder cancer. The next presentation, from Krista Rubin, MS, RN, FNP-BC (Massachusetts General Hospital), under- scored the importance of prompt diagnosis and manage- ment of immune-related AE (irAE). This relies on understanding the mode of action of immune-based agents, which predicts toxicity and differentiates them from chemotherapy. Highlighting the most frequently encountered irAE (fatigue and dermatological, gastrointes- tinal, hepatic, and endocrine system dysfunction), Ms. Rubin proposed approaching symptoms with the adage, ‘it’s inflammatory until proven otherwise’. Toxicities are often reversible if addressed early, hence the value of offer- ing patients a checklist of common symptoms as a resource. Using case studies, Brianna Hoffner, MSN, ANP- BC, AOCNP (University of Colorado, Denver) showed that endocrinopathies are less likely than other irAE to be reversible, hence the importance of early referral to the relevant disease area specialist. In the absence of consensus treatment algorithms, she recommended bringing patients back to the disease specialist’ s clinic for management. Other key takeaways were the value of antibiotic prophy- laxis to prevent infections during high-dose steroid use, and the need to taper steroids slowly; the free app for grad- ing irAE; and the importance of considering the differential diagnosis for atypical symptoms. Long-term (often unusual) irAE can present for the first time even after long- term treatment discontinuation, so continued vigilance is essential.

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Insight in modulation of inflammation in response to diclofenac intervention: a human intervention study

Insight in modulation of inflammation in response to diclofenac intervention: a human intervention study

fenac intervention also altered levels of other plasma oxylipids, increasing 5,6-dihydroxy-eicosatrienoic acid (5,6-DHET) and 20-HETE (20-hydroxyeicosatetraenoic acid) and decreasing the linoleate derived 9,10-dihydrox- yoctadecenoic acid (9,10-DHOME). The arachidonic acid metabolite 5,6-DHET is a stable hydrolysis product of the 5(6)-epoxyeicosatrienoic acid (5(6)-EET) (not measured in current study) which itself is a substrate of cyclooxygenase leading to formation of epoxy prosta- glandins. Levels of arachidonic acid (measured in FFA metabolomics analysis) were increased in the diclofenac group at day 9 compared to day 0, but the responses in diclofenac and placebo treatment groups were not sig- nificantly different (data not shown). Diclofenac inter- vention resulted in decreased levels of TNF-alpha. Furthermore, the selection of genes includes genes with a known role in inflammation like T cell receptor alpha (TRA@), but also genes with unknown function. The network in figure 4 summarizes and visualizes the effect of diclofenac on inflammation: inhibition of prostaglan- din synthesis and changes in associated oxylipids, together with inhibition of TNF-alpha and closely related genes and proteins like caspase 8.

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Ethyleneglycol-Bis-(β-Aminoethylether)Tetraacetate as a Blood Anticoagulant: Preservation of Antigen-Presenting Cell Function and Antigen-Specific Proliferative Response of Peripheral Blood Mononuclear Cells from Stored Blood

Ethyleneglycol-Bis-(β-Aminoethylether)Tetraacetate as a Blood Anticoagulant: Preservation of Antigen-Presenting Cell Function and Antigen-Specific Proliferative Response of Peripheral Blood Mononuclear Cells from Stored Blood

significantly from antigen-specific proliferation. Reconstitution of antigen-specific proliferation of either untreated or EDTA- exposed T-cell lines by fresh untreated monocytes but not by EDTA-treated monocytes established beyond a doubt that presenting cells developed a defect following prolonged expo- sure to EDTA. We also observed that treatment of these af- fected monocytes with bacterial lipopolysaccharide for 24 h failed to make them proficient in presenting antigen. The EDTA effect appeared unlikely to be brought about by a loss of major histocompatibility complex class II molecules on the antigen-presenting cells of the PBMC population, since it was shown that exposure of mouse tissues to 10% EDTA (269 mM) for as long as 14 days to achieve demineralization for histolog- ical examination did not cause any loss of class II molecules (6). The blastogenic response of human or guinea pig T cells to mitogens such as PHA or ConA has been conclusively shown to require the presence of monocytes (9, 17). It is therefore surprising that despite damage to monocytes by EDTA, the response of whole PBMC to the mitogen ConA or to IL-2 was wholly unaffected by EDTA.

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The difference in immune response and IL-12p35 methylation between newborns and adults

The difference in immune response and IL-12p35 methylation between newborns and adults

Otherwise, our result showed neonatal PBMC pro- duced higher IL-10 by LPS stimulation. Several studies have demonstrated higher IL-10 responses in cord blood compared to adult samples [46,47]; however other stud- ies have revealed controversial results [48,49]. Vosters et al. demonstrated that the IL-10 response elicited by LPS was reduced significantly in cord blood mononuclear cells compared to adult PBMCs [49], and the deficient production of IL-10 in response to LPS persisted until 18 months of age. In addition, Belderbos et al. indicated that adult PBMCs produced higher levels of IL-10 in response to LPS when stimulated in the presence of neonatal plasma than in the presence of adult plasma [47]. Therefore, the higher IL-10 responses in cord blood may also reflect the impact of factors present in the plasma [49,50]. In this study, we also showed that isolated PBMCs which stimulated by LPS produced IFN-γ, TNF and IL-6 with no significant difference between newborns and adults.

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Partial Delipidation Improves the T-Cell Antigenicity of Hepatitis B Virus Surface Antigen

Partial Delipidation Improves the T-Cell Antigenicity of Hepatitis B Virus Surface Antigen

carbohydrates, integral parts of glycoproteins, are of viral ori- gin, whereas the lipid moiety, representing approximately 25 to 30% of the particle mass, is of host origin (26, 33). The com- plete removal of lipids from HBsAg destabilizes the particle and precipitates the hydrophobic protein moiety. Partial de- lipidation preserves the particle structure and keeps it in solu- tion but induces minor structural changes that have only been examined at the B-cell level. Some investigators found that lipid extraction did not alter (16) or markedly increased (8, 12, 30) B-cell immunogenicity, whereas others demonstrated that detergent treatment of HBsAg clearly reduced its B-cell anti- genicity (9, 11). The effects of partial delipidation on the T-cell antigenicity of HBsAg have never been examined. We have studied this issue because we estimated that changing the in- teraction of proteins and lipids in HBsAg particles might alter their uptake and processing by antigen-presenting cells (APC) and consequently modify their presentation to and recognition by T lymphocytes. The experiments performed to address this issue are discussed here.

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Expression of immune checkpoints in T cells of esophageal cancer patients

Expression of immune checkpoints in T cells of esophageal cancer patients

Inhibition of immune checkpoint proteins (checkpoints) has become a promising anti-esophageal cancer strategy. We here tested expressions of immune checkpoints in human esophageal cancers. Our results showed the expressions of many immune checkpoints, including CD28, CD27, CD137L, programmed death 1 (PD-1), T cell immunoglobulin mucin-3 (TIM-3), T cell Ig and ITIM domain (TIGIT), CD160, cytotoxic T lymphocyte antigen 4 (CTLA-4), CD200, CD137 and CD158, were dysregulated in peripheral T cells of esophageal cancer patients. Further, the expressions of PD-1, TIM-3 and TIGIT were upregulated in tumor infiltrating lymphocytes (TILs), which might be associated with TILs exhaustion. Meanwhile, the expressions of PD-1 and TIM-3 on CD4+ T cells were closely associated with clinic pathological features of esophageal cancer patients. These results indicate that co-inhibitory receptors PD-1, TIM-3 and TIGIT may be potential therapeutic oncotargets for esophageal cancer.

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Anti-PD-1 monoclonal antibody MEDI0680 in a phase I study of patients with advanced solid malignancies

Anti-PD-1 monoclonal antibody MEDI0680 in a phase I study of patients with advanced solid malignancies

phosphorylated STAT1 positive immune cells in melan- oma tumors have been observed by IHC or gene expres- sion after treatment with pembrolizumab or nivolumab in other trials [50, 51]. In these studies, pretreatment or on-treatment levels of T cells within the tumor or at the tumor margin demonstrated predictive value for response to anti-PD-1 therapy [50, 51]. Other trials with PD-1 inhibitors have also identified pretreatment and post-treatment immune cell correlates with response to therapy [52–62]. For example, Daud et al. showed that patients whose melanoma tumors contained ≥ 20% CD8+ T cells with a CTLA-4 high/PD-1 high phenotype demonstrated a significantly higher ORR to anti-PD-1 blockade compared to those whose tumors contained < 20% of these cells [56]. Inoue et al. described higher pretreatment CD8+/Treg and CD8+/CD4+ expression ratios and higher lytic enzyme (GZMA) and major histocompatibility complex class I (HLA-A) expression correlating with anti-PD-1 mAb response in melan- oma [57]. Likewise, others have demonstrated that pretreatment IFNγ-related immune gene signatures predicted response to anti-PD-1 therapy in head and neck squamous cell carcinoma, gastric cancer, and melanoma [48, 49, 51]. Collectively, these findings suggest that upregulation of CD8+ T cells and markers of effector T-cell function are common phar- macodynamic biomarkers of anti-PD-1 blockade, and pre- or post-treatment intratumoral levels in some settings are associated with clinical response.

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<p>The Exercise Training Modulatory Effects on the Obesity-Induced Immunometabolic Dysfunctions</p>

<p>The Exercise Training Modulatory Effects on the Obesity-Induced Immunometabolic Dysfunctions</p>

More speci fi cally, CD14 ⁺⁺ CD16 ⁺ cells are reported to have a positive association with the cardiometabolic dis- order risk factor, including total cholesterol, LDL, high- density lipoprotein (HDL)/LDL, and TG/HDL ratio. On the contrary, a negative relation of CD14 ⁺ CD16 ⁺⁺ cells with total cholesterol and TG has been demonstrated in individuals with OW and obesity. 91 Additionally, a higher frequency of IM MONs has been shown in individuals who had three or more MetS criteria compared to meta- bolically healthy people. 92 Adiponectin is an anti- in fl ammatory adipokine that contributes to maintaining the anti-in fl ammatory status through the expression of two types of the receptor (adipoR1/R2) on many kinds of cells including liver, MONs, skeletal muscle, and AT cells. 93 – 95 The adiponectin plasma values have inversely correlated with CRP and TNF- α levels. 5 Taking into con- sideration that M Ф s participate in both the lipolysis homeostasis 35 and the foam cells formation in AT, 41 adi- ponectin inhibits the transformation of M Ф s into foam cells and reduces lipid accumulation into MONs-derived M Ф s. 96 Also, it has been reported that reduction of M Ф s in fi ltration, promotion of M2 polarization, and decline of chemokine expression are provoked by adiponectin. 22,97,98 Considering the accumulation of M Ф s in AT during obesity, investigators have demonstrated other pathways resulting in the higher number of ATM. Additionally, the proliferation of tissue-resident M Ф s has been shown to be fully independent from AT MONs in fl ux. In this regard, the populations of hematopoietic progenitor cells have been detected in WAT. These progenitors differentiate into cell lineages, including myeloid and lymphoid. This capacity of local precursors is remarkably similar to hema- topoietic cells derived from bone marrow progenitors. 99,100 In vitro isolated pre-adipocytes are also involved in ATM production. 101 Interestingly, these cells

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Influence of different ex vivo cell culture methods on the proliferation and anti-tumor activity of cytokine-induced killer cells from gastric cancer patients

Influence of different ex vivo cell culture methods on the proliferation and anti-tumor activity of cytokine-induced killer cells from gastric cancer patients

Patients with advanced gastric cancer were recruited for this study in Liaocheng People’s Hospital from May 2012 to June 2015. Patients were enrolled in this study following the listed criteria: 1) patients were diagnosed and confirmed as stages III or IV gastric cancer by histopathological examination, according to the American Joint Committee on Cancer tumor node metastasis Staging Classification (Figure S1); 2) participants were all primary gastric cancer patients without previous tumor history; 3) patients did not have serious heart, lung, liver, and kidney dysfunc- tion; 4) patients were not in pregnancy and lactation; 5) expected survival time was over 6 months; 6) white blood cell counts were over 3 × 10 9 /L; and 7) patients did

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Evaluation of In Vitro Cytotoxic Effects of Three Medicinal Plants on Peripheral Blood Mononuclear Cells (PBMC)

Evaluation of In Vitro Cytotoxic Effects of Three Medicinal Plants on Peripheral Blood Mononuclear Cells (PBMC)

When normal cells are treated with any toxic compound, they may undergo necrosis due to lose of membrane integrity and rapid death occurs as a result of cell lysis. In this process, cells stop dividing and growing, or a decrease in cell viability, or can activate a genetic program to control cell death (apoptosis) [1]. Necrotic cells undergo rapid swelling, loss of membrane integrity, shut down metabolism and tend to release their contents into the surrounding medium [1]. In vitro necrotic cells do not have sufficient time or energy to activate apoptotic machinery. The cytological and molecular events when a cell becomes apoptotic are a change in the refractive index, shrinkage of the cytoplasm, nuclear condensation and DNA fragmentation. The apoptotic cells undergo secondary necrosis and lyse by shutting down the metabolism and by failure of membrane integrity [1].

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Cytometry TOF identifies alveolar macrophage subtypes in acute respiratory distress syndrome

Cytometry TOF identifies alveolar macrophage subtypes in acute respiratory distress syndrome

demonstrated wide diversity of cell-surface marker profiles among CD33 + CD71 + CD163 + AMs. We then used a k-nearest neighbor density estimation algorithm to statistically identify distinct alveolar myeloid subtypes, and we discerned 3 AM subtypes defined by CD169 and PD-L1 surface expression. The percentage of AMs that were classified into one of the 3 AM subtypes was significantly different between healthy and mechanically ventilated subjects. In an independent cohort of subjects with ARDS, PD-L1 gene expression and PD-L1/PD-1 pathway–associated gene sets were significantly decreased in AMs from patients who experienced prolonged mechanical ventilation or death. Unsupervised CyTOF analysis of alveolar leukocytes from human subjects has potential to identify expected and potentially novel myeloid populations that may be linked with clinical outcomes.

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Tumor necrosis factor α release in peripheral blood mononuclear cells of cutaneous lupus and dermatomyositis patients

Tumor necrosis factor α release in peripheral blood mononuclear cells of cutaneous lupus and dermatomyositis patients

elevated levels of IFNa and IFNb (both type I IFNs) [27,28]. Because pDCs are the main cell type producing IFNa and IFNb, they have been implicated in disease pathogenesis. A higher proportion of pDCs are also pre- sent in the skin than in the blood of patients with SLE and CLE, suggesting that pDCs migrate from the circu- lation to the skin and may play a role in the production of skin lesions [29,30]. Our experiments demonstrate that TNFa levels produced by PBMCs vary with the dif- ferent CLE subsets and with DM. We found that pDCs

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The within host dynamics of Mycobacterium avium ssp. paratuberculosis infection in cattle: where time and place matter

The within host dynamics of Mycobacterium avium ssp. paratuberculosis infection in cattle: where time and place matter

3. Mononuclear phagocyte - MAP interactions The mononuclear phagocytes (macrophages, DC) are the primary target cells for MAP in which it is able to persist and replicate. It has become clear that MAP has extensive abilities to subvert the host innate immune system as has recently been reviewed in detail by Arsenault et al. [8] The initial contact between MAP and the mononuclear phagocytes and the receptors used for uptake are important for the subsequent fate of both MAP and the host cell. Selective uptake via certain re- ceptors such as the integrins, mannose receptor and CD14 influences the macrophage response and may lead to suppression of the oxidative burst, and release of pro-inflammatory cytokines [56]. Opsonization of MAP via FcR (specific antibodies) and CR3 (complement re- ceptor 3, CD11b/CD18) can lead to the induction of oxidative burst, changes in intracellular trafficking and phago-lysosomal acidification leading to reduced sur- vival of MAP. However these effects are critically dependent on prior activation of macrophages by IFN-γ /lipopolysaccharide (LPS). In non-activated macro- phages MAP survival and replication is not significantly reduced but rather enhanced [57]. Live MAP, in con- trast to dead MAP, also inhibit the phagolysosome fusion by interfering with the endocytic pathway following phagocytosis enabling survival of MAP indicating active evasive mechanisms [58]. Both in macrophages as well as DC [59], infection with live MAP leads to an upregulation of the production of the suppressive cytokine IL-10 and an arrest in mononuclear phagocyte maturation which also renders them refractory to pro-inflammatory sig- nals from activated γδ and CD4 T cells, most notably IFN-γ [60].

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Inflammatory Responses of Bovine Polymorphonuclear Neutrophils Induced by Staphylococcal Enterotoxin C via Stimulation of Mononuclear Cells

Inflammatory Responses of Bovine Polymorphonuclear Neutrophils Induced by Staphylococcal Enterotoxin C via Stimulation of Mononuclear Cells

formyl-methionyl-leucyl-phenylalanine and opsonized zymo- san, the effect of priming is less than that of receptor-indepen- dent stimulation, such as with PMA (35). Since PMA was used as the stimulator of the oxidative burst, we speculated that priming is slightly involved in the increases of superoxide pro- duction by milk PMN collected from SEC-inoculated mam- mary glands. Therefore, we tested other means to increase oxidative burst activity and confirmed that the viability of S- PBMC sup-stimulated PMN was significantly greater than those of PMN alone and N-PBMC sup-stimulated PMN. A DNA fragmentation assay showed the inhibition of PMN ap- optosis. These results suggest that the difference in cell viability is one of the causes of the difference in oxidative burst activity. Moulding et al. reported that SEA, SEB, and toxic shock syndrome toxin 1 indirectly delay human PMN apoptosis via the production of T-lymphocyte-derived and monocyte-de- rived cytokines (21). Our result that S-PBMC sup-stimulated PMN show a significantly higher survival rate agrees with their report. Although we could not specify the inhibitor of PMN apoptosis, the concentrations of IL-8 and PGE2, which delay apoptosis (1, 9), were significantly higher in S-PBMC sup than in N-PBMC sup. IFN-␥ and GM-CSF, which are reported to contribute to the delayed apoptosis of PMN induced by SEs (21), were induced in SEC-stimulated PBMC. These results suggest that these mediators derived from SEC-stimulated PBMC are responsible for long-time survival of PMN in vitro. On the other hand, Schuberth et al. reported superantigen- dependent accelerated death of bovine PMN in the presence of MNC (30). Although the experimental conditions, such as the concentrations of SEs and the methods of viable cell mea- surement, were different, it is unclear why the findings conflict. However, it was reported that various inflammatory mediators which can be induced by SEs have antiapoptotic effects on PMN (1, 9, 21). Moreover, we have already reported an in- crease of SCC in milk after the intramammary inoculation of SEC (16). Consequently, we believe that the results reported by us and by Moulding et al. are more reasonable than those reported by Schuberth et al.

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