Presence Of Concanavalin Research Articles

COVID-19 vaccination exacerbates ex vivo IL-6 release from isolated PBMCs

Ex vivo culturing of isolated PBMCs from individuals vaccinated with the coronavirus disease 2019 (COVID-19) vaccine BNT162b1 revealed a pronounced T cell response in the presence of the receptor binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein. The latter was 10-fold more pronounced than the ex vivo response of PBMCs from the same individuals to other common pathogen T cell epitope pools, suggesting COVID-19 vaccination to induce RBD-specific T cell responses and not to facilitate T cell (re)activity in general. In the current study we investigated whether COVID-19 vaccination long-lastingly affects plasma interleukin (IL)-6 concentrations, complete blood counts, ex vivo IL-6 and IL-10 secretion of PBMCs cultured under basal conditions or in the presence of concanavalin (Con) A and lipopolysaccharide (LPS), salivary cortisol and α-amylase, mean arterial pressure (MAP), heart rate (HR) as well as mental and physical health status. The study was initially designed to investigate whether the presence vs. absence of own pets during urban upbringing has protective effects against psychosocial stress-induced immune activation during adulthood. However, as COVID-19 vaccines were approved while the study was ongoing and as, therefore, both vaccinated and non-vaccinated individuals have been recruited, we were able to stratify our data set with respect to the COVID-19 vaccination status and to assess the long-lasting effects of COVID-19 vaccination on physiological immunological, cardiovascular and psychosomatic health parameters. This data is presented in the current study. We show that isolated PBMCs from individuals vaccinated against COVID-19 show a ~ 600-fold increase in basal and a ~ 6000-fold increase in ConA-induced proinflammatory IL-6 secretion, and a ~ 2-fold increase in basal and ConA-induced antiinflammatory IL-10 secretion, both in comparison with non-vaccinated individuals. In contrast, LPS-induced ex vivo IL-6 and IL-10 secretions were not affected by vaccination status, as were plasma IL-6 concentrations, complete blood counts, salivary cortisol and α-amylase, cardiovascular measures and psychosomatic health. In summary, our findings are of relevance for many clinical studies ran before/during the pandemic, clearly indicating that consideration of participants’ vaccination status is critical, at least when assessing ex vivo PBMC functionality.

Influence of microbial polyamines on the IL-10 production by peripheral blood leukocytes of healthy donors

As is known, bacterial polyamines, which include cadaverine and putrescine, are capable of influencing the activity of immunocompetent cells in many ways. In particular, this situation is observed in long-term inflammatory diseases, especially with intensive reproduction of microorganisms capable of producing polyamines. It is of interest to study the production of one of the main anti-inflammatory cytokines, IL-10, under the influence of bacterial polyamines. For research, a population of mononuclear leukocytes was isolated from the peripheral blood of healthy donors by gradient centrifugation. The cell suspension was placed in a round-bottomed plates with preliminarily added polyamines at concentrations of 5, 25, 50, 75, and 100 mmol/L. Wells not containing polyamines were used as a control. After incubation for 72 h at 37 °C and 5% CO2, the supernatants were harvested and used to determine the concentration of IL-10. We used a kit for determining the concentration of IL-10 using the enzyme immunoassay method (Russia). Statistical analysis was performed using the Statistica 6.0 software package. In the case of a distribution close to normal, Student’s t-test was used; in the rest, the Mann–Whitney test was used to assess the significance of differences. Studies have shown that leukocytes in the presence of concanavalin A produce IL-10 at a concentration of 17.13±6.08 pg/mL. It has been established that under the influence of polyamines of bacterial origin, the production of IL-10 is enhanced only if putrescine and cadaverine are at concentrations of 50 mmol/L and higher. At low concentrations of polyamines, no significant increase in IL-10 production was detected. Since IL-10 is an anti- inflammatory cytokine, for which the analgesic effect is also known, it should be expected that with an increase in its concentration in the focus of invasion of opportunistic bacteria, the inflammatory process will develop latently, when the symptoms are mild. In general, it can be expected that polyamine-producing bacteria will contribute to the maintenance of few symptomatic inflammation.

Quercetin handles cellular oxidant/antioxidant systems and mitigates immunosenescence hallmarks in human PBMCs: An in vitro study.

Immunosenescence, oxidative stress, and low vaccine efficacy are important symptoms of aging. The goal of our study was to test if quercetin had antiaging and stimulating effects on peripheral blood mononuclear cell(PBMC) immune cells in vitro in the presence of concanavalin a, PBMCs were isolated from healthy elderly and young people and cultured in a complete Roswell Park Memorial Institute 1640 medium supplemented with quercetin. Cell proliferation was assessed using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromidecolorimetric assay after a 48-hincubation period. Spectrophotometricassays were used to assess oxidative biomarkers (proteins carbonyl[PC], malondialdehydes [MDA], and reduced glutathione [GSH]).The enzyme-linkedimmunosorbent assay method was used to determine the amount of interleukin(IL)-2 released. A Griess reagent was used to investigate inducible nitrite oxide synthase(iNOS) activity. When compared to young control cells, aged PBMCs had lower proliferation potency, lower IL-2 and NO release, and higher MDA and PC levels. Importantly, quercetin-treated aged PBMCs have a high proliferative response comparable to young cells, restored iNOS activity, and increased levels of GSH antioxidant defences. In comparison to untreated aged PBMCs, treated PBMCs have lower lipo-oxidative damage but higher PC levels. Quercetin may be used as a promising dietary vaccinal adjuvant in the elderly, it has significant effects in reducing immunosenescence hallmarks, as well as mitigating the lipo-oxidative stress in PBMCs cells.

Effect of prednisolone and saireito co-administration on T-cells of rheumatoid arthritis patients.

Rheumatoid arthritis (RA) is an autoimmune disease. Methotrexate (MTX) and prednisolone (PSL) are used in combination for severe RA therapy. However, it can increase the risk of osteoporosis and osteonecrosis. Saireito (114) can be used to reduce PSL dose owing to its immunosuppressive effects. However, the effect of combination therapy of PSL+114 on the immune system of RA patients remains unknown. This study compared the effect of PSL alone and PSL+114 on peripheral blood mononuclear cell (PBMC) proliferation, T-cell subsets, and cytokine production in adult RA patients receiving MTX monotherapy. We isolated PBMCs from 14 consenting RA patients, and cultured them with PSL (0.0001-1.0 μM) in combination with or without 114 (300 μg/mL) for 96 h in the presence of concanavalin A. We measured the proliferation rates of PBMC, proportions of CD4+, CD8+, and CD4+CD25+Foxp3+T-cells (induced T-regulatory cells), and concentrations of interferon-γ, interleukin (IL)-6, IL-10, IL-17A, and tumour necrosis factor in the culture supernatant. Compared to the blank, the PBMC proliferation rate significantly decreased at a reduced PSL concentration after 114 administration. The 50% inhibition concentration was 0.43 μM PSL for the PSL-only group as compared to 0.29 μM PSL for the PSL+114 co-administration group. The PSL+114 co-administration group had a significantly higher concentration of IL-6 compared to the PSL-only group. The use of 114 in combination with low-concentration PSL intensified its immunosuppressive effect. However, the concentration of IL-6 was elevated in the co-administration group, suggesting exacerbation of RA activity.

Prostaglandin-related immune suppression in cattle

Prostaglandins (PGs) are lipid mediators derived from arachidonic acid by several enzymes including cyclooxygenase (COX)-1 and COX-2. We have previously shown that PGE2 regulates immune responses, such as Th1 cytokine production and T-cell proliferation, in cattle. However, it is still unclear whether other PGs are involved in the regulation of immune responses in cattle.Here, immunosuppressive profiles of PGs (PGA1, PGB2, PGD2, PGE2, PGF1α and PGF2α) were firstly examined using bovine peripheral blood mononuclear cells (PBMCs). In addition to PGE2, PGA1 significantly inhibited Th1 cytokine production from PBMCs in cattle. Further analyses focusing on PGA1 revealed that treatment with PGA1 in the presence of concanavalin A (con A) downregulated CD69, an activation marker, and IFN-γ expression in both CD4+ and CD8+ T cells. Sorted CD3+ T cells stimulated with con A were cultivated with PGA1, and IFN-γ and TNF-α concentrations decreased upon PGA1 treatment. Taken together, these results suggest that the treatment with PGA1in vitro inhibits T-cell activation, especially Th1 cytokine production, in cattle.

Development of Mannose-Modified Carboxylated Curdlan-Coated Liposomes for Antigen Presenting Cell Targeted Antigen Delivery.

Specific delivery to antigen presenting cells (APC) and precise control of the intracellular fate of antigens are crucial to induce cellular immunity that directly and specifically attacks cancer cells. We previously achieved cytoplasmic delivery of antigen and activation of APC using carboxylated curdlan-modified liposomes, which led to the induction of cellular immunity in vivo. APCs express mannose receptors on their surface to recognize pathogen specifically and promote cross-presentation of antigen. In this study, mannose-residue was additionally introduced to carboxylated curdlan as a targeting moiety to APC for further improvement of polysaccharide-based antigen carriers. Mannose-modified curdlan derivatives were synthesized by the condensation between amino group-introduced mannose and carboxy group in pH-sensitive curdlan. Mannose residue-introduced carboxylated curdlan-modified liposomes showed higher pH-sensitivity than that of liposomes modified with conventional carboxylated curdlan. The introduction of mannose-residue to the liposomes induced aggregation in the presence of Concanavalin A, indicating that mannose residues were presented onto liposome surface. Mannose residue-introduced carboxylated curdlan-modified liposomes exhibited high and selective cellular association to APC. Furthermore, mannose residue-introduced carboxylated curdlan-modified liposomes promoted cross-presentation of antigen and induced strong antitumor effects on tumor-bearing mice. Therefore, these liposomes are promising as APC-specific antigen delivery systems for the induction of antigen-specific cellular immunity.

Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets.

The Fusarium mycotoxin deoxynivalenol (DON) contaminates animal feed worldwide. In vivo, DON modifies the cellular protein synthesis, thereby also affecting the immune system. However, the functional consequences of this are still ill-defined. In this study, peripheral blood mononuclear cells from healthy pigs were incubated with different DON concentrations in the presence of Concanavalin A (ConA), a plant-derived polyclonal T-cell stimulant. T-cell subsets were investigated for proliferation and expression of CD8α, CD27, and CD28, which are involved in activation and costimulation of porcine T cells. A clear decrease in proliferation of all ConA-stimulated major T-cell subsets (CD4+, CD8+, and γδ T cells) was observed in DON concentrations higher than 0.4 µM. This applied in particular to naïve CD4+ and CD8+ T cells. From 0.8 μM onwards, DON induced a reduction of CD8α (CD4+) and CD27 expression (CD4+ and CD8+ T cells). CD28 expression was diminished in CD4+ and CD8+ T cells at a concentration of 1.6 µM DON. None of these effects were observed with the DON-derivative deepoxy-deoxynivalenol (DOM-1) at 16 µM. These results indicate that DON reduces T-cell proliferation and the expression of molecules involved in T-cell activation, providing a molecular basis for some of the described immunosuppressive effects of DON.

Biological effects of cyclosporin A on CD3−CD161+ and CD3+CD161+ lymphocytes

Cyclosporin A (CSA) is a widely used drug to prevent the immune cell function. It is well known that CSA blocks transcription of cytokine genes in activated T cells. The connection between T cells and CSA has been well established. However, the effect of CSA on natural killer (NK) cells is not thoroughly understood. Therefore, in the present study, splenocytes and peripheral blood mononuclear cells (PBMCs) were treated with CSA in the presence of concanavalin A (Con A) or interleukin-2 (IL-2). CSA at higher concentrations induces apoptosis and inhibition of proliferation, while lower concentrations showed synergistically enhanced proliferation in splenocytes and PBMCs. Further, CSA favored the in vitro conversion of CD3+CD161+ cells. Splenocytes and PBMC were found to have synergistic proliferation with Con A, and PBMC exhibited significantly higher expression of NKp30, NKp44, and granzyme B along with enhanced cytotoxicity against K-562 cells in CSA-treated animals. Proliferation assay also showed that proliferation of CD161+ cells was higher in CSA-treated animals. Collectively, our results suggest that CSA differentially influences the population, function, and expression of the NK cell phenotype.

Glucose-Sensitive Hydrogels for the Controlled Release of Insulin

Hydrogels with an increased content of fine pores are synthesized via the copolymerization of acrylamide with N-(2-D-glucose) acrylamide and N,N-methylenebis(acrylamide) in the presence of concanavalin A and mercaptoacetic acid. Insulin introduced into the hydrogels is spontaneously released in response to increase in the concentration of glucose in the environment via a two-step mechanism similar to that in the case of the pancreas and the maximum rate is observed at the first stage.

Abstract 4701: Verteporfin inhibits surface PD-L1 expression in triple-negative breast cancer (TNBC) cells

Abstract Background: Programmed death ligand 1 (PDL1) is commonly expressed on the surface of many tumor cells, including breast cancer. The activity of tumor-infiltrating lymphocytes (TIL) is inhibited by PDL1. High PD-L1 basal cells (particularly basal B) overexpress genes involved in invasion, motility, and chemoresistance (Soliman et al., 2014). Targeting and blocking PD-L1 may enhance eradication of aggressive breast cancer cells by the immune system. Verteporfin (VP), a photosensitizer used to treat macular degeneration, was found to inhibit PD-L1 expression in a high-throughput screen. Studies demonstrated VP inhibits YAP activation by disrupting YAP-TEAD interactions and preventing YAP induced oncogenic growth (Zhang et al., 2016). Here we demonstrate the inhibitory effect of VP on PD-L1 expression in TNBC cell lines. Methods: MDA-MB-231 cells co-cultured with human PBMCs were treated with VP (Sigma) in presence of concanavalin A (ConA) and analyzed using flow cytometry to study levels of CD8+IFNg+ cells. TNBC cell lines (MDA-MB-231, BT-20, HCC-1143, Hs-578T) were treated with VP at doses ranging from 1µM-10µM for 24 hrs. The cells were processed for flow cytometry, Western blot and RT-PCR to check PD-L1 in mechanistic studies looking at manipulation of YAP pathway genes. Results: When MDA-MB-231 cells were co-cultured with normal human PBMCs in the presence of ConA, the CD8+IFNg+ stained cells were reduced compared to PBMC + ConA alone. Interestingly, in the group treated with VP, rescue of CD8+IFNg+ cells was observed. Moreover, MDA-MB-231, BT-20, HCC-1143 and Hs-578T TNBC cells treated with VP showed a significant dose-dependent inhibition of PD-L1 expression by flow cytometry. Western blot analysis also showed complete clearance of PD-L1 protein band with the lowest dose (1µM) used. However, RT-PCR analysis did not show a significant fold change in mRNA levels of PD-L1 in MDA-MB-231 treated cells. Surprisingly, mechanistic studies performed by silencing YAP1, E2F1, and TBK1 in MDA-MB-231, BT-20 and HS-578 T showed a decline in PD-L1 in E2F1 silenced cells, highlighting a plausible role of E2F1-PDL1 signaling axis. However, no change in PD-L1 expression was seen in cells silenced with YAP1 and TBK1. Further, chromatin-immunoprecipitation assay demonstrated E2F1 binding to PD-L1 promoter. Conclusion: Our data so far demonstrate that verteporfin treatment leads to inhibition of PD-L1 in TNBC cell lines and improvement in CD8+IFNg+ cells, indicating that VP might have potential for treatment approaches. Our study warrants further attention towards understanding the mechanism of action of VP in inhibiting PD-L1 and the role of E2F1 in the process. Citation Format: Fatema Khambati, Neha Jaiswal, Srikumar Chellappan, Hatem Soliman. Verteporfin inhibits surface PD-L1 expression in triple-negative breast cancer (TNBC) cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4701.

Molecularly imprinted cryogel columns for Concanavalin A purification from jack bean extract

AbstractConcanavalin A, one of most studied lectins for the purification of glycoproteins and sugars, is selected as a model protein. In this study, a Concanavalin A imprinted poly(hydroxyethyl methacrylate) supermacroporous cryogel was prepared for the purification of Concanavalin from jack bean extract. N‐Methacryloyl‐l‐histidine methyl ester with nickel(II) ions was used as the metal ion coordination complex. Concanavalin A imprinted cryogels were characterized by swelling degree, surface area, Fourier transform infrared spectroscopy, scanning electron microscopy and micro computed tomography measurements. Concanavalin A rebinding and desorption on Concanavalin A imprinted and non‐imprinted cryogels were measured using a continuous system. Selective binding studies were carried out in the presence of Concanavalin B and bovine serum albumin. The selectivity studies were confirmed by fast protein liquid chromatography studies using jack bean extract from Canavalia ensiformis.

Structural analysis and mucosal immune regulation of exopolysaccharide fraction from Bifidobacterium animalis RH

ABSTRACTAn exopolysaccharide fraction (EPSa) produced by Bifidobacterium animalis RH was purified to dissect its chemical structure and to investigate its ability to modulate the mucosal immunity. Our results showed that EPSa had a backbone of (1→6)-linked glucose, (1→3)-linked galactose, (1→4, 6)-linked mannose, (1→3, 4)-linked mannose and (1→4)-linked glucose, branched with glucose and arabinose, and terminated with glucose residues. Scanning electron microscopy identified spherical particle to be the basic structural component of EPSa. The Congo red test indicated that EPSa was not in triple-helical conformation. Our in vitro results showed that EPSa was able to stimulate the proliferation of peripheral blood mononuclear cells and intestinal Peyer’s patch lymphocytes in the presence of concanavalin A. Our in vivo results demonstrated that EPSa can enhance the mucosal immunity by maintaining the Peyer’s patch number in small intestine and the IFN-γ level in serum, and balancing the Th1/Th2 responses.

Purification, characterization, and bioactivities of a polysaccharide from mycelial fermentation of Bjerkandera fumosa

In this work, a novel polysaccharide (named DBFM3) was isolated from mycelia of Bjerkandera fumosa by DEAE-32 and Sepharose CL–6B column chromatography. High-performance gel permeation chromatography showed that DBFM3 was homogeneous, with an average molecular weight of 1.8×105Da. Structural characteristics of the purified fraction were investigated by high-performance liquid chromatography (HPLC), FT-IR, and NMR. HPLC analysis indicated that DBFM3 was composed of mannose, galacturonic acid, and galactose at molar ratios of 1:18.16:0.702. Spectral analysis suggested that DBFM3 had (1→6), (1→3,6), (1→3) linkages and pyranose conformation. Antioxidant assay in vitro showed that DBFM3 exhibited 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical scavenging activity, and has protective effect against DNA damage and damage to SH-SY5Y cells induced by H2O2. Immunological tests indicated that DBFM3 significantly increased lymphocyte proliferation in vitro. Furthermore, DBFM3 increased the proliferation of lymphocytes in the presence of concanavalin A or lipopolysaccharide as mitogen.

Pulmonary Surfactant Protein A-Mediated Enrichment of Surface-Decorated Polymeric Nanoparticles in Alveolar Macrophages.

Surfactant protein A (SP-A), a lung anti-infective protein, is a lectin with affinity for sugars found on fungal and micrococcal surfaces such as mannose. We synthesized a mannosylated poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) copolymer and used it to produce nanoparticles with a polyester (PLGA/PLA) core and a PEG shell decorated with mannose residues, designed to be strongly associated with SP-A for an increased uptake by alveolar macrophages. Nanoparticles made of the copolymers were obtained by nanoprecipitation and displayed a size of around 140 nm. The presence of mannose on the surface was demonstrated by zeta potential changes according to pH and by a strong aggregation in the presence of concanavalin A. Mannosylated nanoparticles bound to SP-A as demonstrated by dynamic light scattering and transmission electron microscopy. The association with SP-A increased nanoparticle uptake by THP-1 macrophages in vitro. In vivo experiments demonstrated that after intratracheal administration of nanoparticles with or without SP-A, SP-A-coated mannosylated nanoparticles were internalized by alveolar macrophages in greater proportion than SP-A-coated nonmannosylated nanoparticles. The data demonstrate for the first time that the pool of nanoparticles available to lung cells can be changed after surface modification, using a biomimetic approach.

In vitro evaluation of graphene oxide nanosheets on immune function

HypothesisGraphene oxide (GO) has attracted the scientific community attention due to its novel properties and wide range of potential applications including hyperthermia cancer therapy. However, little is known about the GO effects on the immune function which involves both innate and adaptive defence mechanisms through the activation of different cell populations and secretion of several cytokines. The effect of different GO nanosheets designed for hyperthermia cancer therapy on macrophage and lymphocyte function should be determined before using GO for this application. ExperimentsThe effects of GO nanosheets with 1 (1-GOs) and 6 arms (6-GOs) of polyethylene glycol on RAW-264.7 macrophages and primary splenocytes (as approximation to the in vivo situation) were evaluated through the proinflammatory cytokine secretion and the modulation of cell proliferation in the presence of specific stimuli for either T-lymphocytes (concanavalin A, anti-CD3 antibody) or B-lymphocytes/macrophages (lipopolysaccharide). Findings6-GOs significantly increased the secretion of TNF-α by RAW-264.7 macrophages without alteration of IL-6 and IL-1β levels. The treatment of primary splenocytes with 1-GOs and 6-GOs in the presence of concanavalin A, anti-CD3 antibody and lipopolysaccharide, produced significant dose-dependent decreases of cell proliferation and IL-6 levels, revealing weak inflammatory properties of GOs which are favourable for hyperthermia cancer therapy.

Development of a method for isolating bovine colostrum mononuclear leukocytes for phenotyping and functional studies

The present study reports a method for isolating bovine colostrum mononuclear cells (CMC) for phenotyping and functional studies. As well as being an important source of immunoglobulins, colostrum also contains leukocytes that may be of greater importance for passive immunity than has previously been thought. Different protocols have been reported for isolating leukocytes from bovine colostrum, although none of these have been validated, and phenotypic analysis of cell populations has not always been performed. In this study, bovine CMC were isolated by density gradient centrifugation. Cell populations were identified by flow cytometry using antibodies against selected bovine cell surface markers and the proliferative capacity of these cells was determined using a 3H-thymidine proliferation assay.The mean cell count of isolated CMC was 3 × 104 and 1 × 105 per mL colostrum for the samples used in the flow cytometric assay and the proliferation assay, respectively. A mean of 25.4 ± 17.1% CMC were identified as T lymphocytes, 2.9 ± 3.0% as B lymphocytes and 32.7 ± 13.7% as macrophages. In terms of proliferation, the mean counts per minute were 4.3 × 103 and 1.8 × 104 for cells cultured in medium only or in the presence of concanavalin A, respectively, showing that CMC are viable and capable of responding to mitogen stimulation. Isolation of CMC and the subsequent phenotypic analysis of the different subpopulations were repeatable, with agreement indices varying between 0.5 and 1.0. Agreement indices for the proliferation assay were estimated at 0.8.

Two acid RNases from Dactylis glomerata seeds. Purification, properties and effect of polyamines and lectins on their activity

Two glycoproteidic acid RNases (RNase I and RNase II) were obtained and purified from the seeds of <em>Dactylis glomerata</em> by extraction with acetate buffer, fractionation with ammonium sulfate, ion-exchange chromatography on DEAE-cellulose, DEAE-Sphadex, affinity chromatography on Con A-Sepharose and gel filtration on Bio-Gel P60. RNase I with a specific activity of 2582 U•mg<sup>-1</sup> protein and an optimum pH of 4.9 and RNase II with a specific activity of 1928 U• mg<sup>-1</sup> protein and optimum pH of 4.6, were isolated. They lacked nuclease, phosphodi- and monoesterase activities. Both forms of the enzyme hydrolyzed pyrimidine homopolymers with a preference for poly U and exhibited a low specificity for purine homopolymers (poly G and poly A). RNase I acted with a 3-fold higher hydrolytic activity on poly C homopolymer than RNase IL The hydrolytic activity of both enzymes was inhibited by Zn<sup>+2</sup>, Fe<sup>+2</sup>, Cu<sup>+2</sup> ions when yeast RNA was the substrate. The amines spermine, spermidine and tyramine at a concentration of 0.1 mM increased the enzymatic activity of both RNases by 20 to 60% of the relative activity. The hydrolytic activity of RNases I and II was stimulated by the presence of lentil lectin (LL), soybean lectin (SBA) and potato lectin (STA), and inhibited by the presence of concanavalin A. The 20-200% stimulation and 40-60% inhibition depended on the proportion, on a weight basis, of enzyme to lectin and were reversible in the presence of receptor sugars.

Antioxidant and immunological activity in vitro of polysaccharides from Phellinus nigricans mycelia

Three polysaccharide fractions, namely, PNMP 1, PNMP2, and PNMP3, were isolated from Phellinus nigricans mycelia. The average molecular weights of PNMP1, PNMP2, and PNMP3 were approximately 28.4, 31.5, and 26.1kDa, respectively. PNMP2 and PNMP3 comprised six monosaccharides: arabinose, fucose, glucose, galactose, mannose, and xylose. PNMP1 comprised only four monosaccharides: glucose, galactose, mannose, and xylose. In vitro antioxidant tests showed that PNMP2 and PNMP3 significantly scavenged superoxide radical, hydroxyl radical, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. PNMP1 effectively inhibited DPPH radical. Lymphocyte proliferation was significantly increased by PNMP1, PNMP2, and PNMP3, particularly by PNMP2 and PNMP3 in the presence of concanavalin A or lipopolysaccharide as mitogens for lymphocytes.

Low-level laser reduces the production of TNF-α, IFN-γ, and IL-10 induced by OVA

Delayed, or type IV, hypersensitivity reactions are a useful model to study the effects of new substances on the immune system. In this study, the experimental model of the delayed type hypersensitivity (DTH) reaction to ovalbumin (OVA) was used to evaluate the immunomodulating effects of low-level laser therapy (LLLT), which is used as an adjuvant therapy in medicine, dentistry, and physical therapy because of its potential anti-inflammatory and analgesic effects observed in several studies. The effects of LLLT (λ 780 nm, 0.06 W/cm(2) of radiation, and fluency of 3.8 J/cm(2)) in reaction to ovalbumin in Balb/C mice were examined after the induction phase of the hypersensitivity reaction. The animals treated with azathioprine (AZA), the animals that received a vehicle instead of ovalbumin, and those not immunized served as controls (n = 6 for each group). Footpad thickness measurements and hematoxylin-eosin histopathological exams were performed. Proliferation tests were also performed (spontaneous, in the presence of concanavalin A and ovalbumin) to determine the production in mononuclear cells cultures of tumor necrosis factor-alpha (TNF-α), INF-γ, and IL-10. In the group of animals irradiated with lasers and in the group treated with AZA, footpad thickness measurements were significantly reduced in comparison to the control group (p < 0.05). This reduction was accompanied by a very significant reduction in the density of the inflammatory infiltrate and by a significant reduction in the levels of TNF-α, INF-γ, and IL-10. LLLT radiation was shown to have an immunomodulating effect on DTH to OVA in Balb/C mice.

Antioxidant and immunological activity in vitro of polysaccharides from Gomphidius rutilus mycelium

Two novel polysaccharide fractions, GRMP1 and GRMP2, were isolated from the mycelium of Gomphidius rutilus through submerged fermentation. GRMP1 and GRMP2 had similar average molecular weights (35 and 31kDa, respectively), and were composed of glucose and xylose at molar ratios of 0.46:1 and 0.63:1, respectively. In vitro antioxidant tests showed that GRMP1 and GRMP2 partly scavenged superoxide radical but almost had no scavenging effect on 1,1-diphenyl-2-picrylhydrazyl free radical. The effect of GRMP1 on hydroxyl radicals was stronger than that of GRMP2. Both GRMP1 and GRMP2 had relatively low reducing power and significant lymphocyte proliferation activity. In the presence of concanavalin A or lipopolysaccharide as mitogens for lymphocytes, the lymphocyte proliferation activity increased for GRMP1 but not for GRMP2 within the test dosage range.