West African Sorghum bicolor Leaf Sheaths Have Anti-Inflammatory and Immune-Modulating Properties In Vitro

Kathleen F. Benson,Joni L. Beaman,Boxin Ou,Ademola Okubena,Olajuwon Okubena,Gitte S. Jensen 한국식품영양과학회 2013 Journal of medicinal food Vol.16 No.3

The impact of chronic inflammatory conditions on immune function is substantial, and the simultaneous application of anti-inflammatory and immune modulating modalities has potential for reducing inflammation-induced immune suppression. Sorghum-based foods, teas, beers, and extracts are used in traditional medicine, placing an importance on obtaining an increased understanding of the biological effects of sorghum. This study examined selected anti-inflammatory and immune-modulating properties in vitro of Jobelyn™, containing the polyphenol-rich leaf sheaths from a West African variant of Sorghum bicolor (SBLS). Freshly isolated primary human polymorphonuclear (PMN) and mononuclear cell subsets were used to test selected cellular functions in the absence versus presence of aqueous and ethanol extracts of SBLS. Both aqueous and nonaqueous compounds contributed to reduced reactive oxygen species formation by inflammatory PMN cells, and reduced the migration of these cells in response to the inflammatory chemoattractant leukotriene B4. Distinct effects were seen on lymphocyte and monocyte subsets in cultures of peripheral blood mononuclear cells. The aqueous extract of SBLS triggered robust upregulation of the CD69 activation marker on CD3− CD56+ natural killer (NK) cells, whereas the ethanol extract of SBLS triggered similar upregulation of CD69 on CD3+ CD56+ NKT cells, CD3+ T lymphocytes, and monocytes. This was accompanied by many-fold increases in the chemokines RANTES/CCL5, Mip-1α/CCL3, and MIP-1β/CCL4. Both aqueous and nonaqueous compounds contribute to anti-inflammatory effects, combined with multiple effects on immune cell activation status. These observations may help suggest mechanisms of action that contribute to the traditional use of sorghum-based products, beverages, and extracts for immune support.

A Murine Model of Toluene Diisocyanate-induced Contact Hypersensitivity

Chai, Ok Hee,Park, Sung Gil,Sohn, Jang Sihn,Hwang, Seung Soo,Li, Guang Zhao,Han, Eui-Hyeog,Kim, Hyoung Tae,Lee, Moo Sam,Lee, Hurn-Ku,Lee, Yong Chul,Song, Chang Ho The Korean Association of Immunobiologists 2002 Immune Network Vol.2 No.3

Background: Toluene diisocyanate (TDI) can cause contact allergy and occupational asthma, but the mechanism underlying sensitization to this chemical compound remains controversal. Also the correlation of mast cell with contact hypersensitivity (CHS) and the role of mast cell in the TDI-induced CHS is unknown. This issue was investigated by administrating TDI on the skin of genetically mast cell-deficient WBB6F1/$J-Kit^{W}/Kit^{W-v}$ ($W/W^{V}$) and congenic normal WBB6F1/J-Kit+/+ (+/+) mice. Methods: To development of animal model of TDI-induced CHS and to investigate the correlation of mast cell with CHS and the role of mast cell in the TDI-induced CHS, $W/W^V$ and +/+ mice were sensitized with TDI on the back skin at day 1 and day 8, and then challenged with 1% TDI on the ear at day 15. At 1, 2, 4, 8, and 24 hours after 1% TDI challenge, the ear thicknesses were measured. It was investigated the histologic changes of dermis in the ear of $W/W^V$ and +/+ mice at 24 hours after 1% TDI challenge. Results: TDI induced a significant ear swelling response in $W/W^V$ and +/+ mice. TDI induced the significant infiltrations of polymorphonuclear leukocytes and eosinophils in $W/W^V$ and +/+ mice, but not of mast cells in normal mice. And TDI increased a characteristic extent of mast cell degranulation in normal mice. There were no significant differences in the ear swelling and the infiltrations of polymorphonuclear leukocytes and eosinophils of normal versus $W/W^V$ mice, either at baseline or after TDI-induced CHS. Conclusion: From the above results, TDI can be used as a murine CHS model, and the mast cells may not be essential in TDI-induced CHS.

Effects of Mastitis on Buffalo Milk Quality

Tripaldi, C.,Palocci, G.,Miarelli, M.,Catta, M.,Orlandini, S.,Amatiste, S.,Di Bernardini, R.,Catillo, G. Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.10

The objectives of this study were to compare the effectiveness of different indicators of mammary inflammation in buffalo and to evaluate the association of the indicators with buffalo milk yield, composition, and rennet coagulation properties. This study was carried out at four buffalo farms in central Italy using a total of 50 lactating buffalo. Milk from each buffalo was tested at the beginning, middle, and end of lactation. To evaluate the relationship between mastitis markers and milk components, three classes were defined for each of the following markers: total somatic cell count (TSCC), differential somatic cell count (DSCC), and bacteriological results The regression coefficient for the reference method and the alternative method of determining TSCC was 0.81, indicating that the method routinely used to analyze buffalo milk consistently underestimated actual TSCC. The milk samples positive for udder-specific bacteria also had higher TSCC values than the samples that were negative for bacteria ($872{\times}10^3$/ml vs. $191{\times}10^3$/ml). In samples that were positive for udder-specific bacteria, polymorphonuclear leukocytes (PMN) made up greater than 50% of the cells. Moreover, only 1% of the samples in the lowest TSCC class were positive for bacteria. The correlation between TSCC and PMN was stronger (0.70), and PMN values in buffalo milk increased significantly when the TSCC class changed from low (38%) to medium and high (56% and 64%). Milk yield was negatively related to TSCC. Significant changes in lactose (4.87%, 4.80% and 4.64%) and chloride content (0.650 mg/ml, 0.862 mg/ml and 0.882 mg/ml) were also observed with increasing TSCC values. Higher TSCC was associated with impaired rennet coagulation properties: the clotting time increased, while the curd firming time ($p{\leq}0.05$) and firmness decreased. We concluded that in buffalo as in dairy cows, TSCC is a valid indicator of udder inflammation; we also confirmed that a value of $ 200{\times}10^3 cells/ml should be used as the threshold value for early identification of an animal affected by subclinical mastitis. In addition to its association with significantly decreased milk yield, a TSCC value above this threshold value was associated with changes in milk composition and coagulating properties.

Fucoidan suppresses excessive phagocytic capacity of porcine peripheral blood polymorphonuclear cells by modulating production of tumor necrosis factor-alpha by lipopolysaccharide-stimulated peripheral blood mononuclear cells

Kim, Hyeong-Mok,Ahn, Changhwan,Kang, Byeong-Teck,Kang, Ji-Houn,Jeung, Eui-Bae,Yang, Mhan-Pyo Elsevier 2018 Research in veterinary science Vol.118 No.-

<P><B>Abstract</B></P> <P>We examined the effect of fucoidan, an immune modulator, on the phagocytic capacity of porcine peripheral blood polymorphonuclear cells (PMNs) exposed to culture supernatant from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs). For this purpose, we evaluated the phagocytic capacity of porcine PMNs by flow cytometry and measured levels of tumor necrosis factor-alpha (<I>TNF-α</I>) protein and mRNA in porcine PBMCs by enzyme-linked immunosorbent assay (ELISA) and real time-polymerase chain reaction (PCR), respectively. Fucoidan or LPS alone did not affect the phagocytic capacity of PMNs, but phagocytosis by these cells was increased by exposure to culture supernatant from PBMCs treated with fucoidan or LPS. In particular, the culture supernatant from PBMCs treated with LPS revealed excessive phagocytosis of PMNs. This excessive phagocytic capacity was diminished by co-treatment LPS with fucoidan. Production of <I>TNF-α</I> mRNA and protein increased upon treatment of PBMCs with either fucoidan or LPS, but this effect was also diminished by co-treatment LPS with fucoidan. The ability of culture supernatant from PBMCs treated with LPS and/or fucoidan to increase the phagocytic capacity of PMNs was inhibited by anti-recombinant porcine <I>TNF-α</I> polyclonal antibody. These results suggested that fucoidan suppresses the phagocytic capacity of PMNs by modulating <I>TNF-α</I> production by LPS-stimulated PBMCs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The clinical and pharmacological activities of fucoidan, a cell wall polysaccharide found in the brown seaweed, have been known to modulate the immune function. The objective of this study was to examine the effect of fucoidan on the phagocytic capacity of porcine peripheral blood polymorphonuclear cells (PMNs) in inflammation condition. </LI> <LI> The phagocytic capacity of PMNs was enhanced by the culture supernatant from PBMCs treated with either fucoidan or LPS. In particular, the culture supernatant from PBMCs treated with LPS revealed excessive high phagocytosis of PMNs This excessive phagocytosis of PMNs was reduced by treatment of fucoidan. </LI> <LI> In addition, the production of TNF-α was shown to increase upon the treatment of either fucoidan or LPS in PBMCs. However, the excessive production of TNF-α in PBMCs by LPS treatment was reduced by addition of fucoidan. </LI> <LI> The level of TNF-α mRNA expression in PBMCs was also up-regulated by either the fucoidan or LPS treatment. However, the excessive expression of TNF-α mRNA in PBMCs by LPS treatment was decreased by fucoidan. </LI> <LI> This ability of culture supernatant from PBMCs by LPS and/or fucoidan to increase the phagocytic capacity of PMNs was inhibited by addition of anti-rp TNF-α polyclonal antibody (pAb). </LI> <LI> These results suggested that fucoidan can suppress excessive phagocytic capacity of PMNs through modulation of TNF-α production in inflammation condition. </LI> </UL> </P>

Application of Newly Developed Amniotic Membrane Ointment for Photorefractive Keratectomy in Rabbits

Kim, Tae Hyun,Lee, Dong Yeoul,Rho, Jee Hyun,Rho, Sae Heun,Yoo, Kyung Won,Ahn, Hee Bae,Yoo, Young Hyun,Park, Woo Chan S. Karger AG 2006 Ophthalmic research Vol.38 No.2

<P>We developed amniotic membrane ointment (AMO), and the effect of instilling the AMO after photorefractive keratectomy (PRK) was investigated with respect to inflammatory cell infiltration into the corneal stroma, apoptosis of keratocytes, and suppression of lipid peroxidation of cellular walls. The PRK procedure was performed on both eyes of 10 white rabbits. One eye of each rabbit (the experimental eye) was instilled with the AMO and the other eye of the rabbit (the control eye) with a base ointment 0, 8 and 16 h after the PRK procedure. Corneal specimens were collected 24 h after the PRK procedure. Hematoxylin-eosin stain and TUNEL assay were conducted to demonstrate polymorphonuclear and apoptotic cells, respectively. To assess lipid peroxidation, immunohistochemical staining with an antibody to malondialdehyde was undertaken. Compared to the control, the cornea instilled with the AMO had significantly less polymorphonuclear cells infiltrating into the corneal stroma as well as keratocytes subjected to apoptosis. These corneas also showed a significantly less extent of lipid peroxidation than the control. These data support that instillation of the AMO effectively reduced the recruitment of polymorphonuclear cells, the induction of apoptosis in keratocytes, and lipid peroxidation induced by PRK. Thus, this study could provide basic data on the clinical application of the AMO in the days ahead.</P><P>Copyright © 2006 S. Karger AG, Basel</P>

Zinc Increases Chemotactic Activity of Porcine Peripheral Blood Polymorphonuclear Cells

서동희,강병택,강지훈,양만표 한국임상수의학회 2018 한국임상수의학회지 Vol.35 No.5

Zinc is necessary for normal functions in the immune system. The objective of the study is to examinethe effect of zinc on the chemotactic activity of porcine peripheral blood polymorphonuclear cells (PMNs). A modifiedBoyden chamber was used to determine the directional migration distance of PMNs. Various concentrations of zincshowed no chemotactic activity to PMNs. However, culture supernatant from peripheral blood mononuclear cells(PBMCs) treated with zinc remarkably increased the chemotactic activity of PMNs when compared with culturesupernatant from PBMCs treated without zinc. Culture supernatant from PBMCs treated without zinc also increasedthe migration distance of PMNs relative to vehicle control (medium alone). Increasing effect in chemotactic activityof PMNs by culture supernatant from PBMCs treated with zinc was inhibited by treatment of porcine anti-interleukin(IL)-8 polyclonal antibody (pAb). This effect was not affected by heat treatment (4-85oC). This corresponded withheat stable physical characteristics of IL-8. These results suggest that zinc can upregulate the chemotaxis of PMNs,which is primary mediated by IL-8 chemotactic factor released from PBMCs treated with zinc.

Nickel Increases Chemotactic Activity of Porcine Peripheral Blood Polymorphonuclear Cells

주세휘,김학현,강병택,양만표 한국임상수의학회 2020 한국임상수의학회지 Vol.37 No.2

Nickel is a nutritionally essential trace element that plays an important role in the immune system of several animal species. The aim of this study was to examine the effect of nickel chloride on chemotactic activity of peripheral blood polymorphonuclear cells (PMNs) and whether this effect is associated with interleukin (IL)-8 and a nuclear factorkappa B (NF-B)-dependent pathway. Peripheral blood mononuclear cells (PBMCs) and PMNs were isolated by Percoll solution (Specific gravity; 1.080) and 1.5% dextran treatment, respectively. A modified Boyden chamber assay was used to measure the chemotactic activity of PMNs. The level of IL-8 in culture supernatant from PBMCs was measured by enzyme-linked immunosorbent assay (ELISA). Both of PBMCs and PMNs exhibited a low viability when cultured with concentration of greater than 1,000 M of nickel chloride for 24 h. Thus, nickel chloride was used at concentration of 500 M, which preserved cell viability. Treatment with nickel did not directly affect the chemotactic activity of PMNs. However, the chemotactic activity of PMNs was remarkably increased by culture supernatant from PBMCs treated with nickel chloride (500 M) for 24 h. Recombinant porcine IL-8 polyclonal antibody (pAb) neutralized the enhancing effect on the chemotactic activity of PMNs by culture supernatant from PBMCs treated with nickel and this culture supernatant had higher IL-8 levels than the culture supernatant from untreated PBMCs. In addition, n-tosyll- phenylalanine chloromethyl ketone (TPCK), a NF-B inhibitor, antagonized the enhancing effect on the chemotactic activity of PMNs by the culture supernatant from PBMCs treated with nickel. These results suggested that nickel stimulates porcine PBMCs to produce IL-8, which increases the chemotaxis of PMNs via NF-B-dependent pathway.

Cocoa Flavanols and Procyanidins Can Modulate the Lipopolysaccharide Activation of Polymorphonuclear Cells In Vitro

Kenny, Thomas P.,Shu, Shang-An,Moritoki, Yuki,Keen, Carl L.,Gershwin, M. Eric The Korean Society of Food Science and Nutrition 2009 Journal of medicinal food Vol.12 No.1

Flavanols and procyanidins isolated from cocoa have been reported to possess multiple activities potentially relevant to oxidant defenses, vascular function, and immune function. In a combination of in vivo and in vitro studies, we and others have observed that cocoa can be an anti-inflammatory modulator and that compounds in cocoa are capable of modulating eicosanoid production, platelet aggregation, and the pool size of nitric oxide. The present study extends these findings by examining the in vitro effects of cocoa procyanidins on polymorphonuclear cells (PMNs). PMNs, part of the innate arm of the immune system, represent 50-60% of the total peripheral white blood cells and are the first cells to be recruited to the sites of inflammation or injury secondary to bacterial infections. Herein, we demonstrate that certain flavanols and procyanidins isolated from cocoa can moderate a subset of signaling pathways derived from lipopolysaccharide (LPS) stimulation of PMNs, mainly, PMN oxidative bursts and activation markers, and they can influence select apoptosis mechanisms. We hypothesize that flavanols and procyanidins can decrease the impact of LPS on the N-formyl-Met-Leu-Phe-primed PMN ability to generate reactive oxygen species by partially interfering in activation of the mitogen-activated protein kinase pathway.

Cocoa Flavanols and Procyanidins Can Modulate the Lipopolysaccharide Activation of Polymorphonuclear Cells In Vitro

Thomas P. Kenny,Shang-an Shu,Yuki Moritoki,Carl L. Keen,M. Eric Gershwin 한국식품영양과학회 2009 Journal of medicinal food Vol.12 No.1

Flavanols and procyanidins isolated from cocoa have been reported to possess multiple activities potentially relevant to oxidant defenses, vascular function, and immune function. In a combination of in vivo and in vitro studies, we and others have observed that cocoa can be an anti-inflammatory modulator and that compounds in cocoa are capable of modulating eicosanoid production, platelet aggregation, and the pool size of nitric oxide. The present study extends these findings by examining the in vitro effects of cocoa procyanidins on polymorphonuclear cells (PMNs). PMNs, part of the innate arm of the immune system, represent 50–60% of the total peripheral white blood cells and are the first cells to be recruited to the sites of inflammation or injury secondary to bacterial infections. Herein, we demonstrate that certain flavanols and procyanidins isolated from cocoa can moderate a subset of signaling pathways derived from lipopolysaccharide (LPS) stimulation of PMNs, mainly, PMN oxidative bursts and activation markers, and they can influence select apoptosis mechanisms. We hypothesize that flavanols and procyanidins can decrease the impact of LPS on the N-formyl-Met-Leu-Phe-primed PMN ability to generate reactive oxygen species by partially interfering in activation of the mitogen-activated protein kinase pathway.

Effects of Hyaluronic Acid on the Polymorphonuclear Leukocyte (PMN) Release of Active Oxygen and Protection of Bovine Corneal Endothelial Cells from Activated PMNs

Hyun Soo Lym,Youn Suh,Chan Kee Park 대한안과학회 2004 Korean Journal of Ophthalmology Vol.18 No.1

The goal of this study was to evaluate the function of hyaluronic acid (HA) on the active oxygen release from polymorphonuclear leukocytes (PMNs) and the protective effect of bovine corneal endothelial cells (BCEC) from activated PMNs. We used HA with three different molecular weights (MW 700,000, 2,000,000, and 4,000,000) and five different concentrations (0, 0.1, 1, 2, and 3 mg/ml). We evaluated the amount of released superoxide from activated PMNs by using dismutase-inhibitable ferricytochrome C reduction. To compare the property and protective effect of HA with those of other viscoelastic substances, we used the same concentration of methylcellulose. HA suppressed superoxide release from PMNs and protected BCEC from activated PMNs in a dose-dependent, rather than a molecular weight-dependent, manner. The effect of HA reached almost a plateau at concentration above 2 mg/ml. However, methylcellulose, another viscoelastic substance, showed a similar effect. Therefore, it seems that the suppression of superoxide released from PMNs is not a property that is unique to HA, but is a general property of viscoelastic substances. Our results indicate that the action mechanism of HA proceeds not only through cell surface HA-receptor. We think that HA also acts as a physical barrier and/or a scavenger of superoxide.