Con A-induced Proliferation Research Articles

Uncommon Epoxyquinols Pyrrolocytosporin A and Cytosporin E2 from the Fungus Eutypella sp. D-1.

Two uncommon epoxyquinols, pyrrolocytosporin A (1) and cytosporin E2 (2), along with the known cytosporin Y1 (3), were isolated from the solid defined medium of the Arctic-derived fungus Eutypella sp. D-1. Their structures were established through comprehensive analyses of spectroscopic and electronic circular dichroism data. Structurally, compound 1 represented the first nitrogen-containing epoxyquinol characterized by a pyrrole fused cytosporin framework, while compound 2 contained an uncommon cyclic carbonate functionality. The antibacterial, immunosuppressive, anti-inflammatory, and cytotoxic activities of all compounds were evaluated. Among the three metabolites, only compound 1 exhibited inhibitory effects on nitric oxide production induced by lipopolysaccharide with an IC50 value of 6.55 μM. Additionally, only compound 2 displayed inhibitory activity against ConA-induced T-cell proliferation with an IC50 value of 9.85 μM.

Patumantanes A-D, seco-Polycyclic Polyprenylated Acylphloroglucinols with Diverse Carbon Skeletons from Hypericum patulum.

Patumantanes A-D (1-4), four new seco-polycyclic polyprenylated acylphloroglucinols (PPAPs) were isolated from Hypericum patulum. Patumantane A (1) was an unprecedented 1,2-seco-homoadamantane-type PPAP bearing a new 3,7-dioxatetracyclo[7.7.0.01,6.111,15]heptadecane architecture based on a 6/7/5/6 ring system. Patumantane B (2) was a unique 1,9-seco-adamantane-type PPAP with a tricyclo[4.4.4.0.02,12]tridecane core formed by a 6/6/6 carbon skeleton, and the further breakage between C-5 and C-9 decorated patumantane C (3) with the 9-nor-adamantane skeleton. More importantly, compounds 2 and 3 exhibited moderate immunosuppressive activity on Con A-induced T-lymphocyte proliferation in vitro, with IC50 values of 5.6 ± 1.2 and 11.2 ± 1.2 μM, respectively.

Immunosuppressive steroids quadrilisteroids A−C and derivatives from the terrestrial fungus Aspergillus quadrilineatus

Seven undescribed compounds (1−7) along with six known compounds (8−13) were isolated from Eurotiaceae Aspergillus quadrilineatus. Their structures and absolute configurations were elucidated by NMR, HRESIMS, and ECD calculations. Quadrilisteroids A (1) and B (2) possessed an unprecedented 6/5/6/6/6/5 hexacyclic ring system in conjugation with a highly fused benzene ring, while quadrilisteroid C (3) featured a surprising 6/6/6/5/5-fused carbocyclic skeleton. Quadrilisteroid C (3) exhibited potent inhibitory activity against LPS-induced proliferation of B lymphocyte cells with an IC50 value of 1.03 μM. Compound 4, demonstrated inhibitory activity against Con A-induced proliferation of T lymphocyte cells with IC50 values of 6.42 μM.

Eutypellaolides A–J, Sesquiterpene diversity expansion of the polar fungus Eutypella sp. D-1

Eight new 12,8-eudesmanolide sesquiterpenes, eutypellaolides A–H (1–8), and two new eudesmane-type sesquiterpenes, eutypellaolides I–J (9–10), along with four known 12,8-eudesmanolide compounds 11–14, were isolated from the culture extract of the polar fungus Eutypella sp. D-1 by one strain many compounds (OSMAC) approach. The structures of these compounds were determined through comprehensive spectroscopic data and experimental and calculated ECD analysis. Antibacterial, immunosuppressive, and PTP1B inhibition activities of these compounds were evaluated. Compounds 1 and 11 exhibited strong inhibitory activities against Bacillus subtilis and Staphylococcus aureus, with each showing an MIC value of 2 μg/mL. Compound 9 displayed weak immunosuppressive activity against ConA-induced T-cell proliferation with an inhibitory rate of 61.7% at a concentration of 19.8 μM. Compounds 5, 11, and 14 exhibited weak PTP1B inhibition activities with IC50 values of 44.8, 43.2, and 49.5 μM, respectively.

Discovery of adamantane-type polycyclic polyprenylated acylphloroglucinols that can prevent concanavalin A-induced autoimmune hepatitis in mice

Hyperadamans A–G (1–7), seven new adamantane type polycyclic polyprenylated acylphloroglucinols (PPAPs), were isolated from Hypericum wilsonii N. Robson. Structurally, 1–4 were the first adamantanes bearing an unusual 2,7-dioxabicyclo-[2.2.1]-heptane fragment, and compound 5 was the first adamantane with a rare 1,6-dioxaspiro[4.4]nonane section. Importantly, 1–7 exhibited significant immunosuppressive activity on Con A-induced T-lymphocyte proliferation in vitro, with IC50 values ranging from 3.97 ± 0.10 to 18.12 ± 1.07 μM. Pretreatment with 1 in Con A-challenged autoimmune hepatitis mice could dramatically ameliorate the levels of hepatic injury indexes (ALT and AST) and reduce the product of proinflammatory cytokines (COX-2, IL-6, IL-1β, IL-18, IL-23A and TNF-α). Furthermore, the protective effect of 1 on the Con A-induced liver injury was corroborated by the histological analysis and the immunohistochemistry.

Discovery of bioactive polycyclic polyprenylated acylphloroglucinols with adamantine/homoadamantane skeletons from Hypericum wilsonii

In this work, nine previous undescribed polycyclic polyprenylated acylphloroglucinols with adamantine/homoadamantane skeletons, cumilcinols A−I (1−9), along with six known analogues, were isolated and identified from the stems, leaves and flowers of Hypericum wilsonii. Their structures were determined by HRESIMS, NMR spectroscopic analysis, single-crystal X-ray crystallography as well as electronic circular dichroism calculations and comparisons. Compound 2 formed a unique furan ring bearing a rare acetal functionality. In bioassays, hyperacmosin G (13) could significantly inhibit the production of NO in LPS-stimulated RAW264.7 cell (IC50 = 4.350 ± 1.146 μM), and increased expression of related transcription factors at the gene level, inhibit the nuclear translocation of NF-κBp65, and reduce the protein expression of COX-2. Additionally, compound 5 showed significant inhibitory activity on Con A-induced T-lymphocyte proliferation (IC50 = 4.803 ± 3.149 μM), and treatment of 5 could reduce the increased ratio of CD4 and CD8 subpopulations induced by Con A in vitro. Those results indicated 13 possesses potential anti-inflammatory activity, and 5 exhibits a certain degree of immunosuppressive activity.

Does Panax notoginseng (Sanqi) inhibit H22 tumor growth via improving the host’s cellular immunity?

Ginsenoside Rg3 has many functions, including anti-tumor activity, antioxidant activity, and immune system modulatory activity. Interestingly, There are two stereoisomers exist in Rg3, 20(S)-ginsenoside Rg3 [20(S)-Rg3] and 20(R)-ginsenoside Rg3 [20(R)-Rg3], respectively. These two isomers have different pharmacological effects due to their stereospecificity. In this study, we compared their effect of stimulating the ConA to induce lymphocyte proliferation and increase the cytokines IL-2 and IFN-γ levels in mice and improve the cellular immunity of the host. The study predicted that ginsenoside (Rg3) could increase Th1-type cytokines interleukin-2 and interferon-γ and stimulate ConA-induced lymphocyte proliferation in mice, thereby improving cellular immunity in H22 mice. The effects of the Rg3 R-form were significantly greater than Rg3 S-form.

Does Panax notoginseng (Sanqi) inhibit H22 tumor growth via improving the host’s cellular immunity?

Ginsenoside Rg3 has many functions, including anti-tumor activity, antioxidant activity, and immune system modulatory activity. Interestingly, There are two stereoisomers exist in Rg3, 20(S)-ginsenoside Rg3 [20(S)-Rg3] and 20(R)-ginsenoside Rg3 [20(R)-Rg3], respectively. These two isomers have different pharmacological effects due to their stereospecificity. In this study, we compared their effect of stimulating the ConA to induce lymphocyte proliferation and increase the cytokines IL-2 and IFN-γ levels in mice and improve the cellular immunity of the host. The study predicted that ginsenoside (Rg3) could increase Th1-type cytokines interleukin-2 and interferon-γ and stimulate ConA-induced lymphocyte proliferation in mice, thereby improving cellular immunity in H22 mice. The effects of the Rg3 R-form were significantly greater than Rg3 S-form.

Effects of one hour daily outdoor access on lying and sleeping postures, and immune traits of tethered cows.

We investigated the effects of outdoor access for 1 h per day on the animal welfare (AW) of tethered cows, in terms of lying and sleeping postures, and immune function. A total of five dry cows were tethered all day indoors (tethering) for 30 days and then tethered indoors with 1 h daily outdoor access (ODA-1h) for 30 days. To analyze the effects of ODA-1h, we calculated the total duration and bout frequency per day, and bout duration of lying and sleeping postures during the last five days of each treatment period. We also analyzed the populations of T cells, B cells, and NK cells in peripheral blood mononuclear cells (PBMC) by fluorescence-activated cell sorting and determined the concanavalin A (Con A) -induced proliferation rate of T cells. The mean total time per day of lying during the ODA-1h treatment was significantly shorter than that during the tethering treatment (p<0.001). The Con A-induced proliferation rate of T cells during the ODA-1h treatment was significantly higher than that during the tethering treatment (p = 0.007). The proportion of NK cells in PBMC during the ODA-1h treatment tended to be higher than that during the tethering treatment (p = 0.062). Although ODA-1h may decrease lying time, it increases the available space for tethered cows towards that typically found in grazing and free barn feeding systems. This increased available space promotes the expression of normal behaviors such as walking and social behaviors except lying and may also improve the immune function of tethered dry cows, thereby improving their overall welfare.

An Immune-Suppressive Triterpenoid from the Roots of Kadsura coccinea

Abstract: Kadsura coccinea is a folk medicine from the family Schisandraceae. Clinically, it has been used to treat many diseases like chronic gastritis and rheumatoid arthritis (RA). Lignans and triterpenes are the main constituents of it. In this paper, phytochemical research on the root of K. coccinea was carried out, leading to the isolation of a previously undescribed 3,4-secolanostane (1). Its structure was established by means of comprehensive spectral analysis, including ECD calculation. Furthermore, bioactivity assay revealed that 1 could elicit a moderate immune-suppressive activity, showing an IC50 value of 8.32 ± 0.13 μM against ConA-induced T-cell proliferation and with IC50 value of 15.58 ± 0.25 μM against LPS-induced B-cell proliferation. The results indicate that 1 might be used in the treatment of rheumatic diseases.

Selenium-chitosan alleviates the toxic effects of Zearalenone on antioxidant and immune function in mice.

This study assessed the protective effects of selenium-chitosan (SC) against antioxidant and immune function-related damage induced by zearalenone (ZEN) in mice. In total, 150 female mice were allotted to five groups for a 30-day study. Control mice were fed a basal diet. Mice in the ZEN, ZEN-Se1, ZEN-Se2 and ZEN-Se3 groups were fed the basal diet supplemented with same dose of ZEN (2 mg/kg) and different doses of SC, 0.0, 0.2, 0.4 and 0.6 mg/kg, respectively (calculated by selenium). After 30 days, the total antioxidant capacity (T-AOC) level, glutathione peroxidase (GSH-Px) activity, total superoxide dismutase (T-SOD) activity and malondialdehyde (MDA) content in plasma and liver, as well as Con A-induced splenocyte proliferation, plasma interleukins concentrations and liver interleukin mRNA expression levels were determined. The plasma and liver GSH-Px activities, liver T-AOC levels, Con A-induced splenocyte proliferation, interleukin (IL) contents and mRNA expression levels in the ZEN group were significantly lower than in the control group (P < 0.01 or P < 0.05), whereas plasma and liver MDA contents in the ZEN group were significantly higher than in the control group (P < 0.01 or P < 0.05). Additionally, plasma and liver GSH-Px activities, liver T-AOC levels, Con A-induced splenocyte proliferation, IL-1β, IL-17A, IL-2 and IL-6 contents and mRNA expression levels in ZEN+Se2 and ZEN+Se3 groups were significantly higher than in the ZEN group (P < 0.01 or P < 0.05), whereas plasma and liver MDA contents in the ZEN+Se2 and ZEN+Se3 groups were significantly lower than in the ZEN group (P < 0.01 or P < 0.05). The plasma and liver GSH-Px activities, Con A-induced splenocyte proliferation, IL-1β and IL-6 contents, IL-2 and IL-17A mRNA expression levels in the ZEN+Se1 group were also significantly higher than in the ZEN group (P < 0.01 or P < 0.05), whereas the plasma MDA content in the ZEN+Se1 group was also significantly lower than in the ZEN group (P < 0.01). Thus, SC may alleviate antioxidant function-related damage and immunosuppression induced by ZEN in mice.

Endometrial Regenerative Cell-Derived Conditioned Medium Alleviates Experimental Colitis.

Background Traditional interventions can play a certain role in attenuating ulcerative colitis (UC), known as one type of inflammatory bowel diseases, but sometimes are not effective. Endometrial regenerative cells (ERCs) have been shown to exert immunosuppressive effects in different models of inflammation, and stem cell-derived conditioned media (CM) have advantages over cell therapy in terms of easy access and direct action. However, whether ERC-CM could alleviate colitis remains unclear and will be explored in this study. Methods Menstrual blood was collected from healthy female volunteers to obtain ERCs and ERC-CM. Acute colitis was induced by 3% dextran sodium sulfate (DSS), and ERC-CM was injected on days 4, 6, and 8, respectively, after induction. The disease activity index was calculated through the record of weight change, bleeding, and fecal viscosity during the treatment process. Histological features, macrophage and CD4+ T cell in the spleen and colon, and cytokine profiles in the sera and colon were measured. In addition, an in vitro lymphocyte proliferation assay was measured by using a CCK-8 kit in this study. Results ERC-CM treatment significantly improved the symptoms and histological changes in colitis mice. ERC-CM increased the percentage of Tregs in the spleen and colon but decreased the percentages of M1 macrophages and Th1 and Th17 cells in the spleen and decreased the population of Th17 cells in the colon. In addition, ERC-CM treatment decreased the local expression of TNF-α, IL-6, and iNOS in the colon. Furthermore, ERC-CM increased the levels of anti-inflammatory cytokines IL-10 and IL-27 but decreased proinflammatory cytokines IL-6 and IL-17 in the sera. In addition, ERC-CM significantly inhibited ConA-induced mouse lymphocyte proliferation in vitro. Conclusion The results suggest that ERC-CM can exert similar therapeutic effects as ERCs and could be explored for future application of cell-free therapy in the treatment of colitis.

RETRACTED ARTICLE: Optimization of trypsin extraction technology of Allium cepa L. polysaccharide by response surface methodology and the antitumor effects through immunomodulation

ABSTRACT The trypsin-assisted extraction of polysaccharides from Allium cepa L. was optimized using the response surface methodology (RSM). The optimum extraction conditions were extraction temperature, extraction time, extraction pH, and enzyme amount of 37.16°C, 180 min, 8.57, and 5.16%, respectively. Under the optimized conditions, the yield of A. cepa L. polysaccharides (ACP) reached 9.69%, which was comparable with the predicted yield (9.73%). Mid- and high-dose ACP significantly inhibited the tumor growth (43.93%) and the tumor inhibition percentage (38.05%), which were more than 30%. The ACP could extend the survival time of H22 ascites tumor-bearing mice. Furthermore, the ACP could reduce the thymus and the spleen atrophy and significantly promoted the Con A-induced proliferation of splenocytes and elevated the serum IFN-γ and IL-2 levels. Therefore, the ACP could inhibit the tumor growth in tumor-bearing mice and regulated the immune function of mice. Practical ApplicationsThe trypsin-assisted extraction has high efficiency, is carried out through the polysaccharide extraction and the deproteinization at the same time, and is more convenient and fast than traditional methods. No detailed study on the optimization of the trypsin extraction of onion polysaccharides is available. Thus, this experiment aims to use the BBD (4 factors and 3 levels) to optimize the roles of extraction temperature, extraction time, extraction pH, and amount of enzyme on the yield of polysaccharides obtained from the fruit of A. cepa L. In addition, when looking for high-quality biological functional principles for the pharmaceutical industry, the antitumor activity of ACP was evaluated. A. cepa L. is one of the most widely cultivated and consumed crops worldwide. Polysaccharides are the main active ingredient, and studies have shown that a high intake of Allium vegetables is associated with reduced risk of cancers.

Bioassay-guided isolation of anti-inflammatory constituents from Celtis sinensis leaves.

Ginkgo acids (GAs) in ginkgo products usually lead to allergies or liver toxicity. In this study, the GA-induced toxicity was attenuated and Con A-induced T-lymphocyte proliferation was inhibited by extracts of Celtis sinensis leaves (ECSL). So, the active ingredients in ECSL were studied to solve the problems caused by GAs. First, the eight components of MeOH extracts were determined by HPLC-DAD/LC-MS. Then, the 12 active ingredients were separated based on the anti-inflammatory activity. Lymphocyte conversion showed that the inhibition rates of apigenin, quercetin, and isovitexin at 100μM on Con A-activated proliferation of T cells were up to 82.46%, 62.86% and 42.76%, respectively. The inhibition rate on the LPS-induced NO release in RAW 264.7 cells of quercetin, apigenin, isovitexin, and vitexin were exceeding 80% at 100μM. Taken together, the material foundation for the screen of GAs toxicity-attenuated ingredients were provided here. PRACTICAL APPLICATIONS: Ginkgo biloba extracts (EGBs) have been conducted to develop functional food which could increase blood circulation and enhance memory. Nevertheless, people in consumption of ginkgo products, often caused severe allergic reactions due to the potential allergens identified ginkgolic acids (GAs) of ginkgo products. We first find that the extracts of Celtis sinensis leaves can reduce GAs-induced damage on HepG2 liver cells. Then, the bioactive compounds in C. sinensis leaves were separated and purified based on anti-inflammatory activities against T cells. Quercetin, apigenin, and isovitexin showed well anti-inflammatory activities against Con A-activated T-lymphocytes and LPS activated RAW 264.7 macrophages. However, quercetin and apigenin are flavones O-glycosides which are rich in Ginkgo biloba. To solve the problems in Ginkgo biloba products caused by GAs, flavone C-glycoside (isovitexin) may be used for the further study in GAs toxicity-reduction.

Targeted isolation of terpenoid indole alkaloids from Melodinus cochinchinensis (Lour.) Merr. using molecular networking and their biological activities

Melodinus cochinchinensis (Lour.) Merr. has been used in Traditional Chinese Medicine for the management of various ailments, yet its chemical constituents are rarely studied. This study was aimed to isolate terpenoid indole alkaloids (TIAs) from the aerial parts of M. cochinchinensis by the guidance of ultrahigh-performance liquid chromatography coupled with a tandem mass spectrometry (UHPLC-MS/MS), and to elucidate their cytotoxic, immunosuppressive and anti-inflammatory activities. Eight new TIAs (1 and 5-11) along with 21 previously described analogues were isolated with the basis of molecular networking developed by our previously reported TIAs from the species of Melodinus. The structures of new TIAs were elucidated by the analysis of extensive spectroscopic data. Among the tested isolates, 11-methoxytabersonine (25) exhibited considerable cytotoxic activity against human acute lymphoblastic leukemia (MOLT-4), breast adenocarcinoma (MCF-7), pro-myeloid leukemia (HL-60), lung adenocarcinoma (A-549), and hepatocelluar carcinoma (SMMC-7721) cell lines, which was more effective than the positive control, cisplatin. The 19β-hydroxyvenalstonidine (21) displayed moderate immunosuppressive effect against Con A-induced splenocyte proliferation at 40 μM, while 16,17-dehydro-19S-vindolinine (1) showed potent inhibitory activity on nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) expressions in LPS-induced RAW 264.7 macrophages.

Immune function effect of F3 rats fed with genetically modified maize harboring Cry1Ab and epsps genes

To evaluate the effects of genetically modified maize with Cry1Ab and epsps genes on immune function in F3 rats. A total of 180 weaning SD rats for F0 generation were randomly divided into three groups, which were treated with AIN-93 G feed control diet, parental maize diet and genetically modified maize diet respectively. After three generations of breeding, antibody producing cells determination, concanavalin A(ConA)-induced lymphocyte transformation test, natural killer(NK)cells activities assay, whole blood lymphocyte subtype detection, delayed type hypersensitivity test and immunity organ index were performed. There were no significant differences between parental maize diet and genetically modified maize diet in terms of the number of antibody-producing cells, ConA-induced spleen lymphocyte proliferation, NK cell activity, whole blood lymphocyte subsets, delayed type hypersensitivity and thymus index(P&gt;0. 05). Under the conditions of this experiment, no significant effects were found on immune function of F3 SD rats through the three generation development study of genetically modified maize with CrylAb and epsps genes.

&lt;i&gt;S. pyogenes&lt;/i&gt; M49-16 arginine deiminase inhibits proliferative activity of human peripheral blood lymphocytes

Arginine deiminase is one of three enzymes constituting the arginine deiminase system in bacteria. It was demonstrated that arginine deiminase exerts anti-proliferative effects on some primary and immortalized mouse and human cells. It is assumed that the inhibitory effect of arginine deiminase on cell proliferation might be related to its ability to result in the arginine exhaustion. T-lymphocytes depend on arginine for proliferation, T-cell receptor complex expression, and the differentiation of memory cells. The aim of the current study was to investigate an impact streptococcal arginine deiminase on functions of human peripheral blood lymphocytes. For this, we comparatively analyzed effects of Supernatant of Destroyed Streptococcal Cells (SDSCs) derived from parental strain S. pyogenes M49-16 and its isogenic mutant S. pyogenes M49-16delArcA bearing inactivated arginine deiminase gene (ArcA) on immune cell functions. An impact of supernatants on cell viability was estimated by staining with DAPI dye. Cell proliferation was assessed by MTT-test and flow cytometry by using the method based on intracellular protein staining with vital fluorescent CFSE (carboxyfluorescein succinimidyl ester) dye. In addition, the level of lymphocyte tyrosine phosphatase CD45 expression in various culturing conditions was evaluated. It was demonstrated that S. pyogenes M49-16 SDSCs had no impact on cells viability. Parental strain-derived SDSC exerted virtually no effect on intact cells proliferation, but considerably suppressed ConA-induced cell proliferation. At the same time, mutant strain-derived SDSC significantly stimulated spontaneous cell proliferation, but not that one after mitogen exposure. It was observed that increased proliferation was accompanied by upregulated CD45 expression, although it was not significant in all cases. These data allow to conclude that bacterial arginine deiminase could be one of pathogenicity factors able to limit lymphocyte proliferation and immune response and could be a part of pathogen strategy to suppress immune response in order to improve bacterial growth and dissemination.

Triterpenoids from the stems and leaves of Schisandra incarnata

Six undescribed triterpenoids, schincarins A‒F, categorized as the preschisanartane (schincarin A), 3,4-secocycloartane (schincarins B‒D), 3,4; 9,10-disecocycloartane (schincarin E), and 3,4-secolanostane (schincarin F) classes, along with twelve known analogues, were obtained from the stems and leaves of Schisandra incarnata Stapf. Their structures were elucidated on the basis of extensive spectroscopic analyses. The absolute configuration of schincarin A was corroborated by analysis of the experimental ECD spectrum, and single-crystal X-ray diffraction with Mo Kα irradiation. Schincarin A represented the first example of preschisanartane-type triterpenoid with two oxa-bridged hemiketals. Among the selected compounds which were evaluated for immunosuppressive activities, schincarin C and kadsudilactone A showed potential inhibition against ConA-induced T-cell proliferation with IC50 values of 5.83 and 6.32 μM, and LPS-induced B cell proliferation with IC50 values of 10.21 and 11.49 μM, respectively.

Variability in peripheral blood enrichment techniques can alter equine leukocyte cellularity, viability and function

Peripheral blood is commonly sampled to assess the health status of human and veterinary patients. Venous blood collection is a minimally invasive procedure, and in the horse, the common collection site is the jugular vein. Post blood collection, sample processing for leukocyte enrichment can vary by research laboratory with the potential to yield different effects on the enriched cells and their function. The focus of the present study was to compare a common blood dilution-leukocyte enrichment technique using a Histopaque gradient medium (His) to a modified leukocyte buffy coat syringe-lymphocyte separation medium technique (Syr- LSM) with peripheral blood from 12 healthy horses. The endpoints examined included cell recovery/mL of blood, cell viability, leukocyte enrichment purity, leukocyte cell marker subset phenotype, leukocyte spontaneous and mitogen-induced proliferation and secretory TNFα concentrations. Leukocyte cell recovery/mL of whole blood and cell viability was significantly increased in enriched leukocytes from the Syr-LSM technique. Interestingly, the percentage of CD8+ and CD21+ were significantly increased with the His technique as was Con A-induced proliferation. Still, leukocyte cell purity and TNFα concentrations from the 72 h cell culture supernatants were comparable across the two enrichment techniques. To summarize, the type of whole blood leukocyte enrichment technique employed can affect the results of immunologic assay endpoints possibly altering data interpretation.

Effect of Social Rank on Well-Being and Space Utilization of Dry Sows Kept in a Free Access Stall-Pen Housing Environment

The primary objective of this pilot study was to evaluate the impact of social rank on the well-being of gestating sows housed in a free access stall-pen housing environment. At d 30 post-breeding, 32 multiparous crossbred sows in groups of 4 were allocated to a stall-pen housing environment. Blood samples were taken at gestational d 30 (before mixing) and d 31 (24-h post-mixing), and then again at d 89 and 110; whereas, sow behavior was recorded at gestational d 30, 66, 87, and 102. Social rank was determined based on aggressive encounters, dyadic fighting wins and losses, and displacements between sows by 48-h post-mixing. At d 31 (post-mixing), regardless of social rank, total WBC decreased and, lymphocyte counts increased, but % change was more significant among the subordinates than dominants. The same trend occurred with neutrophil chemotaxis increased more in subordinates, but ConA-induced lymphocyte proliferation increased more in dominants, whereas LPS-induced lymphocyte proliferation was reduced among dominants. Sow behavior was differentially affected by social rank and time-periods or gestational day, with subordinates spending more time sitting and drinking, especially during time-periods 1, 3, and 4. Dominants displayed more oral-nasal-facial behaviors during mid-gestation but more sham-chew during time-period 2, which included feeding time. Social rank also influenced the percentage of time sows spent in either the stall or pen area, with subordinates in stalls and dominants in pen. These data imply that social rank differentially influences physiology and behavior of dry sows housed in a free access stall-pen environment. More specifically, the location of the dominant sow, time of day, and activity may influence when and where the subordinates spend their time within this housing system.