Lymphocyte Proliferative Responses Research Articles

Vacinas Vivas em Crianças com Síndrome de DiGeorge/Deleção 22q11.2

Children with DiGeorge syndrome/chromosome 22q11.2 deletion syndrome might have a variable degree of immunodeficiency, which may limit the use of live vaccines. The aim of this study was to review the adverse effects of live vaccines and possible relation with immune status in patients with DiGeorge Syndrome/partial 22q11.2 deletion syndrome. Retrospective study with analysis of the clinical records of children with chromosome 22q11.2 deletion syndrome and DiGeorge syndrome phenotype, followed in a Primary Immunodeficiency center. Data were collected on: demographic characteristics; medical and vaccination history with live vaccines; T-CD4+ lymphocyte counts and lymphocyte proliferative responses to antigens and mitogens; adverse reactions; vaccine failure. Twenty three children with DiGeorge syndrome/22q11.2 deletion syndrome were included, 65.2% male, with average age at diagnosis of 11.3 months. Eighteen children (78%) received bacillus Calmette-Guérin vaccine: all with evidence of thymic activity; three presented moderate T-CD4+ lymphopenia and abnormal lymphocyte proliferative responses; one had abnormal lymphocyte proliferative responses for mitogens, four for purified protein derivative and one for tetanus toxoid. Measles, mumps and rubella vaccine was administered to 15 children, three of them with moderate immunosuppression and abnormal lymphocyte proliferative responses. Live attenuated polio vaccine was administered to 4 children without immunosuppression and the rotavirus vaccine to three children, one with moderate immunosuppression. No significant adverse reactions were reported. These data are in line with the findings of other international studies. In our sample, live vaccines were well-tolerated, even in children with moderate T-CD4+ lymphopenia and abnormal lymphocyte proliferative responses to antigens/mitogens.

Listeriolysin O immunogenetic adjuvant enhanced potency of hepatitis C virus NS3 DNA vaccine.

Hepatitis C virus (HCV) is a major health problem all over the world. Among HCV proteins, nonstructural protein 3 (NS3) is one of the most promising target for anti-HCV therapy and a candidate for vaccine design. DNA vaccine is an efficient approach to stimulate antigen-specific immunity but the main problem with that is less immunogenic efficiency in comparison with traditional vaccines. Several approaches have been applied to enhance the immunogenicity of DNA. Recently, bacteria-derived substances are considered as one of the most attractive adjuvants for vaccines, which among them, Listeriolysin O (LLO) of Listeria monocytogenes is a toxin with an extremely immunogenic feature. We investigated detoxified form of LLO gene as genetic adjuvant to modulate NS3 DNA vaccine potency. Immunogenic truncated NS3 gene sequence of HCV (1095-1380aa) and detoxified LLO gene region (5-441aa) were amplified by PCR and cloned into the pcDNA3.1 plasmid separately. The expression of recombinant proteins (pc-NS3, pLLO) was confirmed in HEK293T cell line by western blotting. BALB/c mice models received three doses of different formula of plasmids in two-week intervals and two weeks after the final immunization, the immune responses were evaluated by specific total antibody level, lymphocyte proliferation, cytotoxicity, and cytokine levels assays. To evaluate in vivo cytotoxic activity, tumor challenge was performed. The recombinant plasmids were successfully expressed in mammalian cell line, and coadministration of pc-NS3 with pLLO induced the highest titer of total IgG against NS3 antigen compared with other controls. Determination of IgG subclasses confirmed the efficient increase in mixed responses with Th1 dominancy. Furthermore, significant levels of cytokines (p < .05) and lymphocyte proliferation responses (p < .05) indicated the superiority of this regimen. The findings may have important implication for LLO gene application as genetic adjuvant in immune response against HCV.

Intranasal administration with recombinant Bacillus subtilis induces strong mucosal immune responses against pseudorabies

BackgroundPseudorabies caused by pseudorabies virus (PRV) mainly infects the swine and seriously threatens the biosafety of the other animals, including humans. Since 2011, the outbreaks of PRV mutants have caused enormous economic losses in the swine industry, and traditional vaccines cannot offer enough protection. PRV can transmit by direct contact, aerosol transmission and pollutants. PRV mainly transmit through the nasal mucosa. After infecting the nasal epithelial cells, PRV can quickly infect the olfactory nerve and establish a potential infection of sensory neurons. Therefore, nasal immunity can effectively prevent viral colonization infection. Recombinant Bacillus subtilis has been widely used to deliver antigen and achieve adequate protective immune responses.ResultsThe present study successfully constructed recombinant Bacillus subtilis (B. subtilis) expressing the dominant antigen regions of PRV gC and gD proteins (named B. subtilis-gCa and B. subtilis-gDa). Furtherly, we evaluated the immunogenicity of the two recombinant B. subtilis in mice. The mice intranasal administration with B. subtilis-gCa and B. subtilis-gDa effectively stimulated IgA and IgG immune responses, further regulated specific T lymphocytes proliferative response by IFN-γ and IL-10, and ultimately produced high titers of neutralizing antibodies against PRV infection. In particular, B. subtilis-gDa possessed more excellent immune effect than B. subtilis-gCa in mice.ConclusionsThese results suggested that B. subtilis-gCa and B. subtilis-gDa could trigger high levels of mucosal and systemic immune responses and would be potential candidates for developing PRV vaccines.

HBs antigen and mannose loading on the surface of iron oxide nanoparticles in order to immuno-targeting: fabrication, characterization, cellular and humoral immunoassay

Mannosylation of nanovaccine is an appropriate strategy for targeting the mannose receptors on DCs. Here, HBsAg and mannose loaded on the surface of iron oxide nanoparticles to increases HBsAg vaccine potency. Nanoparticles are made by co-precipitation method and bonded to the HBsAg and mannose by chemical bonding. The physicochemical properties of nano-vaccines, their toxicity and antigenicity were determined. The synthesized nano-vaccine showed spherical shape with a mean particle size of 60 nm, a zeta potential of −44 mV, an antigen-binding efficiency of around 100% and for mannose 78%. In vitro release of nanoparticles exhibited about 30% at the first day and about 60% until the third day. SDSPAGE analysis confirmed structural integrity of HBsAg loaded on nanoparticles. The HBsAg-loaded LCMNP and MLCMNP nanoparticles had no toxic effects on HEK293 cell line. The quantification of the intracellular Fe by ICP-OES as a criterion of nano-vaccine uptake revealed mannose intensify uptake of MLCMNP. In addition, mannose in the structure of MLCMNP improved IL-6, TNF-α and IFN-γ (>16 fold) cytokines genes expression by macrophage/dendritic cells after exposure in 12 h. Immunization of experimental mice (subcutaneously, two times with 2-week intervals) with 5 µg of HBsAg loaded on MLCMNP nanoparticles increased specific total IgG and IgG2a/IgG1 ratio. In addition, TNF-α, IL-12, IL-2 and IL-4 cytokines in mannosylated nano-vaccine increased versus nano-vaccine group while lymphocyte proliferation and IFN-γ responses in the targeted nano-vaccine group show a tiny increase versus the nano-vaccine group. The results show that mannosylated nano-vaccine promotes higher level of cellular and humoural immune responses against HBsAg nano-vaccine.

Adjuvanticity of aqueous extracts of Artemisia rupestris L. for inactivated foot-and-mouth disease vaccine in mice

Several methods have been developed to improve the efficacy of foot-and-mouth disease virus (FMDV) vaccine. The study aims to determine whether aqueous extracts of Artemisia rupestris L. (AEAR) as an immunoactivator in combination with inactivated FMDV vaccine can promote immune responses in mice. Intramuscular co-immunization in ICR mice with different doses of AEAR plus FMDV vaccine could substantially improve the FMDV-specific antibody production (IgG, IgG1, and IgG2a) and lead to significant lymphocyte proliferative responses. Th1-type immune responses were also observed, including proliferative responses of CD8+, CD4+, CD4+CD44+, and CD8+CD44+ T cells and the killing efficacy of cytotoxic T lymphocyte (CTL) responses. AEAR also elicited the higher levels of IL-4 and IFN-γ in CD4+ T cells as well as the higher level of IFN-γ in CD8+ T cells. The medium dose of AEAR induced the significant adjuvant activity. Further tests in mice indicated that AEAR could activate DCs maturation by increasing the expression levels of co-stimulatory molecules (CD40, CD86, CD80, and MHC-II) on dendritic cells (DCs) from splenocytes and reduce the activity of regulatory T cells (Treg). Abnormal behaviors, side effects or death were not observed in immunized mice. AEAR could boost humoral and cell-mediated immunity elicited by FMDV vaccine, especially Th1-type immune responses.

Effects of saccharomyces cerevisiae supplementation on milk production, insulin sensitivity and immune response in transition dairy cows during hot season

The objectives were to determine effects of feeding a live yeast supplement on the productive performance, blood metabolic profile, the immune function, and insulin sensitivity traits of transition dairy cows under the hot months of summer. From 21 d before expected date of calving until d 60 postpartum, two groups of multiparous Holstein cows (6 cows per treatment) were fed a diet without or with live yeast supplement (4 g yeast/d/head). Blood was collected on d −14, 0, 14, 28, and 60 d relative to parturition and analyzed for nonesterified fatty acids, β-hydroxybutyrate, glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, creatinine, urea-N, triiodothyronine, and thyroxine. Plasma concentrations of histamine and heat shock protein 70 (Hsp70) were also determined on d –21, 14, 28, and 49 relative to parturition. An intravenous glucose challenge was made on d 14 postpartum. The cell-mediated immune and humoral responses were established through phytohemagglutinin challenge and ovalbumin immunization, respectively. Prepartum DM intake was greater in yeast-fed cows; however, this difference disappeared after parturition. Cows receiving yeast supplement produced more milk (+1.40 kg/d) and had greater concentrations of milk fat and total solid than those receiving no yeast. Loss of body condition score from calving to d 21 postpartum tended to be lower in yeast-fed cows than control cows. Yeast supplementation had no effect on the response variables to the glucose tolerance test. Plasma concentration of Hsp70 was also lower on d 14 and 28 after parturition in yeast-fed cows. Yeast supplementation enhanced cellular immune function; however, it had no effect on immunoglobulin G secretion against ovalbumin immunization. Overall, live yeast supplementation benefited milk production and milk composition, lowered plasma level of Hsp70, and enhanced the lymphocyte proliferative response in transition dairy cows, which may suggest an immunomodulatory effect of yeast supplement.

IMMUNOLOGICAL RESPONSE OF A NEW TRIVALENT CAPRIPOXVIRUS VACCINE IN PREGNANT EWES AND DOES

This study was conducted as a field trial of using a new candidate trivalent Capripoxvirus (CaPVs) vaccine in pregnant ewes and does to detect safety degree and level of immunity, in comparison with commercial Romanian sheep pox viral (RSPPV) vaccine. Monitoring the post-vaccinal reaction and measurement of cellular and humoral immunity by evaluating the lympho­cyte proliferation assay (LPA) and serum neutralization test (SNT), respectively, in addition to detect the acquired maternal immunity in new-born lambs and kids .The post-vaccine reaction in animals vaccinated with trivalent CaPVs vaccine was more noticeable than that vaccinated with RSPPV vaccine without significant difference (P-Value&gt;0.05). In vitro, the evaluation of cel­lular and humoral immunity revealed that the animals vaccinated with trivalent vaccine can significantly induce lymphocyte proliferation response in addition to of high level of antibodies (Abs) in comparison to that vaccinated with RSPPV vaccine (P-Value &lt;0.05) with peak titre of lymphocyte proliferation and Abs titre at 10th and 28th, respectively. Furthermore, the evaluation of maternal immunity in new-born lambs and kids from previously vaccinated dams with trivalent CaPVs and RSPPV vaccines by SNT was shown that the peak level of receiving immunity was appeared at 1st week after birth with increasing its level in lambs and kids of dams vaccinated with trivalent CaPVs vaccine than those of dams vaccinated with RSPPV vaccine with a significant difference in between (P-Value &lt;0.05). The result of this study provide a good profile about a high immune response with a long level of maternal immunity provided with trivalent CaPVs vaccine in comparison to the RSPPV vaccine so toward about the preferable use of new vaccine to control infection with sheep and goat pox diseases. Trivalent CaPVs vaccine could be candidate to be used against lumpy skin disease (LSD) due to antigenic relationship between genus CaPVs strains.Key words: does; ewes; monovalent RSPPV; safety; trivalent CaPVs

Establishing reference ranges for lymphocyte proliferation responses to phytohemagglutinin in patients with T cell dysfunction

Introduction: The evaluation of lymphocyte proliferation responses is a critical component of the clinical work up for patients with suspected immunodeficiencies. Those with severe combined immunodeficiency (SCID) have consistently low to absent responses (stimulation index, SI) to the mitogen phytohemagglutinin (PHA). However, patients with combined immunodeficiency (CID) have more varied proliferative responses, and are open to a wide range of interpretations. Aims: To establish lymphocyte proliferation response reference ranges for patients with T cell defects, especially those with CID as well as healthy controls. Methods: Data was collected retrospectively from charts of patients with a diagnosis of SCID (n = 39), CID (n = 52), or from healthy controls (n = 440). Reference percentiles were calculated using the 95% of the distribution of the test results. Results: The reference ranges for the control group ranged from 134 to 2220.5, whereas those with CID were distributed between 0.81 and 169.1. Patients with typical SCID had profound low proliferative responses, with SI &lt;5. Conclusion: Our results highlight the variability of lymphocyte proliferation responses to PHA in patients with CID as well as healthy controls. These reference ranges will assist with the critical interpretation of assay results, particularly when values fall on the extreme end of the range. Statement of novelty: We provide reference ranges for lymphocyte proliferation responses to PHA from patients with CID and healthy controls.

Transition from metal-DTH resistance to susceptibility is facilitated by NLRP3 inflammasome signaling induced Th17 reactivity: Implications for orthopedic implants.

Metal hypersensitivity has been recognized as an adverse biologic reaction that can compromise total joint arthroplasty (TJA) performance. However, the etiology of metal hypersensitivity responses in TJAs remains unclear. Metal implant debris is known to act as a danger signal that drives NLRP3 inflammasome activation. It remains unknown if implant debris induced inflammasome activation regulates T cell lineage in TJA metal hypersensitivity responses. In this study, we show both in vivo and in vitro that the pathogenesis of metal hypersensitivity responses to implant debris are largely dependent on activation of the inflammasome/caspase-1 pathway and subsequent production of IL-17A/F by CD4+ T cells. Inhibiting either the inflammasome pathway or IL-17A bioactivity in vivo and in vitro (in vivo using NLRP3 and Caspase-1 deficient mice or in vitro using blocking agents such as Capase-1 inhibitor, IL-1Ra and anti-IL-17A), significantly (p<0.05) mitigated metal-DTH paw inflammation as well as lymphocyte cytokine (IFN-γ and IL-17) and proliferation responses in metal-sensitized mice and primary human PBMCs. This study provides mechanistic insight into how in vivo exposure to orthopedic implant debris, and metals in general, elicits NLRP3 inflammasome activation that mediates the generation of IL-17A/F producing CD4+ T cells, leading to metal-delayed type hypersensitivity reactions.

Immunological characterization of onion (Allium cepa) allergy.

IntroductionOnion (Allium cepa) handling can induce contact dermatitis, rhinoconjunctivitis and asthma. However, only sporadic reports exist on allergic reactions to onion consumption.AimWe describe herein a case of a 35-year-old man who had an episode of anaphylaxis following cooked onion ingestion. We evaluated onion-specific IgE, the possible cross-reactivity between onion and peach and lymphocyte proliferation in response to onion.Material and methodsSpecific IgE was evaluated using two techniques: skin test and ImmunoCAP technology. Cross-reactivity between onion and peach was evaluated by IgE-ELISA inhibition test. As for lymphocyte proliferation, blood mononuclear cells were stained with CFSE dye and cultured with an in-house onion extract. Proliferation and phenotype was assessed by flow-cytometry.ResultsThe skin test and ImmunoCAP confirmed the IgE-dependent response towards onion. The incubation of the patient serum with increasing concentrations of the peach extract reduced only scarcely (~30%) onion-specific IgE. Interestingly, B cells but not T cells showed proliferation in response to onion extract.ConclusionsIn conclusion, our report shows that cooked onion can induce severe allergic reactions, suggesting the presence of thermostable components. Moreover, we applied for the first time a B-cell-based approach to the diagnosis of food allergy. This latter approach might also be applied to other allergic conditions.

Effects of dietary Enterococcus faecium NCIMB 11181 supplementation on growth performance and cellular and humoral immune responses in broiler chickens

This study evaluated the effects of dietary Enterococcus faecium NCIMB 11181 on growth performance and immune response in broiler chickens. A total of 360 1-day-old Arbor Acres male birds were randomly assigned to 4 treatments that administered different dosages of E. faecium (0, 5 × 107, 1 × 108, and 2 × 108 CFU E. faecium/kg diet). The results revealed that average daily gain (ADG) changed quadratically, while feed conversion rate (FCR) increased linearly from day 22 to 35 and day 1 to 35 (P < 0.05). Supplementation of E. faecium at 5 × 107CFU/kg diet resulted in increased ADG (P < 0.05) compared with the other groups. Birds fed with 2 × 108 CFU/kg E. faecium exhibited increased peripheral blood lymphocyte proliferation in response to concanavalin A (Con A) (P < 0.05) at day 35 and enhanced skin responses following phytohemagglutinin (PHA) injection (P < 0.05) at 12h. Serum lysozyme activity at day 21 increased linearly with dietary E. faecium concentration (P < 0.05), the highest activity was observed in the 1 × 108 and the 2 × 108 CFU E. faecium groups (P < 0.01). Serum levels of proinflammatory cytokines IL-1β, IL-2, IL-6, IFN-γ, and anti-inflammatory IL-4, IL-10 changed linearly or quadratically both at the initial and final phases (P < 0.05). In addition, BSA antibody titers were significantly increased following both primary and secondary inoculation when birds were fed with 1 × 108 or 2 × 108 CFU/kg E. faecium (P < 0.05). In comparison with other groups, birds received 5 × 107 CFU E. faecium exhibited the highest levels of serum IgG (P < 0.05) at day 35. Together, our results revealed that broiler diet supplemented with 5 × 107 CFU/kg E. faecium NCIMB 11181 was appropriate in relation to growth performance under normal conditions. Upon administration with higher dosages of E. faecium NCIMB 11181, obvious immune-stimulatory effects were observed following both cell-mediated and humoral immunity.

Turkey humoral and cell-mediated immune responses to a Newcastle viscerotropic vaccine and its association with major histocompatibility complex.

Immune responses to vaccines are mainly influenced by the nature of vaccines and host variation in response to vaccination. In this study we aimed to investigate turkey humoral and cell-mediated immune responses to a Newcastle viscerotropic vaccine and its association with major histocompatibility complex (MHC). Turkeys were vaccinated with Villegas–Glisson/University of Georgia (VG/GA) attenuated vaccine against Newcastle disease. The stimulation index of lymphocyte proliferation and antigen-specific local secretory IgA responses in bile, duodenum, ileum, as well as serum IgY and IgA responses were analysed by enzyme-linked immunosorbent assay. The turkey MHC class II B locus was selected as candidate gene for detection of associations with cellular and humoral immune responses. Significant differences were observed between both cellular and humoral responses of vaccinated and unvaccinated groups. A significant positive correlation was also found between ND specific IgY and ND specific IgA titres in serum, intestine (duodenum and ileum) and trachea. Moreover, the correlation between specific IgA titres in ileum and specific bile, duodenum and trachea was positively significant. High resolution melting analysis (HRM) was used to genotype MHC class II B exon 2. Eight melting profiles (A-G) were identified, among which, profile G showed a significant association with cellular response. The profile B revealed significant association with total IgA titres in serum and ileum. These findings help our understanding of the association of turkey MHC types with immune responses. Further correlation analysis between serum and mucosal antibody titres demonstrated that the levels of IgY and IgA in serum can give an impression about the levels of secretory IgA and situation of mucosal immunity. Based on the significant effects, ND specific IgY in serum appears to be a promising indirect marker for specific IgA in serum and trachea.

Immune-enhancing Activity of Potential Probiotic Strains of Lactobacillus Plantarum in the Common Carp (Cyprinus Carpio) Fingerling.

IntroductionImmune-potentiating functions of Lactobacillus plantarum strains in the common carp were evaluated.Material and MethodsFourteen days of feeding fish dry diet supplemented with the bacteria provided parameters of nonspecific humoral immunity (lysozyme, ceruloplasmin, γ-globulin, total protein levels, and serum bactericidal activity) and cellular immunity (pinocytosis, respiratory burst activity, and potential killing activity of organ phagocytes), as well as the proliferative response of organ lymphocytes stimulated with mitogens. The resistance of fish to infection with Aeromonas hydrophila was also determined.ResultsDietary supplementation with L. plantarum had a substantial influence on the activity of organ phagocytes, especially the potential killing activity of head kidney cells. A significant increase in the proliferative activity of LPS-stimulated B lymphocytes and in the levels of γ-globulins and total protein was observed. The supplemented diet conveyed higher resistance than the control diet as the cumulative fish mortalities after infection with A. hydrophila were 65% and 85%, respectively.ConclusionThe results indicate that dietary supplementation with L. plantarum stimulates the antibacterial resistance of common carp and may reinforce defence against bacterial infections, but further studies need to be conducted.

A Subunit Vaccine Based on E2 Protein of Atypical Porcine Pestivirus Induces Th2-type Immune Response in Mice.

An atypical porcine pestivirus (APPV) causing congenital tremor type A-II in piglets was identified in China in 2016. An increased number of cases of APPV have been reported in various countries all over the world since 2015. This study aimed to develop an effective subunit vaccine against APPV based on the E2 protein, which is the main immunogenicity protein of APPV. In this study, E2 protein was successfully expressed by the baculovirus expression system. E2 protein was confirmed by Western blot assay, which showed that E2 protein possesses N-linked glycosylation sites. The immunogenicity of E2 subunit vaccine was evaluated in mice. The E2 protein emulsified with ISA 201VG adjuvant induced significantly higher levels of APPV-specific antibodies and elicited stronger lymphocyte proliferative responses and higher interleukin-10 secretion than those of the E2 protein emulsified with IMS 1313VG adjuvant. This observation indicates that the E2 subunit vaccine induces a Th2-type immune response. Our results showed that E2 protein can be developed as a safe and effective subunit vaccine for the control of APPV infection.

Does proliferation capacity of lymphocytes depend on human blood types?

In vitro human lymphocyte culture methodology is well established yet certain confounding factors such as age, medical history as well as individual's blood type may potentially modulate in vitro proliferation response. These factors have to be carefully evaluated to release reliable test report in routine cytogenetic evaluation for various genetic conditions, radiation biodosimetry, etc. With this objective, the current study was focused on analyzing the proliferation response of lymphocytes drawn from 90 individuals (21-29 years) with different blood types. The proliferation response was assessed in the cultured lymphocytes by cell cycle, mitotic index (MI), and nuclear division index (NDI) after stimulation with phytohaemagglutinin (PHA). To investigate the toxic effect on proliferation, MI was calculated in representative samples of each blood type were X-irradiated. The results showed that there was no significant difference among the cell cycle phases of lymphocytes in different blood types (P > 0.05). Similarly, both MI and NDI of lymphocytes derived from different blood types also did not show significant difference ( P > 0.05). The extensive interindividual variation within and among the blood types is likely responsible for the lack of significant difference in lymphocyte proliferation. Although spontaneous proliferation efficiency of lymphocytes of different blood types after PHA stimulation was grossly similar, the MI observed after radiation exposure showed a significant difference ( P < 0.05) indicating a differential proliferation response among the blood types. Our results suggest that the blood types did not have any impact on PHA-induced proliferation; however, a specific differential lymphocyte proliferation observed after radiation exposure needs to be considered.

Immunomodulatory effect of anise (&lt;i&gt;Pimpinella anisum&lt;/i&gt;) in BALB/c mice

Purpose: The aim of this work was to investigate the effect of anise decoction consumption on lymphocytes activity, complete blood count (CBC) and nitric oxide (NO) production in BALB/c mice.Methods: BALB/c mice were given anise tea instead of drinking water and the effect on selected immune parameters was analyzed after 1 and 2 weeks of treatment.Results: Cell activity of anise treated mice was significantly higher than control group at week 2 as revealed by mixed lymphocyte reaction (MLR2. The spleen cells of anise treated mice showed a significant higher lymphocyte proliferative response to in vitro challenge with phytohaemagglutinin (PHA).compared to controls at week two of treatment. The increase in mouse foot thickness as indicator of delayed type of hypersensitivity (DTH) was less in anise treated mice compared to control group. Nitric oxide production by peritoneal macrophages in response to activation with lipopolysaccharides (LPS) was reduced by anise treatment after 1 and 2 weeks of treatment andno significant changes in CD4 and CD8 were noticed either at week 1 or 2 of treatment.Conclusion: This study provides preclinical evidence that anise possesses immunomodulatory activity when administered orally in mice and selectively activates cell-mediated immune mechanismsKeywords: Immunomodulatory, Anise, Traditional medicine

Effects of repeated social mixing on behavior and blood immune cells of group-housed pregnant sows (Sus scrofa domestica)

Commercial housing of sows often involves frequent regrouping (“mixing”) of animals that poses a potential welfare problem due to social stress. Although mixing is a common management practice worldwide, immunological consequences are poorly investigated in pregnant sows. This study combines both behavioral and endocrine data with detailed analyses of blood cellular immunity to examine whether repeated social mixing affects the immune system in pregnant sows, which none of the studies conducted so far could confirm. Pregnant multiparous sows (German Landrace, n = 40) were housed in groups of five animals. Sows were either assigned to a repeated social mixing treatment with a mutual exchange of two randomly selected sows of two specific groups (2 × 2) twice a week over a period of eight weeks, or remained undisturbed in their original group. Blood samples of all sows were collected at five time points before, during, and after the mixing period to evaluate the distribution of blood lymphocyte subpopulations, mitogen-induced lymphocyte proliferation and plasma cortisol concentrations. Mixing caused higher levels of aggressive behavior (p ≤ 0.05) during the entire mixing period. Compared to pre mixing baseline values, lymphocyte numbers were lower in mixed sows (p ≤ 0.05), which was primarily caused by lower antigen-experienced T helper cells, cytotoxic T cells and natural killer cells. Mixed sows showed a higher proliferative response of lymphocytes while granulocytes and cortisol concentrations were not affected (p > 0.05). Repeated social mixing thus not only resulted in an increase of aggressive behavior, but also in altered immune cell numbers of the adaptive immune system, even though sows became eventually familiar with each other. These findings suggest, in contrast to previous studies, an adverse influence of frequent mixing on the immune system of pregnant sows.

Cardiosphere-derived cells suppress allogeneic lymphocytes by production of PGE2 acting via the EP4 receptor

Cardiosphere-derived cells (CDCs) are a cardiac progenitor cell population, which have been shown to possess cardiac regenerative properties and can improve heart function in a variety of cardiac diseases. Studies in large animal models have predominantly focussed on using autologous cells for safety, however allogeneic cell banks would allow for a practical, cost-effective and efficient use in a clinical setting. The aim of this work was to determine the immunomodulatory status of these cells using CDCs and lymphocytes from 5 dogs. CDCs expressed MHC I but not MHC II molecules and in mixed lymphocyte reactions demonstrated a lack of lymphocyte proliferation in response to MHC-mismatched CDCs. Furthermore, MHC-mismatched CDCs suppressed lymphocyte proliferation and activation in response to Concanavalin A. Transwell experiments demonstrated that this was predominantly due to direct cell-cell contact in addition to soluble mediators whereby CDCs produced high levels of PGE2 under inflammatory conditions. This led to down-regulation of CD25 expression on lymphocytes via the EP4 receptor. Blocking prostaglandin synthesis restored both, proliferation and activation (measured via CD25 expression) of stimulated lymphocytes. We demonstrated for the first time in a large animal model that CDCs inhibit proliferation in allo-reactive lymphocytes and have potent immunosuppressive activity mediated via PGE2.

Evaluation of therapeutic potency of human papillomavirus-16 E7 DNA vaccine alone and with interleukin-18 as a genetic adjuvant

AIMS: Despite the existence of effective preventive vaccines for human papillomavirus (HPV), therapeutic vaccines that trigger cell-mediated immune responses are required to treat established infections and malignancies. The aim of this study was to evaluate the therapeutic potency of HPV-16 E7 deoxyribonucleic acid (DNA) vaccine alone and with interleukin (IL)-18. METHODS: In vitro expressions of IL-18 were performed on human embryonic kidney 293 cells and confirmed it by Western blotting methods. DNA vaccine was available from a previous study. A total of 45 female C57BL/6 mice divided into five groups (DNA vaccine, DNA vaccine adjuvanted with IL-18, pcDNA3.1, and phosphate buffer saline) were inoculated with murine tissue culture-1 cell line of HPV related carcinoma, expressing HPV-16 E6/E7 antigens. They were then immunized subcutaneously twice at a seven-day interval. The antitumor and antigen specific-cellular immunity were assessed by lymphocyte proliferation (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide: MTT) assay, lactate dehydrogenase release assay, IL-4 assay and interferon-gamma (IFN-γ) assay. Tumor size was followed for 62 days.RESULTS: MTT assay, which measures the lymphocyte proliferation in response to the specific antigen, increased in the co-administration and the DNA vaccine groups as compared to control and genetic adjuvant groups (p&lt;0.001). The mice immunized with the co-administration generated significantly more IFN-γ and IL-4 than other immunized mice (p&lt;0.001). Reduction of the tumor size in the co-administration and the DNA vaccine groups was significantly more pronounced than in the control and genetic adjuvant groups (p&lt;0.001), but no statistically significant difference between DNA vaccine and co-administration groups (p=0.15) occurred.CONCLUSIONS: IL-18 as a genetic adjuvant and E7 DNA vaccine alone enhanced immune responses in mouse model systems against cervical cancer. However, using of IL-18 as a genetic adjuvant with E7 DNA vaccine had no significant synergistic effect on the immune responses in vivo.

Short communication: Lymphocyte proliferative responses in cattle naturally infected with digital dermatitis consist of CD8+ and γδ-T cells but lack CD4+ T cells

Digital dermatitis is an infectious disease of cattle and the leading cause of lameness. This disease is complicated by the reoccurrence of the lesions and the observation of lesions on more than one limb at different time points, indicating infection may not result in a protective immune response. The objective of this study was to characterize the peripheral blood cellular response in naturally infected and naïve cattle to bacterial antigens derived from pathogens associated with digital dermatitis lesions. Peripheral blood mononuclear cells were isolated from dairy cattle identified as having active or chronic lesions during routine hoof-trimming. Following bacterial antigen stimulation, cells were analyzed for proliferation and phenotype by flow cytometry, and culture supernatants were analyzed for IFN-γ secretion. Digital-dermatitis-infected animals had greater serum antibody titers to treponemal antigens, higher percentages of proliferating CD8+, γδ-T cells, and B cells, and increased IFN-γ secretion in vitro when compared with responses of naïve animals. No increase in proliferation of CD4+ T cells was detected in infected or naïve cattle. Although CD8+ and γδ-T cell responses may be antigen specific, the memory nature or long-lived response is yet unknown. The lack of responsiveness of CD4+ memory cells to treponemal antigens could explain the high rate of reoccurrence of digital dermatitis in infected animals.