Antigen Sheep Red Blood Cells Research Articles

CD154 Costimulation Shifts the Local T-Cell Receptor Repertoire Not Only During Thymic Selection but Also During Peripheral T-Dependent Humoral Immune Responses.

CD154 is a transmembrane cytokine expressed transiently on activated CD4 T cells upon T-cell receptor (TCR) stimulation that interacts with CD40 on antigen-presenting cells. The signaling via CD154:CD40 is essential for B-cell maturation and germinal center formation and also for the final differentiation of CD4 T cells during T-dependent humoral immune responses. Recent data demonstrate that CD154 is critically involved in the selection of T-cell clones during the negative selection process in the thymus. Whether CD154 signaling influences the TCR repertoire during peripheral T-dependent humoral immune responses has not yet been elucidated. To find out, we used CD154-deficient mice and assessed the global TCRβ repertoire in T-cell zones (TCZ) of spleens by high-throughput sequencing after induction of a Th2 response to the multiepitopic antigen sheep red blood cells. Qualitative and quantitative comparison of the splenic TCZ-specific TCRβ repertoires revealed that CD154 deficiency shifts the distribution of Vβ-Jβ genes after antigen exposure. This data led to the conclusion that costimulation via CD154:CD40 during the interaction of T cells with CD40-matured B cells contributes to the recruitment of T-cell clones into the immune response and thereby shapes the peripheral TCR repertoire.

Immunostimulant activity of standardised extracts of <i>Mangifera indica</i> leaf and <i>Curcuma domestica</i> rhizome in mice

Purpose : To investigate the immunomodulating activity of Mangifera indica. (MI) and Curcuma domestica (CD) extracts in mice. Methods : Mice were randomly divided into 4 groups, namely, vehicle, untreated, MI and CD groups (n = 6). They were treated with MI (160 mg/kg) or CD (200 mg/kg) extracts, and vehicle for control group, for 14 days and sacrificed on day 15. In innate immunity test, the mice were challenged with sheep red blood cells (SRBCs) antigen on day 8, while in adaptive immunity test, mice were immunized and challenged on days 7 and 14, respectively, with SRBC. White blood cells count (WBC), spleen index (SI), and haemagglutination (HA) titer, and delayed type hypersensitivity (DTH) responses were determined. Results : For both plant extracts, adaptive immunity groups showed the highest response compared to innate groups. In adaptive immunity, the WBC count of MI and CD treated animals was significantly higher than in the untreated and vehicle treated groups (p < 0.001). Moreover, the SI of mice from MI and CD treated groups differed significantly that of the untreated group (p < 0.01 and p < 0.05, respectively). HA titer in CD (both non-challenged and challenged) groups was significantly higher than in the non-challenged vehicle group (p < 0.001). HA titer in MI group (non-challenged) was significantly lower than in non-challenged CD and challenged groups (p < 0.01 and p < 0.001, respectively). Conclusion : MI and CD extracts, in appropriate doses, exerted immunostimulant effects in mice by enhancing both innate and adaptive immune systems via increase in WBC, SI, HA titer and DTH responses. Keywords : Mangifera indica, Curcuma domestica, Immunostimulant activity, Spleen index, Haemagglutination, Hypersensitivity response

Evaluation of immunomodulatory effects of lamotrigine in BALB/c mice.

Modulation of the immune system has recently been shown to be involved in the pharmacological effects of old antiepileptic drugs and in the pathogenesis of epilepsy. Therefore, the most recent guidelines for immunotoxicological evaluation of drugs were consulted to investigate the immunomodulatory effects of lamotrigine, a newer antiepileptic drug, in BALB/c mice. These included the in vivo effects of lamotrigine on delayed-type hypersensitivity (DTH) response to sheep red blood cell (SRBC) antigens, hemagglutination titer assays and hematological changes. In vitro effects of lamotrigine on ConA-induced splenocyte proliferation and cytokine secretion were assessed. The results showed that lamotrigine treatment significantly increased the DTH response to SRBC in the mouse model of this study. This was accompanied by a significant increase in relative monocyte and neutrophil counts and in spleen cellularity. Lamotrigine significantly inhibited ConA-induced splenocyte proliferation in vitro and it significantly inhibited IL-2 and TNF-α secretion in ConA-stimulated splenocytes. In conclusion, the results demonstrated significant immunomodulatory effects of lamotrigine in BALB/c mice. These data could expand the understanding of lamotrigine-induced adverse reactions and its role in modulating the immune system in epilepsy.

CP and CP-PGN protect mice against MRSA infection by inducing M1 macrophages

Corynebacterium pyruviciproducens (C. pyruviciproducens, CP), as a newly discovered immunomodulator, has been confirmed to have a stronger immunoregulation than Propionibacterium acnes (P. acnes) of the traditional immune adjuvant, by previous experiments with model antigen ovalbumin and sheep red blood cells. Here, it was designed to assess its ability to resist methicillin-resistant Staphylococcus aureus (MRSA), since MRSA as a vital gram positive pathogen is characterized by high morbidity and mortality. In this report, it was indicated that C. pyruviciproducens and its peptidoglycan (CP-PGN) could help to be against bloodstream infection of MRSA with raised survival rate, decreased bacteria load and alleviated systemic inflammation, and these effects of CP-PGN were more pronounced. However, the whole CP was inclined to prevent localized abdominal infection of MRSA from progressing to a systemic infection. And they showed the potential as a therapeutic drug alone or combined with vancomycin. The diversity of capacity of activating macrophages induced by CP and CP-PGN may result in distinct resistance to MRSA in different infection models. Furthermore, both CP and CP-PGN induced M1 macrophages. In conclusion, CP and its PGN could act as promising immune agents to treat and prevent MRSA infection.

The Small Rho GTPases Rac1 and Rac2 Are Important for T-Cell Independent Antigen Responses and for Suppressing Switching to IgG2b in Mice.

The Rho GTPases Cdc42, Rac1, and Rac2 coordinate receptor signaling to cell adhesion, migration, and proliferation. Deletion of Rac1 and Rac2 early during B cell development leads to failure in B cell entry into the splenic white pulp. Here, we sought to understand the role of Rac1 and Rac2 in B cell functionality and during the humoral antibody response. To circumvent the migratory deficiency of B cells lacking both Rac1 and Rac2, we took the approach to inducibly delete Rac1 in Rac2−/− B cells in the spleen (Rac1BRac2−/− B cells). Rac1BRac2−/− mice had normal differentiation of splenic B cell populations, except for a reduction in marginal zone B cells. Rac1BRac2−/− B cells showed normal spreading response on antibody-coated layers, while both Rac2−/− and Rac1BRac2−/− B cells had reduced homotypic adhesion and decreased proliferative response when compared to wild-type B cells. Upon challenge with the T-cell-independent antigen TNP-conjugated lipopolysaccharide, Rac1BRac2−/− mice showed reduced antibody response. In contrast, in response to the T-cell-dependent antigen sheep red blood cells, Rac1BRac2−/− mice had increased serum titers of IgG1 and IgG2b. During in vitro Ig class switching, Rac1BRac2−/− B cells had elevated germline γ2b transcripts leading to increased Ig class switching to IgG2b. Our data suggest that Rac1 and Rac2 serve an important role in regulation of the B cell humoral immune response and in suppressing Ig class switching to IgG2b.

Nigella sativa (black cumin seed) as a biological detoxifier in diet contaminated with aflatoxin B1.

Aflatoxin B1 (AFB1 ) is a common feed contaminant that adversely affects bird performance and product quality. A total of 600 7-day-old quail chicks were randomly allotted to eight experimental groups in a completely randomized design with five replicate pens and 15 quails per pen. Experimental treatments including two levels of AFB1 (0 and 2.5mg/kg) and 4 levels of Nigella sativa (NS) (0, 0.5, 1.0 and 1.5% of diet) were offered from 7 to 35days of age to quail chicks. Although feeding of AFB1 impaired gain (G) and feed conversion ratio, dietary NS increased G (p<0.05). Relative weight of bursa of Fabricius increased with incremental levels of NS (p<0.05). AFB1 decreased the size of heart, but incremental levels of NS increased the relative weight of heart (p<0.05). The liver hypertrophy was observed in birds receiving AFB1 (p<0.05). The birds in AFB1 group had smaller testes than other groups (p<0.05). Hematocrit value in birds fed AFB1 was lower than that in other groups (p<0.05) and incremental levels of NS increased blood hematocrit (p<0.05). Amount of malondialdehyde (MDA) in meat samples of the birds fed AFB1 was higher than those did not receive toxin but incremental levels of NS decreased the MDA concentration in affected birds (p<0.05). AFB1 suppressed the humoral immunity of the birds while NS augmented the antibody titres against sheep red blood cell and Newcastle disease virus antigens (p<0.05). AFB1 decreased lactic acid bacteria (LAB) and spore-forming bacteria (SFB) but increased Escherichia coli (E.coli; p<0.05). However, NS increased LAB and SFB but decreased the E.coli populations (p<0.05). This study revealed that NS as a biological detoxifier could relatively attenuate the negative effects of AFB1 inquails.

Investigations into the Immunotoxicity and Allergic Potential Induced by Topical Application of N-Butylbenzenesulfonamide (NBBS) in a Murine Model

N-Butylbenzene sulfonamide (NBBS) is a commonly used plasticizer found in numerous products. Due to its extensive use, lack of adequate toxicological data, and suspicion of toxicity based on the presence of structural alerts, it was nominated to the National Toxicology Program for comprehensive toxicological testing. The purpose of this study was to evaluate the potential for hypersensitivity and immune suppression following dermal exposure to NBBS using a murine model. NBBS tested negative in a combined irritancy/local lymph node assay (LLNA), classifying it as nonirritating and nonsensitizing. To estimate the immunosuppressive potential of NBBS, assays that assessed immunotoxicity were performed, including the immumnoglobulin (Ig) M response to T-cell-dependent antigen sheep red blood cells (SRBC), using the plaque-forming cell (PFC) assay and immune cell phenotyping. After a 28-d treatment with NBBS, mice exposed to the lowest concentration (25% NBBS) showed a significant increase in IgM-producing B cells in the spleen. No marked changes were identified in immune cell markers in the lymph node. In contrast to body weight, a significant elevation in kidney and liver weight was observed following dermal exposure to all concentrations of NBBS. These results demonstrate that dermal exposure to NBBS, other than liver and kidney toxicity, did not apparently induce immunotoxicity in a murine model.

Mentha piperita (peppermint) in growing Japanese quails’ diet: Serum biochemistry, meat quality, humoral immunity

Abstract In this experiment, the effect of dietary Mentha piperita L. (MP, peppermint) on biochemical blood parameters, meat quality, and humoral immunity responses of growing Japanese quails were evaluated. A total of 360 seven-day-old quail chicks were fed six experimental diets (with 5 replicates and 12 birds each) including control positive (basal diet + antibiotic), control negative (basal diet), MPI (basal diet + 10 g/kg MP), MPII (basal diet + 20 g/kg MP), MPIII (basal diet + 30 g/kg MP), and MPIV (basal diet + 40 g/kg MP) for 5 weeks. At the end of experiment, two birds in each replicate were sacrificed for blood sampling and assessment of meat quality. The birds were injected with sheep red blood cell (SRBC, at days 18 and 28 for primary and secondary responses) and Newcastle disease virus (NDV, at day 14) antigens. Concentrations of triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL), and alanine aminotransferase (ALT) were decreased in birds received dietary peppermint but the level of high-density lipoprotein (HDL) increased (P

Effects of a multi-strain probiotics against aflatoxicosis in growing Japanese quails

An experiment was conducted to determine the effects of a multi-strain probiotics (PRO) on growth performance and immune status of growing Japanese quails fed aflatoxin B1 (AFB1). A total of 320 quail chicks were assigned to 4 treatments, control (basal diet), AFB1 (basal diet+2.5mg AFB1/kg), PRO (basal diet+150mg PRO/kg), and AFB1+PRO (basal diet+2.5mg AFB1 and 150mg PRO/kg) with 4 replicate pens and 20 quails per pen. The quail chicks had free access to water and experimental diets from 7 to 35 d of age, and feed intake (FI) and body weight gain (BWG) were measured weekly. Overall feed intake and BWG were reduced in AFB1 treatment (P<0.05), but feeding probiotics improved those criteria in quails received dietary AFB1 (P<0.05). During 7–35d of age, AFB1 increased FCR while the PRO supplementation decreased FCR in quails fed AFB1 diet (P<0.05). Moreover, FCR in AFB1+PRO treatment was better than PRO treatment (P<0.05). The most hypertrophy in internal organs (e.g., liver and spleen) and suppressed growth in testes and bursa of Fabricius were observed in AFB1 group but feeding PRO improved the status of these organs (P<0.05). The humoral (e.g., antibody response against sheep red blood cell antigen) and cellular responses (e.g., skin thickness against 2,4-Dinitro 1-chlorobenzene) were suppressed in quails fed AFB1 but feeding PRO boosted those immune systems (P<0.05). The use of AFB1 in diet invariably increased the levels of aspartate transaminase, alanine aminotransferase, and alkaline phosphatase in the sera of Japanese quails (P<0.05). The opposite effect was observed for aforementioned enzymes because of PRO in the diets. This study showed that under aflatoxicosis, the use of dietary multi-strain probiotics could improve the growth performance and immunocompetence in growing Japanese quails.

Impact on antibody responses of B-cell-restricted transgenic expression of a viral gene inhibiting activation of NF-κB and NFAT.

In this work, we have assessed the impact in vivo of the evasion gene A238L of African swine fever virus, an inhibitor of both NF-κB- and NFAT-mediated transcription. The A238L gene was selectively expressed in mouse B lymphocytes using the promoter and enhancer sequences of the mouse Ig μ heavy chain. The IgM primary and IgG2b secondary serological responses and the number of splenic germinal centres in response to the TD antigens DNP-keyhole limpet hemocyanin and sheep red blood cells, respectively, were both lower in the transgenic mice, whereas the response to the TI type-1 and type-2 antigens DNP-Ficoll and DNP-LPS, respectively, were normal, except for the increased levels of IgG3 at day 14 in the DNP-LPS-immunized mice. Thus, it appears that neither p65 (NF-κB) nor NFAT is essential for B-cell development but, in a manner that is still unclear, may be relevant for their function.

Malaria parasites, immune challenge, MHC variability, and predator avoidance in a passerine bird

Several hypotheses predict a relationship between parasite burden and risk-taking behavior, but the underlying causal mechanisms are poorly understood due to the scarcity of experimental studies and the neglected focus on immune defense. Here, in 3 sets of field studies on the collared flycatcher, Ficedula albicollis, we investigated how among-male variation in flight initiation distance (FID, the distance at which an individual flee a potential predator) is linked to among-male variation in health status. First, we correlatively assessed the relationship between FID and the prevalence of haemosporidian blood parasites. We found no difference in risk-taking behavior between parasitized and nonparasitized individuals rejecting a hypothesis that predicts that malaria infection status affects the costs of predator avoidance. Second, we performed an immune challenge experiment, in which randomly chosen birds were injected with a novel antigen (sheep red blood cell) and their change in FID was compared with birds that received a placebo treatment. This experiment revealed no evidence for the immunological treatment affecting risk-taking behavior, thus we failed to obtain support for the hypothesis that posits that immediate health status mediates decisions about when to flee a predator. Finally, we detected a negative relationship between the number of alleles of the major histocompatibility complex and FID. This result, in concordance with the above negative results, supports the “avoidance” hypothesis that states that only individuals with efficient immune defense machinery are able to bear the costs of risk-taking that can emerge through the increased infection rates of risk-taker individuals.

A new mouse anti-mouse complement receptor type 2 and 1 (CR2/CR1) monoclonal antibody as a tool to study receptor involvement in chronic models of immune responses and disease

Although reagents are available to block mouse complement receptor type 2 and/or type 1 (CR2/CR1, CD21/CD35) function in acute or short term models of human disease, a mouse anti-rat antibody response limits their use in chronic models. We have addressed this problem by generating in Cr2−/− mice a mouse monoclonal antibody (mAb 4B2) to mouse CR2/CR1. The binding of murine mAb 4B2 to CR2/CR1 directly blocked C3dg (C3d) ligand binding. In vivo injection of mAb 4B2 induced substantial down regulation of CR2 and CR1 from the B cell surface, an effect that lasted six weeks after a single injection of 2mg of mAb. The 4B2 mAb was studied in vivo for the capability to affect immunological responses to model antigens. Pre-injection of mAb 4B2 before immunization of C57BL/6 mice reduced the IgG1 antibody response to the T-dependent antigen sheep red blood cells (SRBC) to a level comparable to that found in Cr2−/− mice. We also used the collagen-induced arthritis (CIA) model, a CR2/CR1-dependent autoimmune disease model, and found that mice pre-injected with mAb 4B2 demonstrated substantially reduced levels of pathogenic IgG2a antibodies to both the bovine type II collagen (CII) used to induce arthritis and to endogenous mouse CII. Consistent with this result, mice pre-injected with mAb 4B2 demonstrated only very mild arthritis. This reduction in disease, together with published data in CII-immunized Cr2−/− mice, confirm both that the arthritis development depends on CR2/CR1 receptors and that mAb 4B2 can be used to induce biologically relevant receptor blockade. Thus mAb 4B2 is an excellent candidate for use in chronic murine models to determine how receptor blockage at different points modifies disease activity and autoantibody responses.

Resting metabolic rate, stress, testosterone, and induced immune response in spring- and fall-born males of Campbell’s dwarf hamsters: Maintenance in long-day conditions

We have studied morphological and physiological traits of equally aged young males of Campbell’s dwarf hamster (Phodopus campbelli Thomas, 1905) born at the end of summer (fall males) and winter (spring males) in a vivarium with a constant 14-hour day length (LD 14: 10). Animals were removed from parental cages at the age of one month, kept in isolation in the same lighting conditions, and tested at the age of 2 to 3.5 months. Spring males had a higher resting metabolic rate, a higher body mass in the middle of the experiment, bigger testes and seminal vesicles, higher blood testosterone levels, and a stronger T-cell immune response to phytohemagglutinin injected intradermally than fall males. They did not differ significantly from fall males in basal blood cortisol levels or in antibody production in response to the sheep red blood cells (SRBC) antigen challenge, but they showed lower adrenocortical response to a social stressor and to an adrenocorticotropic hormone. A general linear model analysis showed that the blood cortisol level after a 10 min encounter of males in the open arena and the resting metabolic rate were the only factors significantly influencing humoral immune response to SRBC. In the general linear model including the intensity of T-cell immune response or basal testosterone level as dependent variables, birth season was the only factor causing a significant effect.

Effect of Supplementation of Licorice Root (Glycyrrhiza Glabra L.) Extracts on Immune Status in Commercial Broilers

An experimental study was designed with the objective of exploring the effect of supplementation of Glycyrrhiza glabra extracts on immune status in commercial broilers. Fifty four, day old, straight run broiler chicks were randomly allotted to 3 treatment groups (T 1 - unsupplemented; T 2 - supplemented with 1% G. glabra crude extract powder and T 3 - supplemented with 0.1% G. glabra s tandardized extract powder) consisting of three replicates of six chicks each. The birds were reared under standard and uniform management conditions throughout the experimental period. Ranikhet-LaSota strain was used to produce antibodies against Ranikhet disease virus. All the birds were also immunized against Sheep Red Blood Cells (SRBC). HI titre values against Ranikhet (LaSota) and HA titre values against SRBC antigens were estimated to assess the humoral immunity. The Serum Biochemical Parameters viz., serum total protein and albumin were estimated and serum globulin and albumin:globulin ratio were calculated. To assess the cell mediated immunity, total and differential white blood cell counts were performed. The group supplemented with 0.1% licorice standardized extract has shown significantly (p<0.01) higher HI titre value against Ranikhet disease virus, than the other groups, on day 42, whereas, there was no significant differences among the groups on days 7, 14, 21, 28 and 35 in HI titre values. Licorice root standardized extract supplemented group performed better than other groups in increasing HI titre values throughout the study period. Against SRBC antigen, significant differences (p<0.01) were observed among the groups on days 21, 28, 35 and 42 in HA titre values (p<0.01). Licorice root standardized extract supplemented group performed better than other groups in increasing HA titre values throughout the study period. No significant differences were noticed among groups in 2 weeks and 6 weeks except on 4 weeks in which significant (p<0.01) differences were noticed in serum albumin, serum globulin and albumin:globulin ratio. No significant differences were observed in the total WBC and differential cell counts throughout the study period among different treatment groups. This shows that licorice root extracts do not have any direct effect on cell mediated immunity.

UJI AKTIVITAS IMUNOGLOBULIN M (IgM) KELINCI JANTAN (Oryctolagus cuniculus) AKIBAT PENGARUH PEMBERIAN PRODUK MADU

The objective was to study the effect of natural pure honey on the immunoglobulin M (IgM) antibody activity against sheep red blood cells (SRBCs) antigen ini male rabbit (Oryctolagus cuniculus) using the hemoagglutination method. Three different honey products with 2,1 ml/kg body weight given orally twice a day for 5 days. Then each animal was injected intraperitoneally with 2,8 ml of 2% SRBCs. At day 7 after the immunization, serum were collected from the blood samples to know which highest dilution value of rabbit serum still showing agglutination reaction. The result of the the statistical analysis using the Complete Random Device ( CRD) method and than continued with the Duncan’s Multiple Range Test (DMRT) method shows that the administration of three different honey products significantly increase Immunoglobulin M (IgM) activity against SRBCs. Key words : Honey, Hemaglutination, Rabbit

The Histone Demethylase LSD1 Acts As a BCL6 Corepressor In Germinal Center B Cells

The BCL6 transcription repressor (B-Cell Lymphoma 6) is required for B cells to form germinal centers (GC) and undergo antibody affinity maturation. BCL6 mediates its functions mainly through the recruitment of corepressor complexes with distinct enzymatic activities including the SMRT/NCOR, BCOR and MTA3 corepressors. Preliminary chromatin immunoprecipitation (ChIP) studies designed to dissect the biochemical actions of BCL6 on chromatin, showed that BCL6 inhibition results in considerable induction of H3K4me2 levels at selected BCL6 target loci. Consistent with these observations a BCL6 expressing lymphoma cell line had lower levels of H3K4me2 compared to BCL6 negative lymphoma cells. Hence, we hypothesized that in addition to HDAC and PRC1-like complexes BCL6 associates with distinct complexes bearing H3K4me2 demethylase activity. We observed that one of the few H3K4 demethylases known to target H3K4me1/2 residues, Lysine Specific Demethylase-1 (LSD1) is highly expressed in GC B cells. More specifically both LSD1 transcript and protein levels were induced in the transition from naïve to GC B cell state, similar to BCL6 levels. Collectively these results indicate that BCL6 might functionally interact with LSD1 in GC B cells to mediate transcriptional repression of key gene networks. We initially asked if BCL6 and LSD1 proteins interact in GC-derived B cells using coimmunoprecipitation and GST pull down experiments. We found that BCL6 and LSD1 proteins endogenously interact in these cells. Additional GST pull-down experiments using full length GST-LSD1 and in-vitro translated S35 BCL6 confirmed this interaction suggesting it is possibly direct. Moreover, immunoprecitated BCL6 complexes could demethylate H3K4me1 but not H3K4me3 in in vitro assays using histone substrates. To further evaluate the role of LSD1 in the physiological GC response we challenged C57BL/6 mice with T-cell dependent antigen (sheep red blood cells) and we examined GC formation in mice treated with a specific LSD1 inhibitor compared to untreated control mice. LSD1 inhibition resulted in GC formation deficiency indicating that LSD1 enzymatic activity plays key roles in normal immune response. To study the functional role of BCL6-LSD1 complexes we performed Q-ChIP assays for LSD1 at BCL6 target loci and showed that several BCL6 target loci, including MLL2, CDKN1A, BRAF and CCR6 are also bound by LSD1. To extend these findings we performed LSD1 ChIP-seq in a GC-derived lymphoma cell line. These genomic studies revealed an extensive overlap of BCL6 and LSD1 binding sites (1509 common loci). Roughly 560 gene promoters were bound by both BCL6 and LSD1 at the same site. Moreover, about 1,000 BCL6-LSD1 binding sites were outside promoters and might play a role in the regulation of inter/intragenic elements similar to the role of LSD1 in enhancer regulation in embryonic stem cells. Loss of BCL6 at several targeted loci resulted in LSD1 release from the chromatin suggesting that BCL6 mediates tethering of LSD1 at these loci. Genes bound or associated with BCL6-LSD1 complexes were significantly enriched in pathways such as B cell activation, chromatin organization and cell cycle control. Taken together these results suggest that LSD1 acts as a BCL6 corepressor in GC B cells and is an essential transcriptional regulator for the formation of normal GC response. BCL6 and LSD1 possibly cooperate to establish a transcriptional program crucial for the GC stage of B cell differentiation. Because LSD1 is expressed at high levels in lymphoma cells, targeting the LSD1 enzymatic activity could potentially be exploited as a lymphoma targeted therapy. Disclosures:No relevant conflicts of interest to declare.

Characterization of primary immune response in Ghanaian local, Sasso T-44 and broiler chickens to sheep red blood cell antigens

Animals should have the capacity to adapt to changing environmental pressures to ensure sustainable productivity. The easier an animal can adapt to disease burdens without extra cost to the farmer, the more profitable the enterprise becomes. Effective immune response protects organisms against disease insults and contributes to improved productivity. Primary immune response of Ghanaian local, Sasso T-44 and broiler chickens to sheep red blood cell (SRBC) antigens was measured 5 days post-inoculation. Antibody levels were determined by haemagglutination. Local ecotypes and Sasso T-44 chicken were superior to broilers in terms of their ability to respond to antigens. Female chickens produced more antibodies in response to increased concentration of SRBC antigens while the opposite was true in the male population. Further investigation of the genetic correlation between production performance and immune response of various chicken breeds will help establish the effect of, for example, high growth rate of broilers on their immunological status.

Gray Level Co-Occurrence Matrix Texture Analysis of Germinal Center Light Zone Lymphocyte Nuclei: Physiology Viewpoint with Focus on Apoptosis

In our study we investigated the relationship between conventional morphometric indicators of nuclear size and shape (area and circularity) and the parameters of gray level co-occurrence matrix texture analysis (entropy, homogeneity, and angular second moment) in cells committed to apoptosis. A total of 432 lymphocyte nuclei images from the spleen germinal center light zones (cells in early stages of apoptosis) were obtained from eight healthy male guinea pigs previously immunized with sheep red blood cells (antigen). For each nucleus, area, circularity, entropy, homogeneity, and angular second moment were determined. All measured parameters of gray level co-occurrence matrix (GLCM) were significantly correlated with morphometric indicators of nuclear size and shape. The strongest correlation was observed between GLCM homogeneity and nuclear area (p < 0.0001, r(s) = 0.61). Angular second moment values were also highly significantly correlated with nuclear area (r(s)= 0.39, p < 0.0001). These results indicate that the GLCM method may be a powerful tool in evaluation of ultrastructural nuclear changes during early stages of the apoptotic process.

Oral subchronic immunotoxicity study of ethyl tertiary butyl ether in the rat

The potential for immunotoxicological effects of ethyl tertiary butyl ether (ETBE, CAS RN 637-92-3) was studied in young adult female Crl:CD(SD) rats following subchronic oral exposures. Rats were exposed by gavage once daily for 28 consecutive days to 0, 250, 500, or 1000 mg ETBE/kg body weight (BW)/day; a concurrent positive control group received four intraperitoneal injections of at 50 mg cyclophosphamide monohydrate (CPS)/kg/day on study Days 24–27. Immunotoxicity was evaluated using a splenic antibody-forming cell (AFC) assay to assess T-cell-dependent antibody responses in rats sensitized with sheep red blood cells (SRBC). All rats survived to the scheduled necropsy. There were no effects on clinical observations, body weights, feed or water consumption, or macroscopic pathology findings in the ETBE-treated rats. No ETBE-related effects were observed on absolute or relative (to final body weight) spleen or thymus weights, spleen cellularity, or on the specific (AFC/106 spleen cells) or total activity (AFC/spleen) of splenic IgM AFC to the T-cell-dependent antigen SRBC. CPS produced expected effects consistent with its known immunosuppressive properties and validated the appropriateness of the AFC assay. Based on the results of this study, ETBE did not suppress the humoral component of the immune system in female rats. The no-observed-effect level for immunotoxicity was the highest dosage tested at 1000 mg/kg/day.

Evaluation of in vivo immune response activity and in vitro anti-cancer effect by Scrophularia megalantha

Many studies have showed that plant extracts possess immunomodulatory and anti-cancer effects. In this study, these effects of Scrophularia megalantha extract, a native plant in Iran, were investigated in vitro and in vivo. Microculture tetrazolium test (MTT assay) was employed to evaluate the effects of S. megalantha on human Jurkat (lymphoblast-like) cell line cytotoxicity. Furthermore, delayed type hypersensitivity (DTH) and hemagglutination tests in mice employed to evaluate the effects of S. megalantha on cellular and humoral responses, respectively. In vitro exposures of the Jurkat cells with various concentrations of S. megalantha extract (0.05, 0.1 and 0.2 mg/mL) significantly suppressed their growth in a dose-dependent manner. The maximum tumor cell growth inhibition was observed with 0.2 mg/mL of the extract. Moreover, the production of specific antibody to sheep red blood cell (SRBC) antigen in immunized mice significantly increased by different concentrations of S. megalantha extract. In conclusion, our results indicated that S. megalantha extract could be an immunomodulator plant and a good candidate for further investigations in order to develop a natural compound as an immunomodulator and anticancer agent.