Immune Response In Rats Research Articles

Identification of hub genes related to silicone-induced immune response in rats.

Silicone implants are used widely in the field of plastic surgery and are used in a large population. However, their safety profile, especially the silicone-induced immune response, has been a major concern for plastic surgeons for decades. It has been hypothesized that there is a cause and effect relation between silicone and immunity, but this is controversial. The objective of the present study was to determine the hub genes and key pathways related to silicone implant–induced immune responses in a rat model. In addition to cluster and enrichment analyses, we used weighted gene co-expression network analysis (WGCNA) to examine the gene expression profiles in a systematic context. A total five genes (Fes, Aif1, Gata3, Tlr6, Tlr2) were identified as hub genes that are most likely related to the silicone-induced immune response, four of which (Aif1, Gata3, Tlr6, Tlr2) have been associated with autoimmunity as target genes or disease markers. The Toll-like receptor signaling pathway (p < 0.01, fold enrichment: 7.01) and systemic lupus erythematosus signaling pathway (p < 0.05, fold enrichment: 5.01), which are considered strongly associated with autoimmunity, were significantly enriched in the silicone-implanted skin samples. The results indicate that silicone implants might trigger the localized immune response, as various immune reaction genes were detected after silicone implantation. The identified five hub genes will hopefully serve as novel therapeutic targets for silicone-related complications and the associated autoimmune diseases.

Dexmedetomidine ameliorates the inflammatory immune response in rats with acute kidney damage.

It has been demonstrated that dexmedetomidine (Dex) can protect patients with acute kidney injury from experiencing further tissue damage, however its mechanism of action remains unclear. The present study investigated the immune modulatory functions of Dex in rats with acute kidney injury (AKI) induced via injection of lipopolysaccharide into the tail vein. ELISA analysis showed that Dex reduced the levels of inflammatory cytokines in rats with AKI in a dose dependent manner. Furthermore, the regulatory effects of Dex on cytokine production disappeared when the α-2 adrenergic receptor antagonist Yohimbine (YOH) was added. For a detailed investigation on how Dex regulates the immune response in rats with AKI, the impact of Dex on the viability of splenocytes and lymphocytes was determined and it was determined that Dex did not influence splenocyte and lymphocyte viability. In addition, ELISA tests showed that Dex regulated the production of the T-helper (Th) 17 cytokines interleukin (IL)-17 and IL-23, but not the Th1 cytokine tumor necrosis factor α, in splenocytes and lymphocytes. To confirm whether Dex functioned as an α-2-adrenergic receptor in these immune regulations, YOH was administered together with Dex. When Dex and YOH were administered together, the regulatory functions of Dex were reduced, confirming that Dex acted as an agonist on the α-2-adrenergic receptor. Thus the results of the current study may provide novel insights regarding how Dex modulates immune functions in AKI.

Phosphorus protects cardiac tissue by modifying the immune response in rats infected by Trypanosoma cruzi

Aim: This study evaluates and correlates the number of myocarditis focuses and production of cytokines in Rattus norvegicus (Wistar lineage), experimentally infected with T. Cruzi and treated with Phosphorus.Methods: In two blind, controlled and randomized trials, 53 45-day-old, male animals were allocated into groups Control (n=24): Control group infected and treated with 7% hydroalcoholic solution, the preparation vehicle of the test medication; and Phosphorus (n=24 on days 0, 5, 10 and 24 after infection): group infected and treated with Phosphorus 13cH, diluted 10−26 and dynamized (test medication). The animals were inoculated intraperitoneally with 5×106 blood trypomastigotes of T. cruzi-Y strain. The medication was administered overnight (16 consecutive hours), diluted in water (1mL/100mL) in amber water bottles. The animals were treated 2days before and 2, 4, and 6days after infection. Enumeration of inflammatory foci in cardiac tissue (Hematoxylin-Eosin) and dosage of cytokines TNF-α and IFN-γ in the serum were performed on days 0, 5, 10 and 24 after infection, using three animals/group. Mann-Whitney, Friedman ANOVA, Spearman correlation (p<0.05), and Statistica Single User Software version 13.2 were used for data analysis.Results: The animals treated with Phosphorus 13cH had high concentration of INF-ɣ on the 5th day of infection with significant decrease on the 10th and 24th days (p<0.05), and high concentration of TNF-α on the 5th and 10th days of infection with decrease on the 24th day (p<0.05). The treatment with Phosphorus caused a significant increase of INF-ɣ and TNF-α on the 5th day of infection compared with the Control (p<0.05), with reestablishment on the 24th day, as well as in the Control group. The group treated with Phosphorus had 52.5% less number of myocarditis focuses in heart than Control group (p<0.05) on the 10th day of infection. The significant increase in cytokines on the5th day of infection in the Phosphorus group is related to a significant decrease in the number of inflammatory foci in cardiac tissue on the 10th day of infection in this group.Discussion and conclusion: Treatment with Phosphorus 13cH promotes beneficial effects in T. cruzi infection in Wistar rats by modulating the secretion of IFN-γ and TNF-α with decreased inflammation in cardiac tissue. These results reinforce the importance of considering the use of homeopathy for establishing new therapeutic approaches in the management of patients with Chagas disease.

Lymphocyte responses of rats vaccinated with cDNA encoding a phosphoglycerate kinase of Fasciola hepatica (FhPGK) and F. hepatica infection

Lymphocyte responses in the blood, peritoneal fluid and both mesenteric and hepatic lymph nodes of cDNA-FhPGK/pCMV vaccinated and/or Fasciola hepatica infected rats of both sexes were investigated to provide an insight into the immune responses that develop in different body compartments. The immune response that developed in cDNA-FhPGK/pCMV vaccinated females contributed to partial protection against F. hepatica infection (54% reduction in fluke recovery), while more liver flukes were found in the livers and bile ducts of cDNA-FhPGK/pCMV vaccinated male rats than in unvaccinated animals (increase of 13%). Rat sex not only affected the ultimate effectiveness of vaccination but also lymphocyte responses following vaccination and/or infection. Different CD4+ and CD8+ T cell profiles were noted in peritoneal fluid and lymph nodes, but not in blood, during acute and chronic fasciolosis. Moreover, independent lymphocyte responses developed in distinct body compartments. Immune responses of rats were polarized towards Th2/Treg with lymphocytes isolated from male rats showing higher IL-4 and IL-10 production than females. Lymphocyte proliferative capacities in response to mitogen (PHA) or vaccine antigen (FhPGK) were impaired in both sexes with a considerably higher reduction observed for males and restored lymphocyte proliferative capacities reported for females vaccinated with cDNA-FhPGK/pCMV during chronic fasciolosis.

Effects of Coriander Seeds Powder (Coriandrum sativum) as Feed Supplements on Growth Performance Parameters and Immune Response in Albino Rats

Herbs have acquired increasing interest as natural feed supplements and alternatives to antibiotics growth promoters in livestock production. The objectives of the current study to investigate the effects of supplementation by different levels of coriander seeds powder on growth performance indices and immune response of rats. A total of sixty albino rats were randomly assigned to four treatments with five replicates and fed for 30 days. The obtained results revealed that the final body weight, body gain, and body gain % in treatment groups fed diets supplemented with 3% and 5% coriander seeds powder were significantly higher as compared with the control group. Moreover, there were a significant improvement on feed utilization through feed intake and feed conversion ratio values obtained in treatment groups fed on 5%, 3%, and 2% coriander seeds powder in comparison with the control group. Interlukin-6 concentration level appeared significant improvement in rats fed 5% and 3% coriander seeds powder as compared with treatment group fed on 2% coriander seeds powder and the control group. From these result, it is concluded that coriander powder enhanced the growth performance and immune status of rats and can be used as natural feed supplements in their diets.

Physiological Effects of Whey- and Milk-Based Probiotic Yogurt in Rats.

In an in vitro experiment commercially available probiotic products were tested for the survival of bacteria under conditions of simulated human digestion either when used alone or mixed into yogurt. In the in vivo experiment the effects of feeding a whey- and milk-based yogurt prepared with the probiotic strain showing adequate survival in the in vitro experiment, was measured on body weight, feed con¬sumption and immune response of rats (IgG and IgA level after immunisation), on the composition and volatile fatty acid production of the intestinal microbiota and on the structure of intestinal villi. The Lactobacillus acidophilus (LA-15) strain had inadequate surviving ability in rats. Bifidobacterium animalis ssp. lactis (BB-12) improved the composition of the intestinal microflora, whereas whey-containing product had a mild immunostimulating effect and exerted a favourable influence on the morphology of intestinal villi. The consumption of yogurts increased the depth of crypts in the ileum, which resulted in enhanced secretion and thus softer faeces.

Nitric oxide synthase inhibitor, aminoguanidine reduces intracerebroventricular colchicine induced neurodegeneration, memory impairments and changes of systemic immune responses in rats

Intracerebroventricular (i.c.v.) injection of colchicine induces neurodegeneration, memory impairments and changes of some systemic immune responses in rats. Though the role of cox 2 in these colchicine induced changes have been evaluated, the influence of nitric oxide synthase (NOS) remains to be studied. The present study was designed to assess the role of NOS on the i.c.v. colchicine induced neurodegeneration, memory impairments and changes of some systemic immune responses by inhibiting its activity with aminoguanidine. In the present study the impairments of working and reference memories, neurodegeneration (chromatolysis and plaque formation) and changes of neuroinflammatory markers in the hippocampus (increased TNF α, IL 1β, ROS and nitrite) along with changes of serum inflammatory markers (TNF α, IL 1β, ROS and nitrite) and alteration of systemic immune responses (higher phagocytic activity of blood WBC and splenic PMN, higher cytotoxicity and lower leukocyte adhesion inhibition index of splenic MNC) were measured in the intracerebroventricular colchicine injected rats (ICIR). Administration of aminoguanidine (p.o. 30/50mg/kg body weight) to ICIR resulted in recovery of neuroinflammation and partial prevention of neurodegeneration which could be corroborated with the partial recovery of memory impairments in this model. The recovery of serum inflammatory markers and the systemic immune responses in ICIR was also observed after administration of aminoguanidine. Therefore, the present study shows that aminoguanidine can protect the colchicine induced neurodegeneration, memory impairments, and changes of systemic immune systemic responses in ICIR by inhibiting the iNOS.

Trikatu, an herbal compound ameliorates rheumatoid arthritis by the suppression of inflammatory immune responses in rats with adjuvant-induced arthritis and on cultured fibroblast like synoviocytes via the inhibition of the NFκB signaling pathway

The present study was designed to investigate the potential therapeutic effect of trikatu, an herbal compound and its underlying molecular mechanism in rats with adjuvant-induced arthritis (AIA). Our results indicate that trikatu (1000 mg/kg/b.wt. oral) administration suppressed the production of pro-inflammatory cytokines (tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, and monocyte chemoattractant protein (MCP)-1) and downregulated the mRNA expression levels of inflammatory mediators (TNF-α, IL-1β, IL-6, IL-17, MCP-1, receptor activator of nuclear factor kappa B ligand (RANKL), cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS)) and transcription factors (nuclear factor kappa B 65 (NFкB-p65) and activator protein-1 (AP-1)) in cultured AIA-fibroblast like synoviocytes and synovial tissue of AIA rats. Consistently, the protein expression of NFкB-p65, IL-17, TNF-α, COX-2, and RANKL was also dramatically reduced in cultured AIA-fibroblast like synoviocytes and synovial tissue of AIA rats by trikatu treatment. In addition, trikatu suppressed the expression and phosphorylation of NFкB-p65 similar to the Bay 11–7082 (NFкB inhibitor) in cultured AIA-fibroblast like synoviocytes. Furthermore, trikatu alleviated the histopathology of joint of arthritic rats. Overall, these data highlights that trikatu could be a promising alternative modality for the possible treatment of rheumatoid arthritis and other inflammatory diseases.

Effect of systemic injection of heterogenous and homogenous opioids on peripheral cellular immune response in rats with bone cancer pain: A comparative study.

Exogenous and endogenous opioids have been shown to modulate the immune system. Morphine-induced immunosuppression has been investigated extensively. However, the immune-regulating function of endogenous opioid peptides is unclear. The present study aimed to evaluate the difference in effects on cellular immune function between recombinant rat β-endorphin (β-EP; 50 µg/kg) and plant source morphine (10 mg/kg) via intraperitoneal injection treatment in a rat model of bone cancer pain. Walker 256 cells were injected into a tibial cavity injection to establish the bone cancer pain model. The paw withdrawal thresholds and body weights were measured prior to surgery, at 6 days after surgery, and following 1, 3,6 and 8 treatments. The spleen cells were harvested for detection of T cell proliferation, natural killer (NK) cell cytotoxicity, and the relative quantities of T cell subtypes (CD3+, CD4+ and CD8+ cells). Plasma levels of interleukin-2 (IL-2) were also determined. It was found that single or multiple treatments with β-EP (a homogenous opioid peptide) and morphine (a heterogenous opioid) had good analgesic effects on bone cancer pain, while the analgesia provided by morphine was stronger than that of β-EP. Treatment with β-EP 3, 6 and 8 times increased the body weight gain in the rat model of bone cancer pain, while morphine treatment had on effect on it. With regard to immunomodulatory functions, β-EP treatment increased T cell proliferation and NK cell cytotoxicity, and increased the relative quantities of T cell subtypes, but no effect on T cell secretion. However, morphine treatment decreased T cell proliferation and the levels of T cell subtypes. These data indicate that opioids from different sources have different effects on cellular immune function in vivo. A small dose of homogenous opioid peptide exhibited positive effects (analgesia and immune enhancement) on cancer pain. These results provide experimental evidence supporting the exploitation of human opioids for the treatment of cancer pain.

Cholinergic enzymes and inflammatory markers in rats infected by Sporothrix schenckii

The aim of this study was to evaluate the cholinesterase activity in serum, whole blood, and lymphocytes, as well as to verify its relation to immune response in rats experimentally infected by Sporothrix schenckii. For this study, 63 Wistar rats (Rattus norvegicus), male, adult were divided into three groups: the negative control group (GC: n = 21), the group infected subcutaneously (GSC: n = 21), and the group infected intraperitoneally (GIP: n = 21). The groups were divided into subgroups and the following variables were evaluated at 15, 30, and 40 days post-infection (PI): acetylcholinesterase (AChE) activity in lymphocytes and whole blood, butyrylcholinesterase (BChE) activity in serum, cytokines levels (IL-1, IL-6, TNFα, and INF-γ), immunoglobulins levels (IgA, IgG, IgM, and IgE), and protein profile by electrophoresis. Both infected groups showed increased levels of inflammatory parameters (P < 0.05) in tissue and inflammatory infiltrates. The activities of AChE in lymphocytes and BChE in serum increased (P < 0.05) significantly in animals from the GSC group on day 40 PI compared to the GC group. Regarding the GIP, there was a marked increase in the AChE activity in lymphocytes on days 30 and 40 PI, and in whole blood on days 15, 30, and 40 PI compared to GC. Furthermore, IL-10, an anti-inflammatory cytokine, was also present in high levels during chronic systemic S. schenckii infections in animals. Therefore, it is concluded that cholinesterase has an important modulatory role in the immune response during granulomatous infection by S. schenckii.

Cellular immune response in rats with 1,2-dimethylhydrazine-induced colon cancer after transplantation of placenta-derived multipotent cells

We describe the state of the immune system at the late stage of 1,2-dimethylhydrazine (DMH)-induced colon cancer and after administration of placenta-derived multipotent cells (PDMCs). The spleen and thymus indices did not differ among the groups of intact and DMH-treated rats and were not affected by the administration of placenta-derived multipotent cells following the DMH treatment. Moreover, no difference in spontaneous or stimulated phagocytic activity of peritoneal macrophages was observed between healthy rats or the animals with DMH-induced colon cancer (with or without the administration of PDMCs). However, the proliferation of the T cells in the spleen was lower in rats with colon cancer regardless of the administration of PDMCs. Similarly, no changes were observed in the cell cycle distribution of proliferating spleen cells after stimulation by lipopolysaccharide.Conclusion. Our data demonstrate the absence of the active reaction by peritoneal macrophages and spleen cells to a colon cancer at mid/late stage. Additionally, the administration of PDMCs does not result in a measurable anti-tumor immune response.

Enhancement of immunogenic response and protection in model rats by CSTM nanoparticles anticaries DNA vaccine.

To construct anticaries DNA vaccine and evaluate its ability to elicit mucosal and systemic immune responses in rats. wapA fragment was cloned into pVAX1 plasmid to generate pVAX1-wapA. The pVAX1-wapA/trimethyl chitosan nanoparticles were prepared by complex coacervation method. Significantly higher specific IgG antibody titers were observed in rats immunized with nanoparticles compared with rats immunized with naked pVAX1-wapA. Anti-WapA IgA and IgG antibody levels after intranasal immunization were significantly higher than those following intramuscular delivery of nanoparticles or naked pVAX1-wapA. Furthermore, fewer enamel, slight dentin and dentin moderate lesions were observed in rats immunized with nanoparticles. The results implicate WapA as an excellent candidate for anticaries vaccine development and nanoparticles as an effective delivery system.

ВЛИЯНИЕ ОДНОКРАТНОГО ВВЕДЕНИЯ ЭКЗОГЕННОГО ФАКТОРА РОСТА ГЕПАТОЦИТОВ НА ПОКАЗАТЕЛИ НЕСПЕЦИФИЧЕСКОЙ РЕЗИСТЕНТНОСТИ В УСЛОВИЯХ ПОСТРЕЗЕКЦИОННОГО ПОВРЕЖДЕНИЯ ПЕЧЕНИ

HGF is one of the factors taking part in regeneration and recovery of liver after injury. The article is dedicated to the study of influence of single HGF intake on the indices of non-specific immune response in rats after 70% liver resection in the early postoperative period. Research was conducted on 42 six-months white male Wistar rats of 250–300 g. It was revealed that level of segmentonuclear neutrophils and lymphocytes in the group with HGF intake stand at normal level on the 2nd day and normalizes to the 11th day that testifies to the decrease of inflammatory response. Increased level of leukocytes and monocytes is registered in the group with HGF intake. Also integrity of non-specific resistance in the early postoperative period on the 2nd day is registered. Suppression of phagocytosis indices with preservation of functional activity of phagocytes was registered to the 11th day.

Conjugation of ampicillin and enrofloxacin residues with bovine serum albumin and raising of polyclonal antibodies against them.

Aim:The aim of this study is to test the potency of bovine serum albumin (BSA) conjugated ampicillin (AMP) and enrofloxacin (ENR) antigens in eliciting an immune response in rats using indirect competitive enzyme-linked immunosorbent assay (icELISA).Materials and Methods:AMP and ENR antibiotics were conjugated with BSA by carbodiimide reaction using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) as a cross-linker. The successful conjugation was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Sprague-Dawley rats were immunized with the conjugates and blood samples were collected serially at 15 days time interval after first immunization plus first booster, second booster, third booster, and the fourth sampling was done 1½ month after the third booster. The antibody titres in the antisera of each antibiotic in all the four immunization cycles (ICs) were determined by an icELISA at various serum dilutions ranging from 1/100 to 1/6400.Results:Analysis of antibiotic-BSA conjugates by sodium dodecyl sulfate polyacrylamide gel electrophoresis and coomassie blue staining revealed high molecular weight bands of 85 kDa and 74 kDa for AMP-BSA and ENR-BSA respectively when compared to 68 kDa band of BSA. Both the antibiotic conjugates elicited a good immune response in rats but comparatively the response was more with AMP-BSA conjugate than ENR-BSA conjugate. Maximum optical density 450 value of 2.577 was recorded for AMP-BSA antisera, and 1.723 was recorded for ENR-BSA antisera at 1/100th antiserum dilution in third IC.Conclusion:AMP and ENR antibiotics proved to be good immunogens when conjugated to BSA by carbodiimide reaction with EDC as crosslinker. The polyclonal antibodies produced can be employed for detecting AMP and ENR residues in milk and urine samples.

Synergistic effects of glutamine, citrulline, and antioxidant vitamin C and E on modulating splenocytic immune responses in rats with intestinal ischemia and reperfusion

Intestinal ischemia and reperfusion (IIR) is a common event in patients with acute mesenteric ischemia, intestinal obstruction, organ transplantation, and hemorrhagic and septic shock. It has been demonstrated that IIR‐induced intestinal mucosa damage may cause systemic inflammation and multiple organ failure and further lead to death. Our previous study showed that oral glutamine/citrulline and antioxidant vitamins C/E have additive effects on attenuating intestinal damage and systemic inflammation in IIR rats. Herein, we investigated the effects of these supplements on IIR‐induced immune alterations. Male Wistar rats were divided into normal rats and IIR rats that underwent intestinal ischemia for 60 min and fed with placebo, glutamine/citrulline (AA), vitamins C/E (CE), or glutamine/citrulline plus vitamins C/E (AV) at 15 minutes before and 3, 9 and 21 hr after reperfusion. After 24 hr reperfusion, IIR‐increased plasma nitrate/nitrite was significantly reversed in rats with AA and CE and IIR‐increased interleukin‐6 was reversed in rats with AA, CE, and AV. The results of two‐way ANOVA showed that CE was a main factor to reverse IIR‐altered T‐helper leukocytes, B‐leukocytes, and monocytes and to increase tumor necrosis‐factor (TNF)‐alpha production in lipopolysaccharide‐stimulated leukocytes; and AA and CE were main factors to increase granulocytes in the circulation. In concanavalin A‐ and lipopolysaccharide‐stimulated splenocytes, AA was a main factor to increase TNF‐alpha and interleukin‐4 productions, CE was a main factor to increase interleukin‐4 productions, and AA and CE have significant interactions in TNF‐alpha and IL‐4 productions as shown in the significant increases in IIR rats with AV, not with AA or CE alone. In conclusion, oral glutamine/citrulline and vitamins C/E may have synergistic effects on modulating splenocytic immune responses in intestinal ischemia and reperfusion injury. The alleviated intestinal damage and systemic inflammation and the elevated splenocytic activity reveal that oral, combined administration of glutamine, citrulline, and antioxidant vitamins C and E may have beneficial effects in patients with intestinal ischemia and reperfusion.Support or Funding InformationNSC 98‐2320‐B‐030‐003‐MY3

Effects of stimulation of muscarinic acetylcholine receptors in medial septum on some immune responses in rats

ObjectivesThough the immunomodulatory role of medial septum (MS) has been indicated, but the contribution of the muscarinic acetylcholine (mAch) receptors presented in the internal network of the neurons of MS in this regard is not known. The aim of the present study is to assess the contribution of mAch receptors of MS on some immunological parameters. MethodsDifferent immunological parameters i.e. phagocytic activity of peripheral leukocytes, adhesibility and cytotoxicity of splenic mononuclear cells (MNC), delayed type of hypersensitivity (DTH) responses, Total Count (TC) and Differential Count (DC) of WBC with serum corticosterone (CORT) concentration have been measured after stimulation and blocking of mAch receptors of MS in rats. Ach or atropine has been micro infused into MS of separate groups of rats for stimulating or blocking of the mAch receptors respectively. ResultsIn Ach or atropine microinfused rats, the TC of WBC remained unaltered in the present study. The percentage of neutrophil has increased and the percentage of lymphocyte has decreased in Ach microinfused rats, but these parameters remain unaltered in atropine microinfused rats. The observed immunological parameters have increased after microinfusion of 0.60μM and 0.12μM of Ach and serum CORT concentration has decreased in those animals. These immunological parameters have decreased and serum CORT has increased after microinfusion of atropine (2 and 4mM) in MS of rats. ConclusionIt appears that the Ach receptors in MS can modulate the observed immunological parameters, and serum CORT plays an important role for these immune changes.

Cloning and expression of gene FanC-2NT encoding K99-2NT fimbrial antigen of enterotoxigenic Escherichia coli from diarrheic post-weaning piglets

Background and Purpose: The K99 (F5) is one pilus adhesin that mediates the attachment of enterotoxigenic E. coli (ETEC) strains to small intestines to cause to diarrhea in piglets, lambs and newborn calves. In this work, we carried out cloning and expression of the mature peptide of FanC subunit, K99 fimbriae, one of the most common adhesive antigens in E. coli. Materials and Methods: E. coli 2NT strain was isolated from fecal samples of post-weaning piglets with diarrhea. The coding sequence of the mature peptide of K99-2NT subunit was isolated by PCR amplification and cloned into pGEM®-T Easy vector for sequencing using fluorescent dideoxy-terminator method. Expression of K99-2NT protein which was inserted into pET200/D-TOPO vector induced with IPTG. The PCR product and expression level of protein was examined by agarose gel electrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively. Results and Conclusions: We cloned and expressed successfully the mature peptide of K99 subunit with molecular weight of approximately 17.5 kDa from E. coli 2NT strain (named K99-2NT). Nucleotide sequence of the K99-2NT subunit coding region of fanC-2NT gene is 477 bp in length and is 99% similarity with that of fanC gene (accession no: M35282). Highest expression level occurred after 12 h of induction with 0.75 mM IPTG at 37oC. This subunit antigen will be tested for immune response of rat in the next time.

Immune responses in rats and sheep induced by a DNA vaccine containing the phosphoglycerate kinase gene of Fasciola hepatica and liver fluke infection.

Immune responses of rats and sheep following vaccination with cDNA encoding phosphoglycerate kinase of Fasciola hepatica (cDNA-FhPGK/pCMV) and F. hepatica infection were investigated in the present study. cDNA-FhPGK/pCMV vaccinated female Sprague-Dawley rats were better protected by vaccination than their male counterparts - 48% reduction in fluke burden for females and no protection for males when compared with appropriate infection control groups. Moreover, male rats developed marked leukocytosis during the study with higher neutrophil, eosinophil and monocyte responses than females. Additionally, dynamics of eosinophil and monocyte responses varied between sexes. Increased titres of anti-FhPGK IgG1 and IgG2a correlated with the protective effect of vaccination that was observed among female rats. In the case of male sheep, no differences in worm burdens and in the course of the immune response were observed following vaccination. Titres of specific antibodies detected were low, and cellular responses were not significant. Apparently, sheep immune responses induced by cDNA-FhPGK/pCMV vaccination are not effective at controlling F. hepatica infection. Poor immunogenicity of DNA vaccines in large animals is still a major obstacle of this technology that has to be overcome.

Wound healing treatment by high frequency ultrasound, microcurrent, and combined therapy modifies the immune response in rats.

BACKGROUND: Therapeutic high-frequency ultrasound, microcurrent, and a combination of the two have been used as potential interventions in the soft tissue healing process, but little is known about their effect on the immune system. OBJECTIVE: To evaluate the effects of therapeutic high frequency ultrasound, microcurrent, and the combined therapy of the two on the size of the wound area, peritoneal macrophage function, CD4+ and CD8+, T lymphocyte populations, and plasma concentration of interleukins (ILs). METHOD: Sixty-five Wistar rats were randomized into five groups, as follows: uninjured control (C, group 1), lesion and no treatment (L, group 2), lesion treated with ultrasound (LU, group 3), lesion treated with microcurrent (LM, group 4), and lesion treated with combined therapy (LUM, group 5). For groups 3, 4 and 5, treatment was initiated 24 hours after surgery under anesthesia and each group was allocated into three different subgroups (n=5) to allow for the use of the different therapy resources at on days 3, 7 and 14 Photoplanimetry was performed daily. After euthanasia, blood was collected for immune analysis. RESULTS: Ultrasound increased the phagocytic capacity and the production of nitric oxide by macrophages and induced the reduction of CD4+ cells, the CD4+/CD8+ ratio, and the plasma concentration of IL-1β. Microcurrent and combined therapy decreased the production of superoxide anion, nitric oxide, CD4+-positive cells, the CD4+/CD8+ ratio, and IL-1β concentration. CONCLUSIONS:Therapeutic high-frequency ultrasound, microcurrent, and combined therapy changed the activity of the innate and adaptive immune system during healing process but did not accelerate the closure of the wound.

Replacement of inorganic zinc with lower levels of organic zinc (zinc nicotinate) on performance, hematological and serum biochemical constituents, antioxidants status, and immune responses in rats.

Aim:A study was undertaken to investigate the effect of organic zinc (zinc nicotinate, Zn-nic) supplementation (6, 9, and 12 ppm) compared to inorganic zinc (12 ppm) on growth performance, hematology, serum biochemical constituents oxidative stress, and immunity in weaned female Sprague–Dawley rats.Material and Methods:A 48 weaned rats (285.20±1.95 g) were randomly distributed to 4 dietary treatments with 6 replicates in each and reared in polypropylene cages for 10 weeks. Basal diet (BD) was formulated with purified ingredients without zinc (Zn). Four dietary treatments were prepared by adding 12 ppm Zn from ZnCO3 (control) and 6, 9, and 12 ppm Zn from Zn-nic to the BD. On 42nd day, blood was collected by retro-orbital puncture for analyzing hematological constituents, glucose, cholesterol, alkaline phosphatase, total protein, albumin, and globulin and antioxidant enzyme activities. At 43rd day, rats were antigenically challenged with sheep red blood cell (RBC) to assess humoral immune response and on 70th day cell-mediated immune response.Results:Weekly body weight gains, daily feed intake, blood hematological constituents (white blood cell, RBC, hemoglobin concentration, packed cell volume, mean corpuscular volume, lymphocyte, monocyte, and granulocyte concentration) and serum glucose, total protein levels were comparable among the rats feed Zn from ZnCO3 and Zn-nic (6, 9, and 12 ppm). Serum cholesterol reduced with organic Zn supplementation at either concentration (6-12 ppm). Serum globulin concentration reduced (p<0.05) with 6 ppm Zn-nic supplementation compared to other dietary treatments. Lipid peroxidation lowered (p<0.05) reduced with 12 ppm organic Zn; thiobarbituric acid reacting substances and protein carbonyls concentrations in liver reduced (p<0.05) with 9 and 12 ppm levels of organic Zn supplementation compared to 12 ppm Zn supplementation from inorganic source. RBC catalase and glutathione peroxidase enzymes activities were highest (p<0.05) in rats supplemented with 12 ppm Zn-nic, followed by 9 ppm. Comparable immune response (humoral and cell-mediated) was observed between 12 ppm inorganic Zn and 9 ppm organic Zn and higher (p<0.05) immune response was noticed at 12 ppm Zn-nic supplementation.Conclusion:Based on the results, it is concluded that dietary Zn concentration can be reduced by 50% (6 ppm) as Zn nicotinate without affecting growth performance, hemato-biochemical constituents, antioxidant status, and immunity. In addition, replacement of 12 ppm inorganic Zn with 12 ppm organic Zn significantly improved antioxidant status and immune response.