Immune Response In Rats Research Articles

Ferric‐Tannic Nanoparticles Inhibit Early‐Stage Hepatocarcinogenesis by Activating Tumor Immune Responses in Rats

AbstractEarly diagnosis and treatment of hepatocellular carcinoma (HCC) remain major challenges. Significant efforts have been made to find new approaches to address these issues. Ferric‐tannic nanoparticles (FTs) have emerged as promising tools for targeting the early phase of hepatocarcinogenesis due to their preferential accumulation in preneoplastic liver lesions. In this study, the therapeutic potential of FTs is demonstrated in early‐stage hepatocarcinogenesis in rats. FTs inhibit the progression of early hepatocarcinogenesis, reducing hepatic nodules, preneoplastic foci (glutathione S‐transferase placental (GST‐P) form‐positive foci), and HCC cell proliferation. The therapeutic effects of FTs appear to be mediated by inhibiting cell proliferation through the activation of immune responses. FTs show promise as novel immunomodulators or therapeutic agents for the treatment of early‐stage HCC.

Level of Cytokines and C3 Complement in the Blood of Rats under Conditions of Chronic Unpredictable Stress of Different Durations.

The parameters of the cytokine profile and functional activity of the complement system in the blood of rats were studied during different time periods of chronic unpredictable mild stress using a model of sequentially alternating low-intensity stress effects for 1, 2, 3, and 4 weeks. In the dynamics of observation, a general tendency towards multidirectional fluctuations in the concentration of cytokines was revealed: an increase in IL-10, but a decrease in IL-4 in comparison with the control. Statistically significant changes in the level of IL-10 were noted after 2, 3, and 4 weeks, IL-4 - after 2 and 4 weeks of stress loads. The percentage of lysis of the C3 component in rats gradually increased by the 2nd week of chronic stress, but then decreased and practically did not differ from the control values (intact animals) by the end of the study. These results illustrate the specificity of changes in the indicators of the C component of the complement system and the cytokine profile of the blood reflecting activity of the cellular and humoral components of the immune response in rats exposed to repeated stress factors of different origins and duration.

Activation of RIG-I signaling in the early stage of Paragonimus proliferus infection causes lung injury via type I immune response in rat.

Paragonimiasis is a common zoonotic parasitic disease. The retinoic acid-inducible gene I (RIG-I) signaling is very important for the host to recognize invading pathogens (especially viruses and bacteria). However, the role of RIG-I signaling in the early stages of P. proliferus infection remains unclear. Therefore, in this study, Sprague-Dawley (SD) rat models with lung damage caused by P. proliferus were established. Experimental methods including Enzyme-linked Immuno Sorbent Assay (ELISA), real-time fluorescent quantitative polymerase chain reaction (PCR), western blotting, and hematoxylin and eosin (HE) staining were used to explore the mechanisms of lung injury caused by P. proliferus. As a result, the expression of the mRNA and proteins of RIG-I signal-related key target molecules, including RIG-I, tumor necrosis factor (TNF) receptor associated factor 6 (TRAF6), interferon regulatory Factor 7 (IRF7), IPS-1, and downstream C-X-C chemokine ligand 10 (CXCL10), were significantly up-regulated immediately after infection, peaked at 3 or 7 days, and showed a downward trend on after 14 days. The levels of pro-inflammatory cytokines interleukin-1 (IL-1), interferon (IFN)-α, -β, and -γ, which represent type 1 immune response, gradually increased and reached a peak by 14 days, which was consistent with the changes in the degree of inflammatory damage observed under HE staining of lung tissues. In conclusion, RIG-I signaling is activated in the early stage (before 14 days) of P. proliferus infection, it is inferred that the lung injury of the host may be related to the activation of RIG-I like signaling to induce type I immune response.

Propionic acid affects the synaptic architecture of rat hippocampus and prefrontal cortex

It is well documented that propionic acid (PPA) produces behavioral, morphological, molecular and immune responses in rats that are characteristic of autism spectrum disorder in humans. However, whether PPA affects the ultrastructure and synaptic architecture of regions of autistic brain has not been adequately addressed. Earlier we show that single intraperitoneal (IP) injection of PPA (175 mg/kg) produces superficial changes in the spatial memory and learning of adolescent male Wistar rats. However, in neurons, synapses and glial cells of hippocampal CA1 area and medial prefrontal cortex transient (mainly) or enduring alterations were detected. In this study, we used electron microscopic morphometric analysis to test the effect of PPA on different structural parameters of axodendritic synapses of the hippocampus and prefrontal cortex. The animals were treated with a single IP injection of PPA (175 mg/kg). The length and width of synaptic active zone, the area of presynaptic and postsynaptic mitochondria, the distance between presynaptic mitochondria and the synapse active zone, the distance between postsynaptic mitochondria and postsynaptic density and the depth and opening diameter of neuronal porosome complex were evaluated. Our results show that synaptic mitochondria of the hippocampus and prefrontal cortex are the most vulnerable to PPA treatment: in both regions, the area of postsynaptic mitochondria were increased. In general, our results show that even small dose of PPA, which produces only superficial effects on spatial memory and learning is able to alter the synapse architecture in brain regions involved in cognition and autism pathogenesis. Therefore, the microbiome may be involved in the control of neurotransmission in these regions.

Synergistic effect of ferulic acid and caffeic acid on metabolic syndrome and immune response in rats

Synergistic effect of ferulic acid and caffeic acid on metabolic syndrome and immune response in rats

Effect of alcoholic extract of Agaricus Bisporus on blood profiles and immune response in rats with aspergillosis

Effect of alcoholic extract of Agaricus Bisporus on blood profiles and immune response in rats with aspergillosis

Protective efficacy of a recombinant enterotoxin antigen in a maternal immunization model and the inhibition of specific maternal antibodies to neonatal oral vaccination

ETEC (enterotoxigenic Escherichia coli) infection is the leading cause of profuse watery diarrhea in mammals, especially among pre-weaning and post-weaning piglets in swine industry. Maternal immunization is a current rational strategy for providing protection to susceptive piglets and reducing the incidence of ETEC-associated diarrhea. Here we evaluated the protective efficiency of a recombinant antigen (MBP-SLS) fused by major enterotoxin subunits (STa-LTB-STb) via a maternal immunization model, and the impacts of maternal antibodies to neonatal oral vaccination were also investigated in the neonates. The high titers of specific IgG and sIgA in pups shown that the maternal antibodies could be transferred passively. Furthermore, the increases of IL-6 and IL-10 cytokines in breast milk and pup serum indicated that immunization on mother could effectively boost the immune system of neonates. Newborn rats from immunized mothers showed a 70% survival rate after ETEC infection. However, the mucosal immune responses of neonates were inhibited by the high level of maternal specific antibodies. Among the oral-vaccinated neonates, born from mock-immunized rats reached the highest survival after ETEC challenge. Collectively, the fusion MBP-SLS antigen could effectively induce strong immune responses in rats during pregnancy and the pups could receive passive protection through specific antibodies transferred via milk and placenta. However, the specific maternal antibodies exhibited an inhibition effect on the mucosal immune responses in offspring.

The effect of miR-155-5p on M1 polarization of Kupffer cells and immune response during liver transplantation through regulating the expression of KDM5D.

To explore the effect and its specific mechanism of miR-155-5p on M1 polarization of Kupffer cells (KCs) and immune response in liver transplantation (LT) through KDM5D. Primary KCs were isolated from Wistar rats and identified by cell culture, ink-swallowing test and flow cytometry. The cells identified as KCs were induced into LT acute rejection (AR) model cells by LPS/IFN-γ, flow cytometry was used for cell sorting and apoptosis detection. Enzyme-linked immunosorbent assay (ELISA) kit was used to detect the levels of inflammatory factors, macrophages and liver function markers. RT-qPCR detected the expression of miR-155-5p and KDM5D mRNA. The protein expression of KDM5D was detected by Western blot. Dual luciferase reporter gene experiment verified the targeting relationship between miR-155-5p and KDM5D. The separated KCs adhered after being cultured for 24h, had pseudopodia and phagocytosis, and the proportion of F4/80 positive cells was more than 90%. The expression of miR-155-5p was increased in LPS/IFN-γ-induced KCs. And knockdown of miR-155-5p inhibited H3K4me3 and H3K27me3 of TNF-α promoter, M1 polarization of KCs and the immune response of AR model cells by upregulating KDM5D. In animal experiments, knockdown of miR-155-5p was found to inhibit liver damage and immune response in rats with allogeneic orthotopic LT. These results confirmed that miR-155-5p inhibited M1 polarization of KCs induced by LPS/IFN-γ, thereby alleviating AR and liver function impairment after LT by upregulating KDM5D.

Histochemical effects of brodifacoum on rat spleen

In this study, the histochemical effects of Brodifacoum, an anticoagulant used against rodents, on the spleen are examined under a light microscope using CD4 and CD8 histochemical staining methods. A single dose of 0.2 mg Brodifacoum was dissolved in Dimethyl Sulfoxide (DMSO) and was given orally to mature male rats. Spleen samples were collected under ether anesthesia after 24 h, 72 h, 14 days, and 30 days from the rats in the experimental groups and after 14 days from the rats in the control group. In this light microscope study, it was observed that the capsule, white pulp, and red pulp zones in the rat spleen were constructed normally and as their natural structures primary and secondary follicles (germinal center) they were few, and CD4 and CD8 lymphocytes were spherically structured. In the 24 h spleens of the rats, the diameters of germinal centers were expanded and deterioration of the structure of CD4 and CD8 cells was observed. Related to the increase in time (72 h and 14 days) it was determined that primary follicles increased in number and the diameters of germinal centers expanded. In addition to this, after30 days, the rate of CD4:CD8 of the brodifacoum applied rat spleens were approximately the rate of the control group, and the improvement of the structures of the cells was reported as an effect of regeneration. As a result of this study, it was found that Brodifacoum caused immunohistochemical abnormalities in the rat spleen, affected the morphological structure of CD4 and CD8 T lymphocytes and created an immune response in rats. It is thought that the obtained results will be a source for the studies on Brodifacoum.

Immune response strategies of Brucella melitensis and their antigens in rats

Brucella melitensis is an intracellular bacterium and is the main brucella species that cause abortion and placenta retention in sheep and goats. It has many mechanisms to evade the immune response. The current study aimed to investigate Brucella melitensis strategies for producing immune responses in rats after challenging the bacterium. For this purpose, live and killed Brucella melitensis REV1 strain was given to rats subcutaneously, and immunological markers like TLR2, TLR4, IFN- γ, and anti-brucella antibodies were determined. The results showed that the level of immunological markers like TLR2 and TLR4 did not significantly increase in rat groups inoculated with live Brucella melitensis, while it increased in the rats’ groups vaccinated with the sonicated Brucella melitensis; also, the results showed an increase in the level of IFN-γ and anti-brucella antibody titers in all animal groups. The study concluded that the inoculation with killed bacteria and REV1 could protect the animals against challenging doses, as seen when the groups were inoculated with challenge dose of the bacterium.

Preparation, Purification and Performance Evaluation of Polyclonal Antibody Against SARS-CoV-2 Produced in Rat.

Among all known human coronaviruses, some viruses (e.g., SARS-CoV-2) cause severe pneumonia or even death. With the regard to its spread and the importance of its rapid identification/treatment, and because pAbs are relatively cheap, able to bind to more sites on antigens and even neutralize them, this study was done for the production and purification of anti-SARS-CoV-2 polyclonal antibodies (pAb) in rats. Viral antigen purification was performed by PEG/NaCl precipitation. The efficiency of the sucrose cushion method was also investigated to produce a purer antigen. Immunization was done and antibody purification was performed by ammonium sulfate precipitation (33%), dialysis, and ion-exchange chromatography. Western blotting and enzyme-linked immunosorbent assay (ELISA) were performed to verify the antibody specificity. All data were analyzed by SPSS software. The results showed that the amount of concentrated virus increased with the increase of PEG concentration. Moreover, the sucrose cushion method increased its purity. Besides, induction of immune response in rats for pAb production with high titers was reached via these antigens and ELISA/western blot results indicated a suitable antibody-antigen interaction. Additionally, it was shown that ion-exchange chromatography could be a suitable technique for IgG purification. Herein, we presented a simple and cheap method for the purification of whole viral particles with relatively high quality. The results verified that these antigens could elicit a good immune response in the rat. The obtained pAbs showed a good specificity toward SARS-CoV-2 antigens. Accordingly, this study proposes a promising method for viral vaccine development.

Oral Delivery of SARS-CoV-2 DNA Vaccines Using Attenuated Salmonella typhimurium as a Carrier in Rat.

The 2019 novel coronavirus disease (COVID-19) is the disease that has been identified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but the prophylactic treatment of SARS-CoV-2 is still under investigation. The effective delivery of eukaryotic expression plasmids to the immune system's inductive cells constitutes an essential requirement for generating effective DNA vaccines. Here, we have explored the use of Salmonella typhimurium as vehicles to deliver expression plasmids orally. The attenuated Salmonella phoP was constructed by the one-step gene inactivation method, and plasmid-encoded the spike protein of SARS-CoV-2 was transform into the Salmonella phoP by electroporation. Western blot experiment was used for the detection of SARS-CoV-2 expression on 293T cells. Wistar rats were immunized orally with Salmonella that carried a eukaryotic expression plasmid once a week for three consecutive weeks. The ELISA was performed to measure the SARS-CoV-2 specific IgG at rat's serum samples. pSARS-CoV-2 can be successfully expression on 293T cells, and all immunized animals generated immunity against the SARS-CoV-2 spike protein, indicating that a Salmonella-based vaccine carrying the Spike gene can elicit SARS-CoV-2-specific secondary immune responses in rats. Oral delivery of SARS-CoV-2 DNA vaccines using attenuated Salmonella typhimurium may help develop a protective vaccine against SARS-CoV-2 infection.

Newly Method for Preparation of Live Attenuated Vaccine from Local Methicillin-resistant Staphylococcus aureus Isolate and Study its Effect on Immune Response in Rats

Newly Method for Preparation of Live Attenuated Vaccine from Local Methicillin-resistant Staphylococcus aureus Isolate and Study its Effect on Immune Response in Rats

Immunization with SARS-CoV-2 Nucleocapsid protein triggers a pulmonary immune response in rats.

The SARS-CoV-2 pandemic have been affecting millions of people worldwide, since the beginning of 2020. COVID-19 can cause a wide range of clinical symptoms, which varies from asymptomatic presentation to severe respiratory insufficiency, exacerbation of immune response, disseminated microthrombosis and multiple organ failure, which may lead to dead. Due to the rapid spread of SARS-CoV-2, the development of vaccines to minimize COVID-19 severity in the world population is imperious. One of the employed techniques to produce vaccines against emerging viruses is the synthesis of recombinant proteins, which can be used as immunizing agents. Based on the exposed, the aim of the present study was to verify the systemic and immunological effects of IM administration of recombinant Nucleocapsid protein (NP), derived from SARS-CoV-2 and produced by this research group, in 2 different strains of rats (Rattus norvegicus); Wistar and Lewis. For this purpose, experimental animals received 4 injections of NP, once a week, and were submitted to biochemical and histological analysis. Our results showed that NP inoculations were safe for the animals, which presented no clinical symptoms of worrying side effects, nor laboratorial alterations in the main biochemical and histological parameters, suggesting the absence of toxicity induced by NP. Moreover, NP injections successfully triggered the production of specific anti-SARS-CoV-2 IgG antibodies by both Wistar and Lewis rats, showing the sensitization to have been well sufficient for the immunization of these strains of rats. Additionally, we observed the local lung activation of the Bronchus-Associated Lymphoid Tissue (BALT) of rats in the NP groups, suggesting that NP elicits specific lung immune response. Although pre-clinical and clinical studies are still required, our data support the recombinant NP produced by this research group as a potential immunizing agent for massive vaccination, and may represent advantages upon other recombinant proteins, since it seems to induce specific pulmonary protection.

Abatement of cyclophosphamide-induced splenic immunosuppressive indoleamine 2, 3-dioxygenase and altered hematological indices in Wister rats by dietary quercetin

The immunotoxicity mediated by cyclophosphamide (CYP) was earlier reported. Quercetin, due to its anti-oxidative/inflammatory properties elicits a plethora of health benefits. However, the influence of quercetin on the splenic/immunotoxicity linked with CYP-induced indoleamine 2,3-dioxygenase (IDO) is unavailable in the literature. We investigated the effects of quercetin on the splenic immunosuppressive IDO and hematological indices of immune response in rats. Animals were treated with CYP (100 mg/kg) alone or co-treated (CYP + quercetin [100 mg/kg + 50 mg/kg respectively]) at days 1, 3, 5, and 7. Results revealed that CYP treatment alone significantly provoked an oxidative-inflammatory response, increased serum kynurenine concentration, and concomitantly increased immunosuppressive IDO and tryptophan 2, 3-dioxygenase (TDO), in the spleen as well as altering hematological indices. Quercetin co-treatment enhanced activities of antioxidant enzymes, inhibited myeloperoxidase (MPO) activity, lowered levels of nitric oxide, interferon-Υ (IFN-Υ), and interleukin-6 (IL-6), and reduced kynurenine concentration as well as IDO/TDO activities. Quercetin co-treatment augmented white blood cell (WBC), CD4-T cells, and other hematological indices of the immune response. In conclusion, quercetin prevented CYP-induced alterations in immune response in rats by lowering the activities of immunosuppressive IDO and TDO, inhibiting oxidative-inflammatory stress, diminution of kynurenine concentrations, and augmenting hematological parameters.

Irisin Activates M1 Macrophage and Suppresses Th2-Type Immune Response in Rats with Pelvic Inflammatory Disease.

Objective To investigate the mechanism of irisin to treat rats with acute pelvic inflammatory disease (APID). Methods Female rats were established as APID. Firstly, the content of IL-6, IL-8, TNF-α, and NF-kB was tested in rats' serum by enzyme linked immunosorbent assay (ELISA). Interferon-γ (IFN-γ) and IL-4 in the supernatant of pelvic homogenates were also detected. The mRNA expression of the inducible nitric oxide synthase (iNOS), TNF-α, chemokine ligand 1 (CXCL1), arginase-1(Arg1), and chitinase-3-like-3 (Chi313) genes in the pelvic cavity was detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). IFN-γ and IL-4 secreted by spleen CD4+T cells and CD8+T cells were counted by flow cytometry, and the ratio of IFN-γ/IL-4 in CD4+T cells and CD8+T cells in the spleen was also detected by flow cytometry. Results Irisin reduced the levels of IL-6, IL-8, TNF-α, and NF-kB in serum. Compared with the APID group, the expression level of IL-4 in the APID + Irisin group was reduced in the homogenate. At the same time, Irisin promotes the activation of M1 macrophages in the uterus, ovaries, and uterine tubes of rats with APID. Irisin also inhibited Th2-type immune response. Conclusions Irisin activates M1 macrophage and suppresses Th2-type immune response in APID rats.

Immunotoxic Effects of Cypermethrin in Male Wistar Rats: Attenuation by Co-Administration of Zinc and Alpha-Lipoic Acid

Aim The present study investigated the effects of cypermethrin exposure on humoral and cellular immune response in rat and its attenuation by zinc and alpha-lipoic acid. Methods Cypermethrin at the dose levels of 40 mg and 80 mg/kg body weight were orally administered and pre-treatment of zinc (227 mg/L in drinking water) and alpha-lipoic acid (35 mg/kg body wt.) were done. Total leukocyte and differential leukocyte counts (DLC), phagocytic index, serum nitric oxide (NO) activity, total immunoglobulin concentration, quantitative hemolysis, proliferation assay of blood mononuclear cells were estimated and histological examination of spleen was accomplished. Results Total white blood cell (WBC) count and percentage of lymphocyte, serum nitric oxide activity (p<0.001) and quantitative hemolysis were increased significantly increased whereas neutrophil %, total serum immunoglobulin, and blood mononuclear cell proliferation (p<0.001) and the phagocytic function of peritoneal macrophages were significantly reduced in cypermethrin treated rats compared to control group rats at a dose-dependent manner. Zinc and alpha-lipoic acid pre-treatment reversed the results. Conclusion From the findings it can be concluded that the co-administration of zinc and alpha-lipoic acid significantly attenuated the immunotoxic effects in cypermethrin exposed rat.

Astaxanthin Mitigates Thiacloprid-Induced Liver Injury and Immunotoxicity in Male Rats.

Thiacloprid (TCP) is a widely used neonicotinoid insecticide with a probable toxic hazard to animals and human beings. This hazard has intensified the demand for natural compounds to alleviate the expected toxic insults. This study aimed at determining whether astaxanthin (ASX) could mitigate the hepatotoxic effect of TCP and diminish its suppressive effect on immune responses in rats. Animals received TCP by gavage at 62.1 mg/kg (1/10th LD50) with or without ASX at 40 mg/kg for 60 days. Intoxicated rats showed modulation of serum transaminases and protein profiles. The hemagglutination antibody titer to sheep red blood cells (SRBC) and the number of plaque-forming cells in the spleen were reduced. The cell-mediated immunity and phagocytosis were suppressed, while serum interleukins IL-1β, IL-6, and IL-10 were elevated. Additionally, malondialdehyde, nitric oxide, and 8-hydroxy-2′-deoxyguanosine levels were increased in the liver, spleen, and thymus, with depletion of glutathione and suppression of superoxide dismutase and catalase activities. The expressions of inducible nitric oxide synthase and the high mobility group box protein 1 genes were upregulated with histomorphological alterations in the aforementioned organs. Cotreatment with ASX markedly ameliorated the toxic effects of TCP, and all markers showed a regression trend towards control values. Collectively, our data suggest that the protective effects of ASX on the liver and immune system of TCP-treated animals depend upon improving the antioxidant status and relieving the inflammatory response, and thus it may be used as a promising therapeutic agent to provide superior hepato- and immunoprotection.

Preliminary study on serum immunoglobulin G responses following intramuscular inoculation of adjuvanted polyhydroxyalkanoate microparticles with Pasteurella multocida vaccine in white rats

A natural biodegradable polymer, polyhydroxyalkanoate (PHA), was adjuvanted with a vaccine seed to observe the biomaterial's ability in enhancing an immune response in rats. The adjuvant potential of PHA was tested using the whole-killed Pasteurella multocida B:2 (PMB2) vaccine in Sprague Dawley (SD) rats to detect changes in serum immunoglobulin G (IgG) and immunoglobulin M (IgM) responses. A common PHA, poly(3-hydroxybutyrate) [P(3HB)], from Bacillus megaterium UMTKB-1 was constructed into microparticles using the solvent evaporation method. Twelve SD rats were divided into four treatment groups: 1) non-treatment as negative control, 2) P(3HB) adjuvant, 3) PMB2 vaccine, and 4) adjuvanted-P(3HB)/PMB2 vaccine groups, which were intramuscularly vaccinated twice. Immunoglobulins IgG and IgM levels were used as markers of the immune response induced by the adjuvanted-P(3HB)/PMB2 vaccine and analysed over an eight-week study period. The group vaccinated specifically with adjuvanted-P(3HB)/PMB2 vaccine had higher concentrations of immunoglobulins compared to other treatment groups, hence demonstrating the potential of the adjuvant to enhance immune response. Findings showed a need to delay the delivery of the second booster dose to determine the appropriate regime for the adjuvanted-P(3HB)/PMB2 vaccine.

Immunostimulatory Activity of Aqueous Extract of Polyherbal Formulation on Th1/Th2 Cytokines Secretion and Cell Mediated Immune Response in Rats

Immunostimulatory Activity of Aqueous Extract of Polyherbal Formulation on Th1/Th2 Cytokines Secretion and Cell Mediated Immune Response in Rats