Detection of IL-20R1 and IL-20R2 mRNA inC57BL/6 Mice Astroglial Cells and Brain Cortex Following LPS Stimulation

Astrocytes, which comprise 90% of overall brain mass, are involved in brain immunity. These cells represent the non-professional class of CNS-resident APCs and may promote or inhibit CNS inflammation depending on the cytokines they secrete. IL-10 family of cytokines and their receptors, IL-20R1 and IL-20R2, may have a role in shifting astrocytes to a neuroprotective or neurodegenerative function. To address the expression of IL-20R1 and IL-20R2 cytokine receptors in astrocytes and brain cortex of C57BL/6 mice. We investigated the expression of IL-20R1 and IL-20R2 in C57BL/6 mice astroglial cells and brain cortex in response to lipopolysaccharide [LPS], using reverse-transcription polymerase chain reaction [RTPCR] method. Astrocytes were able to express IL-20R1 and IL-20R2 mRNA not only in response to LPS stimulation but also in the absence of LPS. Furthermore, we found the expression of IL-20R1 and IL-20R2 mRNA in the cortex of adult C57BL/6 mice. IL-20R1 and IL-20R2 are constitutively express in the brain. Since most neuropathological processes involve astrocytes and inflammatory cytokines, these findings have important implications for future therapeutic strategies

Study of the expressions of IL-20R1 and IL-20R2 in C57BL/6 mice astroglial cells

Astrocytes are the most abundant glial cell type. They may promote or inhibit CNS inflammation depending on which cytokines are secreted. Astrocytes also have immune roles. IL-19, IL-20, and IL-24 activate a heterodimer receptor composed of the IL-20R1 alpha-chain and the IL-20R2 beta-chain. It has long been considered that signaling by these receptor complexes affects immunological reactions, however the biological functions of IL-20R1 and IL-20R2 in the brain remain unclear. As the first step to address the role of these cytokine receptors in the brain, in this study we have researched the expressions of IL-20R1 and IL-20R2 in C57BL/6 mice astrocytes. We examined expressions of IL-20R1 and IL-20R2 proteins in mice astroglial cells and in the 1321N1 astrocytoma cell line in response to MOG, LPS and GM-CSF by flow cytometry. The effect of LPS on mRNA expression of IL-20R1 and IL-20R2 was investigated by RT-PCR. We provide, for the first time, evidence that astrocytes expressed IL-20R1 and IL-20R2 mRNA not only in response to LPS stimulation but also in unstimulated astrocytes. We did not observe the expressions of IL-20R1 and IL-20R2 proteins in mice astroglial cells and the 1321N1 astrocytoma cell line. IL-20R1 and IL-20R2 mRNA are constitutively expressed in astrocytes. Because the majority of neuropathological processes involve astrocytes and inflammatory cytokines, the results of this study, which are reported for the first time, have important implications for future research

[Study on co-administration of GM-CSF, IL-15 on the enhancement of the immunogenicity of HIV-1 P24-Nef expression vector in BALB/c mouse model]

Today, AIDS is considered as a global problem and many efforts to generate an effective vaccine against this disease have been made, but remain inconclusive. DNA vaccines are a member of the new generation of vaccines that can efficiently stimulate the immune system. However, recent findings indicate low immunogenicity for these vaccines and it is believed that these types of vaccines require strategies that could infer more immunogenicity. The employment of adjuvants could be considered as one of the most important methods involved. In this study, a DNA vaccine candidate for HIV P24-Nef is constructed and then using genetic adjuvants IL-15 and GM-CSF, cellular immune responses have been studied. In this study the gene structure of HIV P24-Nef in eukaryotic expression vector was constructed and expression vectors of IL-15 and GM-CSF were used as adjuvants. After inoculation of the candidate vaccine to BALB/c mice, cytokine patterns, lymphocytes proliferation and cytotoxicity were analyzed. Our findings indicate that candidate vaccine significantly stimulated cellular immune responses. The usage of IL-15 and GM-CSF as DNA adjuvants together and separately with candidate vaccine has strengthened cellular immune responses significantly. Co-administration of DNA adjuvants significantly increased cellular immune responses when the ratio of the vaccine dose was more than the adjuvants. The sequences that we selected as candidate vaccine demonstrated good immunogenicity in mouse model and co-administration of IL-15 and GM-CSF DNA adjuvants increased cellular immune response to DNA vaccine construct

[Evaluation of immune responses from immunization of HIV-1 P24-Nef fusion peptide vaccine with monophosphoryl lipid A [MPL] in BALB/c mice]

Several vaccines against HIV have been investigated but none has been approved as an effective HIV vaccine. An approach that could induce stronger immune response against the pathogen is utilizing a multi-epitopic vaccine. This strategy was used in the design of several vaccines and resulted in improved immune responses. In this study a multi-epitopic fusion peptide including parts of HIV-1 Nef and P24 as a vaccine candidate was injected into mice and immune humoral responses measured with total antibody and IgG sub-classes using ELISA. Also measurement of cellular immune responses through evaluation of spleen cells proliferation response using MTT and cytotoxicity by LDH were performed. Finally, the cytokine pattern of IFN-gamma and IL-4 were also determined with ELISA. The results indicate that candidate vaccine stimulated mouse splenic lymphocyte proliferation response and also induced strong cytotoxicity responses. Analysis of humoral immune response has shown that the candidate vaccine has induced specific antibody production mainly of the IgG2a sub-class. Also cytokine pattern evaluation has shown that IFN-gamma secretion was dominant. The use of immunogen and conserved epitopes from P24 and Nef induced strong humoral and cellular immune responses and this construct could be candidate for further studies in animal models

Immunogenicity of a new HIV-1 DNA Construct in a BALB/c mouse model

Cell mediated immunity, especially cytotoxic T cell responses against HIV-1 infection, plays a critical role in controlling viral replication and disease progression. DNA vaccine is a novel technology which is known to stimulate strong cellular immune responses. Many DNA vaccines have been tested for HIV infection but there is still no effective vaccine against this infection. Construction of a vaccine consisting of multiple conserved and immunogenic epitopes may increase vaccine efficacy. In the present study a DNA vaccine candidate constructed from HIV-1 P24-Nef was evaluated and cellular immune responses were assessed in murine BALB/c model. HIV-1 P24-Nef gene was cloned in PCDNA3.1 expression vector. Mice were immunized with DNA construct and IL-4 and IFN-gamma evaluation was per-formed using ELISPOT. Cytotoxicity response was evaluated with Granzyme B ELIS-POT assay and lymphocyte proliferation was evaluated with LTT assay. Analysis of immune responses showed that, compared to control groups, the candidate vaccine induced production of higher levels of both IL-4 and IFN-gamma [p<0.05]. Cytotoxicity and lymphocyte proliferation responses of mice vaccinated with the candidate vaccine were significantly increased compared to control groups [p<0.05]. HIV-1 P24-Nef DNA construct displayed strong immunogenicity in a murine model

Air pollution induced asthma and alterations in cytokine patterns

In recent decades, clinicians and scientists have witnessed a significant increase in the prevalence of allergic rhinitis and asthma. The factors underlying this phenomenon are clearly complex; however, this rapid increase in the burden of atopic disease has occurred in parallel with rapid industrialization and urbanization in many parts of the world. Consequently, more people are exposed to air pollutants than at any point in human history. Worldwide increases in allergic respiratory disease have mainly been observed in urban communities. Epidemiologic and clinical investigations have suggested a strong link between particulate air pollution and detrimental health effects, including cardiopulmonary morbidity and mortality. The purpose of this review is to provide an evidence-based summary of the effects of air pollutants on asthma, focusing on particulate matter PMs, diesel exhaust particles [DEPs], and ozone as major air pollutants. An overview of observational and experimental studies linking these pollutants with asthma will be provided, followed by consideration of the mechanisms underlying pollutant induced immune response and inflammation. The cytokine response will be viewed in depth and a brief discussion of future research and clinical directions is provided

Immunobiological consequences of sulfur mustard contamination

Sulfur mustard has been employed in chemical warfare in certain regions including Iran. The short and long term biological effects of sulfur mustard contamination have been studied in both basic and clinical aspects. Sulfur mustard has been shown to induce a vast array of pathological effects in affected persons. In addition to skin, lung, eyes and gastrointestinal disturbances, sulfur mustard has been shown to induce hematological complications and a severe suppression of the immune system. The short and long term immunological [both cellular and humoral], hematological, genetic and biochemical consequences of persons exposed to sulfur mustard are extensively reviewed here. The long term complications of these patients indicate the need to develop effective preventive and therapeutic strategies in the clinic. These strategies may be based upon immunopotentiating intervention and therapy