Influenza is a major respiratory viral illness caused by infection of influenza A virus (IAV), which still persists in various global seasonal epidemics every year. the host immune response is a key factor that determines the severity of influenza infection, present an attractive target for the development of new therapies for the treatment.
Among the double track signal transduction that regulates the activation of the host immune response and function in response to infection IAV, the mitogen-activated protein kinase (MAPK) pathway that wicks important signaling, downstream receptors pattern recognition (PRRS), activated by IAVs that regulate various processes cell in the immune cells of both the innate and adaptive immunity.
In addition, aberrant activation of MAPK overexuberant underlying the production of inflammatory mediators, promoting the development of a “cytokine storm”, the characteristics of severe respiratory viral diseases. Therefore, explanation of MAPK regulatory role in the immune response to IAVs not only important for understanding the pathogenesis of severe influenza, but it is also important to develop a MAPK-dependent therapies for the treatment of respiratory viral diseases. In this review, we will summarize the current understanding of the function of MAPK in both the innate and adaptive immune responses to IAVs and discuss their contribution to the cytokine storm caused by the highly pathogenic influenza virus.
Trimeric soluble recombinant influenza A virus hemagglutinins (HA) and tetrameric neuraminidases (NA) has proven to be an excellent tool to decipher biological properties. Receptor binding and cleavage of sialic acid by a recombinant protein correlated satisfactory compared with the whole virus. Expression of HA and NA can be achieved in many different laboratories host.
For the study of immunology and receptor interactions, however, insect and mammalian cell expressed proteins are preferred because of the presence of N-linked glycosylation and disulfide bond formation. Because the expression of mammalian cells is widely applied, yield enhancement expression is an important goal. Here we report that the use of codon-optimized genes and fusion sfGFP, HA expression results can be improved
Regulation of Host Immune Responses against Influenza A Virus Infection by Mitogen-Activated Protein Kinases (MAPKs)
Comparative Analysis of Antiviral Activity of IgG and IgA antibodies to Influenza A Virus M2 Protein
The influenza A virus (IAV) matrix-2 (M2) protein is a viral envelope proteins conserved antigens that plays an important role in virus starters together with other envelope proteins, hemagglutinin (HA). M2-specific mouse monoclonal antibody IgG, rM2ss23, which binds to the ectodomain of the M2 protein, has been shown to be non-neutralizing antibodies, but inhibits plaque formation IAV strains.
In this study, we produce rM2ss23 chimeric (ch-rM2ss23) IgG and IgA antibodies with the same variable region and compared their antiviral activity. Using gel chromatography, ch-rM2ss23 IgA is divided into three subsets of antibodies: IgA monomer (m-IgA), dimeric IgA (d-IgA), and trimeric and tetrameric IgA (t / q-IgA).
Description: For angiogenesis to occur, endothelial cells must escape their stable location and break through the basement membrane. Cells migrate toward an angiogenic stimulus that may be released from nearby tumor cells. These cells proliferate to form new blood vessels. Our Endothelial Tube Formation Assay (In Vitro Angiogenesis) provides an easy, robust system to assess angiogenesis in vitro. The ECM gel matrix very closely resembles an in vivo environment.
Description: The Lactose Assay Kit measures total lactose in milk based food products or biological samples such as blood or urine. Lactose is cleaved into glucose and galactose. Glucose is then oxidized, yielding hydrogen peroxide and D-gluconic acid. The hydrogen peroxide is detected by a fluorometric probe.
Description: Bilirubin, a byproduct of heme breakdown, can exist conjugated to glucuronic acid (direct) and as unconjugated (indirect). The unconjugated form is found in the blood bound to albumin and is transported to the liver. Bilirubin becomes conjugated to glucuronic acid in the liver, making it more soluble and allowing for excretion into bile. High levels of bilirubin have been correlated with jaundice and Gilbert?s syndrome while low levels have been associated with cardiovascular disease and diabetes mellitus.
Description: Our Pyruvate Assay Kit measures pyruvate in biological samples. First, pyruvate is oxidized by pyruvate oxidase, producing hydrogen peroxide. The hydrogen peroxide is then detected at ex. 530-570 nm/em. 590-600 nm using a specific fluorometric probe. Pyruvate levels in unknown samples are determined based on the provided pyruvate standard curve.
Description: Cell Biolabs? Urea Assay Kit is based on the Berthelot reaction. Urea is first degraded into ammonia and carbon dioxide, which further reacts with an alkaline developer to produce a blue-green colored product that can be measured with a standard spectrophotometric plate reader at an optical density between 580-630 nm. Each kit provides sufficient reagents to perform up to 192 assays, including blanks, urea standards and unknown samples.
Description: Our Rac Activation Assays use visible agarose beads to selectively precipitate the active form of Rac1 or Rac2. The precipitated small GTPase is then detected by Western blot using a Rac1- or Rac2-specific antibody included in the kit.
Description: Our Rac Activation Assays use visible agarose beads to selectively precipitate the active form of Rac1 or Rac2. The precipitated small GTPase is then detected by Western blot using a Rac1- or Rac2-specific antibody included in the kit.
Description: Our Cdc42 Activation Assays use visible agarose beads to selectively precipitate the active form of Cdc42 protein. The precipitated small GTPase is then detected by Western blot using a Cdc42-specific antibody included in the kit.
Description: Our Rho Activation Assays use visible agarose beads to selectively precipitate the active form of RhoA, RhoB or RhoC. The precipitated small GTPase is then detected by Western blot using a RhoA-, RhoB- or RhoC-specific antibody included in the kit.
Description: Our Rho Activation Assays use visible agarose beads to selectively precipitate the active form of RhoA, RhoB or RhoC. The precipitated small GTPase is then detected by Western blot using a RhoA-, RhoB- or RhoC-specific antibody included in the kit.
Description: Our Rho Activation Assays use visible agarose beads to selectively precipitate the active form of RhoA, RhoB or RhoC. The precipitated small GTPase is then detected by Western blot using a RhoA-, RhoB- or RhoC-specific antibody included in the kit.
Description: Our Arf Activation Assays use visible agarose beads to selectively precipitate the active form of Arf1 or Arf 6. The precipitated small GTPase is then detected by Western blot using an Arf1- or Arf6-specific antibody included in the kit.
Description: Our Arf Activation Assays use visible agarose beads to selectively precipitate the active form of Arf1 or Arf 6. The precipitated small GTPase is then detected by Western blot using an Arf1- or Arf6-specific antibody included in the kit.
Description: Our Ral Activation Assay uses visible agarose beads to selectively precipitate the active form of Ral protein. The precipitated small GTPase is then detected by Western blot using a Ral-specific antibody included in the kit.
Description: Our Ran Activation Assay uses visible agarose beads to selectively precipitate the active form of Ran protein. The precipitated small GTPase is then detected by Western blot using a Ran-specific antibody included in the kit.
Description: Cell Biolabs? Phosphatidylcholine Assay Kit measures the phosphatidylcholine present within serum, plasma, or tissue samples.Samples are compared to a known concentration of phosphatidylcholine standard within the 96-well microtiter plate format. Samples and standards are incubated for 60 minutes and then read with a standard 96-well fluorometric plate reader.
Description: Cell Biolabs? Sphingomyelin Assay Kit is a simple fluorometric assay that measures the amount of sphingomyelin present in plasma or serum, tissue homogenates, or cell suspensionsin a 96-well microtiter plate format. Each kit provides sufficient reagents to perform up to 96 assays, including blanks, sphingomyelin standards and unknown samples. Sample sphingomyelin concentrations are determined by comparison with a known sphingomyelin standard.
Description: Glutamate is a non-essential amino acid that serves as an important neurotransmitter in the mammalian brain and has a key role in cellular metabolism. Excess glutamate levels in the brain can cause cell injury and death, leading to neurological diseases. Our Glutamate Assay Kit is a quantitative, fluorometric assay that uses glutamate specific enzymes to generate hydrogen peroxide. An ADHP probe is oxidized by hydrogen peroxide to generate fluorescent Resorufin, which correlates to the level of glutamate in the sample. Glutamate levels in an unknown sample are calculated based on a glutamate standard curve.
Description: The Hydroxyproline Assay Kit is a quantitative colorimetric assay for measuring the hydroxyproline concentration in protein samples, including collagen where it is found almost exclusively.
Description: Histamine is naturally occurring in food, with high concentrations associated with spoiled and fermented foods. Exposure to high levels of histamine through the ingestion of food can cause symptoms similar to an allergic response. Our Histamine Assay Kit detects total histamine from food samples using a colorimetric probe. Reduction of the probe yields color development proportional the histamine levels in the sample. Absorbance at 450nm is read after a one hour incubation at 37C and histamine levels are calculated based on a histamine standard curve.
We found that t / q-IgA have significantly higher capacity to reduce plaque size of IAVs of m-IgG and IgA, most likely caused by a decrease in the number of offspring produced virus particles from infected cells. Interestingly, HA-M2 colocalization was greatly reduced on the surface of infected cells with antibodies ch-rM2ss23. These results indicate that the polymeric IgA anti-M2 Limit beginner IAV more efficient than IgG and suggested the role of anti-M2 IgA cross-protective immunity IAVs.