Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: UNIPROT:P43026 (lipopolysaccharide)
62,215 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Dendritic cells (DC) are the initiators of immune responses and are present in most tissues in vivo. To generate myeloid DC from monocytes (MoDC) in vitro the necessary cytokines are granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Using degenerated primers delineated from other species and rapid amplification of cDNA ends reverse transcription-polymerase chain reaction (RACE RT-PCR), the cDNA of equine (eq.) GM-CSF was cloned and found to have a point deletion at the 3'-end of eq.GM-CSF, resulting in a 24-nucleotide extended open reading frame not described in any species thus far. For differentiating eq.MoDC, monocytes were stimulated with eq.GM-CSF and eq.IL-4. The eq.MoDC was analysed by both light and electron microscopy and by flow cytometry and mixed lymphocyte reaction. The eq.MoDC obtained had the typical morphology and function of DC, including the ability to stimulate allogeneic T cells in a mixed lymphocyte reaction. In contrast to the human system, however, monocytes had to be differentiated for 6-7 days before immature DC were obtained. Our data also indicate that lipopolysaccharide or poly(I:C) alone are not sufficient to confer the full phenotypic transition into mature DC. Thus our study contributes to understanding the heterogeneity of immunity and adds important information on the equine immune system, which is clearly distinct from those of mice or man.
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PMID:Monocyte-derived dendritic cells from horses differ from dendritic cells of humans and mice. 1655 60

Pattern recognition proteins (PRPs), such as lipopolysaccharide and beta-1,3-glucan binding protein (LGBP), have been identified in many animals and play a crucial role in invertebrate defense systems. In the current study, an LGBP gene was cloned from fleshy prawn (Fenneropenaeus chinensis, Fc-LGBP) utilizing homology cloning and RACE methods. The full cDNA of the Fc-LGBP gene in fleshy prawn was 1253bp in size with a deduced 366 amino acid protein that includes a glycosyl hydrolase domain. Northern blot and RT-PCR data suggested that Fc-LGBP mRNA was mostly synthesized in haemocytes and that the expression was down-regulated 24h post-injection of bacteria. In situ hybridization demonstrated that Fc-LGBP mRNA was only detected in haemocyte cytoplasm, with no detection in other tissues. The molecular weight of the purified recombinantly expressed Fc-LGBP was approximately 46kDa. Immunohistochemistry of haemocytes revealed that Fc-LGBP protein was localized on the membrane of most cells. Data from bacterial binding assays utilizing purified protein suggested that rFc-LGBP had strong binding activity to Gram-negative bacteria.
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PMID:Molecular cloning and characterization of a lipopolysaccharide and beta-1,3-glucan binding protein from fleshy prawn (Fenneropenaeus chinensis). 1693 Jul 11

Toll-like receptors (TLRs) are an ancient family of pattern recognition receptors, which show homology with the Drosophila Toll protein and play key roles in detecting various non-self substances and then initiating and activating immune system. In this report, the full length of the first bivalve TLR (named as CfToll-1) is presented. CfToll-1 was originally identified as an EST (expressed sequence tag) fragment from a cDNA library of Zhikong scallop (Chlamys farreri). Its complete sequence was obtained by the construction of Genome Walker library and 5' RACE (rapid amplification of cDNA end) techniques. The full length cDNA of CfToll-1 consisted of 4308 nucleotides with a polyA tail, encoding a putative protein of 1198 amino acids with a 5' UTR (untranslated region) of 211bp and a 3'UTR of 500bp. The predicted amino acid sequence comprised an extracellular domain with a potential signal peptide, nineteen leucine-rich repeats (LRR), two LRR-C-terminal (LRRCT) motifs, and a LRR-N-terminal (LRRNT), followed by a transmembrane segment of 20 amino acids, and a cytoplasmic region of 138 amino acids containing the Toll/IL-1R domain (TIR). The deduced amino acid sequence of CfToll-1 was homologous to Drosophila melanogaster Tolls (DmTolls) with 23-35% similarity in the full length amino acids sequence and 30-54% in the TIR domain. Phylogenetic analysis of CfToll-1 with other known TLRs revealed that CfToll-1 was closely related to DmTolls. An analysis of the tissue-specific expression of the CfToll-1 gene by Real-time PCR showed that the transcripts were constitutively expressed in tissues of haemocyte, muscle, mantle, heart, gonad and gill. The temporal expressions of CfToll-1 in the mixed primary cultured haemocytes were observed after the haemocytes were treated with 1microgml(-1) and 100ngml(-1) lipopolysaccharide (LPS), respectively. The expression of CfToll-1 was up-regulated and increased about 2-fold at 6h with the treatment of 1microgml(-1) LPS. The expression of CfToll-1 was down-regulated with the treatment of 100ngml(-1) LPS. The results indicated that the expression of CfToll-1 could be regulated by LPS, and this regulation was dose-dependent.
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PMID:Molecular cloning and expression of a Toll receptor gene homologue from Zhikong Scallop, Chlamys farreri. 1715 63

A subtracted cDNA library from lipopolysaccharide (LPS)-stimulated Epinephelus awoara spleen was constructed using the suppression subtractive hybridization (SSH). Random clones (209) were selected and sequenced. After assembling, 36 contigs and 56 singlets (accession numbers: EB410743-EB410834) were finally obtained, some of which were immune-related genes. A reverse transcription-polymerase chain reaction (RT-PCR) analysis of the expression patterns of eight transcripts showed that seven of them were up-regulated after 24 h of LPS stimulation. Furthermore, full-length cDNAs of homologues of defender against cell death 1 (DAD1) and allograft inflammatory factor-1 (AIF-1) were obtained by RACE-PCR.
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PMID:Identification of differentially expressed genes in lipopolysaccharide-stimulated yellow grouper Epinephelus awoara spleen. 1721 Feb 58

A pattern recognition protein (PRP), lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) cDNA was cloned from the haemocyte of Chinese shrimp Fenneropenaeus chinensis by the techniques of homology cloning and RACE. Analysis of nucleotide sequence revealed that the full-length cDNA of 1,275 bp has an open reading frame of 1,098 bp encoding a protein of 366 amino acids including a 17 amino acid signal peptide. Sequence comparison of the deduced amino acid sequence of F. chinensis LGBP showed a high identity of 94%, 90%, 87%, 72% and 63% with Penaeus monodon BGBP, Litopenaeus stylirostris LGBP, Marsupenaeu japonicus BGBP, Homarus gammarus BGBP and Pacifastacus leniusculus LGBP, respectively. The calculated molecular mass of the mature protein is 39,857 Da with a deduced pI of 4.39. Two putative integrin binding motifs, RGD (Arg-Gly-Asp) and a potential recognition motif for beta-1,3-linkage of polysaccharides were observed in LGBP sequence. RT-PCR analysis showed that LGBP gene expresses in haemocyte and hepatopancreas only, but not in other tissues. Capillary electrophoresis RT-PCR method was used to quantify the variation of mRNA transcription level during artificial infection with heat-killed Vibrio anguillarum and Staphylococcus aureusin. A significant enhancement of LGBP transcription was appeared at 6 h post-injection in response to bacterial infection. These results have provided useful information to understand the function of LGBP in shrimp.
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PMID:Molecular cloning and characterisation of a pattern recognition protein, lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) from Chinese shrimp Fenneropenaeus chinensis. 1816 20

Anti-lipopolysaccharide factors (ALF) are a group of small basic proteins which are released into the hemolymph as a result of rapid degranulation of hemocytes in response to bacterial lipopolysaccharide (LPS). In the present study, using a combined approach of degenerate and RACE PCR, the gene coding for Scylla serrata anti-lipopolysaccharide factor (SsALF) was cloned and characterized. The full-length SsALF cDNA sequence consists of 607 bp and encodes a polypeptide of 97 amino acids, constituting a molecular mass of 11172 Da with an estimated pI of 10.01. The SsALF protein showed upto 92% similarity with ALF from Scylla paramamosain and about 33-53% amino acid sequence identity with other known ALF sequences. SsALF protein sequence demonstrated the presence of two highly conserved cysteine residues and putative LPS binding domain. An in vivo expression study showed that SsALF mRNA was expressed predominantly in hemocytes, heart and muscle of healthy mud crabs. The recombinant form of SsALF protein (rSsALF) was expressed with a Histag, in Escherichia coli, using the pTriEx-4 Ek/LIC vector. The purified rSsALF protein demonstrated antimicrobial activity against both Gram-positive and Gram-negative bacteria. The recombinant protein was able to significantly neutralize LPS-induced expression on SsALF in vivo as demonstrated by real-time PCR. rSsALF was able to permeabilize artificial phospholipid membranes as demonstrated by calcein enclosed liposome model. These studies strongly suggest that SsALF is one among the important antimicrobial factors produced in the crab during a microbial invasion.
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PMID:Identification, cloning, characterization and recombinant expression of an anti-lipopolysaccharide factor from the hemocytes of Indian mud crab, Scylla serrata. 1949 Sep 44

Marijuana cannabinoids, the endocannabinoids, and cannabinoid cell receptors have been shown to play important roles in immune regulation particularly as potent modulators of anti-inflammatory cytokines. The predominant cannabinoid receptor involved in this immune regulation is cannabinoid receptor 2 (CB(2)), which is predominantly expressed in B lymphocytes. However, the promoter region and mechanisms of CB(2) gene regulation are unknown in this immune cell type. Utilizing a combination of bioinformatics, 5' rapid amplification of cDNA ends (5' RACE), real-time reverse transcription-polymerase chain reaction, DNA sequencing, and luciferase reporter assays, we show that human B cells express one CB(2) transcript while mouse B cells express three CB(2) transcripts, with specific transcript selection occurring during B cell activation by lipopolysaccharide. Alignment of our sequenced RACE products to either the mouse or human genome, along with the GenBank submitted mRNA sequences, revealed that the transcripts we isolated contained previously unidentified transcriptional start sites (TSS). In addition, expression construct testing of the genomic region containing the TSSs of the mouse CB(2) exon 1 transcripts showed an eightfold increase of promoter activity over baseline. These data show for the first time that human B cells use only one TSS for CB(2) while mouse B cells use multiple TSSs and that the mouse TSSs are in a genomic area with promoter activity, thus suggesting the location of the gene promoter region. Defining these TSSs also provides clues to the various gene regulatory factors involved in the expression of CB(2) during B cell activation.
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PMID:Identification of transcription start sites and preferential expression of select CB2 transcripts in mouse and human B lymphocytes. 1975 78

Cecropin is a well-studied antimicrobial peptide that is synthesized in fat body cells and hemocytes of insects in response to hypodermic injury or bacterial infection. A 503 bp cDNA encoding for a cecropin-like peptide was isolated by employing annealing control primer (ACP)-based differential display PCR and 5'-RACE with immunized Papilio xuthus larvae. The open reading frame of the isolated cDNA encoded for a 62-amino acid prepropeptide with a putative 22-residue signal peptide, a 2-residue propeptide, and a 38-residue mature peptide with a theoretical mass of 4060.89 Da. The deduced amino acid sequence of the peptide evidenced a significant degree of identity with other lepidopteran cecropins. This peptide was named papiliocin. RTPCR results revealed that the papiliocin transcript was detected at significant levels after injection with bacterial lipopolysaccharide (LPS). On the basis of the deduced amino acid sequence of papiliocin, a 38-mer mature peptide was chemically synthesized via the Fmoc method, and its antimicrobial activity was analyzed. The synthetic papiliocin peptide evidenced a broad spectrum of activity against fungi, Gram-positive and Gram-negative bacteria, and also evidenced no hemolytic activity against human red blood cells.
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PMID:Characterization and cDNA cloning of a cecropin-like antimicrobial peptide, papiliocin, from the swallowtail butterfly, Papilio xuthus. 2021 10

CD29 is the integrin beta1 subunit. Integrin family members are membrane receptors involved in cell adhesion and recognition in a variety of processes including embryogenesis, tissue repair and immune response. In this study, a novel CD29-like gene (LCD29) was identified and characterized in Japanese lamprey (Lethenteron japonicum), an agnathan that occupies a critical phylogenic position between cephalochordates and gnathostomes. After a partial cDNA sequence of LCD29 was found from the leucocyte cDNA library, the full-length cDNA was obtained by means of 3' and 5' RACE, respectively. LCD29 encodes 780 amino acids and shares high sequence homology with other vertebrates. Both real-time PCR and immunohistochemical assays have demonstrated the wide distribution of the LCD29 in lamprey tissues, and FACS analysis has shown that the expression level of this protein is higher in granulocytes than in lymphocytes. Furthermore, after the lampreys were stimulated with lipopolysaccharide, the levels of LCD29 mRNA were obviously up-regulated in the leucocytes, intestine, heart and gill tissues. Most importantly, an interaction between LCD29 and LCD9 in the intestine has been found with co-immunoprecipitation assays for the first time. The results obtained in the present study will help us to understand the function of LCD29 in immune response in jawless vertebrates.
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PMID:A novel CD29-like protein expressed in Japanese lamprey (Lethenteron japonicum) and involved in immune response. 2048 45

The lipopolysaccharide -and beta-1,3-glucan-binding protein (LGBP) is a pattern recognition receptor, which is fundamental for the innate immune response of crustaceans. A LGBP gene was cloned from the haemocytes of Portunus trituberculatus using SMART RACE methods. The full-length LGBP cDNA (1 378 bp) had a 1 095 bp open reading frame encoding a protein of 365 amino acid residues including a 16 amino acid residues signal peptide, a 138 bp 5' untranslated region (UTR) and a 144 bp untranslated region in the 3' UTR with a 29 bp polyA tail. The calculated molecular mass of the mature protein (349 amino acid residues) is 39,825.24 with an estimated pI of 4.49. The gene sequence and secondary structure of LGBP were analyzed by bio-informatics. Additionally, a Glyco hydro 16 domain was identified. The expression of P. trituberculatus in various tissues were detected through RT-PCR methods. The results showed that the LGBP gene expressed in all the tissues detected, including haemocytes, hepatopancreas, heart, gills and muscle. In response to the challenge of Staphyloccocus aureus and Vibrio alginolyticus, the LGBP gene expression in haemocytes of the group challenged with mixed bacteria were higher than the control group within 48 h. It suggested that the LGBP gene plays an active role in immunologic process against bacterial infection.
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PMID:[Cloning and expression pattern analysis of a lipopolysaccharide -and beta-1,3-glucan-binding protein in Portunus trituberculatus]. 2117 47


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