G. Sandra Tjabringa , Joost B. Vos , Diana Olthuis , Dennis K. Ninaber , Klaus F. Rabe , Joost Schalkwijk , Pieter S. Hiemstra , Patrick L.J.M. Zeeuwen
{"title":"Host defense effector molecules in mucosal secretions","authors":"G. Sandra Tjabringa , Joost B. Vos , Diana Olthuis , Dennis K. Ninaber , Klaus F. Rabe , Joost Schalkwijk , Pieter S. Hiemstra , Patrick L.J.M. Zeeuwen","doi":"10.1016/j.femsim.2005.03.004","DOIUrl":"10.1016/j.femsim.2005.03.004","url":null,"abstract":"<div><p><span><span><span>Mucosal secretions contain a range of defense effector molecules including antimicrobial peptides and </span>proteinase inhibitors. These molecules play a central role in host defense against infection, and in a variety of immune and inflammatory reactions. The aim of this study was to analyze the levels of </span>neutrophil<span> defensins, the cathelicidin<span> hCAP-18/LL-37, and the proteinase inhibitors secretory leukocyte proteinase inhibitor, SKALP/elafin and cystatin M/E in various mucosal secretions and urine. We show here that especially </span></span></span>seminal plasma is characterized by high concentrations of hCAP-18/LL-37, SLPI, SKALP/elafin and cystatin M/E. The results of this study demonstrate that each mucosal secretion is characterized by a unique profile of effector molecules, which may supply individual mucosal secretions with specific properties related to the control of local infection and inflammation.</p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 151-158"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.03.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25218519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ross J. Langley , Dervla Kenna , Josefin Bartholdson , Dominic J. Campopiano , John R.W. Govan
{"title":"Temperate bacteriophages DK4 and BcepMu from Burkholderia cenocepacia J2315 are identical","authors":"Ross J. Langley , Dervla Kenna , Josefin Bartholdson , Dominic J. Campopiano , John R.W. Govan","doi":"10.1016/j.femsim.2005.06.001","DOIUrl":"10.1016/j.femsim.2005.06.001","url":null,"abstract":"","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 349-350"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.06.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25183035","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Expression and secretion of the protective antigen of Bacillus anthracis in Bacillus brevis","authors":"Gi-eun Rhie , Young-Mia Park , Jeong-hoon Chun, Cheon-Kwon Yoo, Won-Keun Seong, Hee-Bok Oh","doi":"10.1016/j.femsim.2005.05.008","DOIUrl":"10.1016/j.femsim.2005.05.008","url":null,"abstract":"<div><p>We used the <span><em>Bacillus brevis</em></span>-pNU212 system to develop a mass production system for the protective antigen (PA) of <span><em>Bacillus anthracis</em></span>. A moderately efficient expression-secretion system for PA was constructed by fusing the PA gene from <em>B</em>. <em>anthracis</em> with the <em>B</em>. <em>brevis</em><span> cell-wall protein signal-peptide encoding region of pNU212, and by introducing the recombinant plasmid, pNU212-mPA, into </span><em>B</em>. <em>brevis</em> 47-5Q. The clone producing PA secreted about 300<!--> <!-->μg of recombinant PA (rPA) per ml of 5PY-erythromycin medium after 4 days incubation at 30<!--> <!-->°C. The rPA was fractionated from the culture supernatant of <em>B</em>. <em>brevis</em><span><span><span> 47-5Q carrying pNU212-mPA using ammonium sulfate at 70% saturation followed by anion exchange chromatography on a Hitrap Q, a Hiload 16/60 Superdex 200 </span>gel filtration column and a phenyl </span>sepharose hydrophobic interaction column, yielding 70</span> <!-->mg rPA per liter of culture. The N-terminal sequence of the purified rPA was identical to that of native PA from <em>B</em>. <em>anthracis</em><span><span>. The purified rPA exhibited cytotoxicity towards J774A.1 cells when combined with lethal factor. The rPA formulated in either Rehydragel HPA or MPL-TDM-CWS adjuvant (Ribi-Trimix) elicited the expression of a large amount of anti-PA and </span>neutralizing antibodies in guinea pigs and completely protected them against a 100 LD50 challenge with fully virulent </span><em>B</em>. <em>anthracis</em><span> spores.</span></p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 331-339"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.05.008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25183036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jennifer P. Wang, Tomoko Hayashi, Sandip K. Datta, Richard S. Kornbluth, Eyal Raz, Donald G. Guiney
{"title":"CpG oligonucleotides partially inhibit growth of Mycobacterium tuberculosis, but not Salmonella or Listeria, in human monocyte-derived macrophages","authors":"Jennifer P. Wang, Tomoko Hayashi, Sandip K. Datta, Richard S. Kornbluth, Eyal Raz, Donald G. Guiney","doi":"10.1016/j.femsim.2005.05.007","DOIUrl":"10.1016/j.femsim.2005.05.007","url":null,"abstract":"<div><p><span><span>Immunostimulatory DNA sequences and their synthetic </span>oligonucleotide<span> analogs (CpG-ODN) activate innate immunity and can stimulate antibacterial effects against numerous intracellular pathogens. While it has been shown previously that CpG-ODN inhibit growth of </span></span><span><em>Mycobacterium avium</em></span> in murine and human macrophages, we now report that <span><em>Mycobacterium tuberculosis</em></span> growth can be inhibited by CpG-ODN treatment of human monocyte-derived macrophages (hMDM). This inhibitory effect was reversed by IFN-γ, which has been shown repeatedly to enhance the growth of virulent <em>M. tuberculosis</em> in cultured hMDM. The antibacterial effect of CpG-ODN in human macrophages was specific for <em>M. tuberculosis</em> when compared to other intracellular pathogens including <span><em>Listeria monocytogenes</em></span> and <em>Salmonella enterica</em> serovar Dublin. These data indicate that CpG-ODN can improve the ability of hMDM to contain growth of virulent <em>M. tuberculosis</em>.</p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 303-310"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.05.007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25174883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Charles S. Berenson , Kelly B. Sayles , Jing Huang , Vernon N. Reinhold , Mary Alice Garlipp , Herbert C. Yohe
{"title":"Nontypeable Haemophilus influenzae-binding gangliosides of human respiratory (HEp-2) cells have a requisite lacto/neolacto core structure","authors":"Charles S. Berenson , Kelly B. Sayles , Jing Huang , Vernon N. Reinhold , Mary Alice Garlipp , Herbert C. Yohe","doi":"10.1016/j.femsim.2005.03.007","DOIUrl":"10.1016/j.femsim.2005.03.007","url":null,"abstract":"<div><p>Nontypeable <em>Haemophilus influenzae</em><span> (NTHI) are a major cause of human infections. We previously demonstrated high affinity and high specificity binding of NTHI to minor gangliosides of human respiratory (HEp-2) cells and macrophages, but not to brain gangliosides. We further identified the NTHI-binding ganglioside of human macrophages as α2,3-sialylosylparagloboside (IV</span><sup>3</sup>NeuAc-nLcOse<sub>4</sub><span>Cer, nLM1), which possesses a neolacto core structure that is absent in brain gangliosides. This supported a hypothesis that lacto/neolacto core carbohydrates are critical for NTHI-ganglioside binding. To investigate, we determined the core carbohydrate structure of NTHI-binding gangliosides of HEp-2 cells, through multiple approaches, including specific enzymatic degradation<span>, mass spectral analysis and gas–liquid chromatography. Our analyses denote the following critical structural attributes of NTHI-binding gangliosides: (1) a conserved lacto/neolacto core structure; (2) requisite sialylation<span>, which may be either internal or external, with α2,3 (human macrophages) or α2,6 (HEp-2 cells) anomeric linkages; (3) internalized galactose residues. Mass spectral and gas chromatographic analyses confirm that NTHI-binding gangliosides of HEp-2 cells possess lacto/neolacto carbohydrate cores and identify the structure of the major peak as NeuAcα2-6Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1Cer (α2,6-sialosylparagloboside, nLM1). Collectively, our studies denote NTHI-binding gangliosides as lacto/neolacto series structures.</span></span></span></p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 171-182"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.03.007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25218423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Catherina Eliszbeth Hook, Malgosia K. Matyszak, John S. Hill Gaston
{"title":"Infection of epithelial and dendritic cells by Chlamydia trachomatis results in IL-18 and IL-12 production, leading to interferon-γ production by human natural killer cells","authors":"Catherina Eliszbeth Hook, Malgosia K. Matyszak, John S. Hill Gaston","doi":"10.1016/j.femsim.2005.02.010","DOIUrl":"10.1016/j.femsim.2005.02.010","url":null,"abstract":"<div><p>Control of infection by <span><em>Chlamydia trachomatis</em></span><span> usually requires the production of interferon-γ. Whilst this can be produced by CD4+ and CD8+ T lymphocytes, natural killer (NK) cells are another important source of this cytokine, and are known to be recruited early to the infected genital tract. We show that both IL-12 and IL-18, which synergise to stimulate NK cells to produce interferon-γ, are produced following the infection of dendritic cells and epithelial cells respectively, since supernatants from infected cells could substitute for recombinant cytokines. These results suggest that conditions, which lead to NK cell production of interferon-γ will be present at the site of infection, where epithelial cells are the primary targets of infection and dendritic cells within the epithelium can also access the bacterium.</span></p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 113-120"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.02.010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25218514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Determination of the cytotoxic effect of Clostridium histolyticum culture supernatant on HeLa cells in the presence of protease inhibitors","authors":"Jarosław Jóźwiak , Aldona Komar , Ewa Jankowska , Gayane Martirosian","doi":"10.1016/j.femsim.2005.03.005","DOIUrl":"10.1016/j.femsim.2005.03.005","url":null,"abstract":"<div><p><span><em>Clostridium histolyticum</em></span><span> culture supernatant contains numerous enzymes, which exert a cytotoxic effect on host cells. This includes lethal toxin, clostripain and high-potassium-sensitive toxin. Since the number of </span><em>C. histolyticum</em><span> infections increased during the last several years, it seems worthwhile to evaluate whether protease inhibitors, used for the treatment of many diseases, could influence toxicity, and thus, pathogenicity of </span><em>C. histolyticum</em><span><span>. In this study we evaluated in vitro the influence of four common protease inhibitors: aprotinin, </span>phenylmethylsulphonyl fluoride (PMSF), </span><span>l</span><span>-1-chloro-3-[4-tosylamido]-7-amino-2-heptanone-HCl (TLCK) and chymostatin on the toxicity of </span><em>C. histolyticum</em><span> supernatant towards human epithelial HeLa cells. We show that aprotinin has no effect, while PMSF, TLCK and chymostatin potentiate the cytotoxic activity of </span><em>C. histolyticum</em><span>, probably by hindering natural defence mechanisms of cells. In addition, PMSF and TLCK block clostripain enzymatic activity, while chymostatin leaves it intact. Elevated cytotoxicity of the supernatant is not related to the quantity of high-potassium-sensitive toxin, as was reported previously, since desalted supernatant still exerted its strong toxic effect. Our results show that addition of protease inhibitors for treating diseases complicated by concurrent </span><em>C. histolyticum</em> infection must require special attention.</p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 137-142"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.03.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25218517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Cloning of mce1 locus of Mycobacterium leprae in Mycobacterium smegmatis mc2 155 SMR5 and evaluation of expression of mce1 genes in M. smegmatis and M. leprae","authors":"Ramachandran Sarojini Santhosh, Shunmugiah Karutha Pandian , Nirmala Lini, Abdul Khader Shabaana , Avuthu Nagavardhini, Kuppamuthu Dharmalingam","doi":"10.1016/j.femsim.2005.05.004","DOIUrl":"10.1016/j.femsim.2005.05.004","url":null,"abstract":"<div><p><span>Plasmid pSET152 is a broad host range mobilizable vector which integrates into streptomyces chromosome utilizing </span><em>att</em> site and <em>int</em> function of ∅C31. Transformation of this plasmid into <span><em>Mycobacterium </em><em>smegmatis</em></span> mc<sup>2</sup> 155 SMR5 gave stable transformants carrying the pSET152 as an integrated copy. Integration occurred at the cross over sequence 5′TTG disrupting the <em>gatA</em> gene (Glu-tRNA<sup>Gln</sup> amidotransferase subunitA), which is non-essential under conditions used. Recombinant pSET152 plasmids carrying <em>mce1</em> locus of <span><em>Mycobacterium leprae</em></span> were used to construct <em>M. smegmatis</em> transformants carrying the <em>mce1</em> locus in their chromosome. RT-PCR analysis revealed specific transcripts of <em>M. leprae mce</em> in <em>M. smegmatis</em><span>. The transcribed mRNA carried intergenic regions between genes of </span><em>mce1</em> locus indicating that <em>mce1</em> locus is an operon. Examination of <em>M. leprae</em><span> specific mRNA from lepromatous leprosy patient’s biopsy showed that </span><em>mce</em> locus is transcribed as an operon in the pathogen also.</p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 291-302"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.05.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25131051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mark S. Pearson , Donald P. McManus , Danielle J. Smyth , Fred A. Lewis , Alex Loukas
{"title":"In vitro and in silico analysis of signal peptides from the human blood fluke, Schistosoma mansoni","authors":"Mark S. Pearson , Donald P. McManus , Danielle J. Smyth , Fred A. Lewis , Alex Loukas","doi":"10.1016/j.femsim.2005.03.009","DOIUrl":"10.1016/j.femsim.2005.03.009","url":null,"abstract":"<div><p><span>Proteins secreted by and anchored on the surfaces of parasites are in intimate contact with host tissues. The transcriptome of infective cercariae of the blood fluke, </span><span><em>Schistosoma mansoni</em></span><span><span>, was screened using signal sequence<span> trap to isolate cDNAs encoding predicted proteins with an N-terminal signal peptide. Twenty cDNA fragments were identified, most of which contained predicted signal peptides or transmembrane regions, including a novel putative seven-transmembrane receptor and a membrane-associated mitogen-activated protein kinase. The developmental expression pattern within different life-cycle stages ranged from ubiquitous to a transcript that was highly upregulated in the cercaria. A bioinformatics-based comparison of 100 signal peptides from each of schistosomes, humans, a </span></span>parasitic nematode and </span><em>Escherichia coli</em><span> showed that differences in the sequence composition of signal peptides, notably the residues flanking the predicted cleavage site, might account for the negative bias exhibited in the processing of schistosome signal peptides in mammalian cells.</span></p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 201-211"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.03.009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25218424","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Handan Akbulut , S. Sirri Kilic , Vedat Bulut , Mehmet Ozden
{"title":"Determination of intracellular cytokines produced by Th1 and Th2 cells using flow cytometry in patients with brucellosis","authors":"H. Handan Akbulut , S. Sirri Kilic , Vedat Bulut , Mehmet Ozden","doi":"10.1016/j.femsim.2005.04.001","DOIUrl":"10.1016/j.femsim.2005.04.001","url":null,"abstract":"<div><p><span>The aim of the study was to evaluate intracellular interferon-γ (IFN-γ), and interleukin-4 (IL-4) levels in pre- and post-treatment periods of brucellosis patients and to determine the relationship between these parameters and patients’ clinical findings. Twenty-five patients diagnosed as brucellosis and 11 aged-matched healthy volunteers were included in the study. CD3</span><sup>+</sup>CD4<sup>+</sup> T lymphocytes levels were significantly lower in patients with brucellosis as compared to the control group. CD3<sup>+</sup>CD8<sup>+</sup> T lymphocytes and CD3<sup>+</sup>IFN-γ<sup>+</sup> levels were increased in brucellosis patients compared with the control group. CD4<sup>+</sup>IFN-γ<sup>+</sup> and CD4<sup>+</sup>IL-4<sup>+</sup> levels were no different between patients and healthy individuals. CD3<sup>+</sup>IL-4<sup>+</sup> levels decreased in patients compared with healthy controls. Pre-treatment CD3<sup>+</sup>IFN-γ<sup>+</sup> levels dramatically increased in patients responsive to management compared with the unresponsive ones. In responsive cases, CD3<sup>+</sup>IFN-γ<sup>+</sup><span> levels decreased statistically after the treatment while in unresponsive cases no meaningful change was observed with respect to treatment. Adding IFN-γ to the treatment for improving the depleted levels of IFN-γ can be beneficial in patients with brucellosis who shows a tendency to chronicity or patients who do not respond to the treatment.</span></p></div>","PeriodicalId":12220,"journal":{"name":"FEMS immunology and medical microbiology","volume":"45 2","pages":"Pages 253-258"},"PeriodicalIF":0.0,"publicationDate":"2005-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.femsim.2005.04.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25131572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}