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Drug resistance in the sexually transmitted protozoan Trichomonas vaginalis

Cell Research 2003年第4期13卷 239-249页

作者： rebecca l dunne,linda a dunn,peter upcroft,peterjo’donoghue,jacqueline a upcroft The Queensland Institute of Medical Research The Australian Centre for International and Tropical Health and Nutrition Brisbane Queensland 4029 Australia Tie School of Molecular and Microbial Science University of Queensland Brisbane Queensland 4072 Australia The Queensland Institute of Medical Research The Australian Centre for International and Tropical Health and Nutrition Brisbane Queensland 4029 Australia The Queensland Institute of Medical Research The Australian Centre for International and Tropical Health and Nutrition Brisbane Queensland 4029 Australia The School of Molecular and Microbial Science University of Queensland Brisbane Queensland 4072 Australia The Queensland Institute of Medical Research The Australian Centre for International and Tropical Health and Nutrition Brisbane Queensland 4029 Australia

Trichomoniasis is the most common, sexually transmitted infection. It is caused by the flagellated protozoan parasite Trichomonas vaginalis. Symptoms include vaginitis and infections have been associated with preterm delivery, low birth weight and increased infant mortality, as well as predisposing to HIV/AIDS and cervical cancer. Trichomoniasis has the highest prevalence and incidence of any sexually transmitted infection. The 5-nitroimidazole drugs, of which metronidazole is the most prescribed, are the only approved, effective drugs to treat trichomoniasis. Resistance against metronidazole is frequently reported and cross resistance among the family of 5-nitroimidazole drugs is common, leaving no alternative for treatment, with some cases remaining unresolved. The mechanism of metronidazole resistance in T. vaginalis from treatment failures is not well understood, unlike resistance which is developed in the laboratory under increasing metronidazole pressure. In the latter situation, hydrogenosomal fu

关键词： Trichomonas vaginalis metronidazole pyruvate: ferredoxin oxidoreductase ferredoxin hydrogenase drug resistance.

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High affinity soluble IlT2 receptor:a potent inhibitor of CD8^(+)T cell activation

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Protein & Cell 2010年第12期1卷 1118-1127页

作者： Ruth K.Moysey Yi li Samantha J.Paston Emma E.Baston Malkit S.Sami Brian J.Cameron Jessie Gavarret Penio Todorov Annelise Vuidepot Steven M.dunn Nicholas J.Pumphrey Katherine J.Adams Fang Yuan rebecca E.Dennis Deborah H.Sutton Andy D.Johnson Joanna E.Brewer rebecca Ashfield Nikolai M.lissin Bent K.Jakobsen Immunocore limited 57c Milton ParkAbingdonOxonOX144RXUK Current address:Oxford Nanopore Technologies UK Current address:Geneva Merck Serono S.A. Switzerland Current address:Adaptimmune limited UK

Using directed mutagenesis and phage display on a soluble fragment of the human immunoglobulin superfamily receptor IlT2(synonyms:lIR1,MIR7,CD85j),we have selected a range of mutants with binding affinities enhanced by up to 168,000-fold towards the conserved region of major histocompatibility complex(MHC)class I *** in a dimeric form,either by chemical cross-linking with bivalent polyethylene glycol(PEG)derivatives or as a genetic fusion with human IgG Fc-fragment,the mutants exhibited a further increase in ligand-binding strength due to the avidity effect,with resident half-times(t1/2)on the surface of MHC I-positive cells of many *** novel compounds antagonized the interaction of CD8 co-receptor with MHC I in vitro without affecting the peptide-specific binding of T-cell receptors(TCRs).In both cytokine-release assays and cell-killing experiments the engineered receptors inhibited the activation of CD8^(+)cytotoxic T lymphocytes(CTls)in the presence of their target cells,with subnanomolar potency and in a dose-dependent *** a selective inhibitor of CD8^(+)CTl responses,the engineered high affinity IlT2 receptor presents a new tool for studying the activation mechanism of different subsets of CTls and could have potential for the development of novel autoimmunity therapies.

关键词： CD8^(+)T cells cellular activation autoimmunity cell surface molecules binding affinity phage display

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Investigation of Plasmodium falciparum Resistance Biomarkers among Primary School Children in Western Kenya

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American Journal of Molecular Biology 2022年第2期12卷 43-53页

作者： Otilmoi Poul Stephen Tonny Teya Nyandwaro Robinson Mugasiali Irekwa rebecca Wanjiku Waihenya Matthew Mutinda Munyao peter Kipkemboi Rotich Caroline Wangui Njoroge Anne Wanjiru Mwangi Joanne Jepkemei Yego Nicole Sian Tanchu Dawala Koromtili Oumar Grace Ngendo Kanyita Primrose Muthoni Ndungu Samson Muuo Nzou Department of Molecular Biology and Biotechnology Pan-African University Institute of Basic Sciences Technology and Innovation Nairobi Kenya Innovation Technology Transfer Department Kenya Medical Research Institute (ITTD-KEMRI) Nairobi Kenya Zoology Department School of Biological Sciences College of Pure and Applied Sciences Jomo Kenyatta University of Agriculture and Technology Nairobi Kenya Production Department KEMRI Nairobi Kenya Centre for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Nagasaki University Institute of Tropical Medicine Kenya Medical Research Institute (NUITM-KEMRI) Nairobi Kenya Department of Biochemistry Microbiology and Biotechnology Kenyatta University Nairobi Kenya Department of Pharmacology and Pharmacognosy University of Nairobi Nairobi Kenya

Background: A marked decrease in malaria-related deaths worldwide has been attributed to the administration of effective antimalarials against Plasmodium falciparum. However, the continuous spread of P. falciparum resistance to anti-malarial drugs is raising a serious problem in controlling Malaria to the vulnerable children’s immune system. In recent studies, Plasmodium falciparum Kelch 13 propeller gene (Pfk13) has been reported to develop resistance to artemisinin in South Asia. In this study, we checked Plasmodium falciparum chloroquine resistance transporter gene (Pfcrt) involved in chloroquine (CQ) resistance. Method: In this study, archived 280 samples were collected from Alupe primary school children in Busia, Western Kenya from May, 2016 to November, 2016. Genomic DNA was extracted using the MightyPrep reagent. The samples were investigated for P. falciparum positivity out of which 67 of them tested positive giving a prevalence rate of 24%. The sixty-seven were subjected to PCR amplification for the molecular marker resistance to Pfcrt. After PCR amplification, the amplicons were purified and sequenced using Sanger Sequencing. The sequence data were analyzed using BioEdit software to identify point mutations. Results: 14 samples sequences were analyzed on Bioedit software giving the following amino acid changes F76C, Y66H, l70A, Y58C, T59V, V65I, P67l, T81l, Y60S, Y66S, P67T and I71F). New mutations have been reported at position 76 leading to an amino acid change, one of Pfcrt gold standard biomarkers. However, amino acid changes Y66H, l70A, Y58C, T59V, V65I, P67l, T81l, Y60S, Y66S, P67T and I71F are newly reported giving an increase in Pfcrt prevalence of concern from zero to 5.0%. A phylogenetic evolutionary relationship was constructed as shown below. Generally, the results showed a continuous resistance of *** to Pfcrt which calls for robust continuous monitoring and surveillance. Conclusion: Due to the increase of the resistant Pfcrt gene pre

关键词： P. falciparum Chloroquine Drug Resistance Malaria P. falciparum Chloroquine Resistance Transporter Gene Mutations

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Use of Polyclonal Antibody for the Diagnosis of Human African Trypanosomiasis

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American Journal of Molecular Biology 2023年第2期13卷 127-139页

作者： Dawala Koromtili Oumar Matthew Mutinda Munyao Anne Wanjiru Mwangi rebecca Wanjiku Waihenya peter Kipkemboi Rotich Robinson Mugasiali Irekwa Tonny Teya Nyandwaro Caroline Wangui Njoroge Joanne Jepkemei Yego Primrose Muthoni Ndungu Sharlene Kerubo Mageto Damaris Mutethya Kilei Otilmoi Poul Stephen Nicole Sian Tanchu Grace Ngendo Kanyita Shingo Inoue lucy Gitau Samson Muuo Nzou Department of Molecular Biology and Biotechnology Pan-African University Institute of Basic Sciences Technology and Innovation Nairobi Kenya Centre for Microbiology Research Kenya Medical Research Institute Nairobi Kenya Nagasaki University Institute of Tropical Medicine-Kenya Medical Research Institute Project Nairobi Kenya Production Department Kenya Medical Research Institute Nairobi Kenya Zoology Department Jomo Kenyatta University for Agriculture and Technology Nairobi Kenya Department of Pharmacology and Pharmacognosy University of Nairobi Nairobi Kenya Department of Microbiology Jomo Kenyatta University for Agriculture and Technology Nairobi Kenya Department of Biochemistry Microbiology and Biotechnology Kenyatta University Nairobi Kenya Center for Biotechnology Research and Development Kenya Medical Research Institute Nairobi Kenya

Human African trypanosomiasis (HAT), commonly known as sleeping sickness is one of the neglected tropical diseases (NTDs), which is fatal if left untreated. Its diagnosis is a challenge since the signs and symptoms of the primary phase are not specific, the existing diagnostic methods have low sensitivity and specificity, and the available drugs have some toxicity. New, robust, and cost-effective techniques are needed for the early identification of parasites. This study aimed to assess the sensitivity and specificity of two different types of polyclonal antibodies against T. b. gambiense using antigen detection ElISA. Polyclonal antibodies against the expressed proteins Tbg I2 and Tbg I17 were produced using New Zealand white rabbits. The antibody titer measured was greater than 32 g/l after the 3rd immunization for the expressed protein Tbg I2. For the expressed protein Tbg I17, the antibody titer measured was greater than 32 g/l after the 4th immunization. The sensitivity and specificity of the Tbg I2 polyclonal antibody confirmed with Polymerase Chain Reaction (PCR) as gold standard were respectively 89.5% and 80.6%, while for the Tbg I17 polyclonal antibody, the sensitivity and specificity were respectively 92.1% and 88.9%. The area under the curve for the Tbg I2 polyclonal antibody was 0.90 ± 0.032, while for the Tbg I17 polyclonal antibody, the area under the curve was 0.92 ± 0.0. The Tbg I17 polyclonal antibody produced in New Zealand white rabbits has good sensitivity and good specificity;it can be successfully used in the diagnosis of HAT.

关键词： Human African Trypanosomiasis Polyclonal Antibody Tbg I2 Expressed Protein Tbg I17 Expressed Protein Sensitivity Specificity

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CCR2 enhances CD25 expression by FoxP3^(+) regulatory T cells and regulates their abundance independently of chemotaxis and CCR2^(+) myeloid cells

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Cellular & Molecular Immunology 2020年第2期17卷 123-132页

作者： Yifan Zhan Nancy Wang Ajithkumar Vasanthakumar Yuxia Zhang Michael Chopin Stephen l.Nutt Axel Kallies Andrew M.lew The Walter&Eliza Hall Institute of Medical Research ParkvilleVIC 3052Australia Department of Medical Biology University of MelbourneParkvilleVIC 3010Australia Guangzhou Institute of Paediatrics Guangzhou Women and Children’s Medical CentreGuangzhou Medical University510623 GuangzhouGuangdongChina Department of Microbiology and Immunology Peter Doherty Institute for Infection and ImmunityUniversity of MelbourneParkvilleVIC 3010Australia

A wide array of chemokine receptors,including CCR2,are known to control Treg ***,we report that CCR2 regulates Tregs beyond *** found that CCR2 deficiency reduced CD25 expression by FoxP3^(+) Treg *** a change was also consistently present in irradiation chimeras reconstituted with mixed bone marrow from wild-type(WT)and CCR2−/−***,CCR2 deficiency resulted in profound loss of CD25 ^(hi) FoxP3^(+) Tregs in secondary lymphoid organs as well as in peripheral ***2−/−Treg cells were also functionally inferior to WT ***,these changes to Treg cells did not depend on CCR2+monocytes/moDCs(the cells where CCR2 receptors are most abundant).Rather,we demonstrated that CCR2 was required for TlR-stimulated,but not TCR-or Il-2-stimulated,CD25 upregulation on Treg ***,we propose that CCR2 signaling can increase the fitness of FoxP3^(+) Treg cells and provide negative feedback to counter the proinflammatory effects of CCR2 on myeloid cells.

关键词： CCR2 CD25 organs

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