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1.
Data Brief ; 34: 106663, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33385028

RESUMO

Next-generation sequences (NGS) dataset of nanobody (Nb) clones in a phage display library (PDL) is of immense value as it serves in many different ways, such as: i). estimating the library size, ii). improving selection and identification of Nbs, iii). informing about frequency of V gene families, diversity and length of CDRs, iv). high resolution analysis of natural and synthetic libraries, etc. [1], [2], [3]. We used a fraction of our previously constructed PDL of Nbs derived from an E. coli lipopolysaccharide-immunized Indian desert camel in order to obtain the dataset of NGS reads of Nbs. The cryo-preserved transformants library was revived to extract the Nb-encoding VHH (inserts)-pHEN4 (vector) DNA pool. The DNA sample was used for amplifying VHH pool by PCR [6]. The VHH amplicons band was gel-purified and subjected to NGS using Illumina MiSeqTM platform. 'Nextra XT micro V2 Index' kit was used for the Nb library DNA sample sequencing, with the adaptors: 'i7' (N706: TAGGCATG) and 'i5' (S517: GCGTAAGA). The raw data comprised of a total read count of 182146 (matched= 179591; unmatched=2555), with average read length of 130.33 bases and a total of 23.74 Mb. Of 179591 matched reads, 142004 were paired reads and 37587 broken paired reads. The raw data of NGS reads was submitted to NCBI Sequence Reads Archive accessible at URL: https://www.ncbi.nlm.nih.gov/Traces/study/?acc=PRJNA516512 (dataset ref. [7]), and after analysis deposited in Mendeley Datasets repository, which is accessible at URL: [https://data.mendeley.com/datasets/4rsz3snvk5/3] (dataset ref. [8]). The sequence reads were analyzed by bioinformatics tools [9], [10], [11], [12]. The assembled consensus contigs revealed Nb orthologs of diverse Ag-specificities, including those isolated by conventional panning and Sanger-sequenced functional Nbs. Contig 1 CDR1-3 matched to those of anti-Trypanosoma evansi RoTat1.2 variant surface glycoprotein (VSG), while Contig 2 CDR1-3 matched to those of anti-LPS Nb clones isolated from the library. Contig 3 was however incomplete and lacked CDR3. Despite lacking the depth, the NGS data is a useful guide for selection of antigen-specific Nbs from the library, as demonstrated by anti-T. evansi VSG Nbs, and provides templates for Nb-based diagnostic reagents and therapeutic agents.

2.
Curr Drug Metab ; 18(10): 942-970, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28952435

RESUMO

BACKGROUND: Viruses are the most devastating pathogens of almost all life forms including humans and animals. Viruses can replicate very fast and may affect any metabolic and physiological function of the host cell. Therefore, it has been a challenge to develop a universal and common treatment against viral pathogens, in contrast to bacterial pathogens. Virus-host interaction is a complex phenomenon and often is virus- and host cell-specific. Exciting new insights into the molecular pathogenesis and host-virus interactions have been gained over the past few decades. These advances have enabled researchers to design better antiviral drugs. METHODS: The literature related to various aspects of virus-host interactions: new insights and advances in drug development was collected from several scientific research related databases such as Science Direct, Google Scholar, Scopus, PubMed, AGRICOLA, and Medline, etc. Total number of 319 research papers was used to compile the information regarding drug development against viral pathogens. RESULTS: Clinical adequacy of antiviral drugs and their bioavailability are important parameters for effective treatment of viral infections. The problems associated with effective delivery of a drug in a safe and desired quantity have led to the search for (and design of) better drug delivery systems. In recent past, several new antiviral drugs have been developed, which have high therapeutic effectiveness against life-threatening viral diseases such as HIV, hepatitis B virus, herpes virus, dengue virus and influenza virus infections. The majority of recent advances in antiviral drug discovery were possible due to the developments in allied fields such as in vitro virus cultivation technology, molecular biology of viral-genome-encoded enzymes, complete-genome-sequence-based studies of viruses and identification of suitable targets for antiviral drugs in viral genomes. Recently, several novel drug delivery approaches including small interfering RNAs (siRNAs) have emerged to aid antiviral therapy. CONCLUSION: The present review is aimed at providing an update on research and development efforts being made to create effective antiviral chemotherapeutic agents and approaches to their delivery to appropriate targeted cells or tissues.


Assuntos
Antivirais , Interações Hospedeiro-Patógeno , Fenômenos Fisiológicos Virais , Animais , Antivirais/farmacologia , Antivirais/uso terapêutico , Descoberta de Drogas , Humanos , Anticorpos de Domínio Único/uso terapêutico , Viroses/tratamento farmacológico
3.
Parasite Immunol ; 25(11-12): 553-7, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-15053776

RESUMO

Three-week Mesocestoides corti infections of C57BL/6 mice showed significantly raised parasite counts in animals with a targeted knockout of the IL-4 gene. By contrast, antibody neutralization of IL-5 and inhibition of eosinophilia had no effect on parasite numbers. In SV/129 mice, knockout of the interferon gamma receptor and inducible nitric oxide synthase genes had no significant effect on parasite counts. In IL-4(-/-)mice the dominant IgG1 antibody response was dramatically reduced, with a concomitant increase in IgG2a/b responses and a partial twofold reduction in IgM and IgE responses. We conclude that murine resistance to M. corti is dependent on IL-4, but occurs independently of IL-5 and eosinophils. This provides the first direct evidence for IL-4 mediated immunity of mice to infection with a tissue-dwelling platyhelminth.


Assuntos
Infecções por Cestoides/imunologia , Interleucina-4/metabolismo , Mesocestoides/imunologia , Animais , Anticorpos Anti-Helmínticos/biossíntese , Infecções por Cestoides/parasitologia , Eosinófilos/imunologia , Feminino , Interleucina-4/deficiência , Interleucina-4/genética , Interleucina-5/metabolismo , Mesocestoides/patogenicidade , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células Th2/imunologia
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