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Single Clone (single + clone)
Selected AbstractsDesign and engineering human forms of monoclonal antibodiesDRUG DEVELOPMENT RESEARCH, Issue 3 2004Manuel L. Penichet Abstract The antibody molecule has multiple properties that make it a key component of the immune response. These include its ability to recognize a vast array of different foreign substrates and to interact with and activate the host effector systems. Antibodies with defined specificities may serve as "magic bullets" for the diagnosis and therapy of multiple diseases. With the development of the hybridoma technology, it was possible to produce rodent (mouse or rat) monoclonal antibodies that are the product of a single clone of antibody producing cells and have only one antigen binding specificity. However, the therapeutic use of rodent monoclonals antibodies in humans is limited by their immunogenicity, short circulating half-life, and inability to efficiently trigger human effector mechanisms. However, it proved difficult to produce human monoclonal antibodies using the same methods. To address these problems genetic engineering and expression systems have instead been used to produce chimeric, humanized, and totally human antibodies as well as antibodies with novel structures and functional properties. In addition, the use of yeast and human artificial chromosome vectors for animal transgenesis has allowed the development of animal models that produce antigen specific antibodies that are totally human. As a consequence, recombinant antibody-based therapies are now used to treat a variety of clinical conditions including infectious diseases, inflammatory disorders, and cancer. This article summarizes and compares different strategies for designing and engineering human antibodies and their derivatives. Drug Dev. Res. 61:121,136, 2004. © 2004 Wiley-Liss, Inc. [source] Genotypes of multidrug-resistant Salmonella enterica serotype typhimurium from two regions of KenyaFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 1 2000Samuel Kariuki Abstract A combination of phage typing and pulsed-field gel electrophoresis of XbaI-digested chromosomal DNA has been used to study the epidemiological relationships of multidrug-resistant Salmonella enterica serotype typhimurium from Nairobi (64 isolates) and Kilifi (40 isolates) collected over the period 1994,1997. Isolates from Nairobi belonged to 11 definitive phage types (DTs) encompassing eight different PFGE patterns. In contrast, isolates from Kilifi were mainly DT 56 (60%) and all fell into a single PFGE pattern. The remaining isolates did not conform to a recognisable phage type. We conclude that multidrug-resistant S. typhimurium infections from Nairobi were caused by multiple strains while those from Kilifi were likely to be from a microepidemic caused by a single clone. [source] Limited genetic diversity of Candida albicans in fecal flora of healthy volunteers and inpatients: a proposed basis for strain homogeneity in clinical isolatesMYCOSES, Issue 9-10 2002R. Khatib Candida albicans; Gastrointestinalflora; Molekulare Typisierung Summary. Molecular analysis of Candida albicans isolates from individual patients often yields a single strain at multiple sites. Whether this strain-limitation is due to virulence factors favoring the invasive strain or to lack of genetic diversity in the gastrointestinal reservoir is uncertain. We elected to study C. albicans genotypes in the fecal flora among healthy volunteers and inpatients. Self-obtained stool swabs or stool samples were cultured on inhibitory mold agar. From each subject with C. albicans, nine colonies were randomly selected, individually propagated, and typed utilizing random amplified polymorphic DNA. Colonies were considered identical (all bands matched), related variants (one to three unique bands), or distinct strains (more than three unique bands). Analysis showed a single clone in 33/43 (76.7%) volunteers and 6/18 (33.3%) inpatients (P = 0.018), two to four related variants in eight (18.6%) volunteers and 10 (55.6%) inpatients, and two distinct strains in two volunteers (4.6%) and two inpatients (11.1%). Strain variation was more common in females (33.5 versus 5.6%; P = 0.04) and tended to increase with age (r = 0.245, P = 0.06). These findings illustrate that most healthy subjects harbor a single strain of C. albicans in the fecal flora. This strain may undergo genetic evolution leading to minor clonal variations. The mechanisms for strain selection, maintenance and possible evolution remain to be delineated. Zusammenfassung. Molekularanalysen von Candida albicans -Isolaten von individuellen Patienten zeigen oft einen individuellen Stamm an mehreren Lokalisationen. Ob diese Beschränkung auf einer Förderung durch Virulenzfaktoren des beherbergten Stammes oder auf einem Mangel an genetischer Diversität im Gastrointestinaltrakt beruht, ist unbekannt. Wir untersuchten daher die C. albicans Genotypen in der Fäkalflora von Gesunden und von Krankenhauspatienten. Selbstgewonnene Stuhlabstriche und Stuhlproben wurden auf einem schimmelpilzhemmenden Nährmedium kultiviert. Von jedem Probanden wurden 9 Kolonien randomisiert ausgewählt, individuell subkultiviert und RAPD-typisiert. Die Kolonien wurden wie folgt bewertet: klonal identisch: sämtliche Banden identisch; klonal verwandt: 1,3 Banden nicht identisch; klonal unterschiedlich: >,3 Banden nicht identisch. Die Analyse zeigte Klonidentität bei 33/43 (77%) Gesunden und 6/18 (33%) bei Hospitalisierten (P = 0.018); Klonverwandtschaft wurde bei 8 (19%) Gesunden und 10 (56%) Hospitalisierten gefunden und zwei Hospitalisierten (11%). Klonvariation war häufiger bei Frauen (33.5 vs. 5.6%; P = 0.04) und nahm mit dem Lebensalter zu (r = 0.245, P = 0.06). Diese Resultate belegen, dass die Mehrzahl Gesunder jeweils nur einen Stamm in der Fäkalflora beherbergt. Dieser kann genetisch geringgradig klonal variieren. Die hierbei wirksamen Mechanismen bedürfen noch der Aufklärung. [source] Relationship between incidence and severity of cashew gummosis in semiarid north-eastern BrazilPLANT PATHOLOGY, Issue 3 2004J. E. Cardoso The incidence,severity relationship for cashew gummosis, caused by Lasiodiplodia theobromae, was studied to determine the feasibility of using disease incidence to estimate indirectly disease severity in order to establish the potential damage caused by this disease in semiarid north-eastern Brazil. Epidemics were monitored in two cashew orchards, from 1995 to 1998 in an experimental field composed of 28 dwarf clones, and from 2000 to 2002 in a commercial orchard of a single clone. The two sites were located 10 km from each other. Logarithmic transformation achieved the best linear adjustment of incidence and severity data as determined by coefficients of determination for place, age and pooled data. A very high correlation between incidence and severity was found in both fields, with different disease pressures, different cashew genotypes, different ages and at several epidemic stages. Thus, the easily assessed gummosis incidence could be used to estimate gummosis severity levels. [source] Clonal composition of the peach-potato aphid Myzus persicae (Homoptera: Aphididae) in France and Scotland: Comparative analysis with IGS fingerprinting and microsatellite markersANNALS OF APPLIED BIOLOGY, Issue 3 2003B FENTON Summary Fourteen colonies of the peach-potato aphid, Myzus persicae, were taken either from French peach trees or weeds in 2001. Thirty five apomictic parthenogenetic lineages (APLs) were established. Ribosomal DNA intergenic spacer (IGS) fingerprinting was used to characterise these and 28 fingerprints were duly obtained. Those lineages with different fingerprints were considered different genotypes and those with the same fingerprint as the same. The genetic identity of APLs was further tested using four microsatellite loci. APLs that differed by IGS fingerprint had distinct microsatellite allele combinations and those that had the same IGS fingerprint had the same microsatellite allele combinations. The results confirmed that IGS types corresponded to different aphid genotypes. Independent APLs with identical IGS and microsatellite genotype were therefore considered different representatives of the same clone. APLs from M. persicae found on Scottish crops in 1995, 1996 and 2001, as well as a long-term laboratory line were also examined by the same methods. Their IGS fingerprints were similar or identical suggesting that they all belonged to the same clone. Microsatellite markers also suggested that these lineages were derived from a single clone. Some field lineages exhibited slight modifications to their IGS fingerprints confirming that the IGS evolves more rapidly than these microsatellite alleles. Thus, IGS will continue to provide a useful marker for aphid fieldwork. [source] Primary cutaneous B-cell lymphoma (marginal zone) with prominent T-cell component and aberrant dual (T and B) genotype; diagnostic usefulness of laser-capture microdissectionBRITISH JOURNAL OF DERMATOLOGY, Issue 1 2006F. Gallardo Summary The presence of a dominant B- or T-cell clone is an important diagnostic criterion for distinguishing cutaneous lymphomas from lymphoid reactive infiltrates. Rarely, a combined B- and T-cell rearrangement can be detected from a single sample. In such instances, genotypic analysis does not permit differentiation of the coexistence of a T- and B-cell lymphoma from a single clone harbouring a monoclonal rearrangement for both immunoglobulin heavy chain and T-cell receptor genes. We herein report a case of a skin tumour consistent with a dense cutaneous lymphoid infiltrate showing a double prominent B- and T-cell component. A dual B- and T-cell clonality was detected by polymerase chain reaction from whole-tissue DNA sample. Genotypic analysis with DNA, obtained after laser-assisted microdissection from the B-cell population, again showed both T- and B-cell monoclonal rearrangements. Conversely, the microdissected T-cell population did not reveal a clonal pattern. The diagnosis of cutaneous B-cell lymphoma with a dual B- and T-cell genotype was established. This description illustrates the diagnostic usefulness of laser-capture microdissection in cutaneous lymphomas presenting dual genotype. [source] Functional class switch recombination may occur ,in vivo' in Waldenström macroglobulinaemiaBRITISH JOURNAL OF HAEMATOLOGY, Issue 1 2007Patricia Martín-Jiménez Summary Waldenström macroglobulinaemia (WM) malignant cells have been considered incapable of undergoing class switch recombination (CSR). However, we report a WM patient who developed an IgG M-component 4 years after diagnosis. When the second monoclonal component appeared, reverse transcription-polymerase chain reaction showed the presence of pre (C,) and postswitch (C,) clonotypic isotypes; sequencing of these isotypes demonstrated that both corresponded to the single clone amplified at diagnosis, including the same complementarity-determining region 3 and somatic mutation pattern. This proves that WM cells can undergo a functional in vivo CSR. [source] An epidemiologic analysis of staphylococcus aureus-associated keratitis in BostonACTA OPHTHALMOLOGICA, Issue 2009I BEHLAU Purpose S. aureus is a normal commensal of the human skin and nasopharynx. It is therefore of interest to determine whether S. aureus keratitis is caused by a subset of these organisms. In this study, the phenotypic and genotypic characteristics of S.aureus keratitis isolates were analyzed. Methods All S. aureus clinical isolates were prospectively collected over a 24 month period at the MEEI (2006-2008). The diagnosis of clinical keratitis and associated risk factors was by medical record review. Keratitis-associated S. aureus strains were assessed for: 1) antibiotic susceptibility, 2) biofilm robustness by gentian violet staining using an in vitro microtiter plate assay, and 3) genetic lineage by multi-locus sequence typing (MLST). Results 26 cases of keratitis were identified from the 600 S. aureus clinical isolates. Risk factors associated with S.aureus keratitis included trauma, prior surgery, soft contact lens wear, and the presence of a foreign body. Ocular surface disease does not appear to be an independent risk factor. All 26 isolates were tetracycline- and trimethoprim-sulfamethoxazole- sensitive. All the MRSA strains were found to be ciprofloxacin-resistant (10/26). Nearly one-half of all the S.aureus keratitis-associated isolates were caused by a single clone, ST5. Both methicillin sensitive and resistant S. aureus strains were represented within ST5. Conclusion These results suggest that there may be specific S.aureus lineages which possess phenotypic and genotypic characteristics that enable S. aureus to more effectively cause sight-threatening keratitis. Future work will examine their virulence traits and a comparison to commensal S.aureus strains. [source] Quantitative analysis of antennal mosaic generation in Drosophila melanogaster by the MARCM systemGENESIS: THE JOURNAL OF GENETICS AND DEVELOPMENT, Issue 6 2008Carolina Gomez-Diaz Abstract Mosaics have been used in Drosophila to study development and to generate mutant structures when a mutant allele is homozygous lethal. New approaches of directed somatic recombination based on FRT/FLP methods, have increased mosaicism rates but likewise multiple clones in the same individual appeared more frequently. Production of single clones could be essential for developmental studies; however, for cell-autonomous gene function studies only the presence of homozygous cells for the target recessive allele is relevant. Herein, we report the number and extension of antennal mosaics generated by the MARCM system at different ages. This information is directed to obtain the appropriated mosaic type for the intended application. By applying heat shock at 10 different developmental stages from 0,12 h to 6,7 days after egg laying, more than 50% of mosaics were obtained from 5,028 adults. Single recombinant clones appeared mainly at early stages while massive recombinant areas were observed with late treatments. genesis 46:283,288, 2008. © 2008 Wiley-Liss, Inc. [source] Phage display of human antibodies from a patient suffering from coeliac disease and selection of isotype-specific scFv against gliadinIMMUNOLOGY, Issue 2 2003Claudio Rhyner Summary Coeliac disease (CD), a gastrointestinal illness characterized by intestinal malabsorption, results from gluten intolerance accompanied with immunological responses towards gliadin, an ethanol-soluble protein fraction of wheat and other cereals. The role of gliadin in eliciting immune responses in CD is still partly unclear; however, the occurrence of anti-gliadin in the sera of patients suffering from CD correlates well with clinical symptoms. In this work we report the construction of isotype-specific, phage-displayed scFv libraries from peripheral blood lymphocytes of a patient with CD and from a healthy control individual. VH and VL chains were amplified by reverse transcription,polymerase chain reaction (RT,PCR) using a set of oligonucleotides recognizing all human variable gene families. The three scFv libraries (IgA, IgG and IgM) were selectively enriched for gliadin-binding phage. After four rounds of affinity selection, polyclonal enrichment of gliadin-binding phage was observed in all libraries from the CD patient but in none from the healthy donor. Phagemid particles generated from single clones were demonstrated to be gliadin-specific, as shown by strongly positive enzyme-linked immunosorbent assay (ELISA) and BiaCore signals. The VH and VL chains from samples of these monoclonal isotype-specific phage were sequenced to identify the most common variable regions used by the immune system to elicit antibody responses against gliadin. [source] | |||||||||||||