Current Issue : April - June Volume : 2019 Issue Number : 2 Articles : 7 Articles
Background: Most monoclonal antibodies against mouse antigens have been derived from rat spleen-mouse\nmyeloma fusions, which are valuable tools for purposes ranging from general laboratory reagents to therapeutic\ndrugs, and yet selecting and expressing them remains a time-consuming and inefficient process. Here, we report a\nnovel approach for the rapid high-throughput selection and expression of recombinant functional rat monoclonal\nantibodies with different isotypes.\nResults: We have developed a robust system for generating rat monoclonal antibodies through several processes\ninvolving simultaneously immunizing rats with three different antigens expressing in a mixed cell pools, preparing\nhybridoma cell pools, in vitro screening and subsequent cloning of the rearranged light and heavy chains into a\nsingle expression plasmid using a highly efficient assembly method, which can decrease the time and effort\nrequired by multiple immunizations and fusions, traditional clonal selection and expression methods. Using this\nsystem, we successfully selected several rat monoclonal antibodies with different IgG isotypes specifically targeting\nthe mouse PD-1, LAG-3 or AFP protein from a single fusion. We applied these recombinant anti-PD-1 monoclonal\nantibodies (32D6) in immunotherapy for therapeutic purposes that produced the expected results.\nConclusions: This method can be used to facilitate an increased throughput of the entire process from multiplex\nimmunization to acquisition of functional rat monoclonal antibodies and facilitate their expression and feasibility\nusing a single plasmid....
Background: Dengue Virus (DENV) and Zika Virus (ZIKV) are closely related flaviviruses, circulating in overlapping\ngeographical regions. The recent ZIKV epidemic has been linked to an explosion in reports of microcephaly and\nneurological defects. It is conceivable that our knowledge of DENV might potentiate the development of a ZIKV\nvaccine due to the close phylogenetic relationship between these flaviviruses and cross-reactive antibodies,\nprincipally to the envelope protein (E protein). Alternatively, cross-reactive antibodies that are generated following\nvaccination or infection, might become damaging during subsequent infections.\nMain body: The aims of this review are to collate and analyse data from a recent series of DENV-derived\nmonoclonal antibody (mAb) panels from different research groups. These panels measured DENV-mAb activity\nagainst ZIKV in terms of antibody-dependent enhancement (ADE) and neutralisation. Methodology used across\ngroups was compared and critiqued. Furthermore, the specific antibody targets on E protein were considered and\ntheir therapeutic potential evaluated. Shortcomings of hmAb panels suggest ADE may be over-estimated and\nneutralisation underestimated, as compared to clinical situations. It remains unknown whether preference of\nenhancement or neutralisation by antibodies to ZIKV E protein is dictated by quantitative aspects of antibody\ntitre or epitope specific variation. Additionally, little is known about how duration between flavivirus reinfections\naffect secondary antibody response.\nConclusion: This review concludes that our current knowledge of cross-reactive antibodies to E protein is\ninadequate to anticipate the outcome of deploying an E protein based vaccine to regions co-infected by DENV and\nZIKV....
Determining antibiotic concentration in human blood provides useful pharmacokinetic\ninformation. Commonly used methods such as ELISA require a long time to obtain results and thus\ncannot be applied when information is needed immediately. In this study, a novel antibody-based\nlateral flow technique was developed for tetracycline detection in human serum. Contrary to tests\ndeveloped to analyze food samples, the features of work with serum as analyzed probe were studied\nfor the first time here. The application of labeled and unlabeled specific antibodies was compared.\nFor this purpose, specific and anti-species antibodies were labeled with gold nanoparticles and used\nfor antigenâ??antibody interaction on the membrane surface with observed staining in the test zone.\nFor both schemes, optimal conditions were established to provide the best sensitivity. The developed\nassay has a limit of visual detection as low as 35 and 11 ng/mL for the direct and indirect labeled\nantibodies, respectively. The limit of instrumental detection is from 0.4 to 3.5 ng/mL for diluted and\nundiluted sera. The use of indirect antibody labeling showed a small increase in sensitivity compared\nto traditional direct antibody labeling. The developed method showed no cross-reactivity with\nantibiotics of other classes. The method was used to test samples of serum. The results showed high\ncorrelation with the data obtained by ELISA (R2 = 0.98968). The assay provides a quick assessment\nof the amount of antibiotics in the blood and keeps them under control throughout the duration\nof therapy....
In Japan, herpes zoster is not monitored officially or nationwide. Recently,\nthe databases of all electronic medical claims nationwide (NDBEMC) have\nbeen available for research. We use NDBEMC from April 2011 to March\n2015. To evaluate the effects of initiation of routine immunization for varicella\nin children, we regressed the number of herpes zoster patients on the\ndummy variable for the routine immunization for varicella in children with\nand without a linear time trend. The estimated coefficient for the routine\nimmunization for varicella was 0.5157 and its p-value was 0.001. However, if\nthe time trend was added as an explanatory variable, the estimated coefficient\nfor the routine immunization for varicella changed to be -0.039 and its\np-value was 0.384. It means that the routine immunization for varicella was\n7.8% higher after introduction than before. However, it was presumed to reflect\nsuch an upward trend....
Introduction: Entero bacteria are mainly found in the gut of man and animals.\nThe frequent acquisition of antibiotic resistance mechanisms explains\nwhy they are the bacteria most often implicated in human infectious pathology.\nIt is estimated to be involved in 50% of sepsis, 60% of enteritis, 70% of\nurinary tract infection case. Objective: To determine the prevalence of enterobacterial\ninfections diagnosed at Fann Infectious Diseases Clinic, and describe\ntheir epidemiological, clinical, therapeutic and evolutionary aspects.\nPatients and Methods: This is a retrospective and descriptive study, on patients\nhospitalized from January 2013 to December 2014, at Fann Infectious\nDiseases Clinic, with bacteriological confirmation of an enterobacteria infection.\nResults: A total of 129 cases were collected during the study period. The\naverage age was 41 years, and female were predominant (60%) with a sex ratio\nof 0.67. Comorbidity was found in 88.4% of the cases, most of which were\nHIV infection. The most common clinical signs were infectious syndrome\n(53.49%) and general impairment (40.31%). The main gateway was urinary\n(55.8%). Samples were monomicrobial in 76.7% of cases. Klebsiella and\nEscherichia were the most common and 68.7% of the subjects had probabilistic\ntreatment. Most enterobacterial strains were resistant to third generation\ncephalosporins (C3G), aminoglycosides to ciprofloxacin and cotrimoxazole.\nAside from 4% of them, all were sensitive to imipenem. Conclusion: The advent\nof antibiotics has brought hope in the treatment of enterobacterial infections.\nHowever, an increase in their resistance to the usual antibiotics has\nbeen noted in recent years. As a result, the fight against antibiotic resistance\nmust be a priority....
Background: Disease caused by Bacillus anthracis is often accompanied by high mortality primarily due to toxinmediated\ninjury. In the early disease course, anthrax toxins are secreted; thus, antibiotic use is limited to the early stage.\nIn this regard, antibodies against the toxin component, protective antigen (PA), play an important role in protecting\nagainst anthrax. Therefore, we developed PA21, a fully human anti-PA immunoglobulin G monoclonal antibody.\nMethods: Combining human Fab was screened from a phage library with human heavy constant regions. Enzymelinked\nimmune sorbent assay, Western blot analysis and immunoprecipitation test evaluated the binding ability of\nPA21. Moreover, the affinity and neutralizing activity of the antibody was detected in vitro while the protective\neffectiveness in 60 rats was also examined in vivo.\nResults: The Fischer 344 rats challenged with the lethal toxin can be protected by PA21 at a concentration of 0.067\nmg/kg. All six rats remained alive although PA21 was injected 24 h before the toxin challenge. PA21 did not influence\nthe binding of PA to cell receptors and that of a lethal factor to PA.\nConclusion: The PA21 monoclonal antibody against PA can be used for emergency prophylaxis and anthrax treatment....
Background: Spinal Tuberculosis is the most common and dangerous form\nof skeletal tuberculosis. It has the potential for serious morbidity, including\npermanent neurologic deficits and severe deformity. The aim of this study is\nto review spinal Tuberculosis (TB) cases in our tertiary care center and evaluate\nit from different aspects, which may provide great support to the clinical\ndecisions of this disease. Methods: The study was a retrospective charts review\nof all adult spinal TB patients who were evaluated at King Abdulaziz\nMedical City, Riyadh, from January 2001 to March 2015. The inclusion of\ncases will need to meet a specific case definition. Patients demography, presenting\nsymptoms and signs, and radiological and microbiological data for all\nthe patients were collected and reviewed. Results: A total of 61 cases screened\ncharts were included according to the case definition. 59% were female, and\nthe mean age was 51.3 years. The most presenting symptom was back pain\n(85%), followed by lower limbs weakness and lower limbs pain. The average\nduration of symptoms was 170 days. The average hospital stay was 41.4 days.\nOn physical examination, about half of the patients had bilateral lower limbs\nweakness. Acute phase reactants like C-reactive protein (CRP) and Erythrocyte\nsedimentation rate (ESR) were elevated, 97% and 92% respectively,\nhowever leukocytosis was not common. Out of all the patients who had a biopsy\ndone (59 patients), 51 patients (86%) had positive AFB culture. MTB\nPCR was done from spine biopsy in 16 patients, and 81% were positive. Out\nof 53 tissue specimens, histopathology showed 45% necrotizing granuloma,\n21% non-necrotizing granuloma, and 34% non-specific inflammation. Imaging\n(either CT scan or MRI) was performed on the patients at the time of\npresentation, and the most common signs seen in both were vertebral bone\ndestruction (95%, 80%) and perivertebral collections (70%, 84%). The thoracic\nand lumbar vertebrae were the most common involved locations. Thirty-one\npatients had both modalities done (CT and MRI), and spinal compression\nwas seen on MRI in 68% of the cases compared to 36% on the CT scan. All\nthe patients received the first line therapy with some individual adjustment\nand only 39% (27 patients) underwent surgery. Conclusion: Spinal Tuberculosis\nis a chronic presentation illness with a non-specific clinical picture. Interventional\ndiagnostics is the best current modality to reach a confirmatory\ndiagnosis. The gold standard for confirming TB diagnosis probably will remain\ncultures in addition to early utilization of molecular TB diagnostics.\nTime for diagnosis of Spinal Tuberculosis is delayed despite improvement in\ndiagnostics tests. Medical treatment is the major solution of uncomplicated\nspinal TB....
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