Antibody Screening
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2021 ◽  
Vol 15 (12) ◽  
pp. e0009961
Author(s):  
Elizabeth Ajema Chebichi Luvai ◽  
Aung Kyaw Kyaw ◽  
Nundu Sabiti Sabin ◽  
Fuxun Yu ◽  
Saw Wut Hmone ◽  
...  

Introduction Chikungunya virus (CHIKV) is a mosquito-borne virus known to cause acute febrile illness associated with debilitating polyarthritis. In 2019, several institutions in Myanmar reported a CHIKV outbreak. There are no official reports of CHIKV cases between 2011 and 2018. Therefore, this study sought to determine the seroprevalence of CHIKV infection before the 2019 outbreak. Methods A total of 1,544 serum samples were collected from healthy volunteers and patients with febrile illnesses in Yangon, Mandalay, and the Myeik district in 2013, 2015, and 2018. Participants ranged from one month to 65 years of age. Antibody screening was performed with in-house anti-CHIKV IgG and IgM ELISA. A neutralization assay was used as a confirmatory test. Results The seroprevalence of anti-CHIKV IgM and anti-CHIKV IgG was 8.9% and 28.6%, respectively, with an overall seropositivity rate of 34.5%. A focus reduction neutralization assay confirmed 32.5% seroprevalence of CHIKV in the study population. Age, health status, and region were significantly associated with neutralizing antibodies (NAbs) and CHIKV seropositivity (p < 0.05), while gender was not (p = 0.9). Seroprevalence in 2013, 2015, and 2018 was 32.1%, 28.8%, and 37.3%, respectively. Of the clinical symptoms observed in participants with fevers, arthralgia was mainly noted in CHIKV-seropositive patients. Conclusion The findings in this study reveal the circulation of CHIKV in Myanmar’s Mandalay, Yangon, and Myeik regions before the 2019 CHIKV outbreak. As no treatment or vaccine for CHIKV exists, the virus must be monitored through systematic surveillance in Myanmar.


2021 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Douglas A.E. White ◽  
Erik S. Anderson ◽  
Kellie Basham ◽  
Valerie L. Ng ◽  
Carly Russell ◽  
...  

Author(s):  
Anshika Yadav ◽  
C. S. Joshi ◽  
G. N. Gupta ◽  
Rounak Dubey

Background: Haemolytic disease of the foetus and new-born (HDFN) is a major concern during the antenatal period, especially in countries with low human development index (HDI). The guidelines for antenatal screening and management significantly vary from one geographical region to another. Since the introduction of RhIG immunoprophylaxis, the incidence of HDFN caused by alloimmunization to D antigen has markedly reduced, while that caused by other minor blood group antigens has not been addressed significantly and needs to be given due consideration.Methods: The study was carried out to evaluate the incidence of alloimmunization and analyse various factors associated with HDFN in north-western India. A total of 1700 antenatal cases were evaluated over a period of 20 months, antibody screening and identification was performed on their samples and results were analysed.Results: Out of the 1700 cases, 21 were detected to have the presence of an alloantibody with a prevalence of 1.24%. Out of these, 11 were Rh (D) negative while the remaining 10 were Rh (D) positive. The rate for alloimmunization was higher in females who had a history of blood transfusion (1.24%), bad obstetric history (1.24%), and multigravida status (1.24%).Conclusions: Screening all pregnant females for alloimmunization to RBC antigens, irrespective of their Rh status will help in minimizing the incidence of the HDFN. The practice of providing partial phenotype matched blood to the females of the childbearing age group should be encouraged to reduce the overall incidence of alloimmunization and HDFN.


2021 ◽  
Vol 8 ◽  
Author(s):  
Yaling Zhang ◽  
Huan Xu ◽  
Yi Liu ◽  
Juan Kang ◽  
Hairu Chen ◽  
...  

The clinical manifestations of fascioliasis hepatica in humans are unspecific. Traditional diagnosis relies on evidence of live parasites or eggs in the bile or feces. However, due to similar imaging manifestations, they are often misdiagnosed as malignant tumors. Here, we report a case of a 43-year-old woman with fever and space-occupying liver disease. Liver biopsy, parasite-specific antibody screening, and stool testing did not find any pathogens. Therefore, metagenomic next-generation sequencing (mNGS) and routine microbiological examinations were performed. Finally, Fasciola hepatica was only identified by mNGS. The body temperature of the patient and the eosinophil count remained normal, and the space-occupying liver lesions were significantly absorbed after more than 7 months of treatment with albendazole. The details of this case highlight the timely use of mNGS to identify parasites and judge therapeutic effects after treatment, providing important help for clinical decision-making.


2021 ◽  
pp. postgradmedj-2021-140497
Author(s):  
Saikat Mandal ◽  
Daljit Kaur ◽  
Gita Negi ◽  
Sriparna Basu ◽  
Jaya Chaturvedi ◽  
...  

BackgroundRed blood cell alloimmunisation during the pregnancy is a significant cause for neonatal mortality and morbidity. This study was planned to determine the prevalence and specificity of irregular erythrocyte antibodies in antenatal mothers and their neonatal outcome.MethodsIn this observational study, blood grouping and red cell antibody screening of mothers were performed at first visit and after 28 weeks of gestation and positive cases were identified and followed up monthly till delivery by repeating antibody titre and middle cerebral artery—peak systolic velocity. After delivery of alloimmunised mothers, cord blood haemoglobin, bilirubin and direct antiglobulin tests (DAT) were analysed and further outcome of neonate was recorded.ResultsAmong 652 registered antenatal cases, 18 multigravida women were found to be alloimmunised, accounting to prevalence of 2.8%. Most common alloantibody identified was anti D (>70%) followed by anti-Lea, anti-C, anti-Leb, anti-E and anti-Jka. Only 47.7% Rh D negative women received anti-D prophylaxis during previous pregnancies or whenever indicated. DAT was positive in 56.2% of neonates. Among nine DAT positive neonates, two early neonatal deaths due to severe anaemia were observed following birth resuscitation. Four antenatal mothers required intrauterine transfusion in view of fetal anaemia while three neonates received double volume exchange transfusion and top up transfusions after birth.ConclusionsThis study emphasises importance of red cell antibody screening for all multigravida antenatal women at registration of pregnancy and additionally at 28 weeks or later in high-risk cases irrespective of RhD status.


2021 ◽  
Vol 15 (11) ◽  
pp. e0009925
Author(s):  
Lola Marqué ◽  
Peter Liehl ◽  
Jasper De Boer ◽  
Hans Pottel ◽  
Edward L. Murphy ◽  
...  

Background Human T-Cell Lymphotropic Viruses (HTLV) type 1 and type 2 account for an estimated 5 to 10 million infections worldwide and are transmitted through breast feeding, sexual contacts and contaminated cellular blood components. HTLV-associated syndromes are considered as neglected diseases for which there are no vaccines or therapies available, making it particularly important to ensure the best possible diagnosis to enable proper counselling of infected persons and avoid secondary transmission. Although high quality antibody screening assays are available, currently available confirmatory tests are costly and have variable performance, with high rates of indeterminate and non-typable results reported in many regions of the world. The objective of this project was to develop and validate a new high-performance multiplex immunoassay for confirmation and discrimination of HTLV-1 and HTLV-2 strains. Methodology/Principal findings The multiplex platform was used first as a tool to identify suitable antigens and in a second step for assay development. With data generated on over 400 HTLV-positive blood donors sourced from USA and French blood banks, we developed and validated a high-precision interpretation algorithm. The Multi-HTLV assay demonstrated very high performance for confirmation and strain discrimination with 100% sensitivity, 98.1% specificity and 100% of typing accuracy in validation samples. The assay can be interpreted either visually or automatically with a colorimetric image reader and custom algorithm, providing highly reliable results. Conclusions/Significance The newly developed Multi-HTLV is very competitive with currently used confirmatory assays and reduces considerably the number of indeterminate results. The multiparametric nature of the assay opens new avenues to study specific serological signatures of each patient, follow the evolution of infection, and explore utility for HTLV disease prognosis. Improving HTLV diagnostic testing will be critical to reduce transmission and to improve monitoring of seropositive patients.


2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A934-A934
Author(s):  
Mai Yoshikawa ◽  
Yoji Nagira ◽  
Morio Nagira ◽  
Tetsuya Yoshida ◽  
Shinpei Yoshida ◽  
...  

BackgroundRegulatory T cells (Tregs) are involved in tumor progression and inhibition of anti-tumor immune responses by promotion of immunological tolerance in the tumor microenvironment. On the other hand, Treg cells in peripheral blood are also essential role in preventing autoimmunity and uncontrolled inflammation. So, selective control of tumor infiltrating Treg cells might be an attractive approach of immune-oncology therapies without disrupting their systemic anti-inflammatory functions. Here, we focused on CCR8 (C-C motif chemokine receptor 8) as a target molecule which was selectively and highly expressed on tumor-infiltrating Tregs and developed a novel anti-human CCR8 specific antibody.MethodsWe immunized mice with human CCR8 by our original immunization method which could strongly induce antibodies for membrane proteins, and then constructed hybridoma cells. Anti-human CCR8 (human CCR4 as a negative control) binding assay and human CCL1-CCR8 neutralizing assay were simultaneously performed by using supernatants of hybridoma cells to isolate human CCR8 specific strong-neutralizing antibodies. After humanization and affinity maturation of some selected clones, we selected our lead antibody by binding specificity, neutralizing activity, antibody dependent cellular cytotoxicity (ADCC) and thermodynamic stability as index.ResultsWe rapidly induced human CCR8 specific antibodies in mouse with our unique immunization methods and constructed thousands of hybridomas secreting anti-human CCR8 antibodies. We also successfully humanized some of lead antibodies which show high affinity and specificity and isolated novel anti-human CCR8 specific humanized antibody S-531011 as our development antibody after affinity maturation. S-531011 selectively recognizes human CCR8 on the surface of tumor-infiltrating Tregs and shows strong ADCC. While human CCL1 is known as a dominant ligand of CCR8 which binds extracellular loop2 and N-terminal of CCR8, S-531011 recognizes similar epitopes and effectively neutralizes CCL1-CCR8 signaling. Furthermore, S-531011 also shows favorable blood kinetics in vivo and potently inhibits tumor growth in tumor bearing human CCR8 knock-in mouse model.ConclusionsWe develop S-531011, a novel anti-human CCR8 humanized antibody which could selectively recognize and deplete tumor infiltrating Tregs. Based on our pre-clinical data, S-531011 has strong anti-tumor effect and we expect that it might be a potent novel tumor immuno-therapeutic agent with fewer side effect.Ethics ApprovalThe present study was approved by the Institutional Ethics Committee of Osaka University Hospital (approved number: 13266-15). Animal studies were approved by the Institutional Animal Care and Use Committee (approved number: S20192D, S20197D and S20198D).


2021 ◽  
Vol 5 (1) ◽  
pp. e202101115
Author(s):  
Yueyuan Yin ◽  
Fei Yan ◽  
Ruimin Zhou ◽  
Mingchen Li ◽  
Jinyi Ma ◽  
...  

Single-domain antibody (sdAb) holds the promising strategies for diverse research and translational applications. Here, we describe a method for the adaptation of the in situ proximity ligation assay (isPLA) followed by sequencing (isPLA-seq) to facilitate screening of a high-sensitive, high-throughput sdAb library for a given protein at subcellular and single-cell resolution. Based on the sequence of complementarity-determining region 3 (CDR3), the recombinant sdAb can be produced for in vitro and in vivo utilities. This method provides a general means to identify the functional measure of sdAb and its complementary epitopes and its potential applications to investigate cellular processes.


Author(s):  
Giorgia Borio ◽  
Chiara Terracciano ◽  
Federico Buttafava ◽  
Andrea Vercelli ◽  
Laura Pagani ◽  
...  

We report the case of a 62-year-old male patient fully vaccinated for COVID-19, admitted to our emergency room for persistent fever associated with exertional dyspnoea, skin lesions, diffuse myalgias and arthralgias not responsive to broad-spectrum antibiotic and antiviral therapy, who developed a rapidly progressive refractory to treatment interstitial lung disease due to anti-melanoma differentiation-associated gene 5 (MDA5) antibodies, that required mechanical ventilation and ECMO. Here, we highlight the importance of always considering alternative diagnoses, i.e. viral and autoimmune diseases, including anti-MDA5 antibody screening, when dealing with patients with a skin rash, seronegative polyarthralgias and interstitial pneumonia, or acute respiratory distress syndrome of unknown origin.


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