scholarly journals Epitope Mapping of Exposed Tegument and Alimentary Tract Proteins Identifies Putative Antigenic Targets of the Attenuated Schistosome Vaccine

2021 ◽  
Vol 11 ◽  
Author(s):  
Leonardo P. Farias ◽  
Gillian M. Vance ◽  
Patricia S. Coulson ◽  
Juliana Vitoriano-Souza ◽  
Almiro Pires da Silva Neto ◽  
...  

The radiation-attenuated cercarial vaccine remains the gold standard for the induction of protective immunity against Schistosoma mansoni. Furthermore, the protection can be passively transferred to naïve recipient mice from multiply vaccinated donors, especially IFNgR KO mice. We have used such sera versus day 28 infection serum, to screen peptide arrays and identify likely epitopes that mediate the protection. The arrays encompassed 55 secreted or exposed proteins from the alimentary tract and tegument, the principal interfaces with the host bloodstream. The proteins were printed onto glass slides as overlapping 15mer peptides, reacted with primary and secondary antibodies, and reactive regions detected using an Agilent array scanner. Pep Slide Analyzer software provided a numerical value above background for each peptide from which an aggregate score could be derived for a putative epitope. The reactive regions of 26 proteins were mapped onto crystal structures using the CCP4 molecular graphics, to aid selection of peptides with the greatest accessibility and reactivity, prioritizing vaccine over infection serum. A further eight MEG proteins were mapped to regions conserved between family members. The result is a list of priority peptides from 44 proteins for further investigation in multiepitope vaccine constructs and as targets of monoclonal antibodies.

1997 ◽  
Vol 269 (5) ◽  
pp. 704-718 ◽  
Author(s):  
Laurent Jespers ◽  
Stéphane Jenné ◽  
Ignace Lasters ◽  
Désiré Collen

Vaccine ◽  
2008 ◽  
Vol 26 (35) ◽  
pp. 4585-4593 ◽  
Author(s):  
Daniela M. Resende ◽  
Bráulia C. Caetano ◽  
Míriam S. Dutra ◽  
Marcus L.O. Penido ◽  
Christiane F. Abrantes ◽  
...  

2002 ◽  
Vol 70 (11) ◽  
pp. 6499-6503 ◽  
Author(s):  
N. Sabarth ◽  
R. Hurwitz ◽  
T. F. Meyer ◽  
D. Bumann

ABSTRACT A multiparameter selection of Helicobacter pylori antigens for vaccine development identified 15 candidates, 6 of which are known protective antigens. Two novel antigens with low homology to other organisms (HP0231 and HP0410) were overexpressed and purified with high yields. Both confer protective immunity in the mouse Helicobacter infection model.


2013 ◽  
Vol 2013 ◽  
pp. 1-19 ◽  
Author(s):  
Elena Mata ◽  
Aiala Salvador ◽  
Manoli Igartua ◽  
Rosa María Hernández ◽  
José Luis Pedraz

There is no malaria vaccine currently available, and the most advanced candidate has recently reported a modest 30% efficacy against clinical malaria. Although many efforts have been dedicated to achieve this goal, the research was mainly directed to identify antigenic targets. Nevertheless, the latest progresses on understanding how immune system works and the data recovered from vaccination studies have conferred to the vaccine formulation its deserved relevance. Additionally to the antigen nature, the manner in which it is presented (delivery adjuvants) as well as the immunostimulatory effect of the formulation components (immunostimulants) modulates the immune response elicited. Protective immunity against malaria requires the induction of humoral, antibody-dependent cellular inhibition (ADCI) and effector and memory cell responses. This review summarizes the status of adjuvants that have been or are being employed in the malaria vaccine development, focusing on the pharmaceutical and immunological aspects, as well as on their immunization outcomings at clinical and preclinical stages.


Author(s):  
Sandrine Donadio ◽  
Willy Morelle ◽  
Aurélie Pascual ◽  
Régine Romi-Lebrun ◽  
Jean-Claude Michalski ◽  
...  

AbstractThyroid-stimulating hormone (TSH) is routinely measured in blood to diagnose thyroid disorders using immunoassays. This study used recombinant TSH (recTSH) as a source of hormonal compound exhibiting a serum-type glycosylation and putatively reflecting physiopathological alterations in TSH polymorphism. Mass spectrometry revealed that in recTSH, both subunits display high-molecular-size glycoforms compared to the pituitary hormone (pitTSH), indicating more complex glycosylation. To determine how changes in TSH glycosylation may affect epitope expression, comparative epitope mapping of rec- and pitTSH was carried out using a panel of ten hormone-specific monoclonal antibodies. Three common epitopes, I, II and III, were identified as common to both preparations and allowed the design of six assays as I/II, II/I, I/III, III/I, II/III, and III/II. Highly sialylated recTSHs were produced by enzymatic remodeling to mimic the hormone circulating in blood and revealed limited expression of epitope I, but enhanced recognition of epitope II. Fractionation on a lentil lectin-Sepharose column allowed selection of non-fucosylated recTSH, thought to be associated with primary hypothyroidism. Recognition of epitope I was not modified by TSH core fucosylation, while epitope III expression was increased in non-fucosylated glycoforms. Taken together, our findings demonstrate that changes in both core and terminal glycosylation alter epitope expression in TSH and thereby induce highly variable antibody recognition, resulting in significant discordances among hormone measurements.


1997 ◽  
Vol 5 (7) ◽  
pp. 20-21
Author(s):  
Clare Hasenkampf

I use immunocytochemical detection schemes that employ the use of primary and secondary antibodies for either tissue sections or whole mount preparations in order to study the in situ distribution of proteins and labeled nucleic acids. As the antibodies are either laboriously generated, or represent costly purchases. I have sought a protocol that minimizes the volume of antibody solution needed. Additionally I need a protocol that involves a quick, thorough, and gentle application and removal of the antibody solutions (mm the specimen with no danger of the specimen drying out during the one or two hour incubations. After trying many various schemes 1 have settled into the use of a procedure which very effectively retains the antibody solution on the specimen without damaging the delicate sections or whole mounts, and which is easy to add and remove.


Foods ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1708
Author(s):  
Xingyi Jiang ◽  
Meng Wu ◽  
Jonathan Albo ◽  
Qinchun Rao

Different types of enzyme-linked immunosorbent assays (ELISA) have been widely used to control food safety and quality. To develop an accurate and reproducible ELISA, false immunodetection results caused by non-specific binding (NSB) and cross-reaction must be prevented. During the case study of sandwich ELISA development for the detection of porcine hemoglobin (PHb), several critical factors leading to NSB and cross-reaction were found. First, to reduce the NSB of the target analyte, the selection of microplate and blocker was discussed. Second, cross-reactions between enzyme-labeled secondary antibodies and sample proteins were demonstrated. In addition, the function of (3-aminopropyl)triethoxysilane (APTES) was evaluated. Overall, this study highlights the essence of both antibody and assay validation to minimize any false-positive/negative immunodetection results.


2002 ◽  
Vol 48 (1) ◽  
pp. 121-130 ◽  
Author(s):  
Letizia Mezzasoma ◽  
Tito Bacarese-Hamilton ◽  
Manlio Di Cristina ◽  
Ruggero Rossi ◽  
Francesco Bistoni ◽  
...  

Abstract Background: Progress in robotic printing technology has allowed the development of high-density nucleic acid and protein arrays that have increased the throughput of a variety of assays. We generated protein microarrays by printing microbial antigens to simultaneously determine in human sera antibodies directed against Toxoplasma gondii, rubella virus, cytomegalovirus (CMV), and herpes simplex virus (HSV) types 1 and 2 (ToRCH antigens). Methods: The antigens were printed on activated glass slides with high-speed robotics. The slides were incubated first with serum samples and subsequently with fluorescently labeled secondary antibodies. Human IgG and IgM bound to the printed antigens were detected by confocal scanning microscopy and quantified with internal calibration curves. Both microarrays and commercial ELISAs were utilized to detect serum antibodies against the ToRCH antigens in a panel of characterized human sera. Results: The detection limit (mean + 2 SD) of the microarray assay was 0.5 pg of IgG or IgM bound to the slides. Within-slide, between-slide, and between-batch precision profiles showed CVs of 1.7–18% for all antigens. Overall, >80% concordance was obtained between microarray assays and ELISAs in the classification of sera; for T. gondii, CMV, and HSV1, concordance exceeded 90%. Conclusions: The microarray is a suitable assay format for the serodiagnosis of infectious diseases and can be easily optimized for clinical use. The ToRCH assay performs equivalently to ELISA and may have potentially important advantages in throughput, convenience, and cost.


2014 ◽  
Vol 13 (6) ◽  
pp. 1585-1597 ◽  
Author(s):  
Björn Forsström ◽  
Barbara Bisławska Axnäs ◽  
Klaus-Peter Stengele ◽  
Jochen Bühler ◽  
Thomas J. Albert ◽  
...  

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