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Cells
Special Issues
Recent Advances in ELISPOT Research
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Recent Advances in ELISPOT Research

A special issue of Cells (ISSN 2073-4409).

Deadline for manuscript submissions: closed (30 June 2017) | Viewed by 23221

Special Issue Editor

Prof. Dr. Alexander E. Kalyuzhny

E-Mail Website
Guest Editor
Neuroscience, UMN Twin Cities, 6-145 Jackson Hall, 321 Church St SE, Minneapolis, MN 55455, USA
Interests: investigating the mechanisms underlying the constitutive induced heteromerization of opioid receptors
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

It appears that ELISPOT is a very conservative technique limited, unlike ELISA, to analyzing a handful of proteins. There is some truth to it, because this limitation is due to inherent nature of ELISPOT assay: Only proteins secreted from live cells are to be detected. However, ELISA can be used for both, secreted proteins from live cells, as well as non-secreted intracellular proteins in cell lysates. In spite of detecting a limited number of secreted proteins, ELISPOT offers unsurpassed detection sensitivity that is a hundred times (or even more) sensitive compared to ELISA. In addition to its exceptional sensitivity, ELISPOT allows precise quantification of bona fide secreting cells, which is not possible using any other technique that is currently available to researchers. Initially developed as a research tool, ELISPOT has recently entered a diagnostic market where it firmly holds a unique niche.

This Special Issue “Recent Advances in ELISPOT Research” is the third issue in our ELISPOT series, which has received a very positive feedback from the global research community. Our current Special Issue is welcoming research papers and review articles, as well as entirely technique-related articles. We invite papers on ELISPOT applications for human and animal diagnostics and novel “non-conventional” areas of its application, including environment protection, analysis of non-mammalian cells, and super-sensitive multiplexing.

Dr. Alexander E. Kalyuzhny
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Cells is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • ELISPOT
  • fluorospot
  • diagnostics
  • cancer research
  • vaccine development
  • multiplexing
  • cytokines

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Published Papers (3 papers)

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Research

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14 pages, 2136 KiB  
Article
B Cells and B Cell Blasts Withstand Cryopreservation While Retaining Their Functionality for Producing Antibody
by Philipp Fecher, Richard Caspell, Villian Naeem, Alexey Y. Karulin, Stefanie Kuerten and Paul V. Lehmann
Cells 2018, 7(6), 50; https://doi.org/10.3390/cells7060050 - 31 May 2018
Cited by 15 | Viewed by 6867
Abstract
In individuals who have once developed humoral immunity to an infectious/foreign antigen, the antibodies present in their body can mediate instant protection when the antigen re-enters. Such antigen-specific antibodies can be readily detected in the serum. Long term humoral immunity is, however, also [...] Read more.
In individuals who have once developed humoral immunity to an infectious/foreign antigen, the antibodies present in their body can mediate instant protection when the antigen re-enters. Such antigen-specific antibodies can be readily detected in the serum. Long term humoral immunity is, however, also critically dependent on the ability of memory B cells to engage in a secondary antibody response upon re-exposure to the antigen. Antibody molecules in the body are short lived, having a half-life of weeks, while memory B cells have a life span of decades. Therefore, the presence of serum antibodies is not always a reliable indicator of B cell memory and comprehensive monitoring of humoral immunity requires that both serum antibodies and memory B cells be assessed. The prevailing view is that resting memory B cells and B cell blasts in peripheral blood mononuclear cells (PBMC) cannot be cryopreserved without losing their antibody secreting function, and regulated high throughput immune monitoring of B cell immunity is therefore confined to—and largely limited by—the need to test freshly isolated PBMC. Using optimized protocols for freezing and thawing of PBMC, and four color ImmunoSpot® analysis for the simultaneous detection of all immunoglobulin classes/subclasses we show here that both resting memory B cells and B cell blasts retain their ability to secrete antibody after thawing, and thus demonstrate the feasibility of B cell immune monitoring using cryopreserved PBMC. Full article
(This article belongs to the Special Issue Recent Advances in ELISPOT Research)
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2136 KiB  
Article
Delayed Activation Kinetics of Th2- and Th17 Cells Compared to Th1 Cells
by Andrea Duechting, Anna Przybyla, Stefanie Kuerten and Paul V. Lehmann
Cells 2017, 6(3), 29; https://doi.org/10.3390/cells6030029 - 12 Sep 2017
Cited by 21 | Viewed by 6751
Abstract
During immune responses, different classes of T cells arise: Th1, Th2, and Th17. Mobilizing the right class plays a critical role in successful host defense and therefore defining the ratios of Th1/Th2/Th17 cells within the antigen-specific T cell repertoire is critical for immune [...] Read more.
During immune responses, different classes of T cells arise: Th1, Th2, and Th17. Mobilizing the right class plays a critical role in successful host defense and therefore defining the ratios of Th1/Th2/Th17 cells within the antigen-specific T cell repertoire is critical for immune monitoring purposes. Antigen-specific Th1, Th2, and Th17 cells can be detected by challenging peripheral blood mononuclear cells (PBMC) with antigen, and establishing the numbers of T cells producing the respective lead cytokine, IFN-γ and IL-2 for Th1 cells, IL-4 and IL-5 for Th2, and IL-17 for Th-17 cells, respectively. Traditionally, these cytokines are measured within 6 h in flow cytometry. We show here that 6 h of stimulation is sufficient to detect peptide-induced production of IFN-γ, but 24 h are required to reveal the full frequency of protein antigen-specific Th1 cells. Also the detection of IL-2 producing Th1 cells requires 24 h stimulation cultures. Measurements of IL-4 producing Th2 cells requires 48-h cultures and 96 h are required for frequency measurements of IL-5 and IL-17 secreting T cells. Therefore, accounting for the differential secretion kinetics of these cytokines is critical for the accurate determination of the frequencies and ratios of antigen-specific Th1, Th2, and Th17 cells. Full article
(This article belongs to the Special Issue Recent Advances in ELISPOT Research)
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Review

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2961 KiB  
Review
How Can Elispot Add Information to Improve Knowledge on Tropical Diseases?
by Josué Da Costa Lima-Junior, Fernanda Nazaré Morgado and Fátima Conceição-Silva
Cells 2017, 6(4), 31; https://doi.org/10.3390/cells6040031 - 29 Sep 2017
Cited by 8 | Viewed by 8662
Abstract
Elispot has been used as an important tool for detecting immune cells’ products and functions and has facilitated the understanding of host-pathogen interaction. Despite the incredible diversity of possibilities, two main approaches have been developed: the immunopathogenesis and diagnosis/prognosis of infectious diseases as [...] Read more.
Elispot has been used as an important tool for detecting immune cells’ products and functions and has facilitated the understanding of host-pathogen interaction. Despite the incredible diversity of possibilities, two main approaches have been developed: the immunopathogenesis and diagnosis/prognosis of infectious diseases as well as cancer research. Much has been described on the topics of allergy, autoimmune diseases, and HIV-Aids, however, Elispot can also be applied to other infectious diseases, mainly leishmaniasis, malaria, some viruses, helminths and mycosis usually classified as tropical diseases. The comprehension of the function, concentration and diversity of the immune response in the infectious disease is pointed out as crucial to the development of infection or disease in humans and animals. In this review we will describe the knowledge already obtained using Elispot as a method for accessing the profile of immune response as well as the recent advances in information about host-pathogen interaction in order to better understand the clinical outcome of a group of tropical and neglected diseases. Full article
(This article belongs to the Special Issue Recent Advances in ELISPOT Research)
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