How Can Elispot Add Information to Improve Knowledge on Tropical Diseases?

Lima-Junior, Josué da Costa; Morgado, Fernanda Nazaré; Conceição-Silva, Fátima

doi:10.3390/cells6040031

Cited by 8 publications

(6 citation statements)

References 142 publications

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“…However, ELISpot and Fluorospot are limited to the detection of only 1-2 or 1-4 soluble analytes, respectively, cannot precisely quantify secretion rates, and the ASC identified cannot be recovered for further analysis. ELISpot has seen extensive use in characterizing the ASC response during pathogenic infections (19,20), in autoimmune disorders (21)(22)(23), and following immunization (24). Bonezi et al (25) recently used ELISpot to demonstrate the contribution of altered tryptophan metabolism to the hyperabundance of ASC frequently seen during flavivirid infections, Dengue virus being the most common example.…”

Section: Spot-based Assaysmentioning

confidence: 99%

Single-Cell Technologies for the Study of Antibody-Secreting Cells

Broketa

Bruhns

2022

Front. Immunol.

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Antibody-secreting cells (ASC), plasmablasts and plasma cells, are terminally differentiated B cells responsible for large-scale production and secretion of antibodies. ASC are derived from activated B cells, which may differentiate extrafollicularly or form germinal center (GC) reactions within secondary lymphoid organs. ASC therefore consist of short-lived, poorly matured plasmablasts that generally secrete lower-affinity antibodies, or long-lived, highly matured plasma cells that generally secrete higher-affinity antibodies. The ASC population is responsible for producing an immediate humoral B cell response, the polyclonal antibody repertoire, as well as in parallel building effective humoral memory and immunity, or potentially driving pathology in the case of autoimmunity. ASC are phenotypically and transcriptionally distinct from other B cells and further distinguishable by morphology, varied lifespans, and anatomical localization. Single cell analyses are required to interrogate the functional and transcriptional diversity of ASC and their secreted antibody repertoire and understand the contribution of individual ASC responses to the polyclonal humoral response. Here we summarize the current and emerging functional and molecular techniques for high-throughput characterization of ASC with single cell resolution, including flow and mass cytometry, spot-based and microfluidic-based assays, focusing on functional approaches of the secreted antibodies: specificity, affinity, and secretion rate.

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Section: Spot-based Assaysmentioning

confidence: 99%

Single-Cell Technologies for the Study of Antibody-Secreting Cells

Broketa

Bruhns

2022

Front. Immunol.

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“…Thus, it is less prone to automatization and requires training to work with cellular culture. However, it can also be adapted to different infrastructures and the technique is highly sensitive, being used especially for vaccine investigations [ 68 , 69 ].…”

Section: Cellular Immunitymentioning

confidence: 99%

Enzyme-Linked Immunosorbent Assay: An Adaptable Methodology to Study SARS-CoV-2 Humoral and Cellular Immune Responses

Portilho

Lima

Gaspari

2022

JCM

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The Enzyme-Linked Immunosorbent Assay is a versatile technique, which can be used for several applications. It has enormously contributed to the study of infectious diseases. This review highlights how this methodology supported the science conducted in COVID-19 pandemics, allowing scientists to better understand the immune response against SARS-CoV-2. ELISA can be modified to assess the functionality of antibodies, as avidity and neutralization, respectively by the standardization of avidity-ELISA and surrogate-neutralization methods. Cellular immunity can also be studied using this assay. Products secreted by cells, like proteins and cytokines, can be studied by ELISA or its derivative Enzyme-linked immunospot (ELISpot) assay. ELISA and ELISA-based methods aided the area of immunology against infectious diseases and is still relevant, for example, as a promising approach to study the differences between natural and vaccine-induced immune responses against SARS-CoV-2.

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“…From the described studies it is clear, that IL-21 plays a role in both cellular and humoral rejection. The Elispot assay enables quantification of donor-reactive IL-21 producing cells (pc) and has proven to be a useful monitoring tool due to its sensitive and accurate detection of rare antigen-specific T-cells and its ability to visualize single positive cells within a population of peripheral blood cells (27–29).…”

Section: Introductionmentioning

confidence: 99%

The Number of Donor-Specific IL-21 Producing Cells Before and After Transplantation Predicts Kidney Graft Rejection

et al. 2019

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Interleukin (IL)-21 supports induction and expansion of CD8 + T cells, and can also regulate the differentiation of B cells into antibody-producing plasma cells. We questioned whether the number of circulating donor-specific IL-21 producing cells (pc) can predict kidney transplant rejection, and evaluated this in two different patient cohorts. The first analysis was done on pre-transplantation samples of 35 kidney transplant recipients of whom 15 patients developed an early acute rejection. The second study concerned peripheral blood mononuclear cell (PBMC) samples from 46 patients obtained at 6 months after kidney transplantation of whom 13 developed late rejection. Significantly higher frequencies of donor-specific IL-21 pc were found by Elispot assay in both patients who developed early and late rejection compared to those without rejection. In addition, low frequencies of donor-specific IL-21 pc were associated with higher rejection-free survival. Moreover, low pre-transplant donor-specific IL-21 pc numbers were associated with the absence of anti-HLA antibodies. Donor-reactive IL-21 was mainly produced by CD4 + T cells, including CD4 + follicular T helper cells. In conclusion, the number of donor-specific IL-21 pc is associated with an increased risk of both early and late rejection, giving it the potential to be a new biomarker in kidney transplantation.

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How Can Elispot Add Information to Improve Knowledge on Tropical Diseases?

Cited by 8 publications

References 142 publications

Single-Cell Technologies for the Study of Antibody-Secreting Cells

Single-Cell Technologies for the Study of Antibody-Secreting Cells

Enzyme-Linked Immunosorbent Assay: An Adaptable Methodology to Study SARS-CoV-2 Humoral and Cellular Immune Responses

The Number of Donor-Specific IL-21 Producing Cells Before and After Transplantation Predicts Kidney Graft Rejection

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