ELISpot vs ICS Assays: Perfect Method in Immuno-Oncology Trials

October 29, 2020

Section image

Get a clear understanding of the ELISpot and ICS assays along with key elements to consider for the selection of the appropriate method to investigate your vaccine’s efficacy.

Key takeaways from this webinar:

  • Advantages and limitations of ICS and ELISpot for clinical trials
  • How to design ICS and ELISpot assays for maximal data gathering
  • Critical development and validation approaches to consider with ICS and ELISpot assays in order to fulfill regulatory requirements

To access this webinar, contact us.

Efficacy in a vaccine trial is assessed through the production of protective antibodies against the infectious agent.  Measurement of the neutralizing effect of these antibodies is performed through cell-based assays.

Over the past years, additional assays have been introduced in vaccine trials in order to get a better understanding of the vaccine’s mode of action on the immune system.

These assays allow for the monitoring of T-cell responses in subjects upon vaccination, including the humoral response (supported by CD4 helper T cells) and the elimination of infected cells by CD8 cytotoxic T cells (cellular response).

The mechanism can be further characterized through the profiling of the CD4 T cell polarization: Th1, Th2, Th17 and/or follicular helper T cells (Tfh), all of which can have an impact on the resulting therapeutic effect.

Flow cytometry and multimer detection can be used to follow an antigen specific T-cell response and reveal a specific TCR. However, this method allows to follow only one antigen and the functional state (effector/anergic) of the cells remains unknown.

Given these limitations, most trials now employ ELISpot and ICS to reveal both the antigen-specificity and the functionality of the immune response induced by the treatment.

Performing both ELISpot and ICS assays requires a lot of starting material (PBMC isolated from whole blood), which might not be accessible. The question then arises: which method to use? ELISpot or ICS?

On one hand, the ELISpot is commonly considered more sensitive than the ICS assay while requiring less amount of PBMC.

On the other hand, the ICS allows for the measurement of cytokines as well as surface and intracellular markers to generate a more complete cellular profile but consumes more of the precious PBMC material.

During this webinar, we will present:

(a) characteristics of both methods as well as their advantages and limitations

(b) approaches to define optimal assay conditions during development for robust assay performance and maximal data gathering

(c) validation approaches based on intended use of the data

(d) how to deploy these assays using appropriate criteria for control and study samples during sample analysis in infectious diseases (i.e., COVID-19) and immuno-oncology vaccine trials.

ELISpot vs ICS Assays: Perfect Method in Immuno-Oncology Trials

October 29, 2020

Section image

Get a clear understanding of the ELISpot and ICS assays along with key elements to consider for the selection of the appropriate method to investigate your vaccine’s efficacy.

Key takeaways from this webinar:

  • Advantages and limitations of ICS and ELISpot for clinical trials
  • How to design ICS and ELISpot assays for maximal data gathering
  • Critical development and validation approaches to consider with ICS and ELISpot assays in order to fulfill regulatory requirements

To access this webinar, contact us.

Efficacy in a vaccine trial is assessed through the production of protective antibodies against the infectious agent.  Measurement of the neutralizing effect of these antibodies is performed through cell-based assays.

Over the past years, additional assays have been introduced in vaccine trials in order to get a better understanding of the vaccine’s mode of action on the immune system.

These assays allow for the monitoring of T-cell responses in subjects upon vaccination, including the humoral response (supported by CD4 helper T cells) and the elimination of infected cells by CD8 cytotoxic T cells (cellular response).

The mechanism can be further characterized through the profiling of the CD4 T cell polarization: Th1, Th2, Th17 and/or follicular helper T cells (Tfh), all of which can have an impact on the resulting therapeutic effect.

Flow cytometry and multimer detection can be used to follow an antigen specific T-cell response and reveal a specific TCR. However, this method allows to follow only one antigen and the functional state (effector/anergic) of the cells remains unknown.

Given these limitations, most trials now employ ELISpot and ICS to reveal both the antigen-specificity and the functionality of the immune response induced by the treatment.

Performing both ELISpot and ICS assays requires a lot of starting material (PBMC isolated from whole blood), which might not be accessible. The question then arises: which method to use? ELISpot or ICS?

On one hand, the ELISpot is commonly considered more sensitive than the ICS assay while requiring less amount of PBMC.

On the other hand, the ICS allows for the measurement of cytokines as well as surface and intracellular markers to generate a more complete cellular profile but consumes more of the precious PBMC material.

During this webinar, we will present:

(a) characteristics of both methods as well as their advantages and limitations

(b) approaches to define optimal assay conditions during development for robust assay performance and maximal data gathering

(c) validation approaches based on intended use of the data

(d) how to deploy these assays using appropriate criteria for control and study samples during sample analysis in infectious diseases (i.e., COVID-19) and immuno-oncology vaccine trials.

T-Cell Response in COVID-19 Vaccine Development- A Webinar by CellCarta

October 1, 2020

Discussions on COVID-19 immune responses, including the responses in early vaccine clinical studies, have focused largely on antibody responses.

However, recent data indicate that the cellular immune response also plays an important role and should be considered when evaluating the immune response.

Assessing T-cell responses can provide critical information about vaccine responses and the development and persistence of protective immunity against SARS-CoV-2 that will be critical in the fight against COVID-19.

The webinar is co-sponsored by Caprion-HistoGeneX  and WCG and will feature the following speakers:

author photo

Scott Gottlieb, M.D., Former Commissioner of the Food and Drug Administration (FDA)
Dr. Gottlieb will discuss the implications of immune response testing for overall assessment, regulatory approval, and public acceptance of COVID-19 vaccines.

author photo

James Rothman, Ph.D., Sterling Professor of Cell Biology, Yale University; Recipient, Nobel Prize in Medicine, 2013
Dr. Rothman will provide an overview of the immune response to SARS-CoV-2 and the importance of cellular immunity. The key findings from our previous experiences with SARS-CoV-1 as well as the latest publications will also be discussed.

author photo

Eustache Paramithiotis, Ph.D., Vice President, Research & Development, Caprion-HistoGeneX
Testing methods for understanding complex immune system reactions as part of vaccine development programs will be discussed by Dr. Paramithiotis. Information on the methodology and accuracy of these assays will be provided.

Our speakers will discuss:

  • An overview of the current state of COVID-19 vaccine development
  • The adaptive immune response to SARS-CoV-2 and the importance of cellular immunity
  • Testing methods and assay standardization for understanding complex immune system reactions as part of vaccine development programs

Learn more about CellCarta

T-Cell Response in COVID-19 Vaccine Development- A Webinar by CellCarta

October 1, 2020

Discussions on COVID-19 immune responses, including the responses in early vaccine clinical studies, have focused largely on antibody responses.

However, recent data indicate that the cellular immune response also plays an important role and should be considered when evaluating the immune response.

Assessing T-cell responses can provide critical information about vaccine responses and the development and persistence of protective immunity against SARS-CoV-2 that will be critical in the fight against COVID-19.

The webinar is co-sponsored by Caprion-HistoGeneX  and WCG and will feature the following speakers:

author photo

Scott Gottlieb, M.D., Former Commissioner of the Food and Drug Administration (FDA)
Dr. Gottlieb will discuss the implications of immune response testing for overall assessment, regulatory approval, and public acceptance of COVID-19 vaccines.

author photo

James Rothman, Ph.D., Sterling Professor of Cell Biology, Yale University; Recipient, Nobel Prize in Medicine, 2013
Dr. Rothman will provide an overview of the immune response to SARS-CoV-2 and the importance of cellular immunity. The key findings from our previous experiences with SARS-CoV-1 as well as the latest publications will also be discussed.

author photo

Eustache Paramithiotis, Ph.D., Vice President, Research & Development, Caprion-HistoGeneX
Testing methods for understanding complex immune system reactions as part of vaccine development programs will be discussed by Dr. Paramithiotis. Information on the methodology and accuracy of these assays will be provided.

Our speakers will discuss:

  • An overview of the current state of COVID-19 vaccine development
  • The adaptive immune response to SARS-CoV-2 and the importance of cellular immunity
  • Testing methods and assay standardization for understanding complex immune system reactions as part of vaccine development programs

Learn more about CellCarta

Antigen Presentation: A Critical Step in Immune Therapy Dev.

June 10, 2020

In this webinar you will learn:

  • Successful antigen presentation is required to initiate an immune response
  • The most physiologically relevant way to measure the full breadth of antigen presentation is by direct observation of presented peptides using mass spectrometry
  • The quality and robustness of the mass spectrometry data generation is therefore a critical element

What is antigen presentation?

Antigen presentation is essential for development of a robust immune response.

Binding of an MHC-peptide complex by a T cell receptor is the initial and critical step for starting a new adaptive immune response or for regulating an ongoing one. Identification of effectively presented antigens is therefore a defining element for any successful immune therapy.

Antigen presentation can occur through MHC class I, class II, or non-classical MHC proteins. Each protein complex has characteristic tissue expression, type of antigen peptide presented, and role in the immune response.

Most lymphocytes including all of those with antigen presenting functions can express both MHC I and MHC II, and involvement of both MHC I- and MHC II-restricted responses is considered essential for a full immune response.

Non-classical MHC such as HLA-E and HLA-G are characterized by either a relatively restricted tissue distribution or antigen presentation repertoire and have specific roles to play in the establishment of robust immunity.

Direct observation of presented peptides by mass spectrometry has become required for novel epitope discovery. This approach is the most physiologically relevant way to detect the modified and unmodified peptides presented by any type of MHC.

Additionally, mass spectrometry can be used to quantify the peptide presentation to ensure that attractive targets are sufficiently expressed to trigger an immune reaction. The limiting factor has now become the quality of the mass spectrometry data generated.

This presentation will show examples of direct identification of novel presented peptides as well as survey the requirements for the industrialization of antigen presentation mass spectrometry to generate robust, reliable and quantitative measurements.

With an understanding that antigen presentation is required to initiate an immune response, we will discuss how to use this information in therapeutics development. Designing a good immune therapy requires that the antigen is specific to the targeted pathology, as well as immunogenic.

Those factors are especially relevant for modern immune therapies that seek to engage the host response as part of the complete therapy. Antigen presentation by mass spectrometry should therefore be embedded in a broader, integrated, characterization of host immunity.

Specific characterizations include evaluation of wider host antigen presentation to assess target specificity, monitoring the status of the host microenvironment, as well as host immune exhaustion before and after antigen dosing to better understand the capacity to respond to the therapy.

This presentation will depict case studies addressing each of these components of successful immune therapy candidates that start with successful antigen presentation.

Antigen presentation is the initial critical element, but for successful immune therapy development, antigen presentation should be embedded in an integrated assessment of host immunity

Speaker: Eustache Paramithiotis PhD, Vice-President, Research & Development, Caprion Biosciences Inc.

Antigen Presentation: A Critical Step in Immune Therapy Dev.

June 10, 2020

In this webinar you will learn:

  • Successful antigen presentation is required to initiate an immune response
  • The most physiologically relevant way to measure the full breadth of antigen presentation is by direct observation of presented peptides using mass spectrometry
  • The quality and robustness of the mass spectrometry data generation is therefore a critical element

What is antigen presentation?

Antigen presentation is essential for development of a robust immune response.

Binding of an MHC-peptide complex by a T cell receptor is the initial and critical step for starting a new adaptive immune response or for regulating an ongoing one. Identification of effectively presented antigens is therefore a defining element for any successful immune therapy.

Antigen presentation can occur through MHC class I, class II, or non-classical MHC proteins. Each protein complex has characteristic tissue expression, type of antigen peptide presented, and role in the immune response.

Most lymphocytes including all of those with antigen presenting functions can express both MHC I and MHC II, and involvement of both MHC I- and MHC II-restricted responses is considered essential for a full immune response.

Non-classical MHC such as HLA-E and HLA-G are characterized by either a relatively restricted tissue distribution or antigen presentation repertoire and have specific roles to play in the establishment of robust immunity.

Direct observation of presented peptides by mass spectrometry has become required for novel epitope discovery. This approach is the most physiologically relevant way to detect the modified and unmodified peptides presented by any type of MHC.

Additionally, mass spectrometry can be used to quantify the peptide presentation to ensure that attractive targets are sufficiently expressed to trigger an immune reaction. The limiting factor has now become the quality of the mass spectrometry data generated.

This presentation will show examples of direct identification of novel presented peptides as well as survey the requirements for the industrialization of antigen presentation mass spectrometry to generate robust, reliable and quantitative measurements.

With an understanding that antigen presentation is required to initiate an immune response, we will discuss how to use this information in therapeutics development. Designing a good immune therapy requires that the antigen is specific to the targeted pathology, as well as immunogenic.

Those factors are especially relevant for modern immune therapies that seek to engage the host response as part of the complete therapy. Antigen presentation by mass spectrometry should therefore be embedded in a broader, integrated, characterization of host immunity.

Specific characterizations include evaluation of wider host antigen presentation to assess target specificity, monitoring the status of the host microenvironment, as well as host immune exhaustion before and after antigen dosing to better understand the capacity to respond to the therapy.

This presentation will depict case studies addressing each of these components of successful immune therapy candidates that start with successful antigen presentation.

Antigen presentation is the initial critical element, but for successful immune therapy development, antigen presentation should be embedded in an integrated assessment of host immunity

Speaker: Eustache Paramithiotis PhD, Vice-President, Research & Development, Caprion Biosciences Inc.