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Monitoring Panel

Laboratory Project: “The CIMT Monitoring Panel”

  1. Introduction to the work of the CIMT Monitoring panel

  2. Perspectives of the working group

  3. Interlaboratory testing phases
    3.1 Phase I/2005 (tetramer staining, ELISPOT and ICS)
    3.2 Phase II/2006 (tetramer staining and ELISPOT)
    3.3 Phase III/2007 (tetramer staining)
    3.4 Phase IV/2007 (ELISPOT)
    3.5 Phase V/2007 (ICS)
  4. Active members of the CIMT Monitoring Panel

  5. Contact

  6. Publications and Communications

  7. Supporters of the CIMT Monitoring Panel


1. Introduction to the work of the CIMT Monitoring panel

In February 2005 a working group was founded under the aegis of the Association for Immunotherapy of cancer ("CIMT"). The aims of this group are the critical discussion and practical comparison of the different techniques and various protocols applied for the enumeration and characterization of antigen-specific T-cell responses. As a result of the first meeting of the working group members, a series of inter-laboratory testing projects, called "CIMT Monitoring panels", have meanwhile been initiated. Each panel phase compares individual protocols and aims at identifying distinct critical variables that are predictive for high/low performance in the assays used. Assay-specific recommendations are then formulated in order to improve the sensitivity of the tests. Another aim of the interlaboratory testing projects is the external validation of each laboratory that participates in the panel and this may lead to a certification in the future. The members of the CIMT monitoring panel meet once a year during the CIMT meeting in order to discuss results from the conducted panels, learn more about new developments in the field and to recruit participants for new panel phases. The group is open to all laboratories that have a focus on the monitoring of antigen-specific T-cell responses.

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2. Perspectives of the workgroup

The members of the CIMT Monitoring Panel are dedicated to:
  1. Establish a group of professional developers and users in order to exchange techniques, protocols and reagents and to critically discuss results obtained by our laboratory panels
  2. Organize further interlaboratory testing projects in order to compare various techniques, variables and local protocols for measuring antigen-specific T cells
  3. Share experience with other scientists and working groups that work in the field of immune monitoring (cooperation with the Cancer Vaccine Consortium of the Sabin Vaccine Institute)
  4. Advise in the monitoring of T cell responses in clinical studies, including help for training of staff members
  5. Organize extra-sessions with focus on immunomonitoring during the annual CIMT meetings. International speakers will be invited to present recent developments in the field
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3. Interlaboratory testing phases

3.1 Interlaboratory testing phase I/2005

Conclusions:

Parts of the results from the first phase were presented as a poster during the annual meeting CIMT 2006 (pdf, 156 kb). A manuscript summarizing the results of phases I/2005 and II/2006 has been submitted for publication.

After phase I/2005, we concluded from the tetramer tests that the total number of cells analyzed and the choice of gates and quadrants set for calculating cell frequencies were the most important determinants for the sensitivity and interpretation of the tetramer staining. We therefore proposed to introduce an extended guideline for tetramer staining, specifying setting of the gates and defining a minimum number of cells for each staining. From the ELISPOT tests we concluded that the combination of a high number of cells and a resting phase after thawing should result in a higher sensitivity of the ELISPOT assay. In order to formally show that recommendations deduced from the results of phase I/2005 lead to improved assay performance we repeated the experiments in phase II/2006.

3.2 Interlaboratory testing phase II/2006

Conclusions:

The results of phase II confirmed our previous conclusions and lead to the following recommendations:
Do stain at least 1 x 106 PBMC per test and do acquire all stained cells. The lower the frequency of antigen-specific CD8+ cells is, the higher the number of cell events acquired and analyzed should be.
The combination of more than 400000 PBMC per well and a resting-phase after thawing results in a higher sensitivity of the ELISPOT assay. Allogeneic APC should not be used on a routine basis and tests should be performed in triplicate wells. We also recommend to perform a statistical analysis on all triplicates and to indicate results obtained in the format of 1 spot per x PBMC.

A manuscript summarizing the results of phases I/2005 and II/2006 has been submitted for publication.

3.3 Interlaboratory testing phase III/2007

Conclusions: scheduled for second half of 2008

3.4 Interlaboratory testing phase IV/2007

Conclusions: scheduled for second half of 2008

3.5 Interlaboratory testing phase V/2008

Conclusions: scheduled for second half of 2008

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4. Members of the CIMT Monitoring Panel

Managing committee:

Cedrik M. Britten, MD
Department for Immunohematology and Blood Transfusion
Leiden University Medical Center
Albinusdreef 2
2333 ZA Leiden, The Netherlands
Email:
Phone: +31 71 - 526 3013

Sjoerd H. van der Burg, PhD
Department for Clinical Oncology
Leiden University Medical Center
Albinusdreef 2
2333 ZA Leiden, The Netherlands
Email:
Phone: +31 71 - 526 11 80


gouttefangeas Cecile Gouttefangeas, PhD
Department of Immunology, University of Tuebingen
Auf der Morgenstelle 15
72076 Tuebingen, Germany
Email:
Phone: +49 70 71 - 29 80 994

Participating countries

Participating countries

Participating Members:

Meeting of the group in mai 2005q
Meeting of the group in mai 2005

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5. Contact and questions

If you have any questions about the work of the CIMT Monitoring Panel or if you would like to join the group, do not hesitate to contact the managing committee.

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6. Publications and Communications

Oct. 2005: Report phase I/2005 (available upon request)

Nov. 2006: Report phase II/2005 (available upon request)

July 2007: First guidelines for tetramer staining (pdf 36 kb)

July 2007: First guidelines for IFN-γ ELISPOT (pdf 28 kb)

Results from the CIMT Monitoring panel have been published in the following journals

August 2007: Original Article: "The CIMT-monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8(+) T lymphocytes by structural and functional assays."
Cancer Immunol Immunother. 2007 Aug 25; [Epub ahead of print]

A full text version of the article is available via Springer open choice at:
http://www.springerlink.com/content/y222652657r73810/

August 2007 Commentary: "Toward the harmonization of immune monitoring in clinical trials: Quo vadis?"
Cancer Immunol Immunother. 2007 Aug 25; [Epub ahead of print]

A full text version of the article is available via Springer open choice at:
http://www.springerlink.com/content/p8975670262226nr/

Results from the CIMT Monitoring panel have been presented at the following meetings

Scheduled for 2008:

9th International Conference on Chronic Myeloid and Lymphoid Malignancies, 29 -31 May 2008, Kazimierz Dolny, Poland - Oral Presentation - "Standardization of T cell immunomonitoring - protocol choices and additional sources of variation."

2008

9th International Symposium Biological Therapy of Cancer / From Disease to Targeted Therapy, 12-15 March 2008, Munich, Germany - Oral presentation - "Standardization of T cell immunomonitoring - correct protocol choices and additional sources of variation"

MASIR 2008 - Measurement of Antigen-Specific Immune Responses, 30 January - 2 February 2008, La Plagne, France - Oral presentation - "The CIMT-Monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8+ T lymphocytes."

2007

DC THERA IMMUNOMONITORING WORKSHOP, 6-7 December 2007, Brussels,
Belgium - Oral presentations - "The CIMT-Monitoring panel: efforts towards the harmonization of T-cell monitoring."
Part 1: Harmonizing/Optimizing assay protocols.
Part 2: Harmonization of additional aspects needed?

CANCER VACCINES ADJUVANTS & DELIVERY SYSTEMS - CVADD 2007 Conference, 10-12 October 2007, Heidelberg, Germany - Oral presentation: "The CIMT-Monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8+ T lymphocytes."

CVC Colloquium and Annual Meeting, 27-29 September 2007, Washington, USA -
Oral presentation: "First Harmonization Criteria for Cellular Immune Response Assays for Cancer Vaccine Trials - Results from Two Large Proficiency Panels"

CIMT 2007, 12-14 April, Wuerzburg, Germany - Oral presentation: "Lessons learned from interlaboratory testing in phases I/2005 and II/2006"

2006

Symposium titled: "The need for standardisation of Immune Competence Assessment and Monitoring", 16 October 2006, Cardiff, Wales - Oral presentation: "Defining standardisation for immune monitoring"

CIMT 2006, 4-5 May 2006, Mainz, Germany - Poster abstract P04: "The CIMT Monitoring Panel: Enumeration of antigen-specific CD8+ T lymphocytes by structural and functional assays." Cancer Immunol Immunother. (2007) 56:406-407.

Progress in Vaccination against Cancer 2006 - PIVAC 6 conference, 28-29 September 2006, Granada, Spain - Oral presentation: "The CIMT monitoring panel: Standardization of in vitro methods for enumerating antigen specific CD8+ T lymphocytes".

 

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7. Supporters of the CIMT Monitoring Panel

 

The activities of the CIMT monitoring panel are supported by Beckman Coulter and Immatics. This partnership enabled us to donate traveling grants to presenting group members and to invite external and internationally recognized speakers to the annual meeting of the group ("CIMT 2008 in Mainz - Immunomonitoring session on Friday 16th May 2008"). Both companies thereby help us to foster the aspect of specific education and dissemination of results within the scientific community. Their help is much appreciated.

beckman immatics

 

 

 
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