Results and harmonization guidelines from two large-scale international Elispot proficiency panels conducted by the Cancer Vaccine Consortium (CVC/SVI)

被引：151

作者：

Janetzki, S. ^{[1
]}

Panageas, K. S. ^{[2
]}

Ben-Porat, L. ^{[2
]}

Boyer, J. ^{[3
]}

Britten, C. M. ^{[4
]}

Clay, T. M. ^{[5
]}

Kalos, M. ^{[6
]}

Maecker, H. T. ^{[7
]}

Romero, P. ^{[8
]}

Yuan, J. ^{[9
]}

Kast, W. Martin ^{[10
]}

Hoos, A. ^{[11
]}

机构：

[1] ZellNet Consulting Inc, Ft Lee, NJ USA

[2] Mem Sloan Kettering Canc Ctr, Dept Epidemiol & Biostat, New York, NY USA

[3] Univ Penn, Dept Pathol & Lab Med, Philadelphia, PA USA

[4] Leiden Univ, Med Ctr, Dept Blood Transfus & Immunhematol, Leiden, Netherlands

[5] Duke Univ, Med Ctr, Duke Comprehens Canc Ctr, Dept Surg, Durham, NC USA

[6] City Hope Natl Med Ctr, Div Canc Immunotherapeut & Tumor Immunol, Div Hematol & Hematopoiet Cekk Transplantat, Clin Immunobiol Correlat Studies Lab, Duarte, CA USA

[7] BD Biosci, San Jose, CA USA

[8] CHU Vaudois, Univ Hosp, Ludwig Inst Canc Res, Div Clin Onco Immunol,Lausanne Branch, Lausanne, Switzerland

[9] Mem Sloan Kettering Canc Ctr, Dept Med, New York, NY 10021 USA

[10] Univ So Calif, Kenneth Norris Jr Comprehens Canc Ctr, Dept Mol Microbiol & Immunol, Los Angeles, CA USA

[11] Bristol Myers Squibb Co, Wallingford, CT 06492 USA

来源：

CANCER IMMUNOLOGY IMMUNOTHERAPY | 2008年 / 57卷 / 03期

关键词：

Elispot; proficiency panel; validation; harmonization; immune monitoring;

D O I：

10.1007/s00262-007-0380-6

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

The Cancer Vaccine Consortium of the Sabin Vaccine Institute (CVC/SVI) is conducting an ongoing large-scale immune monitoring harmonization program through its members and affiliated associations. This effort was brought to life as an external validation program by conducting an international Elispot proficiency panel with 36 laboratories in 2005, and was followed by a second panel with 29 participating laboratories in 2006 allowing for application of learnings from the first panel. Critical protocol choices, as well as standardization and validation practices among laboratories were assessed through detailed surveys. Although panel participants had to follow general guidelines in order to allow comparison of results, each laboratory was able to use its own protocols, materials and reagents. The second panel recorded an overall significantly improved performance, as measured by the ability to detect all predefined responses correctly. Protocol choices and laboratory practices, which can have a dramatic effect on the overall assay outcome, were identified and lead to the following recommendations: (A) Establish a laboratory SOP for Elispot testing procedures including (A1) a counting method for apoptotic cells for determining adequate cell dilution for plating, and (A2) overnight rest of cells prior to plating and incubation, (B) Use only pre-tested serum optimized for low background: high signal ratio, (C) Establish a laboratory SOP for plate reading including (C1) human auditing during the reading process and (C2) adequate adjustments for technical artifacts, and (D) Only allow trained personnel, which is certified per laboratory SOPs to conduct assays. Recommendations described under (A) were found to make a statistically significant difference in assay performance, while the remaining recommendations are based on practical experiences confirmed by the panel results, which could not be statistically tested. These results provide initial harmonization guidelines to optimize Elispot assay performance to the immunotherapy community. Further optimization is in process with ongoing panels.

引用

页码：303 / 315

页数：13

共 39 条

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