FIRST in ELISPOT
CTL is the only commercial entity that has extensive, first-hand expertise in all aspects of ELISPOT:
CTL's Founder and CEO, Prof. Paul V. Lehmann, and the scientists working with him were the FIRST to:
- Build an ELISPOT reader (US Patent 6,410,252 B1; filed 12/95) – marking the beginning of the CTL ImmunoSpot® reader line which introduced objectivity, reproducibility, and documentation trails in ELISPOT analysis.
- Introduce and use PVDF plates for ELISPOT assays (Science, 1996, 271:1728-30 and US Patent 6,140,252; filed 12/95), which was one of the critical improvements that transformed an assay that previously had the reputation of “cold fusion” into the robust platform it is today.
- Define which cytokines and spot sizes are T cell-derived (and thus relevant for T cell diagnostics) and which are “bystander” cytokine spots (J. Immunol. 2002,168:545-53 and J. Neuroimmunol. 2005, 170:105-14), introducing the basics for using ELISPOT as a scientifically-validated T cell diagnostic tool.
- Use ELISPOT to understand different qualities of immunity induced by vaccination, delineating different T cell effector classes (Science, 1996, 271:1728-30).
- First to establish that CpG PAMS induce Th1 immunity (J. Exp. Med. 1997, 186:1623-31).
- Establish the scientific principles of ELISPOT analysis, such as how to interpret different spot sizes (J. Immunol. 2001, 167:1353-61), introducing the basics of automated gating strategies that ImmunoSpot® applies.
- Use ELISPOT for measuring T cell avidity for antigen (e.g., J. Exp. Med. 1998, 187:2055-63), introducing an important additional parameter of T cell immunity that ELISPOT assays can provide.
- Apply ELISPOT to comprehensive determinant mapping (e.g., J. Immunol. 2000, 165:1641-51), introducing the high-throughput capabilities of the assay.
- Show that, when following the CTL protocols, PBMC can be cryopreserved without loss of function in ELISPOT assays (J. Immunol Methods 2003, 278:79-93), enabling the testing of clinical samples in specialized central laboratories and to generate cryopreserved PBMC libraries for reference samples and streamlined human research.
- Perform assays with a minute amount of cell material, such as isolates of mouse brain or blood (J. Immunol. 2001,166:4757-64 and J. Immunol.2005, 174:4598-605), showing ELISPOT’s unique efficacy in cell utilization.
- Introduce double-color ELISPOT analysis with the ability to detect cytokine co-expressing/“polyfunctional” T cells with the same sensitivity as ICS (J. Immunol. 2000 164:1862-72).
- Introduce granzyme B (J. Immunol. Methods 240:143-155, 2000) and perforin ELISPOT assays (AIDS Research and Human Retroviruses 2007, 24: 62-71 for detecting effector CD8 cells, and distinguishing them from memory cells that produce IFN-γ only, and TRAIL assays to detect helpless CD8 cells (AIDS Res Hum Retroviruses. 2008 24:1175-83).
- Use ELISPOT to study transplant immunity (Transplantation 1997, 64:292-6, and US Patent 5,939,281; filed 9/96).
- Pioneer autoimmunity using ELISPOT (J. Exp. Med. 1996, 183:1561-7).
- Study adjuvant-guided T cell responses (J. Exp. Med. 1997 186:1623-31).
- Use ELISPOT to study T cell immunity to HIV (J. Immunol. 2000, 164:3723-32).
- Use ELISPOT to study anti-tumor immunology (J. Immunol. 2003, 171:3941-6).
- First to establish that ImmunoSpot® is a robust T cell monitoring platform that generates highly reproducible results in different laboratories (J. Immunotoxicology, 2009, 6:227).
- Introduce ELISPOT to diagnosing Multiple Sclerosis (Clinical Immunol. 2014, 152:20-24)
- Introduce and validate four-color T cell and B cell ELISPOT (Four-color T cell and B cell ELISPOT assay for simultaneous detection of all four human IgG subclasses)
- First to harmonize ELISPOT between laboratories (ELISPOT assays provide reproducible results among different laboratories for T cell immune monitoring) (User vs. software-dependent variability of ELISPOT counts obtained from nine different laboratories)
- To characterize Th1, Th2, Th17 immunity and polyfunctional T cells in HCMV infection (Characterization of the CMV-specific CD4 cell response).
- Introduce and validate 384 well ELISPOT assays (ELISPOT assays in 384-well format allow analysis of 400 independent T cell tests with 10ml blood)
- Study the impact of apoptotic cells within PBMC on ELISPOT results (Are apoptotic cells in a PBMC population bystanders or do they interfere with antigen-specific T cell assays?)
- Systematically study the (lack of) benefit of PBMC resting (Cells 2012, 1:313-324)
- Systematically study interference of peptides within peptide pools (Viruses 2012, 4:2636-2649)
- First to establish a CLIA certified, GLP-compliant (registered 6/09) laboratory.
More of Prof. Lehmann’s publications on ELISPOT are sorted here by theme, providing more information on the basics of ELISPOT and how it can be put to innovative use, possibly offering useful references for you.