Flow Cytometric Assay and Analysis

This assay uses antibodies and to stain cell populations in patient specimens. The MITS Core offers cell staining and data analysis services. Sample acquisition is performed by the UVA Flow Cytometry Core, utilizing their wide range of flow cytometry instruments.

We currently offer services for custom panel design and staining for surface and intracellular targets.

Ready-to-use flow cytometry panels:

• Lymphocyte Panel: Cell Viability (Aqua), CD3, CD4, CD8, CD14, CD19, CD56; CD45RA or RO, CD62L and/or CCR7, CD27, CD28
• Myeloid Panel: Cell Viability (Aqua), CD14, 11b, 11c, CD16, CD123, HLADR, CD68, CD163, CD33, CD206, CD66b, CD141, CD1c, CD80, CD86.
• Regulatory Cell Panel: Cell Viability (Aqua), CD3, CD4, CD8, CD25, CD127, FoxP3, Ki67,
• T Effector Cell/Regulatory Panel: Cell Viability (Aqua), CD3, CD4, CD8, Tbet, EOMES, Ki67, HLADR, PD-1, TIM-3
• NK Panel: Cell Viability (Aqua), CD3, CD4, CD8, CD56, NKp46, NKDG2, CD57
• Companion ELISpot Panel: Cell Viability, CD3, CD8, CD4; these samples will be run on the Guava easyCyte Instrument located in the MITS Core

Brief workflow for flow cytometry assays is as follows:

1. The MITS core offers services to stimulate cells and to stain cells from patient specimens.
2. The MITS core would facilitate sample transfer to the UVA flow cytometry core, where the stained cell samples would run through flow cytometers and cell population data will be acquired.
   The subsequent data would then be returned to the MITS core.
3. The MITS core could then assist with data analysis per the investigators needs to return finalized data to the investigator, or pass the raw flow cytometry data to the investigator for future analysis.

Additionally, investigators have the option to run the samples themselves in the UVA Flow Core, or to contract the Flow or MITS Core to run the samples for them on a cytometer at an additional fee.

For questions regarding flow cytometry services, please contact Kelly Smith at 434-243-6505 or kts4v@virginia.edu.

Phase I/II clinical trial of a helper peptide vaccine plus PD-1 blockade in PD-1 antibody-naïve and PD-1 antibody-experienced patients with melanoma (MEL64). Vavolizza RD, Petroni GR, Mauldin IS, Chianese-Bullock KA, Olson WC, Smith KT, Dengel LT, Haden K, Grosh WW, Kaur V, Varhegyi N, Gaughan EM, Slingluff CL Jr. J Immunother Cancer. 2022 Sep; 10(9):e005424. doi: 10.1136/jitc-2022-005424. PMID: 36100309.

A phase 1 study of NY-ESO-1 vaccine + anti-CTLA4 antibody Ipilimumab (IPI) in patients with unresectable or metastatic melanoma.Clinical Trial OncoImmunology (online). Craig L. Slingluff jr, Hassane M. Zarour, Hussein Abdul-Hassan Tawbi, John M. Kirkwood, Michael A. Postow, Philip Friedlander, Craig E. Devoe, Elizabeth M. Gaughan, Ileana S. Mauldin, Walter C. Olson jr, Kelly T. Smith, Mary J. Macri, Toni Ricciardi, Aileen Ryan, Ralph Venhaus & Jedd D. Wolchok. March 26. Volume 10, Issue 1, 2021. DOI: 10.1080/2162402X.2021.1898105.

Trial to evaluate the immunogenicity and safety of a melanoma helper peptide vaccine plus incomplete Freund’s adjuvant, cyclophosphamide, and polyICLC (Mel63). Slingluff CL Jr, Petroni GR, Chianese-Bullock KA, Wages NA, Olson WC, Smith KT, Haden K, Dengel LT, Dickinson A, Reed C, Gaughan EM, Grosh WW, Kaur V, Varhegyi N, Smolkin M, Galeassi NV, Deacon D, Hall EH.  J Immunother Cancer. Jan; 9(1):e000934, 2021. DOI: 10.1136/jitc-2020-000934.

A Multipeptide Vaccine Plus Toll-like Receptor Agonists LPS or polyICLC in Combination with Incomplete Freund’s Adjuvant in Melanoma Patients. Marit Melssen; Gina Petroni; Kimberly Chianese-Bullock; Nolan Wages; William Grosh; Nikole Varhegyi; Mark Smolkin; Kelly Smith; Nadejda Galeassi; Donna Deacon; Elizabeth Gaughan; Craig Slingluff. Journal for ImmunoTherapy of Cancer; 2019. DOI: 10.1186/s40425-019-0625-x.