In this interview, Cerba Research describes how its teams were able to discover resident memory T cells in solid tumors thanks to multiplex immunofluorescence.
Can you provide an overview of tissue-resident memory T cells and their role in the clinical setting?
tissue resident memory (TRM) cells It is representative of a genus of T cells localized in peripheral tissues that do not return to the circulation. Since they are embedded in tissues, they represent the primary response of T cells in local infections and inflammations.
The TRM is characterized by the expression of CD103, CD69 and CD49a. TRM plays a central role in the development of and effectiveness Cancer vaccines have been associated with improved prognosis and survival in a number of cancers. However, comparisons of TRM and cytotoxic CD8 T cells in the tumor microenvironment are limited.
What is the main aim of your studies?
We set out to develop HISTOPROFILE®Multiplex TRM panel of phenotypic memory T cells in solid tumors showing the ability of the panel to study TRM subpopulations in resected NSCLC FFPE tissue. This was achieved using plate design and validation.
What are your main goals while searching?
The targets were CD3/CD8/CD49a/CD69/CD103; Memory resident T cells – CD8+/ CD103+; and subtypes of memory-resident T cells – CD8+/ CD49a+CD8+/ CD69+CD8+/ CD103+/ CD69+CD8+/ CD103+/ CD69+/ CD49a+.
To achieve this, we obtained three NSCLC tissue excision samples from the Cerba Reseach Montpellier Biobank. The NSCLC tumor area was determined on slides stained with hematoxylin and eosin by a pathologist.
What application methods were applied during the validation of the study?
Once the segments are identified, the multicast protocol is serialized with Opal® (Akoya Biosciences) fluorophores were applied to BOND RX® (Leica) stained slide. Then, multispectral images were taken using VECTRA® PolarisTM slide scanner (Akoya Biosciences) performing full slide image analysis using HALO® (Indica Labs) Highplex unit on unmixed NSCLC images.
Can you describe the detection method used during the research?
Simplex slides were stained for each biomarker in a simple protocol and compared to serial section stained with HISTOPROFILE®-TRM MULTI PLATE. Staining compatibility between multiple slides and simple transmissions was determined using HALO® Without multispectral decoding. Accuracy of the protocol was determined by an intra-run redundancy analysis and an inter-run reproducibility analysis (data not shown).
How do you test the effectiveness of the design of the board?
The power of the plate has been demonstrated on many human solid tumors, including cancers of the prostate, colon, lung, kidney, and Hodgkin’s lymphoma, skin cancer, and serous ovarian adenocarcinoma. Next, multispectral full-scan images were analyzed using HALO to determine the effectiveness of the method.
Can you summarize why a multiplex panel is appropriate for the detection of tissue-resident memory T cells in solid tumors?
histoprofile tissue®The TRM panel consists of markers CD8, CD3, CD69, CD103 and CD49a, and has been developed and used specifically for the detection of resident T cells in human NSCLC samples.
The panel strongly demonstrates the novel ability to phenotype TRM with different combinations of listed targets.
Different subpopulations of TRM in neoplastic and non-neoplastic regions can be identified through image analysis. The strength and durability of the multiplex board make it an excellent tool Investigate the TRM population across a cohort of human solid tumors.
Simplex slides were stained for each individual biomarker in a simple protocol and compared to serial section stained with multiplexed IHC. The coloration concordance between the multiplex slice and the simplex slice was determined by analysis with HALO® After multispectral deconvolution. a) Example images (top) of a single slice of target A (left), the multiplex slice (right), and the corresponding HALO slice® Masks identify positive cells (bottom). B) Results of analysis of human and mouse TRIG plates. The percentage of positive cells from the simplex slide (black bar) is compared to the multiplex slide (white bar). Satisfactory results were obtained for both HISTOPROFILE® -Treg panels. Image credit: Cerba Research
Image credit: Cerba Research
Image credit: Cerba Research
Image credit: Cerba Research
Image credit: Cerba Research
Image credit: Cerba Research
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