Finally, the prognostic impact of the lymphocytic profile change has not been formally evaluated, and the only publication addressing the relevance of post-NCT lymphocytic infiltration in the residual tumor is confined to triple negative breast cancer and does not include data regarding the different lymphocyte sets [ 26].
CD4+ CD25− effector T cells (Teff), were magnetically isolated from PBMC using a CD4 T Lymphocyte Enrichment Set (BD Biosciences).
Mouse CD4+ T cells were purified from a bulk spleen population using a mouse CD4 T lymphocyte enrichment set (BD IMag™).
Cells were analysed using a Coulter Epics XL flow cytometer (Beckman-Coulter, High Wycombe, UK) with a lymphocyte gate set to exclude dead cells and debris.
To address the question of the differential sensitivity to HDACIs of tumor cells as compared to normal cells, and to gain some insight into the pharmacological mechanism of action of these compounds in non-transformed cells, particularly in lymphocytes, we set out to characterize the effects of HDACIs on normal primary murine CD4+ T lymphocytes.
While most immunological studies of lymphocyte sub-sets have failed to show any immune system changes following scrapie infection, recent evidence suggests that B-lymphocytes [13] and in particular the CD21 B-lymphocyte population [14] may be affected.
The genomic positions of start and end points of transcripts, their coding sequences, and their exons and introns in both brain and lymphocyte gene sets were collected from the UCSC genome browser using Genbank accession numbers (e.g. NM_018711) as identifiers.
A ~77% accuracy was reported for the CD4 lymphocytes data set using a nearest neighbor-based classifier [ 9].
Other investigations, including a complete blood count, blood film, chest radiograph, virology screen, lymphocyte sub-sets, Heaf test (Screening for Tuberculosis) and HIV test, were all normal.
Cell cultures of PHA-stimulated lymphocytes were set up according to the standard protocol used for routine karyotype assays described previously [ 26].
