Optimisation of the quantitative analysis of inflammatory cell infiltrates in breast cancer /Optimizarea analizei cantitative a infiltratului celular inflamator în cancerul mamar

In this study we aimed to determine the optimal cut-off point for the quantitative analysis of inflammatory infiltrates in breast cancer, using the HistoQuest system. We used samples of tumour breast tissue which were IHC stained with CD68 and CD8 and subsequently tested with automated systems on three regions: intratumoral, invasive front and peritumoral, using the HistoQuest system. In order to delimit between positive and negative cells on histograms and scattergrams, we need to set a cut-off value. We compared 5 cut-off types for optimisation of the quantitative analysis. The results obtained statistically for the CD8 marker for all 5 types of cut-offs applied on IT, PT and IF regions did not show statistically significant differences (p > 0.05). As for the CD68 marker, we found statistically significant differences (p < 0.05) between manual cut-offs (C2 - manual and C3 - manual, arithmetic mean) and automated cut-offs placed by the software (C1 - automated, C4 - negative region, and C5 - automated, arithmetic mean), which suggests that the use of an automated cut-off should be preferred in order to remove the subjective factor. The automated cut-off setting generates objective and reproducible data and can be used in subsequent quantitative analyses. În acest studiu am analizat modalitatea optimă de poziţionare a cut off-ului în cadrul analizei cantitative a infiltratului inflamator în cancerul mamar cu ajutorul sistemului HistoQuest. Am utilizat specimene de ţesut mamar tumoral, care au fost marcate imunohistochimic cu markerii CD68 (pentru macrofage) şi CD8 (pentru limfocitul T citotoxic) şi ulterior analizate automat pe regiunile IT (intratumoral), PT (peritumoral) şi FI (front de invazie) cu ajutorul sistemului soft HistoQuest. Pentru evidenţierea populaţiei de celule pozitive pe histograme şi scatergrame, este necesară poziţionarea cut off-ului (discriminator). Pentru optimizarea tehnicii, am comparat 5 modalităţi de poziţionare a cut off-ului. Rezultatele obţinute statistic pentru markerul CD8 pentru toate cele 5 tipuri de cutoff aplicate pe regiunile IT, PT şi FI nu au înregistrat diferenţe semnificative statistic (p < 0,05). Pentru markerul CD68 s-au înregistrat diferenţe semnificative statistic (p