Evaluation of PD-L1 expression in various formalin-fixed paraffin embedded tumour tissue samples using SP263, SP142 and QR1 antibody clones

Background & objectives: Cancer cells can avoid immune destruction through the inhibitory ligand PD-L1. PD-1 is a surface cell receptor, part of the immunoglobulin family. Its ligand PD-L1 is expressed by tumour cells and stromal tumour infltrating lymphocytes (TIL). Methods: Forty-four cancer cases were included in this study (24 triple-negative breast cancers (TNBC), 10 non-small cell lung cancer (NSCLC) and 10 malignant melanoma cases). Three clones of monoclonal primary antibodies were compared: QR1 (Quartett), SP 142 and SP263 (Ventana). For visualization, ultraView Universal DAB Detection Kit from Ventana was used on an automated platform for immunohistochemical staining Ventana BenchMark GX. Results: Comparing the sensitivity of two different clones on same tissue samples from TNBC, we found that the QR1 clone gave higher percentage of positive cells than clone SP142, but there was no statistically significant difference. Comparing the sensitivity of two different clones on same tissue samples from malignant melanoma, the SP263 clone gave higher percentage of positive cells than the QR1 clone, but again the difference was not statistically significant. Comparing the sensitivity of two different clones on same tissue samples from NSCLC, we found higher percentage of positive cells using the QR1 clone in comparison with the SP142 clone, but once again, the difference was not statistically significant. Conclusion: The three different antibody clones from two manufacturers Ventana and Quartett, gave comparable results with no statistically significant difference in staining intensity/ percentage of positive tumour and/or immune cells. Therefore, different PD-L1 clones from different manufacturers can potentially be used to evaluate the PD- L1 status in different tumour tissues. Due to the serious implications of the PD-L1 analysis in further treatment decisions for cancer patients, every antibody clone, staining protocol and evaluation process should be carefully and meticulously validated

Therapeutic possibilities of kinesitherapy in progressive muscular dystrophy

Objective: To determine the effectiveness of targeted kinesitherapy in patients with progressive muscular dystrophy. Material and methods: A study was performed in 12 patients, mean age of 45.1 ± 5.8 years with affected proximal muscle groups after being diagnosed with progressive muscular dystrophy (PMD). For this purpose, a Functional Independence Test (FIM) and a Five Times Sit-To-Stand (FTSST) test were used to measure a patient’s functional mobility. Applied kinesitherapy includes exercises to overcome muscle imbalance, through analytical exercises according to the assessment of manual muscle test; improving compensatory self-service capabilities and functional independence; and maintaining cardiorespiratory fitness. The changes in the physical activity were monitored three times after hospital discharge in home rehabilitation. Results: After 1 month of kinesitherapy, an improvement in the functional independence and transverse capabilities of patients with PMD was observed (p <0.001). The improvement in the speed of reactions and muscle strength of the patients is due to the regular kinesitherapy sessions and exercises for strength of the lower limbs and abdominal muscles. Conclusion: The application of 1-month kinesitherapy improves the functional independence of patients with PMD compared to their initial condition. Key words: kinesitherapy, progressive muscular dystrophy, therapeutic possibilitie

Evaluation of PD-L1 expression in tumor tissue

The interaction between the immune system and cancer cells is complex. Cancer cells can avoid immune destruction by suppressing the immune system. Tumors can create an immunosuppressive microenvoirment and therefore cytotoxic T lymphocytes (CTL) can't lysate them. Avoidance of the immune response is through the inhibitory ligand PD-L1, which is found on the surface of cells of the immune system, mesenchymal, epithelial, and endothelial cells as well as tumor cells. PD-1 is a surface cell receptor, part of the immunoglobulin family. Its ligand PD-L1 is expressed by tumor cells and stromal tumor infiltrating lymphocytes (TIL). This information has been used to increase the therapeutic possibilities for many malignant diseases. Objectives: 1.Optimization of immunohistochemical method (IHH) with antibody staining, anti PD-L1. 2. Optimization of the evaluation method while using different clones of antibodies. Material and methods: In this study are included 44 samples from 3 different types of cancer. 24 triple-negative breast cancers (TNBC), 10 non-small cell lung cancer (NSCLC) and 10 malignant melanoma. Monoclonal mouse primary antibodies have been used by manufacturers Quartett (QR1) and Ventana (SP 142, SP263). To visualize the reaction, an ultraView Universal DAB Detection Kit from Ventana was used on an automated platform for immunohistochemical staining Ventana BenchMark GX. The statistical analysis was done with the statistical package SPSS for Windows Version 22.0. Results: With comparing the sensitivity of two different clones on the same tissue samples from triple-negative breast cancer (TNBC), it can be conclude that the Clone QR1 gives more positive results than the Clone SP142, but there is no statistically significant difference. With comparing the sensitivity of two different clones in the same tissue samples from malignant melanoma, it can be conclude that Clone SP263 gives more positive results than Clone QR1, but there is no statistically significant difference. With comparing the sensitivity of two different clones in the same tissue samples from non- small cell lung cancer, we concluded that Clone QR1 gives more positive results than Clone SP142, but there was no statistically significant difference. Conclusion: Use of any of these clones of the manufacturers Quartett and Ventana give satisfactory results and can be used in the determination and evaluation of PD-L1 status in patients, and thus to determine further therapy and outcome of patients. Keywords: breast cancer, malignant melanoma, non-small cell lung cancer, immunohistochemistry, antibodie