ABSTRACT
PD-1/PD-L1-based therapy has been named a revolution in cancer treatment. By the end of 2018, more than 100 anti-PD-1 and anti-PD-L1 antibodies were in various stages of development, and more than 2000 clinical trials with their use have been registered. Characterization of such antibodies requires a bioassay to determine their biological activity. In this study, we developed a cell-based bioassay for analyzing the activity of anti-PD-1 and anti-PD-L1 antibodies. We chose reporter system consisting of two cell lines and compared several approaches for activation of effector cell line based on superantigens, soluble anti-CD3 antibodies, transmembrane anti-CD3 antibodies, chimeric antigenic receptors (CARs) and bispecific T-cell engager antibodies. The bispecific T-cell engager antibodies offer several advantages over the other approaches. We characterized the bioassay and demonstrated its applicability for analyzing the activity of anti-PD-1 and anti-PD-L1 antibodies. The proposed bioassay can be useful in the development of new therapeutic agents and methods for their characterization.
Subject(s)
Antibodies, Monoclonal/immunology , B7-H1 Antigen/immunology , Biological Assay/methods , Luminescent Measurements/methods , Programmed Cell Death 1 Receptor/immunology , T-Lymphocytes/immunology , Antibodies, Bispecific , Antigens, Surface/immunology , B7-H1 Antigen/antagonists & inhibitors , CD3 Complex/metabolism , Genes, Reporter , Genetic Vectors , Humans , Jurkat Cells , Lymphocyte Activation , NFATC Transcription Factors/metabolism , Programmed Cell Death 1 Receptor/antagonists & inhibitors , Receptors, Chimeric Antigen/geneticsABSTRACT
The methylation status of the genes of telomerase reverse transcriptase (tert) and telomerase RNA (terc) was determined in brain tissues of Danio rerio of different age. It is found that, regardless of the age of fish, the regulatory region of the tert gene was completely methylated, whereas the coding region remained unmethylated in all cases. The level of methylation of the region located downstream of the coding region of the terc gene changes with age. This region was analyzed in the samples of other tissues, and its methylation status was also nonuniform. The alteration of the methylation status in the 3'-untranslated region of the terc gene suggests the possibility of transcription of the antisense strand in this region.
Subject(s)
CpG Islands , DNA Methylation , RNA/genetics , Telomerase/genetics , Zebrafish/genetics , Aging/genetics , Aging/metabolism , Animals , Brain/growth & development , Brain/metabolism , Polymerase Chain Reaction , Zebrafish/metabolismABSTRACT
The state of hematopoietic, reproductive and endocrine systems of the organisms of male rats and their offspring in generations (F0-F1-F2) was studied, and the sensitivity of an organism to the action of carcinogen (Af mice) after a stay in the ChNPP exclusion zone was analyzed. It was ascertained that the most significant changes of the morphofunctional state of the animals were observed in the II generation (F2), which remained for a long period under the conditions of radioactive contamination. We have revealed an increased number of leukocytes, neutrophils, lymphocytes, and, especially, monocytes as against while the decrease in the number of erythrocytes and haemoglobin content; the decrease of thyroid function and cortical layer of the adrenals as opposed to while the increase in the relative weight of testes and their epididymides and the decrease in the number of spermatocytes and spermatozoa in the testis tissue. The exposure of Af mice in the exclusion zone increases the processes of mutagenesis and carcinogenesis, and changes the organism response to standardized action of chemical carcinogen. The increase in the exposure time of animals intensifies metabolic processes in a cell and increases their sensitivity to the action of xenobiotics.
