Production of cells with activated immunogenic properties from hematopoietic and non-hematopoietic progenitors by activation specific protective pathways

Abstract

AbstractBackground:The possibility about derivation of immune cells from hematopoietic and non-hematopoietic progenitors was investigated. Pilot studies on the underlining mechanisms in both types of cellular progenitors were performed.Methods:NK cells isolated from mouse spleens were incubatedin the presence of different combinations of cytokines (IL-15/IL-18 + IL-12). Additional oncogene copy in normal mouse embryonic stem cells (mESCs) was inserted by transfection with appropriate recombinant DNA-constructs, based on theAAVDNA-genome. mESCs, both containing and non-containing additional copy of oncogeneDcn1, were pre-incubated in the presence of GM-CSF, and sub-populations of the derived initial myeloid and lymphoid progenitors were then co-cultivated with each other. General features in the protein electrophoretic profile of sperm plasma with these from different cellular, tissue and organ samples were proposed, and the presence of white blood cells in ejaculates from patients with proved inflammatory process (prostatitis) was observed. Several methods for assay were applied: light microscopy, transmission electron microscopy (TEM), electrophoresis in 1% Agarose Gel after standard PCR and Reverse Trancriptase PCR (RT-PCR) and SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) with subsequent Coomasie-blue staining.Results:Despite the established activated proliferative activity of hematopoietic cellular progenitors, incubated in the presence of cytokines and cytokine combinations, statistically significant differences were noted only in the presence of IL-15 and IL-18, compared with the non-treated controls, and with the cells, incubated in the presence of other cytokines and cytokine combinations. The noted signs of initial myeloid and lymphoid progenitors, as well as of further phagocyte and plasmatic cell differentiation, respectively, confirmed additionally the preserved non-malignant characteristics and immunogenic capacity inin vitro-conditions of the so received cells, containing additional oncogene copy. Furthermore, a capability of non-myeloid and non-lymphoid cells to produce membrane receptor glycoproteins was suggested. On the other hand, a possibility for production of recombinant viral vaccines by exchange of nucleotide sequences between the used recombinant DNA-vector and cellular genome were proposed. In analogical way have been proved the preserved non-malignant characteristics and adequate immune response of human embryonic trophoblasts, immortalized by virusSV40. Besides the established morphological similarities, many signs of analogy in the electrophoretic profiles were established in the protein compositions between the separate biological samples, described above. Similarly to seminal plasma, synovia fluid and the extracts of each anatomic organ contain proteins, produced by different types of normal cells in various phases of maturation and differentiation.Conclusions:Besides of cells with hematopoietic origin, a possibility non-hematopoietic cells, tissues and organs to acquire immunogenic properties of in appropriate conditions was also proposed. Future studies are necessary in this direction.