RESUMO
The lack of persistence of infused T cells is a principal limitation of adoptive immunotherapy in man. Interleukin (IL)-15 can sustain memory T cell expansion when presented in complex with IL-15Rα (15Rα/15). We developed a novel in-vitro system for generation of stable 15Rα/15 complexes. Immunologically quantifiable amounts of IL-15 were obtained when both IL-15Rα and IL-15 genes were co-transduced in NIH 3T3 fibroblast-based artificial antigen-presenting cells expressing human leucocyte antigen (HLA) A:0201, ß2 microglobulin, CD80, CD58 and CD54 [A2-artificial antigen presenting cell (AAPC)] and a murine pro-B cell line (Baf-3) (A2-AAPC15Rα/15 and Baf-315Rα/15 ). Transduction of cells with IL-15 alone resulted in only transient expression of IL-15, with minimal amounts of immunologically detectable IL-15. In comparison, cells transduced with IL-15Rα alone (A2-AAPCRα ) demonstrated stable expression of IL-15Rα; however, when loaded with soluble IL-15 (sIL-15), these cells sequestered 15Rα/15 intracellularly and also demonstrated minimal amounts of IL-15. Human T cells stimulated in vitro against a viral antigen (CMVpp65) in the presence of 15Rα/15 generated superior yields of high-avidity CMVpp65 epitope-specific T cells [cytomegalovirus-cytotoxic T lymphocytes (CMV-CTLs)] responding to ≤ 10- 13 M peptide concentrations, and lysing targets cells at lower effector : target ratios (1 : 10 and 1 : 100), where sIL-15, sIL-2 or sIL-7 CMV-CTLs demonstrated minimal or no activity. Both soluble and surface presented 15Rα/15, but not sIL-15, sustained in-vitro expansion of CD62L+ and CCR7+ central memory phenotype CMV-CTLs (TCM ). 15Rα/15 complexes represent a potent adjuvant for augmenting the efficacy of adoptive immunotherapy. Such cell-bound or soluble 15Rα/15 complexes could be developed for use in combination immunotherapy approaches.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Imunoterapia Adotiva , Interleucina-15/metabolismo , Ativação Linfocitária/imunologia , Receptores de Interleucina-15/metabolismo , Apoptose/genética , Apoptose/imunologia , Biomarcadores , Linhagem Celular Transformada , Citocinas/metabolismo , Citomegalovirus/imunologia , Citotoxicidade Imunológica , Epitopos de Linfócito T/imunologia , Humanos , Memória Imunológica , Infecções/imunologia , Infecções/metabolismo , Infecções/terapia , Interleucina-15/genética , Neoplasias/imunologia , Neoplasias/metabolismo , Neoplasias/terapia , Ligação Proteica , Receptores de Interleucina-15/genética , Especificidade do Receptor de Antígeno de Linfócitos T/imunologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismoRESUMO
Use of DNA fingerprinting in population studies is complicated by a number of methodical problems caused by the necessity to work with a large amount of experimental data and insufficient reproducibility of the results. In this work, possible approaches to overcome methodical difficulties faced by the authors during elaboration of the experimental technique are discussed.
Assuntos
Impressões Digitais de DNA , Genética Populacional , Genoma Humano , Humanos , Reprodutibilidade dos TestesRESUMO
The principles of primary treatment of blot-hybridization patterns obtained by means of multilocus DNA fingerprinting method are suggested. Final results, represented as a matrix of fragment sizes, are characterized by sufficiently high fidelity and reproducibility.
Assuntos
Impressões Digitais de DNA , Genética Populacional , Genoma Humano , Southern Blotting , Humanos , Estatística como AssuntoRESUMO
Genetic and demographic information for the Yakut population living in the Republic of Sakha (Yakutia) is presented. The mean number of children per woman constituted 4.605. Crow's index and its components (Im and If) were 0.483, 0.104, and 0.343, respectively.
Assuntos
Genética Populacional , Reprodução , Adulto , Coeficiente de Natalidade , Criança , Demografia , Serviços de Planejamento Familiar , Feminino , Humanos , Masculino , Gravidez , Taxa de Gravidez , Razão de Masculinidade , Sibéria/etnologiaRESUMO
Using multilocus DNA fingerprinting with phage M13 DNA as a probe, we have investigated a heterogeneous group of four human populations from Eastern Europe and Northeastern Asia. These populations belong to two language families: Indo-European (Eastern Slavonic branch: Russians, Belarussians) and Altaian (Turkic branch: Yakuts). The experimental results were treated by different statistical techniques: cluster analysis, multidimensional scaling, and multiple correspondence analysis. Coefficients of genetic differentiation were estimated using similarity indices and heterozygosities. The results of our study demonstrated similarity of Belarussian populations and significant differences between the group of Slavonic populations and Yakuts.
Assuntos
Povo Asiático/genética , Impressões Digitais de DNA/métodos , Genética Populacional , População Branca/genética , Análise por Conglomerados , Interpretação Estatística de Dados , Etnicidade/genética , Variação Genética , Heterozigoto , Humanos , Modelos Genéticos , República de Belarus , Federação Russa/etnologia , SoftwareRESUMO
We used DNA fingerprinting with M13 phage DNA as a probe to estimate the degree of genomic variability and genetic relationships in a heterogeneous group of 13 populations from Eastern Europe and Siberia. The popultaions belong to three language families: Indo-European (Slavonic: Russians, Byelorussians), Uralic (Finno-Ugric: Maris, Mordvinians, Udmurts), and Altaic (Turkic: Bashkirs, Tatars, Chuvashes, Yakuts). Multivariate statistical analyses were used (multidimensional scaling, cluster, and multiple correspondence analyses), and coefficients of gene differentiation ( Gst') were evaluated. The level of interpopulation subdivision in the various ethnic groups appeared to be different: the Byelorussian populations revealed no regional differences, in contrast to the Bashkir populations, which formed a heterogeneous group. The populations subdivided into three general clusters: Slavonic populations formed a separate tight cluster characterized by a minimal level of interpopulation diversity, Bashkir and Yakut populations formed the second cluster, and the Finno-Ugric and several populations of the Turkic linguistic groups formed the third cluster. The robustness of these results obtained by different statistical data treatments reveals that multilocus DNA fingerprinting can be reliably used for population studies.