Cytotoxic T cells and natural killer cells can kill target cells based on their expression and release of perforin, granulysin, and granzymes. Genes encoding these molecules have been only poorly annotated in camelids. Based on bioinformatic analyses of genomic resources, sequences corresponding to perforin, granulysin, and granzymes were identified in genomes of camelids and related ungulate species, and annotation of the corresponding genes was performed. A phylogenetic tree was constructed to study evolutionary relationships between the species analyzed. Re-sequencing of all genes in a panel of 10 dromedaries and 10 domestic Bactrian camels allowed analyzing their individual genetic polymorphisms. The data showed that all extant Old World camelids possess functional genes for two pore-forming proteins (PRF1, GNLY) and six granzymes (GZMA, GZMB, GZMH, GZMK, GZMM, and GZMO). All these genes were represented as single copies in the genome except the GZMH gene exhibiting interspecific differences in the number of loci. High protein sequence similarities with other camelid and ungulate species were observed for GZMK and GZMM. The protein variability in dromedaries and Bactrian camels was rather low, except for GNLY and chymotrypsin-like granzymes (GZMB, GZMH).

• Keywords
• NK cells, camel, cytotoxic T lymphocytes, granulysin, granzymes, perforin, ungulates,
• MeSH
• Killer Cells, Natural metabolism   MeSH
• Pore Forming Cytotoxic Proteins genetics   MeSH
• T-Lymphocytes, Cytotoxic metabolism   MeSH
• Phylogeny MeSH
• Granzymes genetics   MeSH
• Perforin genetics   MeSH
• Camelidae classification   genetics   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Pore Forming Cytotoxic Proteins MeSH
• Granzymes MeSH
• Perforin MeSH

Natural killer cells and cytotoxic T lymphocytes are the main cell populations of the immune system able to directly kill target cells via cytotoxic granules. Different mammalian species may differ in specific features of their pore-forming protein (perforin) and granule-bound serine proteases (granzymes). One perforin gene (PRF1) and four genes encoding granzymes A, B, H, and K (GZMA, GZMB, GZMH, GZMK) were identified in the reference genomes of felids. The objective of this work was to characterize the genes PRF1, GZMA and GZMB in a panel of 17 felid species by next-generation re-sequencing. A search of available felid genomes (17 species) retrieved the coding sequences of these genes for comparison to our data. Both sets of sequences or their combinations (23 species) were used for phylogenetic and selection analyses. Nucleotide PRF1, GZMA and GZMB sequences showed high similarities between felid species (over 95% identity). All trees derived from coding sequences expressed phylogenetic relationships corresponding to the zoological taxonomy of the Felidae, except GZMA. No effects of positive selection were detected in the genes studied, however, effects of purifying selection were observed for PRF1 and GZMA. The conservation of PRF1 is in agreement with its critical biological function. The differentiation observed between granzyme sub-families may reflect an adaptation to pathogen variation. The need to maintain important gene functions and at the same time cope with various pathogens may lead to an equilibrium between positive and negative selective pressures acting on GZMB. The within-species variability in wild felid populations merits further investigation.

• Keywords
• Felidae, NK cell, T cell, cytotoxic granule, granzyme, perforin,
• MeSH
• Alleles MeSH
• Killer Cells, Natural * MeSH
• Pore Forming Cytotoxic Proteins genetics   MeSH
• T-Lymphocytes, Cytotoxic MeSH
• Felidae * genetics   MeSH
• Phylogeny MeSH
• Granzymes genetics   MeSH
• Humans MeSH
• Perforin genetics   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Pore Forming Cytotoxic Proteins MeSH
• Granzymes MeSH
• Perforin MeSH

Investigating HLA-A, -B, -C, -DR and -Dw antigens and MLR, CML, and PLT reactivity in two unrelated persons, it was found that, despite their HLA-D/DR identity, cytotoxic T lymphocytes (CTL) could be induced in the CML assay. The HLA-DP antigens proved to provide the proliferative impetus for the generation of CTL.

• MeSH
• Cell Division MeSH
• T-Lymphocytes, Cytotoxic cytology   immunology   MeSH
• HLA-DP Antigens immunology   MeSH
• Humans MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• HLA-DP Antigens MeSH

Investigating three pairs of HLA-DR (DRB1) identical unrelated persons on the induction of cytotoxic T lymphocytes (CTL) it was found that HLA-DQ antigens provided slight proliferative stimulus which was, however, sufficient for the generation of CTL. The results indicate that the HLA-DQ antigens do play a similar role in clinical bone marrow transplantation as the HLA-DR specificities.

• MeSH
• Lymphocyte Activation MeSH
• Cytotoxicity, Immunologic MeSH
• T-Lymphocytes, Cytotoxic immunology   MeSH
• HLA-DQ Antigens physiology   MeSH
• HLA-DR Antigens immunology   MeSH
• Humans MeSH
• Lymphocyte Culture Test, Mixed MeSH
• Histocompatibility Testing MeSH
• Bone Marrow Transplantation immunology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• HLA-DQ Antigens MeSH
• HLA-DR Antigens MeSH

Multiple myeloma has been considered a weakly immunogenic malignancy that can cause profound defects in the immune system. An important issue for the immunotherapy of myeloma is the identification of appropriate tumor-associated antigens (TAAs). Recently, hTERT (human telomerase reverse transcriptase) was detected on a majority of human malignancies. In the studies reported here, we studied antigen-specific and HLA-A2-restricted cytotoxic activity against an ARH77 myeloma cell line in vitro. An HLA-A2-specific hTERT-derived nonapeptide ((540)ILAKFLHWL(548)) was used as a TAA. Myeloma-specific cytotoxic activity of hTERT-reactive cytotoxic lymphocytes (CTLs) was established by repeated stimulation of the CTLs via dendritic cells loaded with hTERT-derived nonapeptide. These studies were able to demonstrate that hTERT-reactive T-lymphocytes can be identified and expanded using relatively simple in vitro techniques consisting of antigen-specific stimulation, immunomagnetic sorting, and then induction of rapid expansion.

• MeSH
• Lymphocyte Activation * MeSH
• Antigens, Neoplasm immunology   MeSH
• T-Lymphocytes, Cytotoxic immunology   MeSH
• Dendritic Cells immunology   MeSH
• Humans MeSH
• Multiple Myeloma immunology   MeSH
• Cell Line, Tumor MeSH
• Peptide Fragments genetics   immunology   MeSH
• Telomerase genetics   immunology   metabolism   MeSH
• Transduction, Genetic MeSH
• Transfection MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, Neoplasm MeSH
• Peptide Fragments MeSH
• Telomerase MeSH
• telomerase reverse transcriptase (540-548) MeSH Browser
• TERT protein, human MeSH Browser

This review deals with cytotoxic and antiproliferative activity of fifty seven prenylated phenols isolated from Morus alba. Prenyl side chain, which can be variously modified, increases lipophilicity of the substances, thereby improving their penetration through biological membranes and thus results in an increased bioavailability. The objective was to describe the relationship between structure of the prenylated phenols and their cytotoxic effect and to clarify various mechanisms by which cytotoxic prenylated phenols induce apoptosis. The conclusions showed that the cytotoxicity of the substances increases with increasing number of the prenyl side chains and ketal groups. Conversely, modification of the prenyl side chain, such as hydroxylation, reduces cytotoxicity. The cytotoxic activity is also influenced by the presence of prenyl and hydroxyl groups at specific positions.

• Keywords
• <i>Morus alba</i>, anti­proliferative activity, cytotoxicity, prenylated phenols,
• MeSH
• Antineoplastic Agents, Phytogenic isolation & purification   pharmacology   MeSH
• Cytotoxins isolation & purification   pharmacology   MeSH
• Phenols isolation & purification   pharmacology   MeSH
• Molecular Structure MeSH
• Morus chemistry   MeSH
• Prenylation MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Antineoplastic Agents, Phytogenic MeSH
• Cytotoxins MeSH
• Phenols MeSH

The usefulness of cytotoxic T lymphocytes precursors (CTLp) frequency analysis in the search for donors in bone marrow transplantation was studied. The frequency of anti-recipient CTLp was approached by limiting dilution assay in HLA matched unrelated, HLA partially matched related and HLA genotypically identical donors. The majority of patients examined were affected with different hematological malignancies. Alloreactive CTLp recognizing non-HLA gene products were not detected in pretransplant examination of two pairs of HLA identical siblings. However, an increased incidence of allospecific CTLp was identified in HLA matched MLC negative unrelated pairs. Thus, CTLp assay allowed to uncover the residual Class I incompatibilities that remained hidden in standard serotyping. In two matched unrelated pairs with high pretransplant CTLp frequency the severe acute graft-versus-host disease (GVHD) developed after bone marrow transplantation. Examination of other relatives in patients lacking an HLA identical sibling showed the importance of Class I incompatibility for CTLp generation as well. The lack of correlation between CTLp frequency and HLA-D disparity could suggest that Class II antigens do not participate in CTLp induction. With one exception we had good correlation between MLC and DNA analysis of Class II antigens demonstrating that MLC gives interpretable results even in unrelated pairs. Our results demonstrate the significance of CTLp frequency assay in detection of residual Class I incompatibilities in matched unrelated pairs and in assessment of Class I compatibility in related pairs. For that it should be used in the final selection of BMT donors.

• MeSH
• Immunity, Cellular MeSH
• Bone Marrow Cells * MeSH
• T-Lymphocytes, Cytotoxic immunology   MeSH
• Cytotoxicity Tests, Immunologic * MeSH
• Tissue Donors * MeSH
• HLA Antigens immunology   MeSH
• Bone Marrow immunology   MeSH
• Humans MeSH
• Graft vs Host Disease immunology   MeSH
• Bone Marrow Transplantation * MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• HLA Antigens MeSH

The proliferation and development of cytotoxic T cells was investigated in human peripheral blood mononuclear cell (PBMC) cultures stimulated with an antigenic extract from Candida albicans (MPPS), or with the purified protein derivative from Mycobacterium tuberculosis (PPD), or with human recombinant interleukin 2 (rIL-2). Microbial antigen- and rIL-2-induced cytotoxic T cells were able to lyse both natural killer (NK) sensitive and resistant targets. No correlation was observed between the development of T cell cytotoxicity and interferon (IFN) production in vitro. The addition of anti-class II monoclonal antibodies at the beginning of MPPS/PPD-stimulated cultures inhibited the cell proliferation, IFN production and T cell cytotoxicity, while all these cellular activities were not inhibited by anti-class II antibodies in rIL-2-stimulated cultures. Finally, antibodies to class I determinants inhibit T cell cytotoxicity, suggesting a role of such determinants in the development of the non-adaptive immunity to microbial infections.

• MeSH
• Antigens, Bacterial pharmacology   MeSH
• Cell Division drug effects   MeSH
• Candida albicans MeSH
• T-Lymphocytes, Cytotoxic cytology   drug effects   immunology   MeSH
• Adult MeSH
• HLA Antigens metabolism   pharmacology   MeSH
• Interleukin-2 pharmacology   MeSH
• Humans MeSH
• Antibodies, Monoclonal MeSH
• Mycobacterium tuberculosis MeSH
• Recombinant Proteins pharmacology   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• HLA Antigens MeSH
• Interleukin-2 MeSH
• Antibodies, Monoclonal MeSH
• Recombinant Proteins MeSH

The kinase LCK and CD4/CD8 co-receptors are crucial components of the T cell antigen receptor (TCR) signaling machinery, leading to key T cell fate decisions. Despite decades of research, the roles of CD4-LCK and CD8-LCK interactions in TCR triggering in vivo remain unknown. In this study, we created animal models expressing endogenous levels of modified LCK to resolve whether and how co-receptor-bound LCK drives TCR signaling. We demonstrated that the role of LCK depends on the co-receptor to which it is bound. The CD8-bound LCK is largely dispensable for antiviral and antitumor activity of cytotoxic T cells in mice; however, it facilitates CD8+ T cell responses to suboptimal antigens in a kinase-dependent manner. By contrast, the CD4-bound LCK is required for efficient development and function of helper T cells via a kinase-independent stabilization of surface CD4. Overall, our findings reveal the role of co-receptor-bound LCK in T cell biology, show that CD4- and CD8-bound LCK drive T cell development and effector immune responses using qualitatively different mechanisms and identify the co-receptor-LCK interactions as promising targets for immunomodulation.

• MeSH
• CD4 Antigens MeSH
• CD8 Antigens metabolism   MeSH
• T-Lymphocytes, Cytotoxic * metabolism   MeSH
• Mice MeSH
• Receptors, Antigen, T-Cell metabolism   MeSH
• Signal Transduction MeSH
• Lymphocyte Specific Protein Tyrosine Kinase p56(lck) * metabolism   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Names of Substances
• CD4 Antigens MeSH
• CD8 Antigens MeSH
• Receptors, Antigen, T-Cell MeSH
• Lymphocyte Specific Protein Tyrosine Kinase p56(lck) * MeSH

Using exhaustive chromatographic separation we have isolated (-)-tigloyl-deangeloyl-gomisin F as a novel dibenzocyclooctadiene lignan from schisandra chinensis. With the help of HPLC, we further isolated (+)-schisandrin, (+)-deoxyschisandrin, (+)-γ-schisandrin, (-)-gomisin J, (+)-gomisin A, (-)-gomisin N, (-)-tigloyl-gomisin P, (-)-wuweizisu C, (-)-gomisin D, rubrisandrin A, (-)-gomisin G, (+)-gomisin K (3) and (-)-schisantherin C. A full NMR description of (-)-schisantherin C was carried out with the aim to confirm previous reports of its structure. Compounds isolated were identified on the basis of UV, IR, (1)H- and (13)C-NMR and MS. The cytotoxicity of lignans was tested for the BY-2 cell line alone and as a synergistic effect with the cytotoxic agent camptothecin. Lignans showed various toxicity and synergistic and antagonistic effects on camptothecin-induced cytotoxicity. Cytotoxicity against colon cancer cell line LoVo was also tested.

• MeSH
• Apoptosis drug effects   MeSH
• Cell Line MeSH
• Chemical Fractionation MeSH
• Cytotoxins chemistry   isolation & purification   toxicity   MeSH
• Humans MeSH
• Lignans chemistry   isolation & purification   toxicity   MeSH
• Cell Line, Tumor MeSH
• Nuclear Magnetic Resonance, Biomolecular MeSH
• Cell Proliferation drug effects   MeSH
• Plant Extracts chemistry   MeSH
• Schisandra chemistry   MeSH
• Nicotiana drug effects   MeSH
• Chromatography, High Pressure Liquid MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Cytotoxins MeSH
• Lignans MeSH
• Plant Extracts MeSH