Evolution of T-cell clonality in a patient with Ph-negative acute lymphocytic leukemia occurring after interferon and imatinib therapy for Ph-positive chronic myeloid leukemia

<p>Abstract</p> <p>Introduction</p> <p>The development of Philadelphia chromosome (Ph) negative acute leukemia/myelodysplastic syndrome (MDS) in patients with Ph-positive chronic myeloid leukemia (CML) is very rare. The features of restrictive usage and absence of partial T cell clones have been found in patients with CML. However, the T-cell clonal evolution of Ph-negative malignancies during treatment for CML is still unknown.</p> <p>Objective</p> <p>To investigate the dynamic change of clonal proliferation of T cell receptor (TCR) Vα and Vβ subfamilies in one CML patient who developed Ph-negative acute lymphoblastic leukemia (ALL) after interferon and imatinib therapy.</p> <p>Methods</p> <p>The peripheral blood mononuclear cells (PBMC) samples were collected at the 3 time points (diagnosis of Ph-positive chronic phase (CP) CML, developing Ph-negative ALL and post inductive chemotherapy (CT) for Ph-negative ALL, respectively). The CDR3 size of TCR Vα and Vβ repertoire were detected by RT-PCR. The PCR products were further analyzed by genescan to identify T cell clonality.</p> <p>Results</p> <p>The CML patient who achieved complete cytogenetic remission (CCR) after 5 years of IFN-α therapy suddenly developed Ph-negative ALL 6 months following switch to imatinib therapy. The expression pattern and clonality of TCR Vα/Vβ T cells changed in different disease stages. The restrictive expression of Vα/Vβ subfamilies could be found in all three stages, and partial subfamily of T cells showed clonal proliferation. Additionally, there have been obvious differences in Vα/Vβ subfamily of T cells between the stages of Ph-positive CML-CP and Ph-negative ALL. The Vα10 and Vβ3 T cells evolved from oligoclonality to polyclonality, the Vβ13 T cells changed from bioclonality to polyclonality, when Ph-negative ALL developed.</p> <p>Conclusions</p> <p>Restrictive usage and clonal proliferation of different Vα/Vβ subfamily T cells between the stages of Ph-positive CP and Ph-negative ALL were detected in one patient. These changes may play a role in Ph- negative leukemogenesis.</p

Generation of diffuse large B cell lymphoma-associated antigen-specific Vα6/Vβ13+T cells by TCR gene transfer

<p>Abstract</p> <p>Background</p> <p>Our previous study had amplified antigen-specific full-length TCR α and β genes of clonally expanded T cells in the peripheral blood (PB) of patients with diffuse large B-cell lymphoma (DLBCL). The transfer of T cell receptor (TCR) genes endows T cells with new antigen specificity. Therefore, the aim of this study is to generate diffuse large B cell lymphoma (DLBCL)-specific T cells by T cell receptor (TCR) gene transfer.</p> <p>Materials and methods</p> <p>Two different eukaryotic expression plasmids harboring TCR Vα6 and TCR Vβ13 genes specific for DLBCL-associated antigens were constructed and subsequently transferred into human T cells using Nucleofector™ technique. The expression of targeted genes in TCR gene-modified cells was detected by real-time PCR, and western blot using TCR Vβ antibody. The specific cytotoxicity of TCR gene-transferred T cells <it>in vitro </it>was estimated using a lactate dehydrogenase (LDH) release assay.</p> <p>Results</p> <p>Two different eukaryotic expression plasmids harboring TCR Vα6 and TCR Vβ13 genes specific for DLBCL-associated antigens were constructed and subsequently transferred into T cells from healthy donors. Specific anti-DLBCL cytotoxic T lymphocytes (CTL) could be induced by transduction of specific TCR gene to modify healthy T cells. The transgene cassette of TCR Vβ13-IRES-TCR Vα6 was superior to the other in the function of TCR-redirected T cells.</p> <p>Conclusions</p> <p>Specific anti-DLBCL cytotoxic T lymphocyte (CTL) could be inducted by transduction of specific TCR gene to modify healthy T cells.</p

Higher PD-1/Tim-3 expression on IFN-γ+ T cells is associated with poor prognosis in patients with acute myeloid leukemia

ABSTRACTWith the success of immune checkpoint inhibitors (ICI), such as anti- programmed death-1 (PD-1) antibody for solid tumors and lymphoma immunotherapy, a number of clinical trials with ICIs have been attempted for acute myeloid leukemia (AML) immunotherapy; however, limited clinical efficacy has been reported. This may be due to the heterogeneity of immune microenvironments and various degrees of T cell exhaustion in patients and may be involved in the IFN-γ pathway. In this study, we first characterized the percentage of PD-1+ and T cell immunoglobulin mucin-domain-containing-3 (Tim-3) +IFN-γ+ T cells in peripheral blood (PB) in AML compared with healthy individuals (HIs) by flow cytometry and further discussed the possibility of the reversal of T cell exhaustion to restore the secretion capacity of cytokines in T cells in AML based on blockade of PD-1 or Tim-3 (anti-PD-1 and anti-Tim-3 antibody) in vitro using a cytokine protein chip. A significantly increased percentage of PD-1+, Tim-3+, and PD-1+Tim-3+ IFN-γ+ T cells was observed in PB from patients with AML in comparison with HIs. Moreover, higher PD-1+IFN-γ+CD3+/CD8+ T cell levels were associated with poor overall survival in AML patients. Regarding leukemia cells, the percentage of Tim-3 in CD117+CD34+ AML cells was positively correlated with PD-1 in IFN-γ+CD4+ T cells. Furthermore, blocking PD-1 and Tim-3 may involve multiple cytokines and helper T cell subsets, mainly Th1 and Treg cells. Blockade of PD-1 or Tim-3 tends to restore cytokine secretion to a certain extent, a synergistic effect shown by the co-blockade of PD-1 and Tim-3. However, we also demonstrated the heterogeneity of secretory cytokines in ICI-treated T cells in AML patients

Enrichment Factors and Resource Potential Evaluation of Qingshankou Formation Lacustrine Shale Oil in the Southern Songliao Basin, NE China

China shale oil, which is preserved in lacustrine shale with strong heterogeneity and relatively low maturity, has been a research hotspot of unconventional resources. However, controlling factors of shale oil enrichment and resource potential evaluation restricted efficient exploration and development of lacustrine shale oil. On the basis of well logging data, TOC content, Rock-Eval pyrolysis values, thermal maturity, 100 oil saturation data, and pressure coefficient, the core observation, X-ray diffraction analysis, physical property analysis, scanning electron microscopy, CT scan, well logging interpretation, and volumetric genesis method depending on three-dimensional geological modeling were used to determine enrichment factors and evaluate the resource potential of Qingshankou Formation shale oil in the Southern Songliao Basin. Shale oil was mainly enriched in the semideep and deep lake shale of K2qn1, with the high capacity of hydrocarbon generation and favorable petrological and mineralogical characteristics, pore space characteristics, and physical properties in the low structural part of the Southern Songliao Basin. The three-dimensional geological resource model of Qingshankou Formation lacustrine shale oil was determined by the key parameters (Ro, TOC, and S1) of shale oil in the favorable zone of the Southern Songliao Basin, northeast China. The geological resource of shale oil, which was calculated by two grid computing methods (F1 and F2), was, respectively, 1.713×1012 kg and 1.654×1012 kg. The great shale oil resource indicates a promising future in the exploration and development of Qingshankou Formation shale oil of the Southern Songliao Basin

Characteristics and Genetic Mechanism of Pore Throat Structure of Shale Oil Reservoir in Saline Lake—A Case Study of Shale Oil of the Lucaogou Formation in Jimsar Sag, Junggar Basin

The shale oil reservoir of the Lucaogou Formation in the Jimsar Sag has undergone tectonic movement, regional deposition and complex diagenesis processes. Therefore, various reservoir space types and complex combination patterns of pores have developed, resulting in an intricate pore throat structure. The complex pore throat structure brings great challenges to the classification and evaluation of reservoirs and the efficient development of shale oil. The methods of scanning electron microscopy, high-pressure mercury injection, low-temperature adsorption experiments and thin-slice analysis were used in this study. Mineral, petrology, pore throat structure and evolution process characteristics of the shale oil reservoir were analyzed and discussed qualitatively and quantitatively. Based on these studies, the evolution characteristics and formation mechanisms of different pore throat structures were revealed, and four progressions were made. The reservoir space of the Lucaogou Formation is mainly composed of residual intergranular pores, dissolved pores, intercrystalline pores and fractures. Four types of pore throat structures in the shale oil reservoir of the Lucaogou Formation were quantitatively characterized. Furthermore, the primary pore throat structure was controlled by a sedimentary environment. The pores and throats were reduced and blocked by compaction and cementation, which deteriorates the physical properties of the reservoirs. However, the dissolution of early carbonate, feldspar and tuffaceous minerals and a small amount of carbonate cements by organic acids are the key factors to improve the pore throat structure of the reservoirs. The genetic evolution model of pore throat structures in the shale oil reservoir of the Lucaogou Formation are divided into two types. The large-pore medium-fine throat and medium-pore medium-throat reservoirs are mainly located in the delta front-shallow lake facies and are characterized by the diagenetic assemblage types of weak compaction–weak carbonate cementation–strong dissolution, early medium compaction–medium calcite and dolomite cementation–weak dissolution. The medium-pore fine throats and fine-pore fine throats are mainly developed in shallow lakes and semi-deep lakes. They are characterized by the diagenetic assemblage type of strong compaction–strong calcite cementation–weak dissolution diagenesis. This study provides a comprehensive understanding of the pore throat structure and the genetic mechanism of a complex shale oil reservoir and benefits the exploration and development of shale oil

Age-Related Immune Profile of the T Cell Receptor Repertoire, Thymic Recent Output Function, and miRNAs

Background. T cell immunity plays a central role in the body’s defense system, including maintaining homeostasis and preventing tumorigenesis and viral infection. Immune system functions degenerate with age, leading to immune senescence. Physiologically, immune senescence is characterized by a decrease in T cell receptor diversity, naive T cell deficiency, and alterations in T cell immune-related miRNAs. However, little is known about the characteristics of T cell immunosenescence in Chinese individuals. Results. A significant decrease in the miR-17, miR-92a, and miR-181a levels in PBMCs was detected with age. The miR-92a and miR-181a levels were upregulated in CBMCs when comparing healthy individuals to group I (0~9 years), whereas miR-17 was downregulated. The sjTREC level in PBMCs was negatively correlated with age, and a sharp decrease in sjTRECs was found between groups I and II (10~19 years). Twenty-four TCR Vβ subfamilies could be detected in most samples, and most displayed polyclonality, while skewed expression of the Vβ subfamilies as well as an increased oligoclonal tendency was found with age. Similarly, the frequencies of the TCR Vγ and Vδ subfamilies decreased with age, and the alteration in clonality appeared to be stable at different ages. Conclusion. We made the novel observation of T cell immunosenescence with age in Chinese individuals, which may provide information for immune targets to enhance the T cell immune response in immunotherapy settings for elderly patients