Diversity in the T cell receptor (TCR) repertoire provides a miniature defense ability for the T cell immune system that may be related to tumor initiation and progression. Understanding the T cell immune status of leukemia patients is critical for establishing specific immunotherapies. Previous studies have reported abnormal TCR repertoires and clonally expanded TCR Vβ T cells in chronic myeloid leukemia in chronic phase (CP-CML). In this study, we investigated the distribution and clonality of the TCR Vβ repertoire in 4 cases with imatinib-resistant CML in blast crisis (BC-CML) with abelson murine leukemia viral oncogene homolog 1 (ABL1) kinase domain mutations (KDMs). Examination of TCR V expression and clonality was performed by reverse transcription-polymerase chain reaction (RT-PCR) and GeneScan analysis. Significantly skewed TCR Vβ repertoires were observed in BC-CML patients with different KDMs, and 4 to 8 oligoclonally expanded TCR Vβ subfamilies could be identified in each sample. Intriguingly, a relatively highly expanded Vβ9 clone with the same length as complementarity- determining region 3 (CDR3) (139 bp) was found in all three CML patients in lymphoid blast crisis (LBC-CML) who had different KDMs, but the clone was not detected in the only CML patient in myeloid blast crisis (MBC-CML). In conclusion, restricted TCR Vβ repertoire expression and decreased clone complexity was a general phenomenon observed in the BC-CML patients with different KDMs, indicating the T-cell immunodeficiency of these patients. In addition, clonally expanded Vβ9 T cell clones may indicate a specific immune response to leukemia-associated antigens in LBC-CML patients.
In 538 febrile episodes in 188 children enrolled prospectively, 62% of children were neutropenic and 86% had infection-related fever. Respiratory infection was the commonest febrile cause (60%). Bacteremia occurred more often in neutropenic than non-neutropenic episodes (20% vs. 3%) and was accompanied significantly more with shiver, lassitude, and decreased dorsum pedis pulse. About 65% of blood isolates were Gram-negative bacilli, which differs from the observations in western countries.
// Ling Xu 1,* , Jianyu Weng 2,* , Xin Huang 2 , Chengwu Zeng 1 , Shaohua Chen 1 , Suxia Geng 2 , Lijian Yang 1 , Suijing Wu 2 , Suming Huang 4 , Xin Du 2 and Yangqiu Li 1,3,5 1 Institute of Hematology, Jinan University, Guangzhou, China 2 Department of Hematology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China 3 Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China 4 Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida, USA 5 Department of Hematology, First Affiliated Hospital, Jinan University, Guangzhou, China * These authors have contributed equally to this work Correspondence to: Yangqiu Li, email: // Xin Du, email: // Keywords : T-ALL, T cell repertoire, allo-HSCT, MRD, DLI, Immunology and Microbiology Section, Immune response, Immunity Received : April 10, 2016 Accepted : June 13, 2016 Published : June 23, 2016 Abstract The outcome for T-cell acute lymphoblastic leukemia (T-ALL) in relapse after hematopoietic stem cell transplantation (HSCT) is quite poor, while, both donor lymphocytes infusion (DLI) and adoptively infusion of γδ T cells in leukemia patients after HSCT have demonstrated good results in prolonging survival time of patients. Here, we reported a T-ALL case who experienced three relapses and received HSCT and DLI with an overall survival (OS) time lasting for more than seven years. Based on our previous identification of a leukemic and reactive clone in this patient, continual γδ T cell repertoire monitoring affirmed that the same Vδ5 leukemic clone existed in most samples from the patient, particularly including a sample taken at the time of the third T-ALL relapse, while it could not be detected in the donor sample. In addition, an identical Vδ4 monoclonal T cell that proliferated in the recipient for several years was confirmed to come from the donor graft, and its expression level significantly increased in third leukemia recurrence. These results indicate that clonally expanded Vδ4 T cells may represent a reconstituted γδ T cell repertoire after HSCT, which also hints to a relatively better outcome for this case. Based on this case study, we recommend DLI should be as a treatment strategy for patients who achieve CR or relapse from HSCT. Moreover, dynamically monitoring the TCR repertoire in patients who receive HSCT will benefit in supervising of malignant clone evolution and residue, identifying T cell clones mediate anti-infection, GvHD or GvL.
Tumour-infiltrating lymphocytes (TILs) contain T-cell subsets, which are related to immune escape and poor clinical outcomes of cancer patients. Little is known which T-cell receptor (TCR) clones belong to such T cells. Here, we identified that Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 are associated with poor prognosis for epidermal growth factor receptor (EGFR)-mutant stage II/III non-small-cell lung cancer (NSCLC) patients treated with adjuvant gefitinib or chemotherapy VP (vinorelbine/cisplatin) in the ADJUVANT-CTONG1104 trial. EGFR tyrosine kinase inhibitor is the standard targeted therapy for EGFR-mutant NSCLC patients.1, 2 For resectable EGFR-mutant NSCLC patients, the ADJUVANT-CTONG1104 trial showed that the first generation of EGFR-TKI gefitinib could significantly improve disease-free survival (DFS) of patients with N1/N2 lymph node metastasis.3 However, there is still heterogeneity in the clinical response to EGFR-TKIs, which may be related to EGFR co-mutations or immune checkpoint expression.4, 5 Regulatory T cells are correlated with cyclooxygenase-2 expression and are closely associated with adverse clinical outcome of resected NSCLC.6 A recent study reported that TCR clone Vβ6-6 was significantly increased in exhausted T cells at baseline of NSCLC patients treated with immune checkpoint blockade.7 While factors such as quantity of T cells in TILs have been shown to be prognostic, it remains of great interest to investigate whether any specific TCR clones may be prognostic or predictive of treatment efficacy. In this study, we further characterized the TCR repertoire of patients from the ADJUVANT-CTONG1104 trial and investigated the predictive potential of specific TCR-β clones for prognosis as well as benefit from adjuvant gefitinib or chemotherapy in EGFR-mutant NSCLC patients (Figure S1). NSCLC samples from 57 gefitinib-treated patients and 44 chemotherapy-treated patients were collected for TCR β gene sequencing to obtain TCR repertoires (Figure S2 and Supporting Information Materials and Methods). A total of 356 distinct TCR rearrangements were identified.8 Notably, Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 demonstrated statistical significance in predicting poor overall survival (OS) (FDR adjusted p < .05, Figure 1A). Importantly, the combination of Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 was the best model in predicting OS (Figure 1B), which was internally validated by 100 repeated 10-fold cross-validation (Figure S3). Multivariate Cox regression analysis indicated that Vβ6-6Jβ1-3 contributed the greatest to OS prediction (p < .001) (Figure 1C). Overall, the results indicated both Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 associated with poor OS of EGFR-mutant NSCLC patients. Next, the ability of the optimal Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 TCR rearrangement combination model to predict OS or DFS was evaluated. Of note, the risk score was negatively correlated with OS and DFS (p < .001; Figure 1D, Figure S4 and Table S1). Similarly, patients with high frequency either Vβ6-6Jβ1-3 or Vβ6-6Jβ1-6 had significantly poor OS and DFS (p < .01, Figure 1E and Figure S5). Interestingly, hazard ratios from Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 individually were smaller compared to hazard ratios derived from the optimal combination model for both OS and DFS. These results not only show that specific TCR rearrangements (Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6) are prognostic on their own, but the optimal model with a combination of these TCRs have the greatest predictive potential for prognosis of EGFR-mutant NSCLC patients. To confirm the clonotypes of TCRs, we explored the nucleotide (NT) and amino acid (AA) sequences of Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 in the high-frequency TCR groups. The NTs and AAs at both ends other than middle in the CDR3 region were almost conserved (Figure 2A,B). Herein, we identified the top five CDR3 motifs of Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 in the high-frequency group, which might reflect the common CDR3 sequences that contribute to the immune escape of EGFR-mutated NSCLC in this study. The top five CDR3 motifs for Vβ6-6Jβ1-3 were YSGS, YSRS, YSIS, TLPA, and YAGS. For Vβ6-6Jβ1-6, the top five CDR3 motifs were YSESD, DRDGG, YSGGG, YSREG, and PRGSP (Figure 2A,B). In addition, we used TKI-Gefitinib and Chemo-VP arms as training and validation cohorts, and the results indicated that Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 predicted poor OS and DFS for resected EGFR-mutant patients (p ≤ .05, Figure 3A–C). When patients carried high frequency of Vβ6-6Jβ1-3 or Vβ6-6Jβ1-6, the application of Vβ6-6 antibody or other treatment options should be considered. Mutant peptides produced by somatic tumour-specific mutations may create a neoepitope on cancer cells, which can be recognized by T cells. Therefore, the correlation of TCR rearrangements with genes with alteration rates greater than 20% was explored (Figure S6). Higher frequency of Vβ6-6Jβ1-3 rather than Vβ6-6Jβ1-6 was found in patients with NKX2-1 copy number (CN) gain, with marginal significance (p = .058), and they were significantly positively correlated (Cramer's V = 0.27, p = .007) (Figure 4A and Figure S7A). Furthermore, no significant relationship was found between Vβ6-6Jβ1-3 or Vβ6-6Jβ1-6 and TP53 exon 4/5 missense (p > .05, Figure S7B,C). Previously, we found NKX2-1 CN gain was significantly associated with poor prognosis of EGFR-mutant stage II/III NSCLC patients.9 NKX2-1 also serves an essential role in determining the fate of lung cancer cells and shaping the tumour immune microenvironment.10 Hence, results here appear to be consistent with previous findings, and demonstrated cross-talk between mutational and immune landscape. Compared to patients who were Vβ6-6Jβ1-3low and NKX2-1 wild type, patients with either Vβ6-6Jβ1-3high or NKX2-1 CN gain or both have a shorter OS or DFS, especially in Chemo-VP cohort (p < .05, Figure 4B and Figure S8A–C). Taken together, clonally expanded Vβ6-6Jβ1-3 and NKX2-1 CN gain may be an important biomarker for guiding adjuvant chemotherapy decisions in resectable early-stage NSCLC patients. In summary, we identified Vβ6-6Jβ1-3 and Vβ6-6Jβ1-6 in TILs, which were significantly correlated with poor prognosis in EGFR-mutant NSCLC patients in an adjuvant gefitinib or chemotherapy setting using a clinical trial cohort. To our best knowledge, this is the first study to identify specific TCR clone biomarkers related to poor clinical outcomes, as opposed to favourable outcomes. Results here are valuable for future prospective clinical trials and provide information for development of immunotherapy for EGFR-mutant stage II/III NSCLC patients. characterizing poor prognostic TCR clones from intra-tumoural T cells in EGFR-mutant stage II/III NSCLC patients; concurrent high-frequency Vβ6-6Jβ1-3 and NKX2-1 CN gain predicted poor prognosis of EGFR-mutant stage II/III NSCLC patients, especially in the adjuvant chemotherapy setting. This work was supported by grants from the National Natural Science Foundation of China (No. 82002413), Guangdong Provincial Applied Science and Technology Research & Development Program (No. 2016B020237006), China Postdoctoral Science Foundation (No. 2021M701422), and Guangdong Provincial Key Laboratory of Lung Cancer Translational Medicine (No. 2017B030314120). Yi-Long Wu discloses the following personal financial interests: Consulting and advisory services, speaking engagements of Roche, AstraZeneca, Eli Lilly, Boehringer Ingelheim, Sanofi, MSD, and BMS. The other authors have no conflict of interest. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
Natural killer (NK) cells are a key component of the innate immune system as they can attack cancer cells without prior sensitization. However, due to lack of cell-specific receptors, NK cells are not innately able to perform targeted cancer immunotherapy. Aptamers are short single-stranded oligonucleotides that specifically recognize their targets with high affinity in a similar manner to antibodies. To render NK cells with target-specificity, synthetic CD30-specific aptamers are anchored on cell surfaces to produce aptamer-engineered NK cells (ApEn-NK) without genetic alteration or cell damage. Under surface-anchored aptamer guidance, ApEn-NK specifically bind to CD30-expressing lymphoma cells but do not react to off-target cells. The resulting specific cell binding of ApEn-NK triggers higher apoptosis/death rates of lymphoma cells compared to parental NK cells. Additionally, experiments with primary human NK cells demonstrate the potential of ApEn-NK to specifically target and kill lymphoma cells, thus presenting a potential new approach for targeted immunotherapy by NK cells.
Abstract Background Continuous cropping is a significant obstacle to sustainable development in the pea ( Pisum sativum L.) industry, but the underlying mechanisms of this remain unclear. In this study, we used 16 S rDNA sequencing, transcriptomics, and metabolomics to analyze the response mechanism of roots and soil bacteria to continuous cropping and the relationship between soil bacteria and root phenotypes of different pea genotypes (Ding wan 10 and Yun wan 8). Results Continuous cropping inhibited pea growth, with a greater effect on Ding wan 10 than Yun wan 8. Metabolomics showed that the number of differentially accumulated metabolites (DAMs) in pea roots increased with the number of continuous cropping, and more metabolic pathways were involved. Transcriptomics revealed that the number of differentially expressed genes (DEGs) increased with the number of continuous cropping. Continuous cropping altered the expression of genes involved in plant-pathogen interaction, MAPK signal transduction, and lignin synthesis pathways in pea roots, with more DEGs in Ding wan 10 than in Yun wan 8. The up-regulated expression of genes in the ethylene signal transduction pathway was evident in Ding wan 10. Soil bacterial diversity did not change, but the relative abundance of bacteria significantly responded to continuous cropping. Integrative analysis showed that the bacteria with significant relative abundance in the soil were strongly associated with the antioxidant synthesis and linoleic acid metabolism pathway of pea roots under continuous cropping once. Under continuous cropping twice, the bacteria with significant relative abundance changes were strongly associated with cysteine and methionine metabolism, fatty acid metabolism, phenylpropanoid biosynthesis, terpenoid backbone biosynthesis, linoleic acid, and amino sugar and nucleotide sugar metabolism. Conclusion Ding wan 10 was more sensitive to continuous cropping than Yun wan 8. Continuous cropping times and pea genotypes determined the differences in root metabolic pathways. There were common metabolic pathways in the two pea genotypes in response to continuous cropping, and the DEGs and DAMs in these metabolic pathways were strongly associated with the bacteria with significant changes in relative abundance in the soil. This study provides new insights into obstacles to continuous cropping in peas.
The dynamic changes of lymphocyte subsets and cytokines profiles of patients with novel coronavirus disease (COVID-19) and their correlation with the disease severity remain unclear.
Objective T cell dysfunction is a common characteristic of patients with myeloid leukemia and is closely related to clinical efficacy and prognosis. In order to clarify the mechanisms leading to the T cell dysfunction, we characterized the gene expression profile of T cells from chronic myelogenous leukemia (CML) patients by microarray analysis and investigated the related regulating pathway.Methods We employed gene expression profiling, bioinformatics and real-time quantitative reverse transcription PCR (RT-qPCR) to detect genes differentially expressed in CML patients versus healthy donors.Results There were 1704 genes differentially expressed between CD3+ T cells from CML patients and healthy donors, including 868 up-regulated genes and 836 down-regulated genes, which mostly related to T cell functional pathways. In particular, lower expression of NFATC1, a member of the TCR signaling pathway, was detected in CD3+ T cells from CML patients. We further found that the expression of IRF4 and BACH2, transcription factors that potentially regulate NFATC1, in CD3+ T cells from CML patients was significantly lower than that in healthy donors.Conclusion We for the first time observed the altered gene expression profiles of CD3+ T cells from CML patients, and the results suggested that IRF4, BACH2 and NFATC1 may be involved in regulating T cell dysfunction in CML patients in the form of a transcriptional regulatory network. These findings may provide potential targets for tyrosine kinase inhibitors in combination with other targeted immunotherapies .
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