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The honeybees are the most important pollinator in the production of crops and fresh produce. Temperature affects the survival of honeybees, and determines the quality of their development, which is of great significance for beekeeping production. Yet, little was known about how does low temperature stress during development stage cause bee death and any sub-lethal effect on subsequent. Early pupal stage is the most sensitive stage to low temperature in pupal stage. In this study, early pupal broods were exposed to 20°C for 12, 16, 24, and 48 h, followed by incubation at 35°C until emergence. We found that 48 h of low temperature duration cause 70% of individual bees to die. Although the mortality at 12 and 16 h seems not very high, the association learning ability of the surviving individuals was greatly affected. The brain slices of honeybees showed that low temperature treatment could cause the brain development of honeybees to almost stop. Gene expression profiles between low temperature treatment groups (T24, T48) and the control revealed that 1,267 and 1,174 genes were differentially expressed respectively. Functional enrichment analysis of differentially expressed genes showed that the differential expression of Map3k9, Dhrs4, and Sod-2 genes on MAPK and peroxisome signaling pathway caused oxidative damage to the honeybee head. On the FoxO signal pathway, InsR and FoxO were upregulated, and JNK, Akt, and Bsk were downregulated; and on the insect hormone synthesis signal pathway, Phm and Spo genes were downregulated. Therefore, we speculate that low temperature stress affects hormone regulation. It was detected that the pathways related to the nervous system were Cholinergic synapse, Dopaminergic synapse, GABAergic synapse, Glutamatergic synapse, Serotonergic synapse, Neurotrophin signaling pathway, and Synaptic vesicle cycle. This implies that the synaptic development of honeybees is quite possibly greatly affected by low temperature stress. Understanding how low temperature stress affects the physiology of bee brain development and how it affects bee behavior provide a theoretical foundation for a deeper comprehension of the temperature adaptation mechanism that underlies the "stenothermic" development of social insects, and help to improve honeybee management strategies to ensure the healthy of colony.
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INTRODUCTION: Longer follow-up was necessary to determine the exact value of mastoscopic axillary lymph node dissection (MALND). METHODS: From January 1, 2003, to December 31, 2005, 1027 patients with breast cancer were randomly assigned to two groups: MALND and CALND (conventional axillary lymph node dissection); 996 eligible patients were enrolled. RESULTS: The final cohort of 996 patients was followed for an average of 198 months. Events other than death differed significantly between the two cohorts (p = 0.0311; 46.3% in MALND and 53.2% in CALND, respectively). The sum of events other than death and deaths from other causes was much higher in the CALND (59.6%) than MALND (53.4%) group (p = 0.0494). The 17-year disease-free survival DFS rates were 36.7% for the MALND and 33.6% for the CALND group, respectively. There was a significant difference between the groups (p = 0.0306). Overall survival (OS) rates were 53.2% after MALND and 46.0% after CALND (p = 0.0119). MALND patients had much less axillary pain (p = 0.0000), numbness or paresthesia (p = 0.0000), arm mobility (p = 0.0000) and arm swelling on the operated side (p = 0.0000). Aesthetic appearance of the axilla was much better in the MALND than CALND group (p = 0.0000) at an average follow-up of 17 years. CONCLUSIONS: The use of MALND in breast cancer surgery not only decreases the relapse and arm complications but also improves long-term survival of patients. Therefore, MALND should be one of the preferred approaches for breast cancer surgery when ALND is needed. TRIAL REGISTRATION INFORMATION: The comparison of long-term outcomes of mastoscopic and conventional axillary lymph node dissection in breast cancer: a multicenter randomized control trial. ChiCTR-TRC-11001477, CHiCTR. First registration 08/14/2011.
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Neoplasias de la Mama , Axila/patología , Axila/cirugía , Neoplasias de la Mama/patología , Femenino , Estudios de Seguimiento , Humanos , Escisión del Ganglio Linfático/efectos adversos , Recurrencia Local de NeoplasiaRESUMEN
OBJECTIVE: Lung cancer is a disease associated with high levels of morbidity and mortality, with approximately 2.1 million new cases every year. Anlotinib is a new small-molecule multitarget tyrosine kinase inhibitor independently developed in China that can inhibit the formation of tumor blood vessels and has a therapeutic effect on various cancers. However, the application of anlotinib in lung cancer needs further investigation. METHODS: We collected the progress notes of 43 patients with advanced lung cancer treated at the Oncology Department of Guangzhou Chest Hospital from March 2019 to March 2021. Additionally, we assessed the differences between drug combination therapy and single-drug therapy among patients treated with anlotinib. RESULTS: Patients in both the anlotinib-combination and anlotinib-monotherapy groups experienced remission; however, the overall disease control rate in the anlotinib-combination group was higher than that in the anlotinib-monotherapy group. Reexamination via computed tomography showed that patients in the anlotinib-combination group had better recovery than those in the anlotinib-monotherapy group. Although the overall incidence of adverse reactions in the anlotinib-combination group was higher than that in the monotherapy group, most of the adverse reactions were I-II levels and improved after symptomatic treatment. CONCLUSION: Anlotinib combined with other therapies is better than anlotinib alone for the management of patients with advanced lung cancer.
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Antineoplásicos/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Indoles/uso terapéutico , Neoplasias Pulmonares/tratamiento farmacológico , Quinolinas/uso terapéutico , Antineoplásicos/administración & dosificación , Antineoplásicos/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico por imagen , China , Estudios de Cohortes , Biología Computacional , Femenino , Humanos , Indoles/administración & dosificación , Indoles/efectos adversos , Neoplasias Pulmonares/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Inhibidores de Proteínas Quinasas/administración & dosificación , Inhibidores de Proteínas Quinasas/efectos adversos , Inhibidores de Proteínas Quinasas/uso terapéutico , Quinolinas/administración & dosificación , Quinolinas/efectos adversos , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , Estudios Retrospectivos , Seguridad , Tomografía Computarizada por Rayos XRESUMEN
BACKGROUND: It is not a rare clinical scenario to have patients presenting with coexisting malignant tumor and tuberculosis. Whether it is feasible to conduct programmed death-(ligand) 1 [PD-(L)1] inhibitors to these patients, especially those with active tuberculosis treated with concurrent anti-tuberculosis, is still unknown. METHODS: This study enrolled patients with coexisting malignancy and tuberculosis and treated with anti-PD-(L)1 from Jan 2018 to July 2021 in 2 institutions. The progression-free survival (PFS), objective response rate (ORR), and safety of anti-PD-(L)1 therapy, as well as response to anti-tuberculosis treatment, were evaluated. RESULTS: A total of 98 patients were screened from this cohort study, with 45 (45.9%), 21 (21.4%), and 32 (32.7%) patients diagnosed with active, latent, and obsolete tuberculosis, respectively. The overall ORR was 36.0% for anti-PD-(L)1 therapy, with 34.2%, 35.5%, and 41.2% for each subgroup. Median PFS was 8.0 vs 6.0 vs 6.0 months (P=0.685) for each subgroup at the time of this analysis. For patients with active tuberculosis treated with concurrent anti-tuberculosis, median duration of anti-tuberculosis therapy was 10.0 (95% CI, 8.01-11.99) months. There were 83.3% (20/24) and 93.3% (42/45) patients showing sputum conversion and radiographic response, respectively, after anti-tuberculosis therapy, and two patients experienced tuberculosis relapse. Notably, none of the patients in latent and only one patient in obsolete subgroups showed tuberculosis induction or relapse after anti-PD-(L)1 therapy. Treatment-related adverse events (TRAEs) occurred in 33 patients (73.3%) when treated with concurrent anti-PD-(L)1 and anti-tuberculosis. Grade 3 or higher TRAEs were hematotoxicity (n = 5, 11.1%), and one patient suffered grade 3 pneumonitis leading to the discontinuation of immunotherapy. CONCLUSIONS: This study demonstrated that patients with coexisting malignant tumor and tuberculosis benefited equally from anti-PD-(L)1 therapy, and anti-tuberculosis response was unimpaired for those with active tuberculosis. Notably, the combination of anti-PD-(L)1 and anti-tuberculosis therapy was well-tolerated without significant unexpected toxic effects.
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Neoplasias , Tuberculosis , Estudios de Cohortes , Humanos , Inmunoterapia , Neoplasias/complicaciones , Neoplasias/tratamiento farmacológico , Tuberculosis/complicaciones , Tuberculosis/tratamiento farmacológicoRESUMEN
BACKGROUND: Immune checkpoint inhibitors (ICIs) have brought clinical benefits to patients with various histological types of lung cancer. Previous studies have shown an association between mesenchymal-epithelial transition (MET) and the immunotherapy response in non-small cell lung cancer (NSCLC) but there is a lack of clinical data on the correlation of MET amplification with the ICI response in NSCLC. METHODS: Copy number alteration (CNA), somatic mutation, and clinical data from two immunotherapy cohorts (Rizvi et al. cohort and our local cohort) were collected and pooled to further investigate the key role of MET amplification in patients with NSCLC receiving ICIs. The correlations between MET amplification and tumor immunogenicity and antitumor immunity were further investigated in The Cancer Genome Atlas (TCGA)-NSCLC [lung adenocarcinoma (LUAD)/lung squamous cell carcinoma (LUSC)] data-set. RESULTS: In the immunotherapy cohorts, MET amplification was associated with longer progression-free survival (PFS) times in patients receiving ICI treatment (P=0.039; HR =0.37; 95% CI: 0.18-0.73). In the TCGA-NSCLC data-set, MET amplification was associated with high MET mRNA and protein levels, tumor mutation burden (TMB), neoantigen load (NAL), immune-activated cell patterns, immune-related gene expression levels, and the number of gene alterations in the DNA damage response and repair (DDR) pathway. Gene set enrichment analysis (GSEA) results indicated significant up-regulation of the immune response-related pathways in the MET-amplification group. CONCLUSIONS: Our results suggest that MET amplification may be a novel predictive marker for immunotherapy efficacy in NSCLC.
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BACKGROUND: This retrospective study evaluated the safety and efficacy of concurrent anti-tuberculosis (TB) and chemotherapy treatment in patients with advanced lung cancer and active TB. METHODS: We retrospectively analyzed patients who were first diagnosed with advanced lung cancer and received first-line chemotherapy in Guangzhou Chest Hospital from 2015 to 2017. Patients were categorized into two groups (2:1): lung cancer patients without active TB (Group A), and lung cancer patients with active TB (Group B). Primary endpoints included adverse events (AEs), objective response rate (ORR), time to treatment failure, and overall survival (OS). RESULTS: A total of 99 patients were eligible (Group A, n=66; Group B, n=33). Grade ≥3 treatment-related AEs, primarily hematologic toxicity, occurred in 39.4% and 51.5% of patients in Groups A and B, respectively. The hypohepatia in both groups was generally at grade 1 or 2, with similar incidences (26% and 27%, respectively). After two cycles of chemotherapy, the ORR was 42.4% and 33.3% in Group A and B, respectively (P=0.383). The median time to treatment failure (TTF) was 7.0 and 5.6 months for Groups A and B, respectively (P=0.175). The median OS was 17.0 and 14.0 months for Groups A and B, respectively (P=0.312). After 3 months of anti-TB treatment, all patients achieved sputum acid-fast bacilli (AFB) smear conversion and absorption on imaging, and the end of follow-up observed no recurrence. CONCLUSIONS: Concurrent anti-TB and chemotherapy treatment did not increase hematological toxicity or hypohepatia in lung cancer patients with pulmonary TB.
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BACKGROUND: Mesenchymal stem cells (MSCs) can provide therapeutic benefits for myocardial infarction (MI) recovery; however, the molecular mechanism by which MSCs improve the heart function is unclear. METHODS: Microarray analysis was performed to examine the expression profiling of human MSCs (hMSCs) grown as adherent cultures (AC-hMSCs) or nonadherent cultures on ultra-low-adherent plates (nonAC-hMSCs). Real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, and enzyme-linked immunosorbent assays (ELISA) were used to assess VEGFA expression and secretion in the AC-hMSCs and nonAC-hMSCs. The paracrine effect of VEGFA-overexpressing AC-MSCs (AC-VEGFA-hMSCs) or VEGFA-knockdown nonAC-hMSCs (nonAC-shVEGFA-hMSCs) on the angiogenic ability of human umbilical vein endothelial cells (HUVECs) was evaluated using tube formation assay. AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs were transplanted into myocardial infarction rats to investigate the therapeutic effect of AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs. Luciferase reporter assay was used to confirm the association of VEGFA with miR-519d. RESULTS: Microarray analysis revealed that VEGFA is downregulated in AC-hMSCs compared to nonAC-hMSCs. Functional assays revealed that high levels of VEGFA produced from AC-VEGFA-hMSCs increased the tube formation capacity of HUVECs in vitro, improved angiogenesis and cardiac performance, and reduced infarct size in a rat MI model. Low levels of VEGFA secretion from nonAC-shVEGFA-hMSCs had the opposite effects. Mechanistically, we found that miR-519d directly targets VEGFA. High levels of VEGFA secreted from VEGFA-overexpressing nonAC-hMSCs abolished the repressive effect of miR-519d on HUVEC angiogenesis. CONCLUSION: Our findings indicate that nonadherent culture-induced secretion of VEGFA plays an important role in MSCs via the miR-519d/VEGFA pathway and may provide a novel therapeutic strategy for MI treatment.
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Células Madre Mesenquimatosas , MicroARNs , Infarto del Miocardio , Animales , Células Endoteliales de la Vena Umbilical Humana , MicroARNs/genética , Infarto del Miocardio/genética , Infarto del Miocardio/terapia , Neovascularización Patológica , RatasRESUMEN
The purpose of the study is to compare the efficacy and safety between endovascular surgery and open surgery for the treatment of peripheral arterial disease (PAD). Studies were recruited from EMBASE, PubMed, Cochrane Library, and reviewing reference lists of related studies between June 2017 and December 31, 2018. Data of functional outcomes were extracted in this meta-analysis. Odds ratio or standardized mean differences with its 95% confidence interval (95% CI) were used. Pooled data proved to be absent of heterogeneity were tested by a fixed-effects model; otherwise, we used a random-effects model. All analyses were managed with RevMan 5.3 software (Cochrane Collaboration, London, UK). Twenty-seven trials (7 randomized controlled trials and 20 retrospective trials) involving a total of 17,536 participants were included in our meta-analysis. Data showed that endovascular surgery was accompanied with lower complication rate (426/4,496 [9.48%] vs. 551/4,051 [13.60%]; 95% CI, 0.53 [0.34, 0.82]; I2 = 83%; P = 0.004) and shorter hospital stay (95% CI, -4.01 [-4.99, -3.02]; I2 = 76%; P < 0.00001) than open surgery. Higher amputation (204/1,633 [12.49%] vs. 228/1,247 [18.28%]; 95% CI, 0.72 [0.59, 0.89]; I2 = 12%; P = 0.002) and mortality during follow-up (490/4,514 [10.86%] vs. 341/4,520 [7.54%]; 95% CI, 0.73 [0.61, 0.86]; I2 = 0%; P < 0.05) were found in open surgery group. No evidence showed the rate of technical success, survival, or limb salvage reduced in endovascular surgery. In conclusion, our results showed that endovascular surgery is a promising technique for the treatment of PAD. Our data showed that endovascular surgery has less complication, shorter hospital stay, lower amputation, and mortality rate when compared with open surgery, although no significant difference was observed in survival rate, surgery success, and limb salvage.
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Procedimientos Endovasculares , Enfermedad Arterial Periférica/cirugía , Procedimientos Quirúrgicos Vasculares , Amputación Quirúrgica , Procedimientos Endovasculares/efectos adversos , Procedimientos Endovasculares/mortalidad , Humanos , Tiempo de Internación , Recuperación del Miembro , Enfermedad Arterial Periférica/diagnóstico por imagen , Enfermedad Arterial Periférica/mortalidad , Enfermedad Arterial Periférica/fisiopatología , Complicaciones Posoperatorias/mortalidad , Complicaciones Posoperatorias/terapia , Medición de Riesgo , Factores de Riesgo , Factores de Tiempo , Resultado del Tratamiento , Procedimientos Quirúrgicos Vasculares/efectos adversos , Procedimientos Quirúrgicos Vasculares/mortalidadRESUMEN
PURPOSE: Studies on genetic alterations of the heterogenous small cell lung cancer (SCLC) are rare. We carried out the present study to clarify the genomic alterations and TMB levels of Chinese SCLC patients by whole-exome sequencing. MATERIALS AND METHODS: Whole-exome sequencing by next-generation sequencing technique was implemented on twenty SCLC samples. Significant somatic mutations and copy number variations were screened, followed by comparison with the data extracted from COSMIC. Besides, altered signaling pathways were examined in order to figure out actionable targets. RESULTS: A total of 8,062 nonsynonymous mutations were defined. The number of mutations for each case ranged from 98 to 864. As for base substitutions, a total of 15,817 substitutions were detected with C > A conversion which was correlated to smoking occupying 25.57%. The TMB values ranged from 2.51/Mb to 22.1/Mb with a median value of 9.95/Mb. RB1 was the most frequently mutated gene altered in 18 (90%) cases, followed by TP53 altered in 17 (85%) cases. Other commonly changed genes were PTEN, and RBL1, with frequencies of 55% and 50%, respectively. SOX2 significantly amplified in 6 (30%) cases and MYCN amplified in 1 (5%) patient. Notch signaling pathway and PI3K/AKT/mTOR signaling pathway were universally and significantly changed. Major genomic alterations were in consistency with data from COSMIC, but frequencies of less common mutations were different. CONCLUSION: TP53 and RB1 inactivations were universally detected in SCLC. The Notch and PI3K/AKT/mTOR signaling pathways were both significantly altered, implying potential actionable targets.
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Neoplasias Pulmonares/genética , Mutación/genética , Carcinoma Pulmonar de Células Pequeñas/genética , Biomarcadores de Tumor/genética , Variaciones en el Número de Copia de ADN/genética , Femenino , Genómica/métodos , Humanos , Masculino , Persona de Mediana Edad , Fosfohidrolasa PTEN/genética , Fosfatidilinositol 3-Quinasas/genética , Transducción de Señal/genética , Serina-Treonina Quinasas TOR/genética , Proteína p53 Supresora de Tumor/genética , Secuenciación del Exoma/métodosRESUMEN
Targeted delivery of chemotherapeutics by functionalized nanoparticles exhibits a wonderful prospect for cancer treatment. In this paper, selenium nanoparticles (SeNPs) was linked with hyaluronic acid (HA) to prepare tumor-targeted delivery vehicle HA-SeNPs, and HA-SeNPs was loaded with paclitaxel (PTX) to fabricate functionalized selenium nanoparticles HA-Se@PTX. HA-Se@PTX showed greater uptake in A549 cells in comparison with that in HUVEC, verifying HA-mediated specific uptake of HA-Se@PTX. HA-Se@PTX was capable of entering A549 cells via clathrin-associated endocytosis and showed faster drug release in cancer cell microenvironment in comparison with normal physiological environment. HA-Se@PTX could obviously inhibit the proliferation, migration and invasion of A549 cells and trigger A549 cells apoptosis. Moreover, active targeting functionalized selenium nanoparticles HA-Se@PTX showed greater in vivo antitumor activity compared with free PTX or passive targeting delivery system Se@PTX. In addition, HA-Se@PTX exhibited negligible toxicity on the major organs of mice. In a word, HA-Se@PTX may develop into a valuable nanoscale antitumor drug agent for lung cancer treatment.
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Portadores de Fármacos/química , Ácido Hialurónico/química , Neoplasias Pulmonares/patología , Nanopartículas/química , Paclitaxel/química , Paclitaxel/farmacología , Selenio/química , Células A549 , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Liberación de Fármacos , Humanos , Ratones , Invasividad Neoplásica , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The mechanism by which IFN-α regulates the host response to Mycobacterium tuberculosis (M.tb) infection in humans is poorly understood. In the present study, we found that freshly isolated pleural fluid mononuclear cells (PFMCs) from tuberculous pleural effusion but not peripheral blood mononuclear cells (PBMCs) spontaneously expressed IFN-α and IL-1ß in vivo. In addition, exogenous IFN-α significantly inhibited production of IL-1ß in PFMCs after stimulation with Bacillus Calmette-Guérin (BCG). To further evaluate the effect of endogenous IFN-α on BCG-induced IL-1ß production, a neutralizing antibody to IFN-α was added to the cultures of BCG-stimulated PFMCs. As expected, neutralization of IFN-α by antibody significantly enhanced the production of IL-1ß. Notably, we showed that IFN-α inhibited production of IL-1ß through 2 distinct mechanisms: IFN-α signaling, via the STAT1 transcription factor, suppressed caspase-1-dependent IL-1ß maturation, and IFN-α induced the production of IL-10 in a STAT1-dependent manner in which IL-10 reduced the abundance of IL-1ß. In contrast, we found that IFN-α enhanced the production of IFN-γ, and IFN-γ also suppressed IL-1ß production in the PFMCs during BCG stimulation. Our findings demonstrate that IFN-α employs distinct pathways for regulating IL-1ß production and reveal that in the case of M.tb infection, the induction of IFN-α and IFN-γ might be associated with M.tb immune escape and disease progression in infected humans.-Ma, J., Yang, B., Yu, S., Zhang, Y., Zhang, X., Lao, S., Chen, X., Li, B., Wu, C. Tuberculosis antigen-induced expression of IFN-α in tuberculosis patients inhibits production of IL-1ß.
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Antígenos Bacterianos/inmunología , Interferón-alfa/genética , Interferón-alfa/inmunología , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Mycobacterium tuberculosis/inmunología , Tuberculosis/inmunología , Adulto , Anciano , Caspasa 1/genética , Caspasa 1/inmunología , Caspasa 1/metabolismo , Femenino , Humanos , Interferón-alfa/metabolismo , Interferón gamma/genética , Interferón gamma/inmunología , Interferón gamma/metabolismo , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Mycobacterium bovis/inmunología , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT1/inmunología , Factor de Transcripción STAT1/metabolismo , Tuberculosis/genética , Tuberculosis/metabolismo , Adulto JovenRESUMEN
Increasing evidences in animals and humans suggest that CD8(+) T cells contribute significantly to immune defenses against Mycobacterium tuberculosis (Mtb). In the present study, we found that without any stimulation, CD8(+) T cells in pleural fluid cells (PFCs) expressed significantly higher levels of CD69 than PBMCs from patients with tuberculous pleurisy (TBP). CD8(+)CD69(+) T cells expressed significantly higher levels of CD45RO and HLA-DR and lower levels of CD45RA than CD8(+)CD69(-) T cells, demonstrating that CD8(+)CD69(+) T cells were activated memory cells. Furthermore, we found higher expression of CCR6 and lower expression of CCR7 and CD62L on CD8(+)CD69(+) T cells compared with CD8(+)CD69(-) T cells, suggesting that the expression of CCR6 and reduced expression of CCR7 and CD62L might facilitate the migration of circulating CD8(+)CD69(+) T cells into tuberculous pleural space. Importantly, following stimulation with culture filtrate protein of 10 kDa (CFP10) peptides, CD8(+)CD69(+) T cells but not CD8(+)CD69(-) T cells expressed CD107a/b, IFN-γ and TNF-α, demonstrating that CD8(+)CD69(+) T cells were MTB-specific cells. In addition, the majority of CD8(+)CD69(+) T cells were dominated by polyfunctional T cells. In summary, we demonstrated that CD69 as a useful marker for MTB-specific CD8(+) T cells in PFCs from patients with TBP enabled a direct ex vivo estimation of the quantity, as well as the quality, of MTB-specific CD8(+) responses.
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Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos T/metabolismo , Linfocitos T CD8-positivos/metabolismo , Lectinas Tipo C/metabolismo , Tuberculosis Pleural/inmunología , Adulto , Anciano , Biomarcadores/metabolismo , Femenino , Humanos , Interferón gamma/biosíntesis , Antígenos Comunes de Leucocito/metabolismo , Masculino , Persona de Mediana Edad , Mycobacterium tuberculosis/inmunología , Factor de Necrosis Tumoral alfa/biosíntesis , Adulto JovenRESUMEN
We recently reported that pleural fluid mononuclear cells (PFMCs) from tuberculous pleurisy stimulated with Bacillus Calmette-Guérin (BCG) or TB antigens produced high levels of cytokines. However, it was still unclear what mechanism of the PFMCs used to migrate into the pleural fluids in TB infection. In the present study, we found that CD3(+)CD4(+) and CD3(+)CD8(+) T cells from PFMCs expressed significantly high levels of CXCR3 compared to PBMCs. In addition, the levels of CXCL10 (the ligand for CXCR3) in pleural fluids were significantly higher than those in normal serum and cancerous fluids. After stimulation with BCG, PFMCs produced high levels of CXCL10. Importantly, the synthesis of CXCL10 was mainly dependent on the BCG-induced production of IFNs, because the neutralization of endogenous IFN-α or IFN-γ with mAbs significantly reduced the production of CXCL10 from BCG-stimulated PFMCs. In addition, the tubercular pleural fluid (TBPF) or exogenous CXCL10 induced the migration of PFMCs, indicating that IFN-α or IFN-γ modulated the immune response through the expression of CXCL10 to aid the recruitment and selective homing of activated/effector cells to the site of Mycobacterium tuberculosis (M.tb) infection. Taken together, the levels of CXCL10 in pleural fluids were high and BCG-stimulated PFMCs expressed high levels of CXCL10, and CXCL10 induced the migration of PFMCs into the pleural fluids in TB infection.
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Quimiocina CXCL10/inmunología , Interferón-alfa/inmunología , Interferón gamma/inmunología , Leucocitos Mononucleares/inmunología , Neoplasias Pulmonares/inmunología , Mycobacterium tuberculosis/inmunología , Tuberculosis Pleural/inmunología , Adulto , Anciano , Antígenos Bacterianos/inmunología , Movimiento Celular/inmunología , Citocinas/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Linfocitos T/inmunología , Tuberculosis Pleural/tratamiento farmacológicoRESUMEN
CD8(+) T cells are essential for host defense to Mycobacterium tuberculosis (Mtb) infection and identification of CD8(+) T cell epitopes from Mtb is of importance for the development of effective peptide-based diagnostics and vaccines. We previously demonstrated that the secreted 10-KDa culture filtrate protein (CFP10) from Mtb is a potent CD8(+) T cell antigen but the repertoire and dominance pattern of human CD8 epitopes for CFP10 remained poorly characterized. In the present study, we undertook to define immunodominant CD8 epitopes involved in CFP10 using a panel of CFP10-derived 13-15 amino acid (aa) peptides overlapping by 11 aa. Four peptides in CFP10 were observed to induce significant CD8(+) T cell responses and we further determined the size of the epitopes involved in each individual peptide tested. Four 9 aa CD8 epitopes were finally identified and deleting a single amino acid from the N or C terminus of either peptide markedly reduced IFN-γ production, suggesting that they are minimum of CD8 epitopes. In the individuals tested, each epitope represented a single immunodominant response in CD8(+) T cells. The epitope-specific CD8(+) T cells displayed effector or effector memory phenotypes and could upregulate the expression of CD107a/b upon antigen stimulation. In addition, we found that epitope-specific CD8(+) T cells shared biased usage of T cell receptor (TCR) variable region of ß chain (Vß) 12, 9, 7.2 or Vß4 chains. As judged from HLA-typing results and using bioinformatics technology for prediction of MHC binding affinity, we found that the epitope-specific CD8(+) T cells are all restricted by HLA-B alleles. Our findings suggest that the four epitopes in CFP10 recognized by CD8(+) T cells might be of importance for the development of Mtb peptide-based vaccines and for improved diagnosis of TB in humans.
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Alelos , Proteínas Bacterianas/inmunología , Linfocitos T CD8-positivos/inmunología , Antígenos HLA-B/genética , Epítopos Inmunodominantes/inmunología , Mycobacterium tuberculosis/inmunología , Adulto , Anciano , Secuencia de Aminoácidos , Linfocitos T CD4-Positivos/inmunología , Epítopos de Linfocito T/inmunología , Femenino , Prueba de Histocompatibilidad , Humanos , Memoria Inmunológica/inmunología , Interferón gamma/biosíntesis , Proteína 1 de la Membrana Asociada a los Lisosomas/metabolismo , Proteína 2 de la Membrana Asociada a los Lisosomas/metabolismo , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Péptidos/química , Péptidos/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Tuberculosis Pleural/inmunología , Factor de Necrosis Tumoral alfa/biosíntesis , Adulto JovenRESUMEN
OBJECTIVE: To compare the long-term results of mastoscopic axillary lymph node dissection (MALND) and conventional axillary lymph node dissection (CALND). PATIENTS AND METHODS: From January 1, 2003, through December 31, 2005, a group of 1027 consecutive patients with operable breast cancer were randomly assigned to 1 of 2 study groups: MALND and CALND. The median follow-up was 63 months. The primary end points of the study were operative outcomes, complication reduction, function conservation, and cosmetics. The secondary end points were disease-free and overall survival. RESULTS: The mean operative blood loss in the MALND group was less than in the CALND group (P<.001). The patients who underwent MALND had less axillary pain, numbness or paresthesias, and arm swelling (P<.001). The aesthetic appearance of the axilla in the MALND group was much better than that in the CALND group (P=.001 at 6 months and P=.002 at 24 months). A significant difference was found between the 2 groups in distant metastasis (P=.04). The disease-free survival rate was 64.5% in the MALND group and 60.8% in the CALND group (P=.88). The overall survival rate was 81.7% in the MALND group and 78.6% in the CALND group (P=.95). CONCLUSION: Compared with CALND, MALND has advantages in operative outcomes, complication reduction, function conservation, and cosmetics.
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Neoplasias de la Mama/mortalidad , Neoplasias de la Mama/patología , Escisión del Ganglio Linfático/métodos , Axila , Pérdida de Sangre Quirúrgica , Carcinoma Ductal de Mama/mortalidad , Carcinoma Ductal de Mama/patología , Carcinoma Lobular/mortalidad , Carcinoma Lobular/patología , Supervivencia sin Enfermedad , Edema/epidemiología , Estética , Femenino , Humanos , Hipoestesia/epidemiología , Masculino , Persona de Mediana Edad , Metástasis de la Neoplasia , Recurrencia Local de Neoplasia , Dolor Postoperatorio/epidemiología , Parestesia/epidemiología , Tasa de SupervivenciaRESUMEN
Tuberculous pleurisy (TBP) is a frequent extrapulmonary manifestation characterized by the accumulation of inflammatory cells that can sometimes be spontaneously self-cured. To achieve a greater insight into T cells at a local site, we systematically characterized and compared the numbers of antigen-specific T cells responding to BCG- or MTB-specific antigens. Our results showed that significantly higher levels of Th1 cytokines were produced by pleural fluid cells (PFCs) than PBMCs following stimulation with BCG or peptides from Mycobacterium tuberculosis (MTB)-specific antigens, ESAT-6 and CFP-10. The proportions of Th1 cells producing IL-2 alone or IL-2 and TNF-α were higher than those producing IFN-γ alone, following stimulation with ESAT-6 or CFP-10 peptides. The cells responding to BCG, ESAT-6 and CFP-10 displayed a CD45RA(-)CCR7(-)CD62L(-)CD27(-) effector/effector memory phenotype. The percentages and median fluorescence intensity (MFI) of polyfunctional CD4(+) T cells were significantly higher following stimulation with peptides from ESAT-6 or CFP-10 than BCG. Our results demonstrated that significantly higher levels of polyfunctional CD4(+) T cells for the epitopes of ESAT-6 or CFP-10 in PFCs may play an important role in the local control of tuberculosis (TB) infection.
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Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Linfocitos T CD4-Positivos/inmunología , Mycobacterium bovis/inmunología , Mycobacterium tuberculosis/inmunología , Tuberculosis Pleural/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Citocinas/inmunología , Femenino , Humanos , Activación de Linfocitos , Masculino , Persona de Mediana Edad , Mycobacterium bovis/genética , Tuberculosis Pleural/genética , Adulto JovenRESUMEN
Th1 cell-mediated immune responses at the site of active infection are important to restrict the growth of M. tuberculosis (MTB) and for the spontaneous resolution of patients with tuberculous pleurisy (TBP). In the present study, we found that without any stimulation, CD4(+) T cells in pleural fluid cells (PFCs) from patients with TBP expressed significantly higher levels of CD69 than PBMCs from patients with tuberculosis (TB) or healthy donors. CD4(+)CD69(+) T cells expressed T-bet and IL-12Rß2. After stimulation with MTB-specific antigens, CD4(+)CD69(+) T cells expressed significantly higher levels of IFN-γ, IL-2 and TNF-α than CD4(+)CD69(-) T cells, demonstrating that CD4(+)CD69(+) T cells were MTB-specific Th1 cells. In addition, CD4(+)CD69(+) T cells were mostly polyfunctional Th1 cells that simultaneously produced IFN-γ, IL-2, TNF-α and displayed an effector or effector memory phenotype (CD45RA(-)CCR7(-)CD62L(-)CD27(-)). Moreover, the percentages of CD4(+)CD69(+) T cells were significantly and positively correlated with polyfunctional T cells. Interestingly, sorted CD4(+)CD69(+) but not CD4(+)CD69(-) fractions by flow cytometry produced IFN-γ, IL-2 and TNF-α that were significantly regulated by CD4(+)CD25(+) Treg cells. Taken together, based on the expression of CD69, we found a direct quantitative and qualitative method to detect and evaluate the in vivo generated MTB-specific polyfunctional CD4(+) T cells in PFCs from patients with TBP. This method can be used for the potential diagnosis and enrichment or isolation of MTB-specific Th1 cells in the investigations.
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Antígenos CD/biosíntesis , Antígenos de Diferenciación de Linfocitos T/biosíntesis , Lectinas Tipo C/biosíntesis , Mycobacterium tuberculosis/inmunología , Pleura/patología , Células TH1/microbiología , Tuberculosis Pleural/patología , Antígenos Bacterianos/farmacología , Citocinas/inmunología , Humanos , Pleura/inmunología , Especificidad del Receptor de Antígeno de Linfocitos T , Células TH1/inmunologíaRESUMEN
Important advances have been made in the immunodiagnosis of tuberculosis (TB) based on the detection of Mycobacterium tuberculosis (MTB)-specific T cells. However, the sensitivity and specificity of the immunological approach are relatively low because there are no specific markers for antigen-specific Th cells, and some of the Th cells that do not produce cytokines can be overlooked using this approach. In this study, we found that MTB-specific peptides of ESAT-6/CFP-10 can stimulate the expression of CD40L specifically in CD4(+) T cells but not other cells from pleural fluid cells (PFCs) in patients with tuberculous pleurisy (TBP). CD4(+)CD40L(+) but not CD4(+)CD40L(-) T cells express IFN-γ, IL-2, TNF-α, IL-17 or IL-22 after stimulation with MTB-specific peptides. In addition, CD4(+)CD40L(+) T cells were found to be mostly polyfunctional T cells that simultaneously produce IFN-γ, IL-2 and TNF-α and display an effector or effector memory phenotype (CD45RA(-)CD45RO(+)CCR7(-)CD62L(-)ICOS(-)). To determine the specificity of CD4(+)CD40L(+) T cells, we incubated PFCs with ESTA-6/CFP-10 peptides and sorted live CD4(+)CD40L(+) and CD4(+)CD40L(-) T cells by flow cytometry. We further demonstrated that sorted CD4(+)CD40L(+), but not CD4(+)CD40L(-) fractions, principally produced IFN-γ, IL-2, TNF-α, IL-17 and IL-22 following restimulation with ESTA-6/CFP-10 peptides. Taken together, our data indicate that the expression of CD40L on MTB-specific CD4(+) T cells could be a good marker for the evaluation and isolation of MTB-specific Th cells and might also be useful in the diagnosis of TB.
Asunto(s)
Ligando de CD40/inmunología , Mycobacterium tuberculosis/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Tuberculosis Pleural/inmunología , Adulto , Anciano , Secuencia de Aminoácidos , Femenino , Citometría de Flujo , Humanos , Memoria Inmunológica , Masculino , Persona de Mediana Edad , Subgrupos de Linfocitos TRESUMEN
T cell-mediated immunity is critical for the control of Mycobacterium tuberculosis infection. Identifying the precise immune mechanisms that lead to control of initial M. tuberculosis infection and preventing reactivation of latent infection are crucial for combating tuberculosis. However, a detailed understanding of the role of T cells in the immune response to infection has been hindered. In addition, there are few flow cytometry studies characterizing the Vß repertoires of T cell receptors (TCRs) at local sites of M. tuberculosis infection in adult tuberculosis. In this study, we used culture filtrate protein 10 (CFP-10) from M. tuberculosis to characterize T cells at local sites of infection. We simultaneously analyzed the correlation of the production of cytokines with TCR Vß repertoires in CFP-10-specific CD4(+) and CD8(+) T cell subsets. For the first time, we demonstrate that CFP-10-specific CD4(+) or CD8(+) T cells from tubercular pleural fluid can produce high levels of gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) and upregulate the expression of CD107a/b on the cell surface. The CFP-10-specific cells were effector/memory cells with a CD45RO(+) CD62L(-) CCR7(-) CD27(-) expression profile. In addition, we found CFP-10-specific CD4(+) and CD8(+) T cells in tubercular pleural fluid, with biased usage of TCR Vß9, Vß12, or Vß7.2. Our findings of CFP-10-specific CD4(+) and CD8(+) T cells in tubercular pleural fluid are critical for understanding the mechanisms of the local cellular immune response and developing more effective therapeutic interventions in cases of M. tuberculosis infection.
Asunto(s)
Proteínas Bacterianas/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Memoria Inmunológica , Derrame Pleural , Receptores de Antígenos de Linfocitos T alfa-beta/metabolismo , Tuberculosis Pleural/inmunología , Adulto , Linfocitos T CD4-Positivos/química , Linfocitos T CD8-positivos/química , Femenino , Humanos , Interferón gamma/metabolismo , Selectina L/análisis , Antígenos Comunes de Leucocito/análisis , Masculino , Persona de Mediana Edad , Receptores CCR7/análisis , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/análisis , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
AIM: To evaluate cytokine production by CD4(+); T cells and frequency of multifunctional CD4(+); T cells after stimulation with ESAT-6 peptide pool. To understand the role of ESAT-6 specific CD4(+); T cells in the control of local TB infection. METHODS: PFCs were isolated from patients with tuberculous pleurisy, and assessed by flow cytometry for cytokine production, subpopulation, frequency and function of multifunctional CD4(+);T cells after stimulation with ESAT-6 peptide pool. RESULTS: Following stimulation with BCG, ESAT-6 peptide pool and recombinant ESAT-6 protein, CD4(+); but not CD8(+); T cells expressed IFN-γ, IL-2 and TNF-α. Distinct from BCG, ESAT-6 peptide pool and recombinant ESAT-6 protein induced similar frequencies of IFN-γ, IL-2 and TNF-α. High frequency of multifunctional CD4(+);T cells was observed. Analysis of mean fluorescence intensity (MFI) indicated that triple positive cells produced most amounts of cytokines on single cell level compared with double positive and single positive cells. CONCLUSION: ESAT-6 peptide pool predominantly induced Th1 cytokine production by CD4(+);T cells in PFCs. Multifunctional CD4(+); T cells were observed and secreted most amounts of cytokines on single cell level, suggesting that these cells probably played essential role in local TB infection.