Objective
We aimed to evaluate the prognostic value of circulating tumor cells (CTCs) and the impact of intraoperative tumor manipulation on CTCs in colorectal cancer (CRC) patients.
Methods
We ...performed a prospective study on 40 patients with CRC stages I to IV who received curative surgery using the no-touch technique. Flow cytometry was used to identify CTCs in peripheral blood samples (4 mL/sample) collected at two surgical moments: skin incision (T1) and after surgical resection (T2). A threshold of ≥4 CTCs/4 mL blood was established for considering patients CTC positive.
Results
In the univariate analysis, CTC evaluation at T2 was correlated with female sex, vascular invasion, tumor localization in the colon and metastatic lymph nodes. In the multivariate analysis, only female sex and colon cancer maintained statistical significance. At a medium follow-up of 15 months (1–25 months), the mortality rate was 10% (n = 4), with no significant differences between the overall survival of T1 or T2 CTC-positive and CTC-negative patients.
Conclusions
Flow cytometry is a feasible CTC identification technique in CRC, and although surgical manipulation has no influence on CTC numbers, CTCs may serve as a prognostic and predictive factor.
Wide regional differences in the age-related Anti Mullerian hormone (AMH) regression patterns or age at onset of natural menopause have been reported, possibly reflecting genetic, socioeconomic, ...environmental, racial or ethnic peculiarities. Moreover, adaptation of AMH levels from different assays using regression functions may lack accuracy and externally defined references for AMH levels may not fully comply with a specific geographical area. The current study aimed to establish an accurate mathematical relationship between AMH serum values and age in a large group of women from Romania, as any consistent difference from previously reported regression models may aid in building specific profiles for the AMH decline with age in this geographical region. Our study pointed out to the quadratic regression as the most fitted pattern of correlation for all the age groups between 24 and 45. To our knowledge the current manuscript is based on the singular study carried out in this geographical region, generating a particular age-related pattern of association between age and serum AMH levels in women, regardless of their subjacent pathologies.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Acute myeloid leukemia (AML) is generally considered a poorly immunogenic malignancy, displaying a "non-inflamed" leukemia microenvironment (LME), leading to T cell tolerance. However, the immune ...landscape of AML is much more heterogeneous. Since B7 expression is regarded as a consequence of an interferon-mediated "inflammatory" phenotype, we have investigated by flow cytometry the B7 checkpoint ligands B7.1, B7.2, programmed death ligand 1 (PD-L1), PD-L2, ICOS-L, B7-H3, and B7-H4 on the AML blasts of 30 newly diagnosed patients and their corresponding receptors cytotoxic T lymphocyte-associated protein 4 (CTLA-4), programmed death 1 (PD-1), and inducible T cell costimulator (ICOS) on bone marrow (BM) T cell maturation populations. We could thus evidence B7-negative and B7-positive leukemias either with an isolated expression or part of eight different checkpoint ligand "signatures" that always included an inhibitory B7 molecule. B7-positive AMLs encompassed intermediate and adverse European Leukemia Net (ELN) risk cases and displayed mainly central memory CD4+ T cells with high ICOS levels and effector CD8+ T cells with high PD-1 expression. B7-negative cases were rather classified as AML with recurrent genetic anomalies and displayed predominantly naive T cells, with the exception of
mutated AMLs, which expressed B7-H3. These different B7 immune profiles suggest that specific immunotherapies are required to target the distinct immune evasion strategies of this genetically heterogeneous disease.
Virus-specific cellular and humoral responses are major determinants for protection from critical illness after SARS-CoV-2 infection. However, the magnitude of the contribution of each of the ...components to viral clearance remains unclear. Here, we studied the timing of viral clearance in relation to 122 immune parameters in 102 hospitalised patients with moderate and severe COVID-19 in a longitudinal design. Delayed viral clearance was associated with more severe disease and was associated with higher levels of SARS-CoV-2-specific (neutralising) antibodies over time, increased numbers of neutrophils, monocytes, basophils, and a range of pro-inflammatory cyto-/chemokines illustrating ongoing, partially Th2 dominating, immune activation. In contrast, early viral clearance and less critical illness correlated with the peak of neutralising antibodies, higher levels of CD4 T cells, and in particular naïve CD4+ T cells, suggesting their role in early control of SARS-CoV-2 possibly by proving appropriate B cell help. Higher counts of naïve CD4+ T cells also correlated with lower levels of MIF, IL-9, and TNF-beta, suggesting an indirect role in averting prolonged virus-induced tissue damage. Collectively, our data show that naïve CD4+ T cell play a critical role in rapid viral T cell control, obviating aberrant antibody and cytokine profiles and disease deterioration. These data may help in guiding risk stratification for severe COVID-19.
The diagnostic spectrum for AML patients is increasingly based on genetic abnormalities due to their prognostic and predictive value. However, information on the AML blast phenotype regarding their ...maturational arrest has started to regain importance due to its predictive power for drug responses. Here, we deconvolute 1350 bulk RNA-seq samples from five independent AML cohorts on a single-cell healthy BM reference and demonstrate that the morphological differentiation stages (FAB) could be faithfully reconstituted using estimated cell compositions (ECCs). Moreover, we show that the ECCs reliably predict ex-vivo drug resistances as demonstrated for Venetoclax, a BCL-2 inhibitor, resistance specifically in AML with CD14+ monocyte phenotype. We validate these predictions using LUMC proteomics data by showing that BCL-2 protein abundance is split into two distinct clusters for NPM1-mutated AML at the extremes of CD14+ monocyte percentages, which could be crucial for the Venetoclax dosing patients. Our results suggest that Venetoclax resistance predictions can also be extended to AML without recurrent genetic abnormalities and possibly to MDS-related and secondary AML. Lastly, we show that CD14+ monocytic dominated Ven/Aza treated patients have significantly lower overall survival. Collectively, we propose a framework for allowing a joint mutation and maturation stage modeling that could be used as a blueprint for testing sensitivity for new agents across the various subtypes of AML.
: Colorectal cancer (CRC) is the third most common neoplasia in the world. Circulating tumor cells (CTC) have a prognostic value and can be useful in monitoring solid neoplasia. Only one method for ...CTC identification has received the approval and this is the CellSearch® system based on the immunomagnetic separation. Multiple markers are used in CTC identification, as epithelial markers and cytokeratines. CTC identification in peripheral blood is associated with a worse prognostic and reduced free survival in CRC.
: We performed a systematic search in PubMed database for articles that reports the circulating tumor cells in CRC until July 2019. We selected studies in English and French and the main words used for search were ‘circulating tumor cells’, ‘colorectal cancer’, ‘colon cancer’, ‘rectal cancer’, ‘flow cytometry’, ‘peripheral blood’. We included studies with more than 10 patients, where samples were collected from the blood in relation with surgery and flow cytometry was used as analyzing technique.
: We included 7 studies in final analysis, that showed in flow cytometry analysis a cut-off value of CTC that can vary from 2-4 CTC/ 7.5 ml peripheral blood with a sensitivity of 50.8% and specificity of 95%. Patients with positive CTC were associated with higher T stage and positive lymph nodes, with a worse overall survival (OS) and disease free survival (DFS) comparing with negative patients.
CTC are considered to be a prognostic factor who needs more validation studies in order to be included in the clinical practice.
Several alternative methods to peripheral blood DNA extraction have been implemented so far. Saliva seems to represent a very advantageous type of sample, easy to harvest and able to generate DNA ...yields comparable to those extracted from blood mononuclear cells.
8 patients suspected of ankylosing spondylitis, 9 patients with various hematological malignancies, displaying post-chemotherapy leucopenia and 30 healthy volunteers were included in our study. DNA was extracted with various commercially available kits and used for HLA typing either by PCR amplification, or by PCR followed by hybridization.
Our data regarding HLA typing support already published results regarding the good DNA quality that allows its use in various molecular biology techniques. However, when attempting to use saliva from immunosuppressed patients for DNA extraction we have generated very low yields, comparable again with the ones obtained from peripheral blood. Flow cytometry and immunocytochemistry investigations confirmed the low number of leukocytes present in the saliva of these patients, while the number of epithelial cells was virtually unchanged.
The main source of saliva DNA seems to be represented by leukocytes present in this fluid and not by the epithelial cells. Under these circumstances, for immunosuppressed patients saliva cannot represent an alternative to blood when attempting DNA extraction.
Background: Acute basophilic leukemia is a rare subtype of acute myeloid leukemia, as categorized by the 2008 World Health Organization classification of myeloid neoplasms. Acute basophilic leukemia ...diagnosis requires thorough morphological, cytochemical, immunophenotypic, molecular, and cytogenetic studies and exclusion of other hematological neoplasms associating basophilia. The disease course is defined by histamine driven, occasionally life-threatening respiratory, cardiovascular, cutaneous or digestive complications, as well as primary refractoriness to standard therapy. Clinical presentation: We herein report a case of a 63-year-old asthmatic female patient diagnosed with acute basophilic leukemia, associated with previously unpublished cytogenetic features and FLT-3 ITD mutation, pulmonary leukostasis and spontaneous pulmonary capillary leak syndrome, which worsened immediately following chemotherapy initiation. Respiratory complications were successfully managed, but recrudesced upon emergence of refractory disease and were ultimately fatal. We highlight the likelihood of pulmonary complications induced by basophil degranulation and tumor lysis in hypercellular acute basophilic leukemia and the potential benefit of histamine receptor blockade in this setting.
Systemic immune cell dynamics during coronavirus disease 2019 (COVID-19) are extensively documented, but these are less well studied in the (upper) respiratory tract, where severe acute respiratory ...syndrome coronavirus 2 (SARS-CoV-2) replicates
. Here, we characterized nasal and systemic immune cells in individuals with COVID-19 who were hospitalized or convalescent and compared the immune cells to those seen in healthy donors. We observed increased nasal granulocytes, monocytes, CD11c
natural killer (NK) cells and CD4
T effector cells during acute COVID-19. The mucosal proinflammatory populations positively associated with peripheral blood human leukocyte antigen (HLA)-DR
monocytes, CD38
PD1
CD4
T effector (T
) cells and plasmablasts. However, there was no general lymphopenia in nasal mucosa, unlike in peripheral blood. Moreover, nasal neutrophils negatively associated with oxygen saturation levels in blood. Following convalescence, nasal immune cells mostly normalized, except for CD127
granulocytes and CD38
CD8
tissue-resident memory T cells (T
). SARS-CoV-2-specific CD8
T cells persisted at least 2 months after viral clearance in the nasal mucosa, indicating that COVID-19 has both transient and long-term effects on upper respiratory tract immune responses.
Background/aim: T lymphocytes are important players of the immune response. B-CLL is characterized by several immune defects. Our study aims to characterize the distinct maturational and functional ...T/NK cell subsets within B-cell chronic lymphocytic leukemia disease Rai stages. Patients and methods: Peripheral blood mononuclear cells from 43 patients enrolled in the study (16 females and 27 males, aged 68±10, 8 Rai 0, 22 Rai 1/2 and 13 Rai 3/4) were analyzed by multiparameter flow cytometry. Distinct subsets within the CD4+ (naive, central memory, effector/peripheral memory, regulatory-Tregs, follicular-TFH, CXCR3+ and/or CCR4+), CD8+ (naive+memory, effector, senescent) and NK (CD57+ and/or CD94+) were identified and compared between disease Rai stages. Results: Total numbers of T lymphocytes increase with disease stage. Both CD4+ and CD8+ T cells are elevated in absolute counts. The majority of CD4+ T cells are antigen-experienced, with increased Tregs, TFH and CXCR3+ (Th1-associated profile) T cell counts. The CD8+ T cells expansion is due mostly to the senescent CD57+ subset. No significant difference within NK subsets was observed among different disease stages. Conclusions: B-CLL behaviour seems to be associated with increased numbers of TFH and Tregs. The therapeutic modulation of T cell response in B-CLL patients may play an important role in the disease behaviour and may be a key event compensating for the immunodeficiency occurring mostly in advanced stages of the disease.
Obiectiv: Limfocitele T se constituie în factori celulari importanţi ai răspunsului imun. Leucemia limfocitară cronică B se caracterizează prin defecte ale sistemului imun. Studiul nostru îşi propune caracterizarea diferitelor subseturi maturative şi funcţionale limfoide T şi NK la pacienţi aflaţi în stadii distincte de boală. Material şi metoda: au fost utilizate celule mononucleate separate în gradient de densitate de la 43 de pacienţi cu LLC-B (16 femei, 27 bărbaţi, vârsta 68±10 ani, 8 în Rai0, 22 în Rai1/2 şi 13 în Rai3/4) în vederea analizei prin citometrie in flux multiparametrică. Au fost identificate şi comparate între diferite stadii Rai, subseturile limfoide T CD4+ (naive, memorie centrală, memorie periferică/efector, reglatorii, foliculare-TFH, CXCR3 şi/sau CCR4+), CD8+ (naive+memorie, efectorii, senescente) şi NK (CD57 şi/sau CD94+). Rezultate: Limfocitele T totale circulante se găsesc în numere crescute la pacienţi în stadii avansate ale bolii. Majoritatea limfocitelor T CD4+ sunt celule ce au întâlnit antigenul, cu subseturi crescute de tip reglator, folicular şi CXCR3+ (asociat profilului Th1). Expansiunea limfocitelor citotoxice CD8+ este corelată cu un subset diferenţiat terminal, senescent, CD57+. În ceea ce priveşte subseturile limfocitare NK, nu s-au identificat diferente semnificative între diferite stadii ale bolii. Concluzii: Comportamentul leucemiei limfocitare cronice B pare a fi asociat cu o creştere în valori absolute a limfocitelor T CD4+ reglatorii şi foliculare circulante, dar şi cu expansiunea limfocitelor T citotoxice senescente. Modularea răspunsului T la pacienţii cu LLC-B joacă un rol important în evoluţia bolii şi se constituie în eveniment cheie al compensarii imunodeficientei caracteristice stadiilor avansate ale bolii.
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