scholarly journals Maturation of T and B Lymphocytes in the Assessment of the Immune Status in COVID-19 Patients

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2615
Author(s):  
Iwona Kwiecień ◽  
Elżbieta Rutkowska ◽  
Krzysztof Kłos ◽  
Ewa Więsik-Szewczyk ◽  
Karina Jahnz-Różyk ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Lymphocytes ◽  
Cd8 T Cells ◽  
X Ray ◽  
Healthy Control ◽  
Chest X Ray ◽  
Novel Coronavirus ◽  
And Control ◽  
B And T Lymphocytes

Cell response to novel coronavirus disease 19 (COVID-19) is currently a widely researched topic. The assessment of leukocytes population and the maturation of both B and T lymphocytes may be important in characterizing the immunological profile of COVID-19 patients. The aim of the present study was to evaluate maturation of B and T cells in COVID-19 patients with interstitial lesions on chest X-ray (COVID-19 X-ray (+)), without changes on X-ray (COVID-19 X-ray (−)) and in healthy control. The study group consisted of 23 patients divided on two groups: COVID-19 X-ray (+) n = 14 and COVID-19 X-ray (−) n = 9 and control n = 20. The flow cytometry method was performed. We observed a significantly higher percentage of plasmablasts and lower CD4+ lymphocytes in COVID-19 X-ray (+) patients than in COVID-19 X-ray (−) and control. In the COVID-19 X-ray (+) patients, there was a lower proportion of effector CD4+ T cells, naïve CD8+ T cells and higher central memory CD4+ cells and effector CD8+ T cells than control. The above results showed that the assessment of selected cells of B and T lymphocytes by flow cytometry can distinguish patients with COVID-19 and differentiate patients with and without changes on chest X-ray.

189 A clear increase in TILs and modest tumor growth inhibition by pembrolizumab in prostate cancer tumors growing in bone of CD34+ engrafted NOG mice

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A203-A203
Author(s):  
Suominen Suominen ◽  
Justyna Zdrojewska ◽  
Jenni Mäki-Jouppila ◽  
Philip Dube ◽  
Ivan Gladwyn-Ng ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Flow Cytometry ◽  
T Cells ◽  
Tumor Growth ◽  
Cancer Patients ◽  
Cd8 T Cells ◽  
List Type ◽  
X Ray ◽  
Bone Changes ◽  
Nog Mice

BackgroundThe recent KEYNOTE-199 trial raises hope for new treatment options for prostate cancer patients with the encouraging results of checkpoint inhibitor activity in a subset of prostate cancer patients, also including patients with bone-predominant disease. However, the patient subset that benefited from the treatment was small, needing identification predictive biomarkers1. Proper preclinical models can help in the biomarker quest as well as in the search and selection of the best possible combination partners for further clinical trials.MethodsIn this study the bone-metastatic disease was modeled by intratibial inoculation of LNCaP human prostate cancer cells to male CIEA NOG® (NOG) mice and NOG mice engrafted with human CD34+ hematopoietic stem cells (huNOG, Taconic Biosciences). Tumor growth was followed by serum PSA measurements and tumor-induced bone changes by X-ray images. At study week 4, the PSA positive mice were stratified to two groups (n=10) treated with IgG4 isotype control or pembrolizumab (5 mg/kg, i.p., Q5D) until the end of the study. Tumor-induced bone changes were followed by X-ray 4, 8 and 10 weeks after inoculation. The study was terminated 10 weeks after inoculation and tumors were processed for histological and immunohistochemical (IHC) analysis of tumor infiltrating lymphocytes (TILs). Changes in blood cell counts were assessed by flow cytometry and hematology (n=5/group).ResultsAt sacrifice, tumor-induced bone changes were observed in all mice, and there was no difference between the groups. Even though the PSA was not significantly lower in the pembrolizumab-treated group, the average histological tumorous surface was lower. In flow cytometry of peripheral blood, increases in the portions of CD3+ leukocytes and double positive CD4+CD8+ cells were observed, but no differences were found in CD4+ nor CD8+ T-cells. However, CD8+ T-cells were radically increased within the tumor as analyzed by IHC.ConclusionsThe model successfully mimicked the prevalent clinical situation, where clear responses in PSA or target lesions are not observed. However, a dramatic increase of cytotoxic T-cells in the tumor was observed, revealing the effects of pembrolizumab in a model of prostate cancer growth in bone of huNOG mice. The model presents a suitable platform for studying combination partners with pembrolizumab, that would boost or unlock the anti-tumor activity of the increased TILs.Ethics ApprovalThis study was approved by the National Animal Experiment Board in Finland; license number ESAVI-2331-04 10 07-2017.ReferenceAntonarakis ES, Piulats JM, Gross-Goupil M, et al. Pembrolizumab for Treatment-Refractory Metastatic Castration-Resistant Prostate Cancer: Multicohort, Open-Label Phase II KEYNOTE-199 Study. J Clin Oncol 2020;38:395–405.

scholarly journals Automated COVID-19 Detection from Chest X-Ray Images : A High Resolution Network (HRNet)Approach

2020 ◽  
Author(s):  
Sifat Ahmed ◽  
Tonmoy Hossain ◽  
Oishee Bintey Hoque ◽  
Sujan Sarker ◽  
Sejuti Rahman ◽  
...  
Keyword(s):  
High Resolution ◽  
Well Being ◽  
X Rays ◽  
X Ray ◽  
Pcr Diagnosis ◽  
Chest X Ray ◽  
Novel Coronavirus ◽  
And Control ◽  
Learning Architectures ◽  
Global Population

The pandemic, originated by novel coronavirus 2019 (COVID-19), continuing its devastating effect on the health, well-being, and economy of the global population. A critical step to restrain this pandemic is the early detection of COVID-19 in the human body, to constraint the exposure and control the spread of the virus. Chest X-Rays are one of the noninvasive tools to detect this disease as the manual PCR diagnosis process is quite tedious and time-consuming. In this work, we propose an automated COVID-19 classifier, utilizing available COVID and non-COVID X-Ray datasets, along with High Resolution Network (HRNet) for feature extraction embedding with the UNet for segmentation purposes. To evaluate the proposed dataset, several baseline experiments have been performed employing numerous deep learning architectures. With extensive experiment, we got 99.26% accuracy, 98.53% sensitivity, and 98.82% specificity with HRNet which surpasses the performances of the existing models. Our proposed methodology ensures unbiased high accuracy, which increases the probability of incorporating X-Ray images into the diagnosis of the disease.

scholarly journals P06.14 Characterization of tumor-infiltrating T cells by highly multiplexed immunofluorescence imaging

2020 ◽  
Vol 8 (Suppl 2) ◽  
pp. A47.2-A48
Author(s):  
E Criado-Moronati ◽  
A Gosselink ◽  
J Kollet ◽  
A Dzionek ◽  
B Heemskerk
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
T Lymphocytes ◽  
Tumor Cells ◽  
Cd8 T Cells ◽  
T Cell Subsets ◽  
Imaging Technology ◽  
Part Time

BackgroundThe adoptive cell transfer (ACT) of tumor-infiltrating T lymphocytes (TILs) has shown remarkable results in patients with different cancer types. The antitumor effect of this therapy is mainly attributed to a small fraction of tumor-reactive T lymphocytes (TRLs) that recognize mutated peptides as well as overexpressed self-antigens. Therefore, the enrichment and expansion of TRLs constitutes a promising immunotherapy approach. However, the specific targeting of individual mutated antigens represents a daunting challenge for widespread therapeutic application. Alternatively, we hypothesize that TRLs could be identified and enriched by a surface marker (or combination thereof) in an antigen-independent manner as a result of the chronic antigen exposure and other factors present in the tumor microenvironment (TME).Materials and MethodsWe screened T cell activation and exhaustion markers, among others, on different tumor tissues using the MACSima™ Imaging Platform, an instrument for the highly multiplexed immunofluorescence imaging technology MICS (Multiparameter Imaging Cell Screen), enabling investigation of hundreds of markers on a single section. Moreover, flow cytometry and single-cell RNA sequencing analyses of T cells from tumor digests were performed to complement the characterization of TILs.ResultsThe MICS results highlighted the complexity of the TME, mainly composed of tumor cells, fibroblasts and endothelial vessels. In some cases, an extensive immune infiltrate consisted of T cells, plasma cells, some B cells and distinct myeloid cells was observed. Particularly, CD8 T cells from different tumor areas exhibited a tissue-resident memory phenotype with the expression of CD69, CD45RO or CD103. Activated/exhausted CD8 T cells were homogenously found across the imaged tumor areas. However, there was a tendency to find them in close proximity to tumor cells, especially for CD8 subsets expressing CD39 and other relevant markers, which may suggest the identification of tumor-reactive CD8 T cell populations. Flow cytometry data revealed the presence of similar T cell phenotypes in the patient´s TILs from tumor digests.ConclusionsThis imaging technology offers the possibility to study multiple parameters—including the localization—of relevant cells in the TME such as T cells. The phenotypic and functional characterization of different T cell subsets will allow the further investigation of their anti-tumor reactivity. Ultimately, the enrichment and expansion of the identified tumor-reactive T cell population hold great promises to improve the efficiency of T cell therapy against cancer.Disclosure InformationE. Criado-Moronati: A. Employment (full or part-time); Significant; Miltenyi Biotec B.V. & Co. KG. A. Gosselink: A. Employment (full or part-time); Significant; Miltenyi Biotec B.V. & Co. KG. J. Kollet: A. Employment (full or part-time); Significant; Miltenyi Biotec B.V. & Co. KG. A. Dzionek: A. Employment (full or part-time); Significant; Miltenyi Biotec B.V. & Co. KG. B. Heemskerk: A. Employment (full or part-time); Significant; Miltenyi Biotec B.V. & Co. KG.

scholarly journals Automated COVID-19 Detection from Chest X-Ray Images : A High Resolution Network (HRNet) Approach 

2020 ◽  
Author(s):  
Sifat Ahmed ◽  
Tonmoy Hossain ◽  
Oishee Bintey Hoque ◽  
Sujan Sarker ◽  
Sejuti Rahman ◽  
...  
Keyword(s):  
High Resolution ◽  
Well Being ◽  
X Rays ◽  
X Ray ◽  
Non Invasive ◽  
Pcr Diagnosis ◽  
Chest X Ray ◽  
Novel Coronavirus ◽  
And Control ◽  
Learning Architectures

Abstract Background/ introduction: The pandemic, originated by novel coronavirus 2019 (COVID-19), continuing its devastating effect on the health, well-being, and economy of the global population. A critical step to restrain this pandemic is the early detection of COVID-19 in the human body to constraint the exposure and control the spread of the virus. Chest X-Rays are one of the non-invasive tools to detect this disease as the manual PCR diagnosis process is quite tedious and time-consuming. Our intensive background studies show that, the works till now are not efficient to produce an unbiased detection result.Method: In this work, we propose an automated COVID-19 classification method, utilizing available COVID and non-COVID X-Ray datasets, along with High Resolution Network (HRNet) for feature extraction embedding with the UNet for segmentation purposes.Results: To evaluate the proposed method, several baseline experiments have been performed employing numerous deep learning architectures. With extensive experiment, we got a significant result of 99.26% accuracy, 98.53% sensitivity, and 98.82% specificity with HRNet which surpasses the performances of the existing models.Conclusions: Finally, we conclude that our proposed methodology ensures unbiased high accuracy, which increases the probability of incorporating X-Ray images into the diagnosis of the disease.

scholarly journals T Lymphocyte Maturation Profile in the EBUS-TBNA Lymph Node Depending on the DLCO Parameter in Patients with Pulmonary Sarcoidosis

Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3404
Author(s):  
Elżbieta Rutkowska ◽  
Iwona Kwiecień ◽  
Joanna Bednarek ◽  
Rafał Sokołowski ◽  
Agata Raniszewska ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Lymphocytes ◽  
Cd8 T Cells ◽  
Pulmonary Sarcoidosis ◽  
Unknown Etiology ◽  
Median Proportion ◽  
Normal Diffusion ◽  
Granulomatous Disorder ◽  
Inflammatory Changes

Sarcoidosis (SA) is a systemic granulomatous disorder of unknown etiology with lung and mediastinal lymph nodes (LNs) as the main location. T lymphocytes play important role in the formation of granulomas in SA, but still little is known about the role of maturation profile in the development of inflammatory changes. The aim of this study was to determine the CD4+ and CD8+ T cells maturation profile in LNs and in peripheral blood (PB) and its relation to disease severity expressed by diffusing capacity of the lung for carbon monoxide (DLCO). 29 patients with newly pulmonary SA were studied. Flow cytometry was used for cells evaluation in EBUS-TBNA samples. We observed lower median proportion of T lymphocytes, CD4+ T and CD8+ T cells in patients with DLCO< 80% than in patients with normal diffusion (DLCO > 80%). Patients with DLCO < 80% had lower median proportion of effector and higher median proportion of central memory CD4+ and CD8+ T cells than patients with DLCO > 80%. We reported for the first time that LNs CD4+ and CD8+ T cells maturation differs depending on the DLCO value in sarcoidosis. Lymphocytes profiles in LNs may reflect the immune status of patients with SA and can be analysed by flow cytometry of EBUS-TBNA samples.

Tumor Localization and Quantitation of Adoptively Transfered T Lymphocytes in a Murine Model.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1343-1343
Author(s):  
Marianne Hokland ◽  
Mikkel S. Petersen ◽  
Charlotte C. Fleischer ◽  
Hans Stødkilde-Jørgensen ◽  
Søren B. Hansen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Lymphocytes ◽  
Cd8 T Cells ◽  
Tumor Tissue ◽  
Cellular Therapy ◽  
Mean Value ◽  
Tumor Localization ◽  
Toxic Substances

Abstract Tracking adoptively transferred antigen-specific T lymphocytes is an important prerequisite for devising better protocols for cellular therapy. To this end we have developed a highly sensitive method for “in situ” visualization of labelled lymphocytes in vivo by combined PET and magnetic resonance imaging (MRI) to monitor the distribution of adoptively transferred tumour-specific T cells in a mouse model system. Moreover, quantitation of the adoptively transferred cells in tumor was performed by flow cytometry. C57BL/6J mice carrying subcutaneous tumours of the ovalbumin (OVA)-expressing malignant melanoma cell line B16-OVA were adoptively transferred with OVA-specific CD8+ T cells labelled with 124IdU. Five days after transfer of T cells, mice were killed and subjected to PET and MR imaging. Using a newly developed method for co-registration of the two image modalities, the anatomical localisation of the transferred cells was visualised and the amount of radioactivity in various anatomical locations very accurately determined. For quantitation of tumor infiltrating non-labelled OVA-specific CD8+ T cells by flow cytometry (using AbsoluteCount Beads), tumors were removed from mice day 1 until day 8 following adoptive transfer (6 mice/group) and prepared for single cell suspension before labeled with anti-CD8-FITC and SIINFEKL-Tetramer-PE. Results showed a clear tumor localization of the adoptively transferred OVA-specific T cells in the tumours. In two independent experiments comprising 12 and 13 evaluable mice, respectively, we found a mean value of 0.909 +/− 0.468 Bq and 0.926 +/− 0.553 Bq in the tumours, and only 0.182 +/− 0.479 Bq and 0.026 +/− 0.480 Bq in the corresponding contralateral control volumes. The difference in activity between the tumour regions and the control regions was statistically highly significant with 2p-values of 0.002 and 0.006 for the two experiments. Using flow cytometry it was shown that the number of OVA specific T lymphocytes accumulating in tumor gradually increased until day 5 after transfer when an average of 3.3 million SIINFEKL-specific cells per gram tumor tissue was found. From day 5 until day 8 the number of SIINFEKL-specific cells per gram tumor tissue fluctuated at a fairly constant level. This method presented for tracking adoptively transfered tumor specific T lymphocytes represent a significant advancement for studies of adoptively transferred specific T cells, and could potentially be developed for diagnostic purposes. Moreover, since these studies show that tumor-specific T cells home to subcutaneous tumours in substantial numbers, we suggest that these migrating cells could be employed in a new form of therapy as carriers of toxic substances to tumors.

scholarly journals CD27 Expression Promotes Long-Term Survival of Functional Effector–Memory CD8+Cytotoxic T Lymphocytes in HIV-infected Patients

2004 ◽  
Vol 200 (11) ◽  
pp. 1407-1417 ◽  
Author(s):  
Adrian F. Ochsenbein ◽  
Stanley R. Riddell ◽  
Michele Brown ◽  
Lawrence Corey ◽  
Gabriela M. Baerlocher ◽  
...  
Keyword(s):  
T Cells ◽  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Cd8 T Cells ◽  
Interleukin 2 ◽  
Tumor Necrosis Factor Receptor ◽  
Cell Receptor ◽  
Effector Memory ◽  
Term Survival ◽  
Functional Consequences

Human immunodeficiency virus (HIV)-specific CD8+ T cells persist in high frequencies in HIV-infected patients despite impaired CD4+ T helper response to the virus, but, unlike other differentiated effector cytotoxic T lymphocytes, most continue to express the tumor necrosis factor receptor family member CD27. Because the ligand for CD27 (CD70) is also overexpressed in HIV-infected hosts, we examined the nature of expression and potential functional consequences of CD27 expression on HIV-specific CD8+ T cells. Analysis of CD27+ and CD27− T cells derived from the same HIV-specific clone revealed that retention of CD27 did not interfere with acquisition of effector functions, and that after T cell receptor stimulation, CD27+ cells that concurrently were triggered via CD27 exhibited more resistance to apoptosis, interleukin 2 production, and proliferation than CD27− T cells. After transfer back into an HIV-infected patient, autologous HIV-specific CD27− T cells rapidly disappeared, but CD27+ T cells derived from the same clone persisted at high frequency. Our findings suggest that the CD27–CD70 interaction in HIV infection may provide CD27+ CD8+ T cells with a survival advantage and compensate for limiting or absent CD4+ T help to maintain the CD8 response.

scholarly journals Modulation of let-7 miRNAs controls the differentiation of effector CD8 T cells

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Alexandria C Wells ◽  
Keith A Daniels ◽  
Constance C Angelou ◽  
Eric Fagerberg ◽  
Amy S Burnside ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Cd8 T Cells ◽  
T Cell Responses ◽  
Cd8 T Cell ◽  
Cell Receptor ◽  
Activated T Cells ◽  
Cell Responses

The differentiation of naive CD8 T cells into effector cytotoxic T lymphocytes upon antigen stimulation is necessary for successful antiviral, and antitumor immune responses. Here, using a mouse model, we describe a dual role for the let-7 microRNAs in the regulation of CD8 T cell responses, where maintenance of the naive phenotype in CD8 T cells requires high levels of let-7 expression, while generation of cytotoxic T lymphocytes depends upon T cell receptor-mediated let-7 downregulation. Decrease of let-7 expression in activated T cells enhances clonal expansion and the acquisition of effector function through derepression of the let-7 targets, including Myc and Eomesodermin. Ultimately, we have identified a novel let-7-mediated mechanism, which acts as a molecular brake controlling the magnitude of CD8 T cell responses.

Abstract 001: Axl Expression by CD4+ T Lymphocytes Promotes Salt-Dependent Hypertension

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Kristine M Wadosky ◽  
Sri N Batchu ◽  
Angie Hughson ◽  
Kathy Donlon ◽  
Craig N Morrell ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
T Lymphocytes ◽  
Cd4 T Cells ◽  
White Blood Cells ◽  
Interleukin 4 ◽  
Th2 Cells ◽  
Salt Hypertension ◽  
Ifn Γ

Introduction: Our laboratory has shown that Axl, a receptor tyrosine kinase, is important in both vascular and immune functions during deoxycorticosterone acetate (DOCA)-salt hypertension. We hypothesized that Axl activity specifically in T lymphocytes could explain the dependence of hypertension on Axl. Methods and Results: We did adoptive transfers of either Axl+/+ or Axl-/- CD4+ T cells to RAG1-/- mice that lack mature T cells. Once CD4+ T cell repopulations were confirmed, we induced DOCA-salt hypertension for 6 weeks. Systolic blood pressure (BP, mmHg) increased by 20±5 in Axl+/+RAG-/- mice after DOCA-salt, but Axl-/- RAG-/- mice had increases in BP by only 6+3 after 6 weeks of DOCA-salt. We isolated naïve CD4+ T cells from both Axl+/+ and Axl-/- littermates and primed them under either Th1 or Th2 polarizing conditions in culture. Production of interferon gamma (IFN-γ ng/mL) was significantly decreased (-23%, p<0.05) in Axl-/- (396±23) compared to Axl+/+ (512±42) under Th1-priming. However, Axl had no effect on interleukin 4 (IL-4, ng/mL) production under Th2 polarizing conditions. Intracellular staining of the Th1/Th2 cells with IFN-γ and IL-4 antibodies by flow cytometry confirmed expression of cytokines in culture media. Complete blood counts showed that Axl-/- mice had significantly lower white blood cells due to decreased numbers of lymphocytes (4.5±0.7x10 9 ) compared to Axl+/+ mice (7.8±0.7x10 9 ). We found a higher population of AnnexinV (marker of early apoptosis)-positive peripheral leukocytes in Axl-/- mice (10±1%) compared to Axl+/+ (4±1%) by flow cytometry; while the percentages of dead cells (~10%) were similar between Axl+/+ and Axl-/- mice. Conclusions: Altogether we show that expression of Axl by T cells drives salt-induced hypertension. The mechanism of Axl-dependent effects on T cells occurs via T-cell-dependent expression of the pro-inflammatory cytokine IFN-γ. In addition, Axl plays a role in inhibiting lymphocyte apoptosis in the circulation. Future work will focus on how Axl expression in T cells affects T cell-dependent vascular remodeling during hypertension.

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