Purification of viable peripheral blood mononuclear cells for biobanking using a robotized liquid handling workstation

Coppola, Luigi; Smaldone, Giovanni; Cianflone, Alessandra; Baselice, Simona; Mirabelli, Peppino; Salvatore, Marco

doi:10.1186/s12967-019-2125-7

Cited by 11 publications

(9 citation statements)

References 25 publications

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“…For the flow cytometry experiments, a sample aliquot of 200 μL of fresh EDTA-collected whole blood was used, while the remaining part was banked for plasma purification and collection of mononuclear cells as previously described. 7 Flow cytometry experiments were performed as follows: 100 μL of whole blood were distributed into 2 tubes containing the following antibody mixes:…”

Section: Methodsmentioning

confidence: 99%

Impact of Breast Tumor Onset on Blood Count, Carcinoembryonic Antigen, Cancer Antigen 15-3 and Lymphoid Subpopulations Supported by Automatic Classification Approach: A Pilot Study

et al. 2021

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Background Recent observations showed that systemic immune changes are detectable in case of breast cancer (BC). In this preliminary study, we investigated routinely measured peripheral blood (PB) parameters for malignant BC cases in comparison to benign breast conditions. Complete blood count, circulating lymphoid subpopulation, and serological carcinoembryonic antigen (CEA) and cancer antigen 15-3 (CA15-3) levels were considered. Methods A total of 127 female patients affected by malignant (n = 77, mean age = 63 years, min = 36, max = 90) BC at diagnosis (naïve patients) or benign breast conditions (n = 50, mean age = 33 years, min = 18, max = 60) were included in this study. For each patient, complete blood count and lymphoid subpopulations (T-helper, T-cytotoxic, B-, NK-, and NKT-cells) analysis on PB samples were performed. Hormonal receptor status, Ki-67 expression, and serological CEA and CA15-3 levels were assessed in the case of patients with malignant BC via statistical analysis. Results Women with malignant BC disclosed increased circulating T-helper lymphocytes and CD4/CD8 ratio in PB when compared to those affected by benign breast conditions (2.345 vs 1.894, P < .05 Wilcoxon rank-sum test). In the case of malignant BC patients, additive logistic regression method was able to identify malignant BC cases with increased CA15-3 levels (CA15-3 >25 UI/mL) via the hematocrit and neutrophils/lymphocytes ratio values. Moreover, in the case of women with aggressive malignant BC featured by high levels of Ki-67 proliferation marker, an increasing number of correlations were found among blood count parameters and lymphocytes subpopulations by performing a Spearman’s correlation analysis. Conclusions This preliminary study confirms the ability of malignant BC to determine systemic modifications. The stratification of malignant BC cases according to the Ki-67 proliferation marker highlighted increasing detectable alterations in the periphery of women with aggressive BC. The advent of novel and more sensitive biomarkers, as well as deep immunophenotyping technologies, will provide additional insights for describing the relationship between tumor onset and peripheral alterations.

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Section: Methodsmentioning

confidence: 99%

Impact of Breast Tumor Onset on Blood Count, Carcinoembryonic Antigen, Cancer Antigen 15-3 and Lymphoid Subpopulations Supported by Automatic Classification Approach: A Pilot Study

et al. 2021

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“…PBMCs were characterized phenotypically by flow cytometry for T and B lymphocytes [ 12 ]. CD3 (APC-H7), CD4 (Percp Cy 5.5), and CD8 (FITC) for T lymphocyte and CD20 (PE) for B lymphocyte were characterized [ 13 ]. Appropriate isotype-matched, nonreactive fluorochrome-conjugated antibodies were employed as controls.…”

Section: Methodsmentioning

confidence: 99%

Association of Lower Antispike Antibody Levels with Mortality in ICU Patients with COVID-19 Disease

Yelle

Amte

Gella

et al. 2023

Critical Care Research and Practice

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Background. Though vaccines have been reported as highly efficacious in preventing severe COVID-19 disease, there is emerging data of severe infections, albeit a small number, in vaccinated individuals. We have conducted a retrospective observational study to assess the clinical characteristics, immunological response, and disease outcomes among the vaccinated and unvaccinated patients admitted to the ICU with severe COVID-19 disease. Methods. Study Design and Participants. We conducted a retrospective observational study in COVID ICU of a tertiary care hospital. Data were collected from the month of 1 April 2021 to 31 November 2021. All adult patients admitted to the ICU having severe COVID-19 disease were included in the study. Data were collected from the medical records database which included demographics, a clinical course in the ICU, laboratory and radiological parameters, and disease outcomes. In a subset of patients, cell-mediated immunity and S1S2-neutralising antibody assessment was done. Results. A total of 419 patients with severe COVID-19 were included in the study. Of the 419 patients, 90 (21.5%) were vaccinated, and 329 (78.5%) were unvaccinated. There was a significantly higher mortality in unvaccinated severe COVID 19 patients as compared to vaccinated severe COVID patients (46.2% vs 34.4%; P < 0.0455 ). The neutralizing antibody titre was significantly higher in survivors as compared to nonsurvivors (2139.8, SE ± 713.3 vs 471, SE ± 154.4); P < 0.026 . Conclusion. Our study suggests the association of lower neutralizing antibody levels with mortality in ICU patients admitted with COVID-19 breakthrough infections.

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“…The biological samples and cell lines included in this study were provided and processed by the Biobank of the SDN institute [39,40].…”

Section: Methodsmentioning

confidence: 99%

KCTD15 Protein Expression in Peripheral Blood and Acute Myeloid Leukemia

et al. 2020

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Leukocytes are major cellular components of the inflammatory and immune response systems. After their generation in the bone marrow from hematopoietic stem cells, they maturate as granulocytes (neutrophils, eosinophils, and basophils), monocytes, and lymphocytes. The abnormal accumulation and proliferation of immature blood cells (blasts) lead to severe and widespread diseases such as leukemia. We have recently shown that KCTD15, a member of the potassium channel tetramerization domain containing protein family (KCTD), is remarkably upregulated in leukemic B-cells. Here, we extend our investigation by monitoring the KCTD15 expression levels in circulating lymphocytes, monocytes, and granulocytes, as well as in leukemia cells. Significant differences in the expression level of KCTD15 were detected in normal lymphocytes, monocytes, and granulocytes. Interestingly, we also found overexpression of the protein following leukemic transformation in the case of myeloid cell lineage. Indeed, KCTD15 was found to be upregulated in K562 and NB4 cells, as well as in HL-60 cell lines. This in vitro finding was corroborated by the analysis of KCTD15 mRNA of acute myeloid leukemia (AML) patients reported in the Microarray Innovations in Leukemia (MILE) dataset. Collectively, the present data open interesting perspectives for understanding the maturation process of leukocytes and for the diagnosis/therapy of acute leukemias.

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Purification of viable peripheral blood mononuclear cells for biobanking using a robotized liquid handling workstation

Cited by 11 publications

References 25 publications

Impact of Breast Tumor Onset on Blood Count, Carcinoembryonic Antigen, Cancer Antigen 15-3 and Lymphoid Subpopulations Supported by Automatic Classification Approach: A Pilot Study

Impact of Breast Tumor Onset on Blood Count, Carcinoembryonic Antigen, Cancer Antigen 15-3 and Lymphoid Subpopulations Supported by Automatic Classification Approach: A Pilot Study

Association of Lower Antispike Antibody Levels with Mortality in ICU Patients with COVID-19 Disease

KCTD15 Protein Expression in Peripheral Blood and Acute Myeloid Leukemia

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