Flow Cytometry and Surface-Marker Phenotyping Using Monoclonal Antibodies: A Combined Approach to Precisely Define the State of the Immune System

2015 ◽  
pp. 120-146 ◽  
Author(s):  
Martin R. Hadam
Keyword(s):  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Immune System ◽  
The State ◽  
Surface Marker ◽  
Combined Approach

scholarly journals The use of poly- and monoclonal antibodies to assess the state of the immune system

2020 ◽  
Vol 421 ◽  
pp. 052002
Author(s):  
I Yu Ezdakova ◽  
O V Kapustina ◽  
E V Popova ◽  
A G Grigoriev ◽  
V M Kovaikina
Keyword(s):  
Monoclonal Antibodies ◽  
Immune System ◽  
The State

scholarly journals The expression and modulation of human myeloid-specific antigens during differentiation of the HL-60 cell line

Blood ◽  
1983 ◽  
Vol 61 (6) ◽  
pp. 1215-1221
Author(s):  
RF Graziano ◽  
ED Ball ◽  
MW Fanger
Keyword(s):  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Retinoic Acid ◽  
Cell Surface ◽  
Cell Line ◽  
Surface Marker ◽  
Myeloid Differentiation ◽  
Dimethyl Formamide ◽  
Specific Antigens ◽  
Chemical Mediators

Antigenic changes detected by myeloid-specific monoclonal antibodies on HL-60 cells induced to differentiate by various chemical mediators were investigated using flow cytometry. Antigen levels detected by monocyte- granulocyte-specific monoclonal antibodies AML-2–23, 61D3, and 63D3 increased dramatically after differentiation of HL-60 cells along the granulocytic pathway by the addition of dimethyl formamide (DMF), dimethylsulfoxide (DMSO), or cis-retinoic acid. The expression of these same antigens also increased in conjunction with monocytoid differentiation when HL-60 cells were treated with supernatants from leukocytes stimulated with phytohemagglutinin (PHA-LCM) or with mixed lymphocyte conditioned medium (MLC). In contrast, treatment of HL-60 cells with phorbol 12-myristate 13-acetate (PMA), which also induced differentiation along the monocyte pathway, had no effect on the expression of these monocyte-associated antigens. The expression of antigens on HL-60 cells recognized by the granulocyte-specified monoclonal antibodies PMN 6 and PMN 29 decreased after treatment of HL- 60 cells with PMA, but remained constant after treatment with DMF, DMSO, cis-retinoic acid, PHA-LCM, or MLC. These results suggest that normal myeloid differentiation may be dependent on various signals and that morphological and cell surface marker maturity may, under some conditions, be separable. The utility of the HL-60 cell line as a model of myeloid differentiation and for evaluation of inductive signals is discussed.

Leukocyte counts and lymphocyte responsiveness associated with repeated bouts of strenuous endurance exercise

2001 ◽  
Vol 91 (1) ◽  
pp. 425-434 ◽  
Author(s):  
Ola Ronsen ◽  
Bente Klarlund Pedersen ◽  
Tone Rasmussen Øritsland ◽  
Roald Bahr ◽  
Jens Kjeldsen-Kragh
Keyword(s):  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Immune System ◽  
Endurance Exercise ◽  
Bed Rest ◽  
Cycle Ergometer ◽  
Single Bout ◽  
Leukocyte Counts ◽  
Carry Over ◽  
Carry Over Effect

This study compared leukocyte counts and lymphocyte responsiveness during and after a second bout of high-intensity endurance exercise on the same day with the response to a similar but single bout of exercise. Nine athletes participated in three 24-h trials: 1) rest in bed (Rest); 2) one bout of exercise (One); and 3) two bouts of exercise (Two). All bouts consisted of 75 min at ∼75% of maximal O2uptake on a cycle ergometer. Lymphocytes in whole blood were stimulated with monoclonal antibodies against CD2 and assessed by flow cytometry for expression of the early activation molecule CD69. The second bout of exercise in the Two trial was associated with significantly increased concentrations of total leukocytes, neutrophils, lymphocytes, CD4+, CD8+, and CD56+cells and a significantly decreased percentage of CD56+cells expressing CD69 compared with a single bout. Additionally, there was a significantly decreased CD69 fluorescence in CD56+cells postexercise. These differences suggest a “carry-over” effect in the immune system from a first to a second bout of exercise on the same day.

scholarly journals The expression and modulation of human myeloid-specific antigens during differentiation of the HL-60 cell line

Blood ◽  
1983 ◽  
Vol 61 (6) ◽  
pp. 1215-1221 ◽  
Author(s):  
RF Graziano ◽  
ED Ball ◽  
MW Fanger
Keyword(s):  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Retinoic Acid ◽  
Cell Surface ◽  
Cell Line ◽  
Surface Marker ◽  
Myeloid Differentiation ◽  
Dimethyl Formamide ◽  
Specific Antigens ◽  
Chemical Mediators

Abstract Antigenic changes detected by myeloid-specific monoclonal antibodies on HL-60 cells induced to differentiate by various chemical mediators were investigated using flow cytometry. Antigen levels detected by monocyte- granulocyte-specific monoclonal antibodies AML-2–23, 61D3, and 63D3 increased dramatically after differentiation of HL-60 cells along the granulocytic pathway by the addition of dimethyl formamide (DMF), dimethylsulfoxide (DMSO), or cis-retinoic acid. The expression of these same antigens also increased in conjunction with monocytoid differentiation when HL-60 cells were treated with supernatants from leukocytes stimulated with phytohemagglutinin (PHA-LCM) or with mixed lymphocyte conditioned medium (MLC). In contrast, treatment of HL-60 cells with phorbol 12-myristate 13-acetate (PMA), which also induced differentiation along the monocyte pathway, had no effect on the expression of these monocyte-associated antigens. The expression of antigens on HL-60 cells recognized by the granulocyte-specified monoclonal antibodies PMN 6 and PMN 29 decreased after treatment of HL- 60 cells with PMA, but remained constant after treatment with DMF, DMSO, cis-retinoic acid, PHA-LCM, or MLC. These results suggest that normal myeloid differentiation may be dependent on various signals and that morphological and cell surface marker maturity may, under some conditions, be separable. The utility of the HL-60 cell line as a model of myeloid differentiation and for evaluation of inductive signals is discussed.

Internalization of an Anti-Glycoprotein IIb/IIIa Antibody by HEL Cells

1995 ◽  
Vol 73 (02) ◽  
pp. 291-296 ◽  
Author(s):  
Kenjiro Hamamoto ◽  
Shosaku Nomura ◽  
Masahiko Suzuki ◽  
Shigetoshi Ohga ◽  
Shirou Fukuhara
Keyword(s):  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Immunoelectron Microscopy ◽  
Surface Membrane ◽  
Intracellular Pool ◽  
Adhesion Proteins ◽  
Hel Cells

SummaryPlatelets are known to internalize monoclonal antibodies directed against the glycoprotein (GP) IIb/IIIa complex. We investigated whether an antibody directed against this complex (NNKY 2-11) was transported from the surface membrane to the intracellular pool in HEL cells. Flow cytometry showed that the percent binding of NNKY 2-11 to the surface membrane of HEL cells was decreased after incubation for 24 h compared with 1 h, while the binding of an anti-GPIb antibody (NNKY 5-5) did not change. It did not seem likely that the GP Ilb/IIIa complex antibody was shed from the surface membrane of the HEL cells during incubation, because the medium conditioned by incubation with these cells for 24 h showed almost no binding to washed platelets. In addition, immunoelectron microscopy demonstrated that GP IIb/IIIa complex antibodies were incorporated into the intracellular pool of HEL cells and were associated with alpha granules. These findings indicated that an anti-GP IIb/IIIa antibody could be internalized by megakaryocytes, as has been previously shown with platelets, suggesting that megakaryocyte GP IIb/IIIa may act as a carrier for various adhesion proteins.

scholarly journals The Role of Monoclonal Antibodies in Smoldering and Newly Diagnosed Transplant-Eligible Multiple Myeloma

2020 ◽  
Vol 13 (12) ◽  
pp. 451
Author(s):  
Elena Zamagni ◽  
Paola Tacchetti ◽  
Paola Deias ◽  
Francesca Patriarca
Keyword(s):  
Multiple Myeloma ◽  
Monoclonal Antibodies ◽  
Immune System ◽  
Survival Outcomes ◽  
Newly Diagnosed ◽  
Cellular Targets ◽  
Recent Introduction

The recent introduction of monoclonal antibodies (MoAbs), with several cellular targets, such as CD-38 (daratumumab and isatuximab) and SLAM F7 (elotuzumab), differently combined with other classes of agents, has significantly extended the outcomes of patients with multiple myeloma (MM) in different phases of the disease. Initially used in advanced/refractory patients, different MoAbs combination have been introduced in the treatment of newly diagnosed transplant eligible patients (NDTEMM), showing a significant improvement in the depth of the response and in survival outcomes, without a significant price in terms of toxicity. In smoldering MM, MoAbs have been applied, either alone or in combination with other drugs, with the goal of delaying the progression to active MM and restoring the immune system. In this review, we will focus on the main results achieved so far and on the main on-going trials using MoAbs in SMM and NDTEMM.

scholarly journals Contribution of Flow Cytometry to Acute Leukemia Classification in Tunisia

2000 ◽  
Vol 16 (3-4) ◽  
pp. 131-133 ◽  
Author(s):  
S. Feki ◽  
H. El Omri ◽  
M. A. Laatiri ◽  
S. Ennabli ◽  
K. Boukef ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Age Distribution ◽  
Tunisian Population ◽  
Acute Leukemias ◽  
Flow Cytometric Method ◽  
Technical Innovations ◽  
Definition Of ◽  
Sahel Region ◽  
Cluster Of Differentiation

The precision of immunological characterization of leukemias was improved by a certain number of technical innovations, particularly hybridoma production and standardization, resulting in monoclonal antibodies and definition of recognised cellular antigens (designated by CD: Cluster of Differentiation).The aim of this work was to determine the immunophenotyping profile of patients with leukemia, by means of a flow cytometric method: 66 blood samples coming from leukemic persons in the Sahel region were studied by flow cytometry, using about thirty monoclonal antibodies all marked with a fluorochrome, in one or two colour systems to assess their distribution according to type (lymphoid B or T / myeloid) and age, and to search for possible co-expressions of markers of different lineages.The marked preponderance of childhood B-ALL in our series is, at least partly, attributable to the age distribution of the Tunisian population. In agreement with studies from other countries, the majority of AML cases occurred among adults. A high proportion of AML cases in our series co-expressed markers of other lineages. Overall, accurate classification of acute leukemias was possible from a simple peripheral blood sample in 62 of 66 cases (93.9%).

Surface chemistry modification of silica nanoparticles alters the activation of monocytes

2021 ◽  
Author(s):  
Romina Mitarotonda ◽  
Martín Saraceno ◽  
Marcos Todone ◽  
Exequiel Giorgi ◽  
Emilio L Malchiodi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune System ◽  
Cell Membrane ◽  
Cell Activation ◽  
Membrane Damage ◽  
Living Cells ◽  
Cell Membrane Damage ◽  
Study Materials ◽  
Therapeutic Molecules ◽  
Pro Inflammatory Cytokines

Aim: Nanoparticles (NPs) interaction with immune system is a growing topic of study. Materials & methods: Bare and amine grafted silica NPs effects on monocytes/macrophages cells were analyzed by flow cytometry, MTT test and LIVE/DEAD® viability/cytotoxicity assay. Results: Bare silica NPs inhibited proliferation and induced monocyte/macrophages activation (increasing CD40/CD80 expression besides pro-inflammatory cytokines and nitrite secretion). Furthermore, silica NPs increased cell membrane damage and reduced the number of living cells. In contrast, amine grafted silica NPs did not alter these parameters. Conclusion: Cell activation properties of bare silica NPs could be hindered after grafting with amine moieties. This strategy is useful to tune the immune system stimulation by NPs or to design NPs suitable to transport therapeutic molecules.

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