A novel EGFP-CEM-NKr flow cytometric method for measuring antibody dependent cell mediated-cytotoxicity (ADCC) activity in HIV-1 infected individuals

2006 ◽  
Vol 315 (1-2) ◽  
pp. 1-10 ◽  
Author(s):  
Wannee Kantakamalakul ◽  
Kovit Pattanapanyasat ◽  
Surat Jongrakthaitae ◽  
Vatcharain Assawadarachai ◽  
Silawun Ampol ◽  
...  
Keyword(s):  
Adcc Activity ◽  
Flow Cytometric Method ◽  
Flow Cytometric ◽  
Dependent Cell ◽  
Hiv 1

A flow cytometric method for measuring neutralization of HIV-1 subtype B and E primary isolates

Cytometry ◽  
2000 ◽  
Vol 40 (2) ◽  
pp. 141-150 ◽  
Author(s):  
Janice M. Darden ◽  
Victoria R. Polonis ◽  
Mark S. deSouza ◽  
Somsak Chantakulkij ◽  
Arthur E. Brown ◽  
...  
Keyword(s):  
Flow Cytometric Method ◽  
Subtype B ◽  
Flow Cytometric ◽  
Hiv 1

scholarly journals Development of antibody-dependent cell cytotoxicity function in HIV-1 antibodies

2020 ◽  
Author(s):  
Laura E. Doepker ◽  
Sonja Danon ◽  
Elias Harkins ◽  
Duncan Ralph ◽  
Zak Yaffe ◽  
...  
Keyword(s):  
Hiv Vaccine ◽  
Developmental Pathways ◽  
Antigen Binding ◽  
Antibody Dependent Cellular Cytotoxicity ◽  
Adcc Activity ◽  
Sequencing Data ◽  
Protective Antibodies ◽  
Dependent Cell ◽  
Complementarity Determining Regions ◽  
Hiv 1

AbstractA prerequisite for the design of an HIV vaccine that elicits protective antibodies is understanding the developmental pathways that result in desirable antibody features. The development of antibodies that mediate antibody-dependent cellular cytotoxicity (ADCC) is particularly relevant because such antibodies have been associated with HIV protection in humans. We reconstructed the developmental pathways of six human HIV-specific ADCC antibodies using longitudinal antibody sequencing data. Most of the inferred naïve antibodies did not mediate detectable ADCC. Gain of antigen binding and ADCC function typically required mutations in complementarity determining regions of one or both chains. Enhancement of ADCC potency often required additional mutations in framework regions. Antigen binding affinity and ADCC activity were correlated, but affinity alone was not sufficient to predict ADCC potency. Thus, elicitation of human ADCC antibodies may require mutations that first enable high affinity antigen recognition, followed by mutations that optimize factors contributing to functional ADCC activity.

scholarly journals Development of antibody-dependent cell cytotoxicity function in HIV-1 antibodies

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Laura E Doepker ◽  
Sonja Danon ◽  
Elias Harkins ◽  
Duncan K Ralph ◽  
Zak Yaffe ◽  
...  
Keyword(s):  
Hiv Vaccine ◽  
Developmental Pathways ◽  
Antigen Binding ◽  
Antibody Dependent Cellular Cytotoxicity ◽  
Adcc Activity ◽  
Sequencing Data ◽  
Protective Antibodies ◽  
Dependent Cell ◽  
Complementarity Determining Regions ◽  
Hiv 1

A prerequisite for the design of an HIV vaccine that elicits protective antibodies is understanding the developmental pathways that result in desirable antibody features. The development of antibodies that mediate antibody-dependent cellular cytotoxicity (ADCC) is particularly relevant because such antibodies have been associated with HIV protection in humans. We reconstructed the developmental pathways of six human HIV-specific ADCC antibodies using longitudinal antibody sequencing data. Most of the inferred naïve antibodies did not mediate detectable ADCC. Gain of antigen binding and ADCC function typically required mutations in complementarity determining regions of one or both chains. Enhancement of ADCC potency often required additional mutations in framework regions. Antigen binding affinity and ADCC activity were correlated, but affinity alone was not sufficient to predict ADCC potency. Thus, elicitation of broadly active ADCC antibodies may require mutations that enable high affinity antigen recognition along with mutations that optimize factors contributing to functional ADCC activity.

A sensitive flow cytometric method for measuring the oxidative burst

1997 ◽  
Vol 202 (2) ◽  
pp. 105-111 ◽  
Author(s):  
Michael A Model ◽  
Mark A KuKuruga ◽  
Robert F Todd
Keyword(s):  
Oxidative Burst ◽  
Flow Cytometric Method ◽  
Flow Cytometric

scholarly journals Flow cytometric method for the routine follow‐up of red cell populations after bone marrow transplantation

1997 ◽  
Vol 97 (1) ◽  
pp. 141-145 ◽  
Author(s):  
E. C. M. Hendriks ◽  
A. J. M. De Man ◽  
Y. C. M. Van Berkel ◽  
S. Stienstra ◽  
T. De Witte
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Transplantation ◽  
Marrow Transplantation ◽  
Cell Populations ◽  
Red Cell ◽  
Flow Cytometric Method ◽  
Flow Cytometric

scholarly journals A 5-color flow cytometric method for extended 8-part leukocyte differential

2017 ◽  
Vol 92 (6) ◽  
pp. 498-507 ◽  
Author(s):  
Julien Guy ◽  
Orianne Wagner-Ballon ◽  
Olivier Pages ◽  
François Bailly ◽  
Jessica Borgeot ◽  
...  
Keyword(s):  
Color Flow ◽  
Flow Cytometric Method ◽  
Flow Cytometric

scholarly journals Novel compound inhibitors of HIV-1NL4-3 Vpu

2021 ◽  
Author(s):  
Carolyn A Robinson ◽  
Terri D Lyddon ◽  
Hwi Min Gil ◽  
David T. Evans ◽  
Yury V Kuzmichev ◽  
...  
Keyword(s):  
Host Cell ◽  
Particle Production ◽  
Viral Glycoprotein ◽  
New Class ◽  
Host Cell Proteins ◽  
Viral Particles ◽  
Dependent Cell ◽  
A Cell ◽  
Hiv 1

HIV-1 Vpu targets the host cell proteins CD4 and BST-2/Tetherin for degradation, ultimately resulting in enhanced virus spread and host immune evasion. The discovery and characterization of small molecules that antagonize Vpu would further elucidate the contribution of Vpu to pathogenesis and lay the foundation for the study of a new class of novel HIV-1 therapeutics. To identify novel compounds that block Vpu activity, we developed a cell-based 'gain of function' assay that produces a positive signal in response to Vpu inhibition. To develop this assay, we took advantage of the viral glycoprotein, GaLV Env. In the presence of Vpu, GaLV Env is not incorporated into viral particles, resulting in non-infectious virions. Vpu inhibition restores infectious particle production. Using this assay, a high throughput screen of >650,000 compounds was performed to identify inhibitors that block the biological activity of Vpu. From this screen, we identified several positive hits but focused on two compounds from one structural family, SRI-41897 and SRI-42371. It was conceivable that the compounds inhibited the formation of infectious virions by targeting host cell proteins instead of Vpu directly, so we developed independent counter-screens for off target interactions of the compounds and found no off target interactions. Additionally, these compounds block Vpu-mediated modulation of CD4, BST-2/Tetherin and antibody dependent cell-mediated toxicity (ADCC). Unfortunately, both SRI-41897 and SRI-42371 were shown to be specific to the N-terminal region of NL4-3 Vpu and did not function against other, more clinically relevant, strains of Vpu.

scholarly journals A flow-cytometric method for continuous measurement of intracellular Ca2+ concentration

2010 ◽  
Vol 77A (11) ◽  
pp. 1091-1097 ◽  
Author(s):  
Alice Vines ◽  
Gethin J. McBean ◽  
Alfonso Blanco-Fernández
Keyword(s):  
Continuous Measurement ◽  
Flow Cytometric Method ◽  
Flow Cytometric

scholarly journals Fluorescence-Activated Cell Sorting of EGFP-Labeled Neural Crest Cells From Murine Embryonic Craniofacial Tissue

2005 ◽  
Vol 2005 (3) ◽  
pp. 232-237 ◽  
Author(s):  
Saurabh Singh ◽  
Vasker Bhattacherjee ◽  
Partha Mukhopadhyay ◽  
Christopher A. Worth ◽  
Samuel R. Wellhausen ◽  
...  
Keyword(s):  
Neural Crest ◽  
Cell Sorting ◽  
Present Report ◽  
Neural Crest Cells ◽  
Egfp Expression ◽  
Marker Genes ◽  
Flow Cytometric Method ◽  
Flow Cytometric ◽  
Polymerase Chain ◽  
Craniofacial Tissue

During the early stages of embryogenesis, pluripotent neural crest cells (NCC) are known to migrate from the neural folds to populate multiple target sites in the embryo where they differentiate into various derivatives, including cartilage, bone, connective tissue, melanocytes, glia, and neurons of the peripheral nervous system. The ability to obtain pure NCC populations is essential to enable molecular analyses of neural crest induction, migration, and/or differentiation. CrossingWnt1-CreandZ/EGtransgenic mouse lines resulted in offspring in which theWnt1-Cretransgene activated permanent EGFP expression only in NCC. The present report demonstrates a flow cytometric method to sort and isolate populations of EGFP-labeled NCC. The identity of the sorted neural crest cells was confirmed by assaying expression of known marker genes by TaqMan Quantitative Real-Time Polymerase Chain Reaction (QRT-PCR). The molecular strategy described in this report provides a means to extract intact RNA from a pure population of NCC thus enabling analysis of gene expression in a defined population of embryonic precursor cells critical to development.

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