Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based Assay
The accurate assessment of immune competence through ex vivo analysis is paramount to our understanding of those immune mechanisms that lead to protection or susceptibility against a broad range of human pathogens. We have developed a flow cytometry-based, whole blood phagocyte functional assay that...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fimmu.2017.01222/full |
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| author | Ankur Gupta-Wright Ankur Gupta-Wright Dumizulu Tembo Kondwani C. Jambo Kondwani C. Jambo Elizabeth Chimbayo Leonard Mvaya Shannon Caldwell David G. Russell Henry C. Mwandumba Henry C. Mwandumba |
| author_facet | Ankur Gupta-Wright Ankur Gupta-Wright Dumizulu Tembo Kondwani C. Jambo Kondwani C. Jambo Elizabeth Chimbayo Leonard Mvaya Shannon Caldwell David G. Russell Henry C. Mwandumba Henry C. Mwandumba |
| author_sort | Ankur Gupta-Wright |
| collection | DOAJ |
| description | The accurate assessment of immune competence through ex vivo analysis is paramount to our understanding of those immune mechanisms that lead to protection or susceptibility against a broad range of human pathogens. We have developed a flow cytometry-based, whole blood phagocyte functional assay that utilizes the inflammatory inducer zymosan, coupled to OxyBURST-SE, a fluorescent reporter of phagosomal oxidase activity. The assay measures both phagocytic uptake and the superoxide burst in the phagocyte populations in whole blood. We utilized this assay to demonstrate impaired superoxide burst activity in the phagocytes of hospitalized HIV-positive patients with laboratory-confirmed tuberculosis. These data validate the use of the assay to assess the immune competence of patients in a clinical setting. The method is highly reproducible with minimal intraindividual variation and opens opportunities for the rapid assessment of cellular immune competence in peripheral blood in a disease setting. |
| first_indexed | 2024-12-11T09:34:45Z |
| format | Article |
| id | doaj.art-f326c7e323a342daa73374371642c844 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-11T09:34:45Z |
| publishDate | 2017-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-f326c7e323a342daa73374371642c8442022-12-22T01:12:56ZengFrontiers Media S.A.Frontiers in Immunology1664-32242017-09-01810.3389/fimmu.2017.01222302441Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based AssayAnkur Gupta-Wright0Ankur Gupta-Wright1Dumizulu Tembo2Kondwani C. Jambo3Kondwani C. Jambo4Elizabeth Chimbayo5Leonard Mvaya6Shannon Caldwell7David G. Russell8Henry C. Mwandumba9Henry C. Mwandumba10College of Medicine, Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, MalawiClinical Research Department, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, United StatesCollege of Medicine, Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, MalawiDepartment of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, United KingdomCollege of Medicine, Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, MalawiCollege of Medicine, Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, MalawiDepartment of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, United StatesDepartment of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, United StatesCollege of Medicine, Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, MalawiDepartment of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, United KingdomThe accurate assessment of immune competence through ex vivo analysis is paramount to our understanding of those immune mechanisms that lead to protection or susceptibility against a broad range of human pathogens. We have developed a flow cytometry-based, whole blood phagocyte functional assay that utilizes the inflammatory inducer zymosan, coupled to OxyBURST-SE, a fluorescent reporter of phagosomal oxidase activity. The assay measures both phagocytic uptake and the superoxide burst in the phagocyte populations in whole blood. We utilized this assay to demonstrate impaired superoxide burst activity in the phagocytes of hospitalized HIV-positive patients with laboratory-confirmed tuberculosis. These data validate the use of the assay to assess the immune competence of patients in a clinical setting. The method is highly reproducible with minimal intraindividual variation and opens opportunities for the rapid assessment of cellular immune competence in peripheral blood in a disease setting.http://journal.frontiersin.org/article/10.3389/fimmu.2017.01222/fullphagocytosiszymosaninflammationmonocytesneutrophilsHIV |
| spellingShingle | Ankur Gupta-Wright Ankur Gupta-Wright Dumizulu Tembo Kondwani C. Jambo Kondwani C. Jambo Elizabeth Chimbayo Leonard Mvaya Shannon Caldwell David G. Russell Henry C. Mwandumba Henry C. Mwandumba Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based Assay Frontiers in Immunology phagocytosis zymosan inflammation monocytes neutrophils HIV |
| title | Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based Assay |
| title_full | Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based Assay |
| title_fullStr | Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based Assay |
| title_full_unstemmed | Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based Assay |
| title_short | Functional Analysis of Phagocyte Activity in Whole Blood from HIV/Tuberculosis-Infected Individuals Using a Novel Flow Cytometry-Based Assay |
| title_sort | functional analysis of phagocyte activity in whole blood from hiv tuberculosis infected individuals using a novel flow cytometry based assay |
| topic | phagocytosis zymosan inflammation monocytes neutrophils HIV |
| url | http://journal.frontiersin.org/article/10.3389/fimmu.2017.01222/full |
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