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A new cell line for assessing HIV-1 antibody dependent cellular cytotoxicity against a broad range of variants
- Source :
- J Immunol Methods
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Human immunodeficiency virus type 1 (HIV-1) studies suggest that antibody-dependent cellular cytotoxicity (ADCC) influences both virus acquisition and subsequent disease outcome. Technical issues with currently available assays, however, have limited the ability to comprehensively assess the impact of ADCC on transmission and disease progression. Commonly used ADCC assays use a target cell line, CEM.NKr-CCR5-Luc, that often does not support replication of relevant HIV-1 variants. Thus, the extent of ADCC responses against a large panel of HIV-1 strains often cannot be assessed using the currently available methods. We developed two new reporter cell-lines (MT4-CCR5-Luc and PM1-CCR5-Luc) to overcome these issues. MT4-CCR5-Luc cells are resistant, whereas PM1-CCR5-Luc cells are susceptible, to killing by a natural killer cell line, CD16(+)KHYG-1, in the absence of antibody. Polyclonal HIVIG gave similar ADCC estimates against HIV-1 isolate, NL4–3, regardless of which of the three cell lines were used as the targets. In contrast to CEM.NKr-CCR5-Luc and PM1-CCR5-Luc, however, MT4-CCR5-Luc target cells produce significantly higher luciferase after exposure to various HIV-1 strains, including transmitted founder variants and viruses incorporating specific envelopes of interest. This higher luciferase expression does not yield spurious results because ADCC estimates are similar when killing is assessed by both reporter protein expression and flow cytometry. Furthermore, ADCC estimates derived from MT4-CCR5-Luc cells are not skewed by non-antibody contents present in human plasma. In aggregate, the MT4-CCR5-Luc cell line can be used to estimate monoclonal antibody or plasma-induced ADCC responses against a diverse range of HIV-1 envelopes relevant for transmission and disease progression studies.
- Subjects :
- 0301 basic medicine
HIV Antigens
medicine.drug_class
viruses
Immunology
HIV Infections
HIV Antibodies
CD16
Biology
Monoclonal antibody
Article
Virus
Cell Line
Natural killer cell
Flow cytometry
03 medical and health sciences
0302 clinical medicine
Genes, Reporter
medicine
Humans
Immunology and Allergy
Lymphocytes
Luciferases
Antibody-dependent cell-mediated cytotoxicity
medicine.diagnostic_test
Antibody-Dependent Cell Cytotoxicity
env Gene Products, Human Immunodeficiency Virus
virus diseases
Virology
Coculture Techniques
Killer Cells, Natural
030104 developmental biology
medicine.anatomical_structure
Cell culture
Host-Pathogen Interactions
HIV-1
biology.protein
Antibody
030215 immunology
Subjects
Details
- ISSN :
- 00221759
- Volume :
- 480
- Database :
- OpenAIRE
- Journal :
- Journal of Immunological Methods
- Accession number :
- edsair.doi.dedup.....0a081858c4913726a8acd2d46d6a22e3