Sensing of HIV-1 Infection in Tzm-bl Cells with Reconstituted Expression of STING

Tumor necrosis factor (TNF) is a key cytokine in HIV replication and pathogenesis. For reasons that are not entirely clear, the cytokine remains upregulated despite anti-retroviral therapy (ART). Here we demonstrate that HIV Nef induces an alternative TNF secretion mechanism that remains active in chronic infection. Ingestion of Nef-containing plasma extracellular vesicles (pEV) from ART patients by primary immune cells, but also Nef expression, induced intracellular proTNF cleavage and secretion of vesicular TNF endosomes. Key event was the Nef-mediated routing of the TNF-converting enzyme ADAM17 into Rab4 + early endosomes and the Rab27 + secretory pathway. Analysis of lymph-node tissue by multi-epitope-ligand-cartography (MELC) confirmed a vesicular TNF secretion phenotype that co-localized with persistent Nef expression, and implicated Notch1 as an essential co-factor. Surprisingly Notch1 had no transcriptional effect but was required for the endosomal trafficking of ADAM17. We conclude that Nef expression and Nef-containing pEV mobilize TNF from endosomal compartments in acute and chronic infection.

show abstract

“…or HIV reverse transcription activity by the SG-PERT assay (over 16 days p.i.) (Trotard et al, 2016). …”

Section: Methodsmentioning

confidence: 99%

HIV Nef- and Notch1-dependent Endocytosis of ADAM17 Induces Vesicular TNF Secretion in Chronic HIV Infection

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, in mice, enhanced activation of STING suppresses the magnitude of Lm-induced CD8 þ T-cell priming specific for encoded antigens and diminishes protection against lethal wildtype (WT) Lm challenge, and conversely, mice lacking functional STING are more resistant to infection with intracellular bacteria, including Lm (18). The cGAS-STING signaling axis is also activated by dsDNA viruses, including pox viruses, adenoviruses, gamma herpes viruses such as HSV, and retroviruses such as HIV and HTLV (5,15,(19)(20)(21)(22)(23)(24). In response to infection with dsDNA viruses, induced cGAMP can be transported to neighboring cells through gap junctions and inhibit virus spread, supporting the central role of this pathway as a major antimicrobial defense mechanism (25,26).…”

Section: Introductionmentioning

confidence: 99%

TNFα and Radioresistant Stromal Cells Are Essential for Therapeutic Efficacy of Cyclic Dinucleotide STING Agonists in Nonimmunogenic Tumors

Francica

Ghasemzadeh

Desbien

et al. 2018

Cancer Immunology Research

View full text Add to dashboard Cite

The cGAS-STING cytosolic DNA sensing pathway may play an integral role in the initiation of antitumor immune responses. Studies evaluating the immunogenicity of various cyclic dinucleotide (CDN) STING agonists administered by intratumoral (i.t.) injection showed potent induction of inflammation, tumor necrosis, and, in some cases, durable tumor-specific adaptive immunity. However, the specific immune mechanisms underlying these responses remain incompletely defined. The majority of these studies have focused on the effect of CDNs on immune cells but have not conclusively interrogated the role of stromal cells in the acute rejection of the CDN-injected tumor. Here, we revealed a mechanism of STING agonist-mediated tumor response that relied on both stromal and immune cells to achieve tumor regression and clearance. Using knockout and bone marrow chimeric mice, we showed that although bone marrow–derived TNFα was necessary for CDN-induced necrosis, STING signaling in radioresistant stromal cells was also essential for CDN-mediated tumor rejection. These results provide evidence for crosstalk between stromal and hematopoietic cells during CDN-mediated tumor collapse after i.t. administration. These mechanistic insights may prove critical in the clinical development of STING agonists.

show abstract

“…This transmittance percentage difference observed could be hypothetically attributed to the difference in index of refraction of the two cells (infected and uninfected); hence in their scattering coefficient as other parameters that play a major role in transmission measurement are constants. Several lines of evidence support that HIV‐1 infection can trigger a wide range of reactions in the host cell including TZM‐bl cells . These reactions involve the processing of viral protein into peptides molecules that change the biochemical composition of the TZM‐bl cells.…”

Section: Resultsmentioning

confidence: 99%

Label‐free differentiation of human immunodeficiency virus‐1 infected from uninfected cells using transmission measurement

Ombinda‐Lemboumba

Malabi

Lugongolo

et al. 2019

Journal of Biophotonics

View full text Add to dashboard Cite

Transmission measurement has been perceived as a potential candidate for label‐free investigation of biological material. It is a real‐time, label‐free and non‐invasive optical detection technique that has found wide applications in pharmaceutical industry as well as the biological and medical fields. Combining transmission measurement with optical trapping has emerged as a powerful tool allowing stable sample trapping, while also facilitating transmittance data analysis. In this study, a near‐infrared laser beam emitting at a wavelength of 1064 nm was used for both optical trapping and transmission measurement investigation of human immunodeficiency virus 1 (HIV‐1) infected and uninfected TZM‐bl cells. The measurements of the transmittance intensity of individual cells in solution were carried out using a home built optical trapping system combined with laser transmission setup using a single beam gradient trap. Transmittance spectral intensity patterns revealed significant differences between the HIV‐1 infected and uninfected cells. This result suggests that the transmittance data analysis technique used in this study has the potential to differentiate between infected and uninfected TZM‐bl cells without the use of labels. The results obtained in this study could pave a way into developing an HIV‐1 label‐free diagnostic tool with possible applications at the point of care .

show abstract

Sensing of HIV-1 Infection in Tzm-bl Cells with Reconstituted Expression of STING

Cited by 32 publications

References 81 publications

HIV Nef- and Notch1-dependent Endocytosis of ADAM17 Induces Vesicular TNF Secretion in Chronic HIV Infection

HIV Nef- and Notch1-dependent Endocytosis of ADAM17 Induces Vesicular TNF Secretion in Chronic HIV Infection

TNFα and Radioresistant Stromal Cells Are Essential for Therapeutic Efficacy of Cyclic Dinucleotide STING Agonists in Nonimmunogenic Tumors

Label‐free differentiation of human immunodeficiency virus‐1 infected from uninfected cells using transmission measurement

Contact Info

Product

Resources

About