RESUMO
BACKGROUND/AIMS: Immune cells are reported to upregulate CD47 during infection, however, the role of CD47 in innate and adaptive immune cells remains unclear. METHODS: To bridge this knowledge gap, we analysed our single cell (sc)-RNA dataset along with other publicly available sc-RNA datasets from healthy controls, people with HIV-1 (PWH) and COVID-19 patients. We characterized each immune cell based on low, intermediate, and high expression of CD47 . RESULTS: Our analyses revealed that CD47 high pDCs and monocytes exhibited relatively higher expression of IFN-α regulatory genes, antiviral interferon-stimulated genes (ISGs) and MHC-I associated genes compared to CD47 inter. and CD47 low cells. Furthermore, CD47 high NK and CD8+ T cells showed higher expression of antiviral ISGs, as well as genes encoding for cytotoxic markers like granzyme B, perforin, granulysin, interferon gamma and NKG7. Additionally, CD47 high CD8+ T cells expressed higher levels of PD-1 and LAG-3 genes. Lastly, we found that CD47 high B cells had enriched expression of genes involved in cell activation and humoral responses. CONCLUSION: Overall, our analyses revealed that innate and adaptive immune cells expressing elevated activation and functional gene signatures also express higher CD47 levels.
Assuntos
Antígeno CD47 , Linfócitos T CD8-Positivos , Granzimas , HIV-1 , Células Matadoras Naturais , Perforina , Receptor de Morte Celular Programada 1 , RNA Mensageiro , Análise de Célula Única , Humanos , Antígeno CD47/metabolismo , Antígeno CD47/genética , Linfócitos T CD8-Positivos/metabolismo , Linfócitos T CD8-Positivos/imunologia , Granzimas/metabolismo , Granzimas/genética , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Perforina/metabolismo , Perforina/genética , HIV-1/imunologia , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , Receptor de Morte Celular Programada 1/metabolismo , Receptor de Morte Celular Programada 1/genética , COVID-19/imunologia , COVID-19/virologia , COVID-19/genética , Infecções por HIV/imunologia , Infecções por HIV/virologia , Infecções por HIV/genética , Proteína do Gene 3 de Ativação de Linfócitos , Antígenos de Diferenciação de Linfócitos T/metabolismo , Antígenos de Diferenciação de Linfócitos T/genética , SARS-CoV-2/imunologia , Interferon gama/metabolismo , Interferon gama/genética , Monócitos/metabolismo , Monócitos/imunologia , Antígenos CD/metabolismo , Antígenos CD/genética , Linfócitos B/metabolismo , Linfócitos B/imunologia , Imunidade InataRESUMO
Many immunological treatment strategies for reducing the HIV-1 reservoir and enhancing adaptive immunity aim at activating the human plasmacytoid dendritic cells (pDCs). Here, we present a protocol for pDC enrichment, single-cell analysis, and development of a pDC transcriptomic database from healthy individuals and people with HIV-1 before and after Toll-like receptor 9 agonist treatment. For complete details on the use and execution of this protocol, please refer to Cham et al.1.
Assuntos
HIV-1 , Humanos , HIV-1/genética , Interferon-alfa , Imunidade Adaptativa , Perfilação da Expressão Gênica , Células DendríticasRESUMO
There are two known mechanisms by which natural killer (NK) cells recognize and kill diseased targets: (i) direct killing and (ii) antibody-dependent cell-mediated cytotoxicity (ADCC). We investigated an indirect NK cell activation strategy for the enhancement of human NK cell killing function. We did this by leveraging the fact that toll-like receptor 9 (TLR9) agonism within pools of human peripheral blood mononuclear cells (PBMCs) results in a robust interferon signaling cascade that leads to NK cell activation. After TLR9 agonist stimulation, NK cells were enriched and incorporated into assays to assess their ability to kill tumor cell line targets. Notably, differential impacts of TLR9 agonism were observed-direct killing was enhanced while ADCC was not increased. To ensure that the observed differential effects were not attributable to differences between human donors, we recapitulated the observation using our Natural Killer-Simultaneous ADCC and Direct Killing Assay (NK-SADKA) that controls for human-to-human differences. Next, we observed a treatment-induced decrease in NK cell surface CD16-known to be shed by NK cells post-activation. Given the essential role of CD16 in ADCC, such shedding could account for the observed differential impact of TLR9 agonism on NK cell-mediated killing capacity.