bioRxiv Science⌕ Search

Biology subjects

Singer, B. B.

Publications and source records attributed to Singer, B. B..

2 recordsLinked to original sources

LILRA5 functions to induce ROS production on innate immune cells

Activating immune receptors provide mechanisms for phagocytes to elicit important effector functions that promote the killing of microbes. Leukocyte immunoglobulin-like receptor A5 (LILRA5) an orphan immune receptor expressed by human phagocytes and co-localising with FcR{gamma}, remains poorly characterised. To address this, we developed a highly-specific anti-LILRA5 monoclonal antibody that has agonistic properties. Using a specific anti-LILRA5 antibody, we show LILRA5 expression on naive monocytes and neutrophils, and that ligation of LILRA5 stimulates ROS production. While increased LILRA5 transcript copy numbers have been associated with sepsis, we also observed increased levels in patients with systemic infection but without sepsis complications. Ex vivo bacterial infection of whole blood did not alter surface LILRA5 expression, but LPS stimulation changed expression levels. This indicates that surface LILRA5 expression is dynamic and likely regulated posttranscriptionally, changing responses to different stimuli or over time. Soluble (s)LILRA5 was enhanced in sera from sepsis patients and in supernatants of monocytes that were LPS-stimulated, indicative that shedding of LILRA5 from cell surfaces or that expression of sLILRA5 isoforms provides a mechanism to regulate surface LILRA5 expression levels. Finally, we show that altered surface LILRA5 expression influences LILRA5-induced ROS production capacity. We propose that LILRA5 is a dynamically regulated activating receptor expressed on phagocytes that can stimulate ROS production.

immunology↗

Regulation of Human Neutrophil Responses to Candida albicans by Carcinoembryonic Antigen-Related Cell Adhesion Molecule 1 (CEACAM1), CEACAM3 and CEACAM6

Invasive candidiasis, often caused by Candida albicans, is an important healthcare-associated fungal infection that results in a high mortality rate of up to 40%. Neutrophils are the first line of defense during Candida infections. They can initiate various killing mechanisms and release cytokines to attract further immune cells to the site of infection. These responses are tightly controlled, since they can also lead to severe tissue/organ damage. We hypothesized that the regulation of C. albicans-specific neutrophil functions by the immunoregulatory C. albicans receptors CEACAM1, CEACAM3, and CEACAM6 are involved in the immune pathology of candidemia. Here, we analyzed the effects of the specific antibodies B3-17, 308/3-3, and 1H7-4B, respectively, targeting the three CEACAM receptors on C. albicans-induced neutrophil responses. We show that ligation of CEACAM6 significantly enhanced the response to C. albicans, as evidenced by the increased CXCL8/IL-8 secretion. By assessing the transcriptional responses, we found that CEACAM6 ligation and to some extent CEACAM1 ligation, but not CEACAM3 ligation, resulted in altered gene regulation of the C. albicans-stimulated neutrophils. Genes that were differentially regulated by the different CEACAM-targeting antibodies were analyzed for affected cellular processes and signaling pathways using various bioinformatics methods, including integrated network analyses and dynamic simulations of signaling cascades. Predicted changes in cellular pathways and cellular functions included CEACAM-specific alterations in apoptosis and cytokine secretion. In particular, we verified predicted changes in IL-1{beta}/IL-6 expression in response to the antibody ligation of all three targeted CEACAMs and apoptosis induction by anti-CEACAM6 antibody treatment in presence of C. albicans stimulation. Specifically, CEACAM6 ligation by 1H7-4B enhanced neutrophil apoptosis and increased immediate and long-term cytokine release in responses to C. albicans. CEACAM1 ligation by B3-17 mainly enhanced the immediate secretion of pro-inflammatory cytokines, and CEACAM3 ligation by 308/3-3 increased the long-term release of pro-inflammatory cytokines. Taken together, we demonstrated for the first time that CEACAM receptors have an important and differential impact on the regulation of C. albicans-induced immune functions in human neutrophils.

immunology↗