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. 2017 Feb 24;46(2):301–314. doi: 10.1016/j.immuni.2017.01.010

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© 2017 Elsevier Inc.

Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

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HRSV Infects nBreg Cells via CX3CR1-G Protein Interaction

(A–C) FACS analysis of CX3CR1 expression in nBreg cells.

(A) Representative FACS plot of freshly isolated nBreg cells (MFI: 193 for Ab ctrl and 237 for CX3CR1 Ab) as compared to CD14⁺ monocytes (MFI: 9971).

(B) nBreg cells were infected with HRSV-A (MOI = 2.5) for 48 hr. MFI are shown for CX3CR1 Ab as compared to isotype control. Results are expressed as means ± SD of duplicates from three donors.

(C) nBreg cells were stimulated with R848, agM, or HRSV-A (MOI = 2.5) for 48 hr and their CX3CR1 expression was assessed by FACS. Results are expressed as means ± SD of duplicates from two independent experiments. nBreg cells were infected for 48 hr with WT or ΔG strains of rHRSV-Ch (MOI = 2.5).

(D) IL-10 production was measured by ELISA. Results are expressed as means ± SD of four independent experiments.

(E) Phopshorylation of CD79a was assessed intracellularly by FACS in nBreg cells after 30 min of exposure to ΔG RSV or WT RSV. Results are expressed as means ± SD of triplicates and are representative of three experiments.

(F) Representative FACS plot (left) of nBreg cell infection with ΔG RSV or WT RSV, as compared to non-infected cells (none). Histogram plot shows the percentage of infected nBreg cells, as measured by FACS as percent of mCherry⁺ cells (right), and is representative of three independent experiments.

(G) Viral replication measured through the detection of the mCherry fluorescence for 48 hr with ΔG as compared to WT counterpart. Results are expressed as means ± SD of triplicates and are representative of three independent experiments.

(H) nBreg cells were cultured on CX3CL1-coated plates and infected with rHRSV-Ch (MOI = 2.5). The percentage of infected nBreg cells and IL-10 secretion were measured at 48 hr. Results are expressed as means ± SD and represent three independent experiments. Unpaired t test was used for comparison. Results are expressed as the means ± SD.

^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001.