PolyICLC Exerts Pro- and Anti-HIV Effects on the DC-T Cell Milieu In Vitro and In Vivo

Abstract

Myeloid dendritic cells (mDCs) contribute to both HIV pathogenesis and elicitation of antiviral immunity. Understanding how mDC responses to stimuli shape HIV infection outcomes will inform HIV prevention and treatment strategies. The long double-stranded RNA (dsRNA) viral mimic, polyinosinic polycytidylic acid (polyIC, PIC) potently stimulates DCs to focus Th1 responses, triggers direct antiviral activity in vitro, and boosts anti-HIV responses in vivo. Stabilized polyICLC (PICLC) is being developed for vaccine adjuvant applications in humans, making it critical to understand how mDC sensing of PICLC influences HIV infection. Using the monocyte-derived DC (moDC) model, we sought to describe how PICLC (vs. other dsRNAs) impacts HIV infection within DCs and DC-T cell mixtures. We extended this work to in vivo macaque rectal transmission studies by administering PICLC with or before rectal SIVmac239 (SIVwt) or SIVmac239ΔNef (SIVΔNef) challenge. Like PIC, PICLC activated DCs and T cells, increased expression of α4β7 and CD169, and induced type I IFN responses in vitro. The type of dsRNA and timing of dsRNA exposure differentially impacted in vitro DC-driven HIV infection. Rectal PICLC treatment similarly induced DC and T cell activation and pro- and anti-HIV factors locally and systemically. Importantly, this did not enhance SIV transmission in vivo. Instead, SIV acquisition was marginally reduced after a single high dose challenge. Interestingly, in the PICLC-treated, SIVΔNef-infected animals, SIVΔNef viremia was higher, in line with the importance of DC and T cell activation in SIVΔNef replication. In the right combination anti-HIV strategy, PICLC has the potential to limit HIV infection and boost HIV immunity.

Griffithsin carrageenan fast dissolving inserts prevent SHIV HSV-2 and HPV infections in vivo

“…Virus growth was monitored by p27 enzyme linked immunosorbent assay (ELISA). The harvested virus stock was characterized for p27 content by the ELISA, SIV gag RNA content by quantitative reverse transcription PCR (qRT-PCR) 53 , TCID 50 in rhesus macaque PBMCs by Reed and Muench method 54 , and focus forming units in TZM-bl cells by MAGI assay for repeated titration of the same stock over time. qRT-PCR was performed on viral RNA isolated from the stock with the Qiagen RNeasy kit and amplified by the standard curve method using the One-step RT-qPCR Kit (KAPA Biosystems, Wilmington, MA) on a ViiA-7 Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA) with the following primers: SIVgag FW (5′-GGTTGCACCCCC TATGACAT-3′), SIVgag RV (5′-TGCATAGCCGCTTGATGGT-3′).…”

T Cells Modified with CD70 as an Alternative Cellular Vaccine for Antitumor Immunity

“…Furthermore, administration of peptidebased booster containing anti-CD40 Abs after CD70-T priming was substantially effective in producing potent secondary expansion of primed CD8 T cells. Of those, good manufacturing practice (GMP)-grade poly-IC is commercially available and costimulatory agonistic antibodies including anti-CD40 have been developed, which have undergone clinical studies in some cancer patients [32][33][34][35]. More importantly, numerous human T-cell epitopes have been identified for decades, and recent progress on application of neo-antigen-derived peptide vaccines to cancer patients is rising [36][37][38] because GMP-grade peptides containing T-cell epitopes are relatively easy to synthesize cost-effectively.…”

Experimental Oral Herpes Simplex Virus-1 (HSV-1) Co-infection in Simian Immunodeficiency Virus (SIV)-Infected Rhesus Macaques

“…Blood, swabs, and tissues in L-15 were shipped cold, and snap frozen tissues were shipped on dry ice. Plasmas were isolated and stored at −80°C as previously described (Aravantinou et al, 2016 ). PBMCs, isolated as previously described (Aravantinou et al, 2016 ), were used immediately for flow cytometry or were cryopreserved in freezing medium consisting of Roswell Park Memorial Institute medium (RPMI) 1640 (Thermo Fisher) with 50% FBS and 10% DMSO (Sigma).…”

“…Plasmas were isolated and stored at −80°C as previously described (Aravantinou et al, 2016 ). PBMCs, isolated as previously described (Aravantinou et al, 2016 ), were used immediately for flow cytometry or were cryopreserved in freezing medium consisting of Roswell Park Memorial Institute medium (RPMI) 1640 (Thermo Fisher) with 50% FBS and 10% DMSO (Sigma). Splenocytes were isolated by manual dissociation and passage through a 40 μm cell strainer and then cryopreserved in freezing medium.…”

“…For plasma, RNA copies of SIV were determined by reverse transcriptase qPCR (RT-qPCR) as previously described (Cline et al, 2005 ; Aravantinou et al, 2016 ). The lower limit of quantification of the assay was 30 RNA copies/ml.…”