Fig 2: Modulation of early and integrated HIV-1 DNA production by SLPI (A) or SERPIN C1 (B); MDMs (5 × 105 cells/well) were pre-treated with (A) SLPI 1 µg/mL and 10 µg/mL or (B) SERPIN C1 0.05 µg/mL and 0.1 µg/mL for 3 h then infected with HIV-1 BaL 5 ng/mL p24 units. After 2 h, the virus was removed, and fresh culture media added with 1 µg/mL and 10 µg/mL SLPI or 0.05 µg/mL and 0.1 µg/mL SERPIN C1 and cultured. MDMs were collected 16 h after infection, and total DNA was extracted with the DNeasy tissue kit. Real-time PCR was performed with equal amounts of DNA and measured the amounts of early and late reverse transcription DNA products, as well as 2-LTR circle DNA. Levels of integrated viral DNA were measured by Alu-PCR. Effect of SLPI and SERPIN C1 on HIV-1 DNA production was determined by calculating the ratios of viral DNA amounts produced without SLPI or SERPIN C1 versus treatment with SLPI or SERPIN C1 in MDMs. Results expressed as mean ± SEM from three independent donors. Asterisk (*) over the bars indicates significant difference with control, ** p < 0.001; * p ≤ 0.05 and ns p > 0.05. p-values were generated by one-way ANOVA with multiple comparisons.

Fig 3: Chronic obstructive pulmonary disease (COPD) small airway epithelial cells (SAECs) exert a delayed antimicrobial peptide (AMP) response to non-typeable Haemophilus influenzae (NTHi). mRNA expression of AMPs (DEFB1, DEFB4A (gene name for β-defensin 2 protein), CAMP, SLPI, S100A7, S100A8, S100A9, PI3) in non-COPD-derived (nC, grey) and COPD-derived (C, red) SAECs exposed to NTHi for 3 h or 24 h relative to PBS-exposed SAECs. Gene expression comparative crossing point change (ΔΔCp) is presented as a mean (n=11–18). Differences between NTHi and corresponding PBS-exposed SAECs were calculated with paired t-test; *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001. Differences between means of non-COPD and COPD SAECs were calculated with a Mann–Whitney test; #: p<0.05.

Fig 4: In Trans HIV-1 transmission Assay; HeLa cells (0.05 × 106 cells/well in a 24-well plate) were incubated for 2 h at 37 °C with SLPI (1 µg/mL and 10 µg/mL) or SERPIN C1 (with 0.05 µg/mL and 0.1 µg/mL). Heparinase I and III (20 IU/mL) was used as a negative control. HIV-1 BaL (10 ng of p24) was added to HeLa cells target cells for 1 h at 37 °C in a final volume of 100 µL of serum free DMEM to facilitate virus adsorption. Cells were washed five times with 100 µL of PBS to remove unbound material and overlaid with TZM-bl indicator cells (0.05 × 106 cells/well). After sixty hours post infection productive HIV-1infection and replication was quantitated using the Luciferase Assay System. Results are expressed as mean ± SEM from three independent experiments. Asterisk (*) over the bars indicates significant difference with control, ** p < 0.001 and * p ≤ 0.05. p-values were generated by one-way ANOVA with multiple comparisons.

Fig 5: Effects of SLPI and SERPIN C1 on MDMs infection by VSV-G pseudo typed HIV-1 construct.; MDMs (5 × 104 cells/well) were pre-treated with 1 µg/mL and 10 µg/mL SLPI or 0.05 µg/mL and 0.1 µg/mL SERPIN C1 for 2 h and then infected with VSV-G pseudo typed HIV-1 (5 ng/mL p24 units). After 2 h, the unbound virus was removed and replenished with medium containing the same concentrations of SLPI (1 µg/mL and 10 µg/mL) or SERPIN C1 (0.05 µg/mL and 0.1 µg/mL). MDMs were collected 72 h after infection, and total RNA extracted, and real-time PCR was performed to quantitate HIV-1 replication. Results expressed as mean ± SEM from three independent donors. Mark over the bars indicates non-significant difference with control, ns p > 0.05. p-values were generated by one-way ANOVA with multiple comparisons.