Human rhinovirus induced cytokine/chemokine responses in human airway epithelial and immune cells.

Infections with human rhinovirus (HRV) are commonly associated with acute upper and lower respiratory tract disease and asthma exacerbations. The role that HRVs play in these diseases suggests it is important to understand host-specific or virus-specific factors that contribute to pathogenesis. Since species A HRVs are often associated with more serious HRV disease than species B HRVs, differences in immune responses they induce should inform disease pathogenesis. To identify species differences in induced responses, we evaluated 3 species A viruses, HRV 25, 31 and 36 and 3 species B viruses, HRV 4, 35 and 48 by exposing human PBMCs to HRV infected Calu-3 cells. To evaluate the potential effect of memory induced by previous HRV infection on study responses, we tested cord blood mononuclear cells that should be HRV naïve. There were HRV-associated increases (significant increase compared to mock-infected cells) for one or more HRVs for IP-10 and IL-15 that was unaffected by addition of PBMCs, for MIP-1α, MIP-1β, IFN-α, and HGF only with addition of PBMCs, and for ENA-78 only without addition of PBMCs. All three species B HRVs induced higher levels, compared to A HRVs, of MIP-1α and MIP-1β with PBMCs and ENA-78 without PBMCs. In contrast, addition of CBMCs had less effect and did not induce MIP-1α, MIP-1β, or IFN-α nor block ENA-78 production. Addition of CBMCs did, however, increase IP-10 levels for HRV 35 and HRV 36 infection. The presence of an effect with PBMCs and no effect with CBMCs for some responses suggest differences between the two types of cells possibly because of the presence of HRV memory responses in PBMCs and not CBMCs or limited response capacity for the immature CBMCs relative to PBMCs. Thus, our results indicate that different HRV strains can induce different patterns of cytokines and chemokines; some of these differences may be due to differences in memory responses induced by past HRV infections, and other differences related to virus factors that can inform disease pathogenesis

HRV species associated cytokine/chemokine production in calu-3 cells.

<p>Fig. 1A, 1B, 1C and 1D. Levels of cytokines/chemokines (pg/ml) in the supernatant of calu-3 cells infected with species A (HRV 25, HRV 31 and HRV 36) or species B (HRV 4, HRV 35 and HRV 48) HRVs and uninfected control cells with or without PBMCs. Data are mean from duplicate infection using three PBMC donors (n = 3).</p

HRV-associated responses in calu-3 cells with or without PBMCs.

<p>Shown are the median values of cytokines and chemokines (pg/ml) produced in response to various HRVs and uninfected control infections at 24 hours after the time inserts with PBMCs were added (n = 3). Significant differences in cytokine/chemokine levels for HRV infected cells when compared to uninfected controls were calculated using Wilcoxen rank sum tests. * indicates significantly different from control (p<0.05).</p><p>HRV-associated responses in calu-3 cells with or without PBMCs.</p

Comparison of CBMC vs. PBMC.

<p>Comparison of HRV-associated response in calu-3 cells with CBMCs and PBMCs. Shown are the median values of cytokines and chemokines (pg/ml) produced in response to various HRVs and uninfected control infections at 24 hours after the time inserts with PBMCs or CBMCs were added (n = 3). Significant differences in cytokine/chemokine levels for HRV infected cells comparing CBMCs and PBMCs were calculated using Wilcoxen rank sum tests. * indicates significantly different in values with CBMCs compared to those with PBMCs (p<0.05).</p><p>Comparison of CBMC vs. PBMC.</p

HRV-associated responses in calu-3 cells with or without CBMCs.

<p>Shown are the median values of cytokines and chemokines (pg/ml) produced in response to various HRVs and uninfected control infections at 24 hours after the time inserts with CBMCs were added (n = 3). Significant differences in cytokine/chemokine levels for HRV infected cells when compared to uninfected controls were calculated using Wilcoxen rank sum tests. * indicates significantly different from control (p<0.05).</p><p>HRV-associated responses in calu-3 cells with or without CBMCs.</p