Structure-Function Analysis of CCL28 in the Development of Post-viral Asthma

Abstract

Background: CCL28 is associated with the pathogenesis of acute post-viral asthma. Results: In the absence of viral infection, natively folded CCL28 can induce asthma pathology, whereas unfolded CCL28 cannot. Conclusion:The structure of CCL28 is critical for its role in asthma pathogenesis. Significance: Inhibition of CCL28 presents a novel therapeutic option for the prevention of post-viral asthma.

POL7085 or anti-CCL28 treatment inhibits development of post-paramyxoviral airway disease

“…Furthermore, we have shown that CCL28 in the absence of a viral infection is sufficient to drive AHR and MCM . Administering an anti‐CCL28 mAb during the viral infection significantly reduced post‐viral MCM and AHR at a late time point (day 49 PI) . These data reiterate the importance of CCL28 to the development of post‐viral lung disease in this animal model.…”

Production of Recombinant Chemokines and Validation of Refolding

“…Additionally, tandem mass spectrometry (MS-MS) can in some instances be used to define the pairing of cysteines, like the non-conserved disulfide bond in CCL28 that links C30 to C80 (Thomas et al, 2015), or confirm the locations of posttranslational modifications, like pyroglutamate formation in CCL2 as shown in Figures 4 and 5, respectively.…”

“…Classic in vitro assays for biological activity can include calcium flux assays or chemotaxis assays (Kar, Srivastava, Andersson, Baratelli et al, 2011; Love et al, 2012; Miller & Krangel, 1992; Neote, DiGregorio, Mak, Horuk et al, 1993; Thomas et al, 2015). Calcium flux assays are those that detect the increase in the cytosol of the secondary messenger calcium that is released as a result of chemokine receptor activation (Love et al, 2012; Miller et al, 1992; Neote et al, 1993).…”

“…Calcium flux assays are those that detect the increase in the cytosol of the secondary messenger calcium that is released as a result of chemokine receptor activation (Love et al, 2012; Miller et al, 1992; Neote et al, 1993). Chemotaxis assays monitor chemokine recruitment of cells expressing chemokine receptor across a barrier that is impermeable to cells without the chemokine functioning as an attractant (Kar et al, 2011; Miller et al, 1992; Thomas et al, 2015). An example chemotaxis assay protocol for CCL21 is presented here.…”

“…The metamorphic chemokine XCL1 has only two cysteines that form one disulfide bond, which permits unfolding and interconversion between two unrelated folded structures, the canonical chemokine domain or an all β-sheet structure (Tyler, Murray, Peterson, & Volkman, 2011). Other chemokines, like CCL21 or CCL28, contain six cysteines corresponding to a novel third disulfide in addition to the two conserved disulfides (Love, Sandberg, Ziarek, Gerarden et al, 2012; Thomas, Buelow, Nevins, Jones et al, 2015). …”