Fig. 7

Distribution of sCD11/CD18 oligomers in MDDCs culture supernatants. a GPC followed by TRIFMA detection of sCD11/CD18 in a “sandwich-type” assay with the monoclonal Ab KIM185 to CD18 as a capture Ab and KIM127. The plot shows the TRIFMA signal (S _TRF, read on the left ordinate) and the optical density (OD) at λ = 280 nm (read on the right ordinate) in the fractions plotted as a function of the elution volume, V _e. The elution volumes of oligomeric recombinant soluble CD11a/CD18 (established in a previous study [13] using the same chromatography equipment) are indicated hatched lines together with the elution volumes of the hydrodynamic radius calibration markers thyroglobulin, ferritin, IgG, and albumin. b-d Distribution of sCD11/CD18 oligomers in cell culture supernatant from MDDCs. The analysis was carried out with culture supernatants derived from two Donors (#67 and #68), either left untreated or treated with 1 or 5000 ng/ml IL-20 (Incubation VII in Fig. 1b) The sCD11/CD18 signal in the fractions was measured with the KIM185-KIM127 assay and plotted as a function of V _e. (e,f) Cumulative distribution of the CD18 signal in the GPC fractions. The cumulative distribution profile was calculated by plotting \( 100\%\ \frac{{\displaystyle {\sum}_{V_e=30 ml}^{V_e=i}{S}_{TRF}\left({V}_e\right)}}{{\displaystyle \sum {S}_{TRF}}} \) as a function of V _e, where S _TRF(V _e) is the CD18 signal for the fraction with elution volume V _e and ∑S _TRF the sum of signals for all fractions eluting from 30 ml to 100 ml, both end points included. Black arrows indicate a feature in the distribution profile shared between the donors