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To determine the effect of moisture regain of wool on atmospheric pressure plasma treatment results, wool fibers and fabrics conditioned in 100% relative humidity (RH) and 65% RH were treated by an atmospheric pressure plasma jet with pure helium and helium/oxygen mixed gas, respectively. Scanning electron microscope (SEM) indicated that scales of wool fiber were smoothened for fibers conditioned in the 100% RH. X-ray photoelectron spectroscopy (XPS) showed that carbon content decreased substantially after the plasma treatment. Surface chemical composition of 100% RH conditioned groups changed more significantly than the 65% RH conditioned groups. Water contact angle decreased significantly after the plasma treatments. In shrinkage test, plasma-treated wool fabrics preconditioned in 100% RH showed the lowest shrinkage ratios of 5% and 6%, below 8% is required for machine-washable wool fabrics according to ISO standard.