Background: Leukocyte subtypes have unique biological roles, but no study has investigated them in relation to incident nonalcoholic fatty liver disease (NAFLD). Few prospective studies revealed a positive association between total leukocyte count and NAFLD. Whereas the results suggested that their association may not be linear, this possibility has never been tested. Here we investigated the association between peripheral total and differential leukocyte counts and incidence of NAFLD and to detect possible non-linearity. Methods: A prospective cohort study (n=33,618; median: 2.00 y, 95 % CI: 2.87-2.90 y) was conducted in a general population sample from Tianjin, China. Dose-response relations of total and differential leukocyte counts as well as neutrophil to lymphocyte ratio (NLR) with incident NAFLD were estimated using restricted cubic spline analysis with Cox proportional hazards model. Results: During the follow-up period, 5,971 participants had a first-ever NAFLD. Total leukocyte count, neutrophil count and NLR were all positively and nonlinearly related to NAFLD (P-non-linearity <0.05). Before reaching certain threshold levels, each one standard deviation increases in them were respectively associated with 19%, 27% and 22% greater risk of NAFLD, which then remained stable thereafter. In addition, a weak positive and linear association was observed between lymphocyte count and NAFLD (P-non-linearity=0.30). Conclusions: We demonstrated for the first time that total leukocyte and neutrophil counts, but not lymphocyte count, were nonlinearly related to NAFLD. Further studies are needed to investigate whether these different association patterns represent different effects of leukocyte subtypes in the pathogenesis of NAFLD. Funding Statement: This study was supported by grants from the National Natural Science Foundation of China (No. 91746205, 81673166, 81372118, 81372467 and 81302422), the key technologies R&D program of Tianjin (Key Project: No. 11ZCGYSY05700, 12ZCZDSY20400, 13ZCZDSY20200, and 15YFYZSY00020), the National Science and Technology Support Program (No. 2012BAI02B02), 2012 and 2016 Chinese Nutrition Society Nutrition Research Foundation—DSM Research Fund (No. 2014-071, 2016-046 and 2016-023), the Technologies development program of Beichen District of Tianjin (No. bcws2013-21, bcws2014-05 and 2015-SHGY-02), the technologies project of Tianjin Binhai New Area (No. 2013-02-04 and 2013-02-06), the Science Foundation of Tianjin Medical University (No. 2010KY28 and 2013KYQ24), the Key Laboratory of Public Health Safety (Fudan University), Ministry of Education (No. GW2014-5), and the National Training Programs of Innovation and Entrepreneurship for Undergraduates (No. 201510062013), China. Declaration of Interests: The authors declare that they have no conflict of interest. Ethics Approval Statement: The Institutional Review Board of the Tianjin Medical University approved this study. Informed consent was obtained from all individual participants included in the study.
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