Abstract Per- and polyfluoroalkyl substances (PFAS) are persistent organic pollutants, some immunotoxic, that are detectable in the serum of most U.S. adults. Some studies of highly-exposed individuals have suggested positive associations between PFAS and non-Hodgkin lymphoma (NHL); however, it is unknown whether associations exist at lower exposure levels that are more common in the general population. To address this knowledge gap, we conducted a nested case-control study investigating serum PFAS concentrations and NHL within the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. We measured pre-diagnostic serum concentrations (collected between 1993-2001) of five PFAS among 706 cases and 706 controls individually matched on the basis of age at baseline, sex, self-reported race and ethnicity, study center, calendar year of blood collection, and number of prior serum thaws. We estimated odds ratios (ORs) and 95% confidence intervals (CIs) for PFAS concentrations in relation to NHL, both overall and for selected histologic subtypes [diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), lymphoplasmacytic lymphoma (LPL), and marginal zone lymphoma (MZL)] using conditional logistic regression. We found no evidence of a positive association with NHL for any of the five PFAS in analyses adjusting for all measured PFAS. In analyses of histologic subtypes, we observed a positive association between perfluorohexane sulfonate (PFHxS) and DLBCL (OR per doubling in concentration=1.27, 95% CI=1.07, 1.50) which remained for cases diagnosed ≥10 years after blood collection (OR=1.34, 95% CI=1.09, 1.64). We also observed inverse associations with PFNA for all subtypes (DLBCL, OR=0.84, 95% CI=0.72, 0.98; FL, OR=0.86, 95% CI=0.69, 1.08; LPL/MZL, OR=0.86, 95% CI=0.66, 1.12), although these associations were null among participants with blood drawn prior to 1997 (DLBCL: OR<1997=0.95, OR≥1997=0.67, Pinteraction=0.001; FL: OR<1997=1.01, OR≥1997=0.77, Pinteraction =0.08; LPL/MZL: OR<1997=1.26, OR≥1997=0.88, Pinteraction =0.03). In conclusion, our findings from a cohort study with PFAS serum concentrations comparable to that of the general population do not support an association with increased risk of NHL overall. The suggestive evidence of a positive association between PFHxS and DLBCL warrants further investigation. Citation Format: Jongeun Rhee, Jani Koponen, Joshua N. Sampson, Alexander P. Keil, Mary H. Ward, Jonathan N. Hofmann, Wen-Yi Huang, Debra T. Silverman, Panu Rantakokko, Mark P. Purdue. Serum concentrations of per- and polyfluorinated substances and risk of non-Hodgkin lymphoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 840.