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Supplementary Table 2 from Elevated Systemic Levels of Inflammatory Cytokines in Older Women with Persistent Cervical Human Papillomavirus Infection

Supplementary Tables 1-3. Supplementary Table 1 - a) Correlations between selectb serum inflammation markers among lung cancer controls. b) Correlations between select and serum inflammation markers among NHL controls. c) Correlations between selectb serum inflammation markers among ovarian cancer controls. Supplementary Table 2. Odds ratios (ORs)a and 95% confidence intervals (CIs) for selectb circulating inflammation markers and body mass index. Supplementary Table 3. Odds ratios (ORs)a and 95% confidence intervals (CIs) for select circulating inflammation markers and body mass index (per 5-kg/m2) by case-control study

Background: Coffee drinking has been inversely associated with mortality as well as cancers of the endometrium, colon, skin, prostate, and liver. Improved insulin sensitivity and reduced inflammation are among the hypothesized mechanisms by which coffee drinking may affect cancer risk; however, associations between coffee drinking and systemic levels of immune and inflammatory markers have not been well characterized.Methods: We used Luminex bead-based assays to measure serum levels of 77 immune and inflammatory markers in 1,728 older non-Hispanic Whites. Usual coffee intake was self-reported using a food frequency questionnaire. We used weighted multivariable logistic regression models to examine associations between coffee and dichotomized marker levels. We conducted statistical trend tests by modeling the median value of each coffee category and applied a 20% false discovery rate criterion to P values.Results: Ten of the 77 markers were nominally associated (P trend < 0.05) with coffee drinking. Five markers withstood correction for multiple comparisons and included aspects of the host response namely chemotaxis of monocytes/macrophages (IFNγ, CX3CL1/fractalkine, CCL4/MIP-1β), proinflammatory cytokines (sTNFRII), and regulators of cell growth (FGF-2). Heavy coffee drinkers had lower circulating levels of IFNγ [odds ratios (OR), 0.35; 95% confidence intervals (CI), 0.16–0.75], CX3CL1/fractalkine (OR, 0.25; 95% CI, 0.10–0.64), CCL4/MIP-1β (OR, 0.48; 95% CI, 0.24–0.99), FGF-2 (OR, 0.62; 95% CI, 0.28–1.38), and sTNFRII (OR, 0.34; 95% CI, 0.15–0.79) than non-coffee drinkers.Conclusions: Lower circulating levels of inflammatory markers among coffee drinkers may partially mediate previously observed associations of coffee with cancer and other chronic diseases.Impact: Validation studies, ideally controlled feeding trials, are needed to confirm these associations. Cancer Epidemiol Biomarkers Prev; 24(7); 1052–60. ©2015 AACR.

SupplementaryTable S5. Odds ratios (OR) for high versus low levels e for eight circulating inflammatory markers (P-trend

Supplementary Materials and Methods: Details on development of sampling weights and their use in analysis.

Supplementary Table S4. Categorization of markers by weighted percent of values below the lower limit of detection (LOD)

Background: Chronic inflammation is etiologically related to several cancers. We evaluated the performance [ability to detect concentrations above the assay's lower limit of detection, coefficients of variation (CV), and intraclass correlation coefficients (ICC)] of 116 inflammation, immune, and metabolic markers across two Luminex bead–based commercial kits and three specimen types.Methods: From 100 cancer-free participants in the Prostate, Lung, Colorectal, and Ovarian Cancer Trial, serum, heparin plasma, and EDTA plasma samples were utilized. We measured levels of 67 and 97 markers using Bio-Rad and Millipore kits, respectively. Reproducibility was assessed using 40 blinded duplicates (20 within-batches and 20 across-batches) for each specimen type.Results: A majority of markers were detectable in more than 25% of individuals on all specimen types/kits. Of the 67 Bio-Rad markers, 51, 52, and 47 markers in serum, heparin plasma, and EDTA plasma, respectively, had across-batch CVs of less than 20%. Likewise, of 97 Millipore markers, 75, 69, and 78 markers in serum, heparin plasma, and EDTA plasma, respectively, had across-batch CVs of less than 20%. When results were combined across specimen types, 45 Bio-Rad and 71 Millipore markers had acceptable performance (>25% detectability on all three specimen types and across-batch CVs Conclusions: Inflammation and immune markers can be measured reliably in serum and plasma samples using multiplexed Luminex-based methods.Impact: Multiplexed assays can be utilized for epidemiologic investigations into the role of inflammation in cancer etiology. Cancer Epidemiol Biomarkers Prev; 20(9); 1902–11. ©2011 AACR.

Supplementary Table S3. Description of three case-control studies nested within the PLCO Cancer Screening Trial

Supplementary Table S1. Odds ratios (OR) for high versus low levels d for ten circulating inflammatory markers (P-trend

Supplementary Table S8. Odds ratios (OR) for high versus low levels e for the top five circulating inflammatory markers in three case-control studies nested within the PLCO Cancer Screening Trial by case-control study of origin

Supplementary Figures 1 and 2 from Evaluation of Multiplexed Cytokine and Inflammation Marker Measurements: a Methodologic Study

Background: Regular aspirin use may decrease cancer risk by reducing chronic inflammation. However, associations between aspirin use and circulating markers of inflammation have not been well studied.Methods: Serum levels of 78 inflammatory markers were measured in 1,819 55- to 74-year-old men and women in the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial. Data were combined from three completed case–control studies and reweighted to the PLCO screening arm. Self-reported aspirin and ibuprofen use (number of tablets taken per day/week/month) over the previous 12 months was collected at baseline. Associations between (i) nonregular (Results: Aspirin use was nominally associated with (Ptrend across categories ≤ 0.05) decreased levels of chemokine C-C motif ligand 15 [CCL15; OR, 0.5; 95% confidence intervals (CI), 0.3–0.8; moderate versus nonregular use]; soluble vascular endothelial growth factor receptor 2 (sVEGFR2; OR, 0.7; 95% CI, 0.4–1.0); soluble tumor necrosis factor receptor 1 (sTNFR1; OR, 0.6; 95% CI, 0.4–0.9) and increased levels of CCL13 (OR, 1.3; 95% CI, 0.8–2.1); CCL17 (OR, 1.1; 95% CI, 0.7–1.9) and interleukin 4 (IL4; OR, 1.6; 95% CI, 0.9–2.8). Trends were not statistically significant following correction for multiple comparisons. Likewise, no statistically significant associations were observed between ibuprofen use and marker levels.Conclusions: No significant associations were observed between regular aspirin use and the inflammatory markers assessed.Impact: Additional studies are needed to better understand the relationship between aspirin use, chronic inflammation, and cancer risk. Cancer Epidemiol Biomarkers Prev; 24(5); 825–32. ©2015 AACR.

Background: Epidemiologic studies examining circulating levels of inflammatory markers in relation to obesity and physical inactivity may aid in our understanding of the role of inflammation in obesity-related cancers. However, previous studies on this topic have focused on a limited set of markers.Methods: We evaluated associations between body mass index (BMI) and vigorous physical activity level, based on self-report, and serum levels of 78 inflammation-related markers. Markers were measured using a bead-based multiplex method among 1,703 men and women, ages 55–74 years, and with no prior history of cancer at blood draw, and selected for case–control studies nested within the Prostate, Lung, Ovarian, and Colorectal Cancer Screening Trial. Analyses were adjusted for age, sex, smoking, case–control study, physical activity, and BMI.Results: Twelve markers were positively associated with BMI after FDR correction. ORs and 95% confidence interval (CI) for highest versus lowest levels of CCL2/MCP-1, CXCL5/ENA-78, sTNFRII, CXCL10/IP-10, CXCL6/GCP2, CCL13/MCP-4, amylin, CRP, C-peptide, CCL19/MIP-3b, insulin, and leptin were: 1.50 (1.14–1.98), 1.52 (1.12–2.05), 1.61 (1.17–2.20), 1.69 (1.25–2.28), 1.74 (1.24–2.44), 1.75 (1.22–2.50), 1.91 (1.31–2.78), 2.41 (1.36–4.25), 2.78 (1.83–4.24), 3.30 (2.28–4.78), 4.05 (2.51–6.55), and 50.03 (19.87–125.99) per 5 kg/m2, respectively. Only CXCL12/SDF-1a was associated with physical activity (≥3 vs. Conclusions: BMI was associated with a wide range of circulating markers involved in the inflammatory response.Impact: This cross-sectional analysis identified serum markers could be considered in future studies aimed at understanding the underlying mechanisms linking inflammation with obesity and obesity-related cancers. Cancer Epidemiol Biomarkers Prev; 23(12); 2840–9. ©2014 AACR.

Background: Defects in lymphoproliferative responses to mitogens/antigens in women >45 years old with a persistent type-specific human papillomavirus (HPV) infection have been reported.Methods: To determine whether these defects were associated with altered cytokine profiles, plasma and peripheral blood mononuclear cell (PBMC) culture supernatants from 50 cases (oversampled for their reduced lymphoproliferative ability) and 50 uninfected controls (oversampled for their robust lymphoproliferative ability) were examined for 24 cytokines using multiplexed bead–based immunoassays and ELISA.Results: The following plasma cytokines were significantly increased in cases relative to controls (cases versus controls; median pg/mL): interleukin (IL)-6, 393.1 versus 14.5; IL-8, 1,128.5 versus 43.9; tumor necrosis factor-α (TNF-α), 164.1 versus 9.2; macrophage inflammatory protein-1α (MIP-1α), 1,368.9 versus 25.5; granulocyte macrophage colony-stimulating factor (GM-CSF), 13.8 versus 7.3; IL-1β, 8.3 versus 1.6 (all P < 0.0001); and IL-1α, 218.2 versus 169.5 (P = 0.02). We focused our analysis on the cytokines IL-6, IL-8, TNF-α, and MIP-1α due to their high fold change (>10) and highly statistically significant difference between cases and controls. Length of persistence or type of infection (high risk and low risk) did not affect these differences. IL-6, TNF-α, and MIP-1α levels were also increased in unstimulated PBMC culture supernatants from cases compared with controls (P < 0.05), however, the cytokine levels from phytohemagglutinin-stimulated PBMC culture supernatants were significantly lower in the cases (P < 0.0001).Conclusions: Persistent HPV infection in older women with evidence of immune deficit is associated with an increase in systemic inflammatory cytokines.Impact: Future studies are needed to determine whether the inflammatory profile is age dependent and to examine the role that inflammatory cytokines play in HPV-induced progression from infection to cervical cancer. Cancer Epidemiol Biomarkers Prev; 19(8); 1954–9. ©2010 AACR.

Supplementary Table 1 from Elevated Systemic Levels of Inflammatory Cytokines in Older Women with Persistent Cervical Human Papillomavirus Infection

Supplementary Table S2. Multiplex immune panel markers measured in the PLCO Cancer Screening Study

Supplemental Tables 1-7 and Methods. Details on development of sampling weights and their use in analysis. Supplemental Table 1. Description of three case-control studies nested within PLCO. Supplemental Table 2. Multiplex immune panel markers measured in the PLCO Cancer Screening Study. Supplemental Table 3: Percent detection and median levels for each inflammatory marker by original study. Supplemental Table 4: Participant characteristics for i) the 1,819 individuals with inflammatory marker data, ii) the weighted population and iii) those in the PLCO screening arm that met the study eligibility criteria. Supplemental Table 5: Associations between regular aspirin use and circulating inflammatory markers2, restricted to individuals without a history of chronic heart disease, stroke or arthritis. Supplemental Table 6: Associations between regular aspirin use and circulating inflammatory markers2, by study. Supplemental Table 7: Associations between regular aspirin use and circulating inflammatory markers.

Supplementary Table 1 from Evaluation of Multiplexed Cytokine and Inflammation Marker Measurements: a Methodologic Study

Supplementary Table 2 from Evaluation of Multiplexed Cytokine and Inflammation Marker Measurements: a Methodologic Study

Supplementary Table S6. Weighted Pearson correlation (r) matrix between dichotomized marker variables ({greater than or equal to} weighted mean vs. < weighted mean or in the case of analytes with

Background: Previous prospective studies of associations between circulating inflammation-related molecules and pancreatic cancer risk have included limited numbers of markers. Methods: We conducted a case–cohort study nested within the Japan Public Health Center-based Prospective Study Cohort II. We selected a random subcohort (n = 774) from a total of 23,335 participants aged 40 to 69 years who returned a questionnaire and provided blood samples at baseline. During the follow-up period from 1993 to 2010, we identified 111 newly diagnosed pancreatic cancer cases, including one case within the subcohort. Plasma concentrations of 62 inflammatory markers of chemokines, cytokines, and growth factors were measured by a Luminex fluorescent bead-based assay. Cox regression models were applied to estimate HR and 95% confidence intervals (CI) for pancreatic cancer risk for quartiles of marker levels adjusted for potential confounders. Results: The HR (95% CI) for the highest versus the lowest category of C–C motif ligand chemokine 8/monocyte chemoattractant protein 2 (CCL8/MCP2) was 2.03 (1.05–3.93; Ptrend = 0.048). After we corrected for multiple comparisons, none of the examined biomarkers were associated with pancreatic cancer risk at P-value Conclusions: We found no significant associations between 62 inflammatory markers and pancreatic cancer risk. Impact: The suggestive association with circulating levels of leukocyte recruiting cytokine CCL8/MCP2 may warrant further investigation.

Background: Metabolic syndrome (MetS) is a systemic inflammatory state. Low physical activity (PA) could modify this patho-physiology or act as an independent contributor to inflammation. Previous studies of both conditions have identified altered levels of inflammation- and immune-related proteins based on limited sets of candidate markers. Methods: We investigated associations of MetS and low PA with circulating inflammation markers in a stratified random sample of Japanese adults (N = 774, mean age 60.7 years) within the Japan Public Health Center-based Prospective Study (JPHC) Cohort II. AHA/NHLBI criteria were used to define MetS (19%) and the bottom quartile of PA was considered low. 110 circulating biomarkers, including cytokines, chemokines, and soluble receptors were measured by multiplex bead-based and proximity-extension assays. Associations of MetS and low PA with marker quantiles were adjusted for each other and for age, sex, study site, cigarette smoking, alcohol consumption, and blood sample fasting state by ordinal logistic regression. P values were corrected for FDR. Results: MetS was significantly associated with levels of six markers: IL18R1 [odds ratio 2.37; 95% confidence interval (CI), 1.45–3.87], CRP (2.07; 95% CI, 1.48–2.90), SAP (2.08; 95% CI, 1.47–2.95), CCL19/MIP3β (2.06; 95% CI, 1.48–2.88), CXCL12/SDF1α+β (0.48; 95% CI, 0.32–0.65), and CCL28 (0.44; 95% CI, 0.27–0.71). Low PA had no significant marker associations. Conclusions: Positively associated markers with MetS are mostly Th1 immune response–related and acute phase proteins, whereas negatively associated markers are generally Th2-related. Impact: MetS is associated with a broad range of alterations in immune and inflammatory biomarkers that may contribute to risks of various chronic diseases, independent of low PA.

Background: Circulating inflammation proteins may be important mediators or markers of carcinogenic mechanisms. There have been few studies with limited numbers of analytes in patients with upper gastrointestinal (GI) tract tumors. We therefore evaluated risk associations of gastric and esophageal cancers with prediagnostic levels of a wide range of these molecules. Methods: We performed a case–cohort analysis within the Japan Public Health Center–Based Prospective Study Cohort II, including incident cases of gastric (n = 446) and esophageal (n = 68) cancers and a random subcohort (n = 774). A total of 64 biomarkers were measured in baseline plasma using Luminex bead-based assays. The median time between blood collection and diagnosis was 8.1 years for gastric cancer and 9.4 years for esophageal cancer. HRs for association with each marker were adjusted for potential confounders using Cox regression. Results: In separate models, sEGFR and TSLP were nominally associated with gastric cancer risk, and CRP, CXCL11/ITAC, and CCL15/MIP1D were associated with esophageal cancer. However, no association satisfied statistical significance after FDR correction. Associations did not differ by time from blood collection to cancer ( Conclusions: Our study failed to identify associations of circulating inflammation markers with risk of upper GI tract tumors. Impact: To date, this is the largest assessment of inflammation-related proteins with gastric and esophageal cancer risks. However, the evaluated molecules may not fully represent the complex inflammation processes preceding malignant transformation. Further investigation of other markers in prospective studies is warranted, as demonstration of associations may have important implications for prevention and treatment of these cancers.

Background: Evidence suggests that natural antibodies developed after HPV16 infection may protect some women but not men against subsequent HPV16 reacquisition. Less is known whether antibodies developed following HPV16 infection are protective among men who have sex with men (MSM). Methods: Four hundred seventy-five MSM from the Human Papillomavirus Infection in Men (HIM) study were tested for serum antibodies to HPV16 L1 using enzyme-linked immunosorbent assays, and for anal and genital HPV16 DNA using PCR consensus primer system (PGMY 09/11). Adjusted Cox regression was used to evaluate whether baseline HPV16 seropositivity impacts subsequent genital or anal HPV16 DNA. Results: The risk of subsequent genital HPV16 [aHR = 1.05, 95% confidence interval (CI) = 0.66–1.68] and anal HPV16 infections among MSM (aHR = 2.34, 95% CI = 0.92–5.98) was similar or nonsignificantly higher in HPV16-seropositive than HPV16-seronegative MSM. The risk of genital HPV16 was also similar between HPV16-seronegative and HPV16-seropositive MSM in the highest tertile of HPV16 antibody levels and when restricting to those with new sex partners during follow-up (P > 0.20). Among the 118 MSM who were HPV16 seropositive, 90% remained HPV16 seropositive up to 4 years later. When tested together, MSM with the highest antibody titers (top tertile) had similar levels to females (mean = 130.3 vs. 134.5 EU/mL, P = 0.84). Conclusions: Despite years of HPV16 seropositivity persistence and antibody titers comparable with females, this study suggested no evidence of HPV16 natural antibodies protecting against subsequent genital or anal HPV16 infection in MSM. Impact: This could help partially explain the high incidence of genital and anal HPV16 infection and related anal cancer seen in middle-aged and older MSM. Cancer Epidemiol Biomarkers Prev; 27(4); 496–502. ©2018 AACR.

Background: Blood group antigens may play a role in immune and inflammatory processes. The aim of this analysis was to identify specific inflammatory pathways associated with blood group antigens. Methods: We investigated associations between blood type and circulating inflammatory markers in data for 3,537 non-Hispanic white participants selected from previous case-control studies nested in the Prostate, Lung, Colorectal, and Ovarian cancer screening trial. Whole genome scans were used to infer blood types for twelve common antigen systems (ABO, Duffy, Lewis, RhE, Kidd, Kell, Secretor, Lutheran, Colton, Dombrock, Auberger, and Knops) based on well-characterized single nucleotide polymorphisms. Circulating levels of 96 immune- and inflammation-related biomarkers were measured on multiplex fluorescent bead-based panels. We estimated associations between blood type and markers using design-weighted linear or logistic regression adjusted for age, sex, smoking status, and principal components of population substructure, and using a Bonferroni correction to control for multiple comparisons. Results: Among the 1,152 blood type-biomarker associations tested, we identified 10 which were significant after correction for multiple comparisons. ABO subtype was associated with levels of sVEGFR2, sVEGFR3, and sGP130, and Duffy subtype was associated with levels of CXCL6/GCP2, CXCL5/ENA78, CCL11/EOTAXIN, CXCL1/GRO, CCL2/MCP1, CCL13/MCP4, and CCL17/TARC. Post-hoc pairwise t-tests showed that individuals with type A blood had lower levels of all ABO-associated markers than individuals with type O or B blood, while individuals with type Fy(a+b-) had lower levels of all Duffy-associated markers than individuals with type Fy(a+b+) or Fy(a-b+). Conclusion: ABO blood type is associated with levels of three circulating angiogenic markers, while Duffy blood type is associated with eight chemokines. Additional work to understand the clinical implications of these differences is needed. Citation Format: Sarah C. Van Alsten, John Aversa, Loredana Santo, M. Constanza Camargo, Wen-Yi Huang, Troy Kemp, Jia Liu, Joshua Samspon, Charles S. Rabkin. Association between ABO and Duffy blood types and circulating chemokines and cytokines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 834.

Background: Coffee drinking has been inversely associated with mortality as well as cancers of the endometrium, colon, skin, prostate, and liver. Improved insulin sensitivity and reduced inflammation are among the hypothesized mechanisms by which coffee drinking may affect cancer risk; however, associations between coffee drinking and systemic levels of immune and inflammatory markers have not been well characterized. Methods: We used Luminex bead-based assays to measure serum levels of 77 immune and inflammatory markers in 1,728 older non-Hispanic Whites. Usual coffee intake was self-reported using a food frequency questionnaire. We used weighted multivariable logistic regression models to examine associations between coffee and dichotomized marker levels. We conducted statistical trend tests by modeling the median value of each coffee category and applied a 20% false discovery rate criterion to P values. Results: Ten of the 77 markers were nominally associated (P trend < 0.05) with coffee drinking. Five markers withstood correction for multiple comparisons and included aspects of the host response namely chemotaxis of monocytes/macrophages (IFNγ, CX3CL1/fractalkine, CCL4/MIP-1β), proinflammatory cytokines (sTNFRII), and regulators of cell growth (FGF-2). Heavy coffee drinkers had lower circulating levels of IFNγ [odds ratios (OR), 0.35; 95% confidence intervals (CI), 0.16–0.75], CX3CL1/fractalkine (OR, 0.25; 95% CI, 0.10–0.64), CCL4/MIP-1β (OR, 0.48; 95% CI, 0.24–0.99), FGF-2 (OR, 0.62; 95% CI, 0.28–1.38), and sTNFRII (OR, 0.34; 95% CI, 0.15–0.79) than non-coffee drinkers. Conclusions: Lower circulating levels of inflammatory markers among coffee drinkers may partially mediate previously observed associations of coffee with cancer and other chronic diseases. Impact: Validation studies, ideally controlled feeding trials, are needed to confirm these associations. Cancer Epidemiol Biomarkers Prev; 24(7); 1052–60. ©2015 AACR.

Background: Chronic inflammation is etiologically related to several cancers. We evaluated the performance [ability to detect concentrations above the assay's lower limit of detection, coefficients of variation (CV), and intraclass correlation coefficients (ICC)] of 116 inflammation, immune, and metabolic markers across two Luminex bead–based commercial kits and three specimen types. Methods: From 100 cancer-free participants in the Prostate, Lung, Colorectal, and Ovarian Cancer Trial, serum, heparin plasma, and EDTA plasma samples were utilized. We measured levels of 67 and 97 markers using Bio-Rad and Millipore kits, respectively. Reproducibility was assessed using 40 blinded duplicates (20 within-batches and 20 across-batches) for each specimen type. Results: A majority of markers were detectable in more than 25% of individuals on all specimen types/kits. Of the 67 Bio-Rad markers, 51, 52, and 47 markers in serum, heparin plasma, and EDTA plasma, respectively, had across-batch CVs of less than 20%. Likewise, of 97 Millipore markers, 75, 69, and 78 markers in serum, heparin plasma, and EDTA plasma, respectively, had across-batch CVs of less than 20%. When results were combined across specimen types, 45 Bio-Rad and 71 Millipore markers had acceptable performance (>25% detectability on all three specimen types and across-batch CVs Conclusions: Inflammation and immune markers can be measured reliably in serum and plasma samples using multiplexed Luminex-based methods. Impact: Multiplexed assays can be utilized for epidemiologic investigations into the role of inflammation in cancer etiology. Cancer Epidemiol Biomarkers Prev; 20(9); 1902–11. ©2011 AACR.

Objective: Severe immune dysregulation is a strong risk factor for multiple myeloma (MM), and experimental studies have implicated chemokines and pro-angiogenic cytokines in MM pathogenesis. We investigated whether circulating immune markers were prospectively associated with MM and its precursor, monoclonal gammopathy of undetermined significance (MGUS), in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. Methods: We characterized levels of 43 immune markers in banked sera from 243 MM cases, 506 subjects with MGUS (including those with light-chain disease) who did not progress to MM, and 258 MGUS-free controls using Luminex-based multiplex assay panels. Odds ratios (ORs) and 95% confidence intervals (CIs) for MM and MGUS were estimated using multivariate-adjusted polytomous logistic regression models. Results: Relative to MGUS-free controls, we observed statistically significant associations with MM risk for six markers using a false discovery rate of 10%, including several chemokines (MCP-2/CCL8, fourth quartile vs first: OR 2.1; 95% CI 1.2-3.7; Ptrend=0.017 and SDF-1/CXCL12, 2.6; 1.5-4.4; Ptrend=2.9x10-4), angiogenesis markers (ANG-2, 3.0; 1.7-5.3; Ptrend=1.3x10-4 and HGF, 3.0; 1.7-5.1; Ptrend=2.8x10-5), and growth factors (EGF, 3.2; 1.9-5.5; Ptrend=1.9x10-6 and BMP-9, 2.3; 1.4-4.0; Ptrend=3.3x10-4). Associations with these markers remained after restricting to MM cases diagnosed ≥8 years after blood collection although risk estimates were somewhat attenuated (Ptrend≤0.07). MCP-2/CCL8, SDF-1/CXCL12, and HGF were also significantly elevated among those with MGUS compared with MGUS-free controls (Ptrend≤0.04). Four markers were associated with progression from MGUS to MM after adjusting for clinical characteristics such as M-protein concentration, serum free light-chain ratio, and immunoglobulin type (ANG-2, fourth quartile vs first: OR 2.5, 95% CI 1.4-4.6; HGF, 2.6, 1.4-5.0; EGF, 3.3, 1.8-6.0; and BMP-9, 1.9, 1.0-3.4); dose-response trends were statistically significant for all four markers (Ptrend≤0.03). Conclusions: This is, to our knowledge, the first prospective study to investigate circulating immunologic markers in relation to MM risk and progression from MGUS to MM. Our findings provide insights into potential biological mechanisms associated with MM development and may have clinical utility for improving risk stratification models assessing the likelihood of progression from MGUS to clinically manifest MM. Citation Format: Jonathan N. Hofmann, Ola Landgren, Hormuzd Katki, Troy Kemp, Loredana Santo, Charlene McShane, Qing Lan, Nathaniel Rothman, Ligia Pinto, Ruth Pfeiffer, Allan Hildesheim, Mark Purdue. Circulating immunologic markers and risk of multiple myeloma and its precursor disease: A nested case-control study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4943.

We compared systemic and cervical total IgA and IgG during the menstrual cycle among 154 women who attended clinic visits at follicular/early, periovulatory/mid, and luteal/late phases of menstrual cycle. Paired serum and cervical secretions were tested at each visit for total IgA and IgG using ELISA. Geometric mean titers for systemic IgA and IgG were 1.92 and 8.25 mg/mL, respectively. There were no differences in titers by menstrual cycle phase, neither were they correlated to cervical titers (ρ = 0.17 and 0.16, respectively). The lack of correlation between systemic and cervical total IgA and IgG suggests that systemic concentrations are not reflective of cervical levels. (Cancer Epidemiol Biomarkers Prev 2009;18(10):2672–6)

Recent prospective studies conducted in Western populations among mostly Caucasians have suggested that higher levels of soluble CD27 (sCD27) and soluble CD30 (sCD30), two markers indicative of B-cell activation, are associated with risk of non-Hodgkin lymphoma (NHL), with significant associations persisting in individuals who were diagnosed with NHL >10 years after blood collection. However, there are currently no molecular epidemiologic data evaluating whether these biomarkers are associated with NHL risk in East Asian populations, in whom the descriptive characteristics of NHL in terms of subtype distributions and incidence rates are quite different from those in Western countries. To explore potential mechanistic commonalities for NHL in these populations, we conducted a pooled nested case-control study from three prospective studies of Chinese men and women including 218 NHL cases and 218 individually matched controls. Levels of sCD27 and sCD30 were measured in all study subjects at the same time using ELISA. Conditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) according to quartile levels of sCD27 or sCD30 in all study controls. An increased risk of NHL in the pooled population was observed for elevated levels of both sCD27 and sCD30. Compared to the lowest quartile, ORs (95% CIs) for the 2nd, 3rd, and 4th quartiles of sCD27 were 1.60 (0.83-3.09), 1.94 (0.98-3.83), and 4.45 (2.25-8.81), respectively (ptrend = 0.000005). The corresponding ORs (95% CIs) for sCD30 were 1.74 (0.85-3.58), 1.86 (0.94-3.67), and 5.15 (2.62-10.12) (ptrend = 0.0000002). These associations remained statistically significant in individuals diagnosed with NHL 10 or more years after blood draw. Notably, the magnitude of the associations with risk of NHL was very similar to those in Western populations in previous studies. These findings of the similar association between sCD27 or sCD30 and NHL risk across different populations support an important underlying mechanism of B-cell activation in lymphomagenesis. Citation Format: Bryan A. Bassig, Xiao-Ou Shu, Woon-Puay Koh, Yu-Tang Gao, Mark P. Purdue, Lesley M. Butler, Jennifer Adams-Haduch, Yong-Bing Xiang, Troy J. Kemp, Renwei Wang, Ligia A. Pinto, Tongzhang Zheng, Bu-Tian Ji, H. Dean Hosgood, Wei Hu, Gong Yang, Heping Zhang, Wong-Ho Chow, Christopher Kim, Wei Jie Seow, Wei Zheng, Jian-Min Yuan, Qing Lan, Nathaniel Rothman. Soluble levels of CD27 and CD30 are associated with risk of non-Hodgkin lymphoma in a pooled analysis of three prospective cohorts of Chinese men and women in Shanghai and Singapore. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 846. doi:10.1158/1538-7445.AM2015-846

Background: Formaldehyde has been proposed to be a human myeloid leukemogen. However, the exact mechanism for the basis for this link is still debated. We aimed to evaluate whether circulating immune/inflammation markers were different in workers occupationally exposed to formaldehyde. Methods: Using a multiplexed bead-based assay, we measured serum levels of 38 immune/inflammation markers in a cross-sectional study of 43 formaldehyde-exposed and 51 unexposed factory workers in Guangdong, China. Linear regression models adjusting for potential confounders were used to compare marker levels in exposed and unexposed workers. Results: We found significantly lower circulating levels of four markers among exposed factory workers compared to unexposed controls that remained significant after adjusting for multiple comparisons, including thymus and activation regulated chemokine (TARC;52pg/ml in exposed versus 75pg/ml in controls,P = 0.0015), TNF-related apoptosis-inducing ligand (TRAIL;11.7pg/ml in exposed versus 22.1pg/ml in controls,P = 0.011), C-reactive protein (CRP;797.3ng/ml in exposed versus 3269.1ng/ml in controls,P = 0.013) and chemokine (C-X-C motif) ligand 11 (CXCL11;36.2pg/ml in exposed versus 48.4pg/ml in controls,P = 0.017), suggesting immunosuppression among formaldehyde-exposed workers. Conclusions: Our findings are consistent with recently emerging understanding that immunosuppression might be associated with myeloid diseases. These findings, if replicated in a larger study, may provide insights into the mechanism by which formaldehyde promotes leukemogenesis. Citation Format: Qing Lan, Wei Jie Seow, Luoping Zhang, Roel Vermeulen, Xiaojiang Tang, Wei Hu, Bryan A. Bassig, Zhiying Ji, Meredith S. Shiels, Troy J. Kemp, Min Shen, Chuangyi Qiu, Boris Reiss, Laura Beane Freeman, Aaron Blair, Christopher Kim, Weihong Guo, Cuiju Wen, Laiyu Li, Ligia A. Pinto, Hanlin Huang, Martyn T. Smith, Allan Hildesheim, Nathaniel Rothman. Circulating immune/inflammation markers in Chinese workers occupationally exposed to formaldehyde. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 829. doi:10.1158/1538-7445.AM2015-829

Background: Few epidemiologic studies have investigated the role of inflammation in lung cancer etiology. We have carried out 2 independent studies nested within the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial to assess associations between circulating inflammation markers and lung cancer risk, after carefully controlling for smoking. In the discovery study, we showed associations between 11 circulating inflammation markers (CRP, SAA, sTNFRII, IL-1RA, IL-7, TGF-A, ENA 78/CXCL5, MIG/CXCL9, BCA-1/CXCL13, TARC/CCL17 and MDC/CCL22) and prospective lung cancer risk (Shiels, JNCI 2013). To assess the reproducibility of these associations, we have carried out a large, independent replication study. Methods: We designed a replication case-control study nested within the PLCO in 526 lung cancer cases and 625 controls, matched by gender, age, smoking (status, pack-years and time since quit), study visit and years of blood draw and exit. Serum levels of 51 inflammation markers were measured using Luminex bead-based assays. We combined the data from the discovery and replication studies (1052 cases and 1217 controls) and created study-specific marker categories. Conditional logistic regression was used to estimate odds ratios (ORs) overall and stratified by latency and histology in the pooled dataset. Results: In our prior study, 11 inflammation markers were associated with lung cancer risk with ORs ranging from 1.41 to 2.27. Of these 11 markers, C-reactive protein (CRP), serum amyloid A (SAA), soluble tumor necrosis factor receptor 2 (sTNFRII) and monokine induced by gamma interferon (MIG/CXCL9) were associated with prospective lung cancer risk in the replication study with ORs ranging from 1.70 to 2.09 (p-trend across quartiles3 years) (all p-interactions>0.2). Further, each marker was associated with both lung adenocarcinoma and squamous cell carcinoma (all p-trends Conclusions: With results from two independent case-control studies, we have confirmed that pre-diagnostic circulating levels of two acute phase proteins (CRP and SAA), a receptor for a pro-inflammatory cytokine (sTNFRII), and a chemokine (MIG/CXCL9) are each associated with prospective lung cancer risk, after carefully controlling for cigarette smoking. These results provide evidence implicating inflammation in the development of lung cancer. Citation Format: Meredith S. Shiels, Hormuzd A. Katki, Allan Hildesheim, Ruth M. Pfeiffer, Eric A. Engels, Marcus C. Williams, Troy J. Kemp, Neil E. Caporaso, Ligia A. Pinto, Anil K. Chaturvedi. Circulating inflammation markers and subsequent lung cancer risk: A discovery and replication study. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2920. doi:10.1158/1538-7445.AM2014-2920

Background: We and other investigators have recently reported associations between elevated serum concentrations of selected immune markers and subsequent risk of non-Hodgkin lymphoma (NHL). While these findings support a role for subtle immunologic effects in lymphomagenesis, the relatively short length of follow-up after phlebotomy in these studies (10-13 years) limits the ability to rule out reverse causation as an explanation for these findings. To clarify the temporal nature of these associations, we conducted a nested case-control study investigating serum levels of six immune markers and NHL diagnosed up to 23 years after phlebotomy within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study, a cohort with banked fasting serum from 29,133 male Finnish smokers. Methods: We selected 272 first-primary NHL cases diagnosed 2+ years after phlebotomy, identified through linkage with the Finnish Cancer Registry, and 325 controls individually matched to cases on baseline age, phlebotomy date, and number of previous specimen thaws. Serum from these individuals were tested for soluble CD23 (sCD23), sCD27, sCD30, B cell attracting chemokine-1 (BCA-1), soluble tumor necrosis factor receptor-2 (sTNFR-2), and soluble vascular endothelial growth factor receptor-2. Odds ratios (OR) and 95% confidence intervals (CI) relating categories of analyte concentration to NHL risk were computed through conditional logistic regression with adjustment for cigarette pack-years. Polytomous regression models were also fit to compute ORs for the two most common NHL histologic subtypes, chronic lymphocytic leukemia (CLL) and diffuse large B cell lymphoma (DLBCL). Results: We observed NHL associations with elevated pre-diagnostic concentrations of the B cell activation markers sCD23 (highest analyte category vs. lowest: OR 2.4, 95% CI 1.5-3.8, Ptrend = 0.0005) and sCD30 (OR 3.3, 95% CI 1.9-5.5, Ptrend < 0.0001). The associations with sCD23 and sCD30 were apparent for cases diagnosed as far as 15-23 years after blood collection (OR 5.1, 95% CI 1.8-14.6, Ptrend = 0.002 and OR 3.7, 95% CI 1.5-9.3, Ptrend = 0.01 respectively). In subtype-specific analyses, both sCD23 and sCD30 were significantly associated with DLBCL, both overall and in later follow-up periods. sCD23 was significantly associated with CLL, particularly for earlier follow-up periods, while sCD30 was not. Associations with NHL were observed for sCD27, BCA-1, and sTNFR-2 but became weaker or null for cases diagnosed >8 years post-phlebotomy. Conclusions: Our study provides strong evidence that elevated serum sCD23 and sCD30 are associated with future risk of NHL many years after blood collection. Our findings for these B cell activation markers suggest that B cell activation is an important mechanism in NHL development in the general population. Citation Format: Mark P. Purdue, Qing Lan, Troy J. Kemp, Allan Hildesheim, Stephanie J. Weinstein, Demetrius Albanes, Ligia A. Pinto, Nathaniel Rothman. Serum sCD23 and sCD30 associated with non-Hodgkin lymphoma risk as far as 15 to 23 years after blood collection. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5073. doi:10.1158/1538-7445.AM2014-5073

Background: Although severe immune dysregulation is an established risk factor for non-Hodgkin lymphoma (NHL), the etiologic relevance of subclinical immunologic effects is unclear. Few prospective studies have investigated the relationship between circulating immune markers and NHL, and those conducted to date have been limited in sample size, the number of evaluated analytes, or in their ability to examine markers of risk many years preceding disease diagnosis. To better understand this question, we conducted a nested case-control study investigating pre-diagnostic serum levels of 51 immune markers within the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. Methods: Fifty-one immune markers (including cytokines, chemokines, extracellular matrix proteins, growth factors and soluble products of immune activation) were measured in baseline serum from 301 NHL cases diagnosed 5+ years after blood collection (median 8.0 years, range 5.0-13.9) and 301 individually matched controls using Luminex bead-based assay panels. Odds ratios (ORs) and 95% confidence intervals (CIs) relating marker levels with NHL risk were computed using conditional logistic regression modeling. Results: We observed strong associations with NHL for elevated levels of several analytes: a pro-inflammatory marker, soluble tumor necrosis factor receptor 2 (sTNFRII; fourth quartile vs. first: OR 3.4, 95% CI 2.0-5.8; Ptrend = 5.9 x 10−6); a regulator of B-cell traffic, B-cell attracting chemokine 1 (BCA-1; 2.7, 1.7-4.2; Ptrend = 1.1 x 10−5); a regulator of angiogenesis, soluble vascular endothelial growth factor receptor 2 (sVEGFR2; 2.3, CI 1.4-3.9; Ptrend = 0.0005); and the pro-inflammation and apoptotic cytokine TNF-related apoptosis inducing ligand (TRAIL; 1.6, 1.0-2.5; Ptrend = 0.02). These associations remained significant in analyses of cases diagnosed 8-13 years after blood collection and, with the exception of TRAIL, following Bonferroni correction and simultaneous model adjustment. Conclusions: Our findings suggest that elevated levels of sTNFRII, BCA-1, sVEGFR2, and, possibly, TRAIL may reflect biologic processes contributing to lymphomagenesis. Additional research is needed to replicate these findings, identify cell sources of these markers, and elucidate the underlying biologic mechanisms. Citation Format: Mark P. Purdue, Jonathan N. Hofmann, Troy J. Kemp, Anil K. Chaturvedi, Qing Lan, Ju-Hyun Park, Ruth M. Pfeiffer, Allan Hildesheim, Ligia A. Pinto, Nathaniel Rothman. A prospective study of 51 serum immune markers and risk of non-Hodgkin lymphoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2880. doi:10.1158/1538-7445.AM2013-2880

Background: Based on bone marrow (BM) biopsies, increased angiogenesis has been reported in multiple myeloma (MM) and smoldering multiple myeloma (SMM) patients compared with individuals with monoclonal gammopathy of undetermined significance (MGUS). In this prospective clinical trial open for MGUS, SMM and MM patients, we conducted BM biopsies and used immunohistochemistry to define microvessel density (MVD) in every patient; results were compared to results obtained from dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and multiplex ELISA-based serum angiogenic cytokine assays. Patients and Methods: We included 10 MGUS, 10 SMM and 10 MM patients. We performed DCE-MRI of entire lumbosacral spine to compare exchange rate constants (contrast agent transit from the extravascular compartment to the intravascular compartment, Kep; movement of contrast agent from plasma to extravascular extracellular space, Ktrans) with bone marrow biopsies immunostained with CD34 as measures of MVD, and serum angiogenic markers By ELISA to include EGF, Ang2, GCSF, BMP9, endoglin, Leptin, Follistatin, IL8, HGF, HBEGF, PLGF, VEGFC, VEGFD, FGF2, VEGFA. The association among continous parameters was determined by Spearman rank correlation. Trends in continuous parameters according to ordered categorical parameter were determined by Jonckheere-Terpstra test for trend. The parameters between two groups were compared using an exact Wilcoxon rank sum test. Results: Both MVD and Kep increased along the myeloma spectrum (MGUS < SMM < MM). MVD and Kep were strongly correlated (r=0.93 and p=0.002). Ktrans was weakly to moderately well correlated with MVD (r=0.43 p=0.03). Higher Kep values were seen in MM/SMM vs. MGUS patients (median 7.1 vs. 3.9; p=0.08). Similarly, higher MVD values were seen in MM/SMM patients versus MGUS (median 20 vs. 15; p=0.01). As regards serum angiogenic markers, levels of HGF (r=0.45; p=0.02), Ang2(r=0.37 and p=0.06), and VEGFD (r=-0.35 and p=0.07), correlated with Kep. The levels of Ang2 (p=0.02), GCSF (p=0.06), follistatin (p=0.06), HGF (p=0.01), and VEGFA (p=0.02), were elevated in MM/SMM patients in comparison to MGUS. Other angiogenic cytokines did not correlate with MVD or Kep. Conclusions: MVD and Kep (measured by DCE-MRI) were highly statistically correlated. MVD increased along the disease spectrum from MGUS to SMM to MM (p=0.008). Some, but not all, angiogenic biomarkers detected in peripheral blood were associated with increased vascularity in the bone marrow. Citation Format: Manisha Bhutani, Esther Mena, Irina Maric, Esther Tan, Neha Korde, Alexandra R. Berg, Alex R. Minter, Brendan M. Weiss, Liza Lindenberg, Baris Turkbey, Omer Aras, Seth Steinberg, Troy Kemp, Katherine R. Calvo, Peter L. Choyke, Karen Kurdziel, Ola Landgren. Role of bone marrow angiogenesis in myeloma and its precursor disease: a prospective clinical trial. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 369. doi:10.1158/1538-7445.AM2013-369

Background: Ovarian cancer is linked to pro-inflammatory mechanisms through its association with increased lifetime number of ovulations, pelvic inflammatory disease, endometriosis, and exposure to talc and asbestos. Exposures that directly reduce inflammation, such as non-steroidal anti-inflammatory drugs and factors that may reduce exposure to irritants, including tubal ligation and hysterectomy, have been shown to reduce ovarian cancer risk. However, limited data are available linking ovarian cancer to measurements of circulating inflammatory markers. Methods: To evaluate whether circulating levels of cytokines and other immune markers are associated with ovarian cancer risk, we conducted a nested case-control study within the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. Four Luminex based multi-plex panels were used to measure circulating levels of 60 inflammation-related biomarkers in all 150 incident ovarian cancer cases with pre-diagnostic serum samples available and 149 controls matched on age at baseline, race, date and time of day of sample collection. Serum samples were collected between 2 and 13 years before cancer diagnosis. We limited analysis to the 24 markers with at least 67% of values above the lower limit of detection and categorized the markers based on the tertile (n=2 markers) or quartile (n=22 markers) distribution in controls. Coefficients of variation for these 24 markers ranged from 1% to 13%. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using conditional logistic regression in unadjusted models and with further adjustment for parity, oral contraceptive use, menopausal hormone therapy use, cigarette smoking and body mass index. Results: In preliminary analyses, elevated serum levels of IL-8 were associated with increased risk of ovarian cancer [Q4 vs. Q1: adjusted OR 2.62 95% CI (1.15-5.95)]. Elevated serum levels of CRP and TNF-alpha were marginally associated with an increased risk [CRP Q4 vs. Q1: 2.11 (0.97-4.59); TNF-alpha Q4 vs. Q1: 2.27 (0.98-5.28)]. In analyses restricted to serous ovarian cancer, the associations with IL-8 and CRP were strengthened [IL-8 Q4 vs. Q1: 3.47 (1.04-11.55); CRP Q4 vs. Q1: 5.62 (1.54-20.56)]. Elevated IL-8, CRP and TNF-alpha levels were statistically significantly related to increased ovarian cancer risk in analysis restricted to specimens collected at least 5 years before diagnosis. Conclusions: Our preliminary results suggest a positive association between IL-8, CRP and TNF-alpha and increased ovarian cancer risk. Our findings confirm an increased risk of ovarian cancer reported for CRP. Associations between IL-8 and TNF-alpha with ovarian cancer risk have not been previously reported, but both markers have been shown to be involved in ovarian function, specifically follicle rupture and repair, and are detectable in tumor tissue from ovarian cancer patients. These findings suggest that increased inflammation is etiologically important in ovarian carcinogenesis, and argue for additional research to confirm and extend these findings. Citation Format: Britton Trabert, Ligia Pinto, Patricia Hartge, Troy Kemp, Amanda Black, Mark E. Sherman, Louise A. Brinton, Ruth Pfeiffer, Anil Chaturvedi, Allan Hildesheim, Nicolas Wentzensen. Prediagnostic serum levels of inflammation markers and risk of ovarian cancer in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial: Preliminary results. [abstract]. In: Proceedings of the AACR Special Conference on Post-GWAS Horizons in Molecular Epidemiology: Digging Deeper into the Environment; 2012 Nov 11-14; Hollywood, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2012;21(11 Suppl):Abstract nr 22.