02407nas a2200253 4500008004100000022001400041245016800055210006900223260001500292300000700307490000700314520158100321100002401902700001901926700002201945700002101967700001901988700002302007700002002030700002202050700002002072700002502092856003602117 2020 eng d a1471-228800aIncorporating sampling weights into robust estimation of Cox proportional hazards regression model, with illustration in the Multi-Ethnic Study of Atherosclerosis.0 aIncorporating sampling weights into robust estimation of Cox pro c2020 03 14 a620 v203 a

BACKGROUND: Cox proportional hazards regression models are used to evaluate associations between exposures of interest and time-to-event outcomes in observational data. When exposures are measured on only a sample of participants, as they are in a case-cohort design, the sampling weights must be incorporated into the regression model to obtain unbiased estimating equations.

METHODS: Robust Cox methods have been developed to better estimate associations when there are influential outliers in the exposure of interest, but these robust methods do not incorporate sampling weights. In this paper, we extend these robust methods, which already incorporate influence weights, so that they also accommodate sampling weights.

RESULTS: Simulations illustrate that in the presence of influential outliers, the association estimate from the weighted robust method is closer to the true value than the estimate from traditional weighted Cox regression. As expected, in the absence of outliers, the use of robust methods yields a small loss of efficiency. Using data from a case-cohort study that is nested within the Multi-Ethnic Study of Atherosclerosis (MESA) longitudinal cohort study, we illustrate differences between traditional and robust weighted Cox association estimates for the relationships between immune cell traits and risk of stroke.

CONCLUSIONS: Robust weighted Cox regression methods are a new tool to analyze time-to-event data with sampling, e.g. case-cohort data, when exposures of interest contain outliers.

1 aSitlani, Colleen, M1 aLumley, Thomas1 aMcKnight, Barbara1 aRice, Kenneth, M1 aOlson, Nels, C1 aDoyle, Margaret, F1 aHuber, Sally, A1 aTracy, Russell, P1 aPsaty, Bruce, M1 aDelaney, Joseph, A C uhttps://chs-nhlbi.org/node/840502512nas a2200229 4500008004100000022001400041245012000055210006900175260001200244300001000256490000800266520180100274100001902075700002402094700002302118700002002141700002002161700002202181700002002203700002302223856003602246 2020 eng d a1879-148400aInnate and adaptive immune cell subsets as risk factors for coronary heart disease in two population-based cohorts.0 aInnate and adaptive immune cell subsets as risk factors for coro c2020 05 a47-530 v3003 a

BACKGROUND AND AIMS: Cell-mediated immunity is implicated in atherosclerosis. We evaluated whether innate and adaptive immune cell subsets in peripheral blood are risk factors for coronary heart disease.

METHODS: A nested case-cohort study (n = 2155) was performed within the Multi-Ethnic Study of Atherosclerosis (MESA) and the Cardiovascular Health Study (CHS). Cases of incident myocardial infarction (MI) and incident angina (n = 880 total cases) were compared with a cohort random sample (n = 1275). Immune cell phenotypes (n = 34, including CD14 monocytes, natural killer cells, γδ T cells, CD4, CD8 and CD19 lymphocyte subsets) were measured from cryopreserved cells by flow cytometry. Cox proportional hazards models with adjustment for cardiovascular disease risk factors were used to evaluate associations of cell phenotypes with incident MI and a composite phenotype of incident MI or incident angina (MI-angina) over a median 9.3 years of follow-up. Th1, Th2, Th17, T regulatory (CD4CD25CD127), naive (CD4CD45RA), memory (CD4CD45RO), and CD4CD28 cells were specified as primary hypotheses. In secondary analyses, 27 additional cell phenotypes were investigated.

RESULTS: After correction for multiple testing, there were no statistically significant associations of CD4 naive, memory, CD28, or T helper cell subsets with MI or MI-angina in MESA, CHS, or combined-cohort meta analyses. Null associations were also observed for monocyte subsets, natural killer cells, γδ T cells, CD19 B cell and differentiated CD4 and CD8 cell subsets.

CONCLUSIONS: The proportions of peripheral blood monocyte and lymphocyte subsets are not strongly related to the future occurrence of MI or angina in adults free of autoimmune disease.

1 aOlson, Nels, C1 aSitlani, Colleen, M1 aDoyle, Margaret, F1 aHuber, Sally, A1 aLanday, Alan, L1 aTracy, Russell, P1 aPsaty, Bruce, M1 aDelaney, Joseph, A uhttps://chs-nhlbi.org/node/840302649nas a2200265 4500008004100000022001400041245016700055210006900222260001600291300000700307490000700314520178800321100002502109700001902134700002402153700002202177700002002199700002002219700002302239700002202262700002002284700002002304700002302324856003602347 2021 eng d a1471-226100aNatural killer cells, gamma delta T cells and classical monocytes are associated with systolic blood pressure in the multi-ethnic study of atherosclerosis (MESA).0 aNatural killer cells gamma delta T cells and classical monocytes c2021 Jan 22 a450 v213 a

BACKGROUND: Hypertension is a major source of cardiovascular morbidity and mortality. Recent evidence from mouse models, genetic, and cross-sectional human studies suggest increased proportions of selected immune cell subsets may be associated with levels of systolic blood pressure (SBP).

METHODS: We assayed immune cells from cryopreserved samples collected at the baseline examination (2000-2002) from 1195 participants from the multi-ethnic study of atherosclerosis (MESA). We used linear mixed models, with adjustment for age, sex, race/ethnicity, smoking, exercise, body mass index, education, diabetes, and cytomegalovirus titers, to estimate the associations between 30 immune cell subsets (4 of which were a priori hypotheses) and repeated measures of SBP (baseline and up to four follow-up measures) over 10 years. The analysis provides estimates of the association with blood pressure level.

RESULTS: The mean age of the MESA participants at baseline was 64 ± 10 years and 53% were male. A one standard deviation (1-SD) increment in the proportion of γδ T cells was associated with 2.40 mmHg [95% confidence interval (CI) 1.34-3.42] higher average systolic blood pressure; and for natural killer cells, a 1-SD increment was associated with 1.88 mmHg (95% CI 0.82-2.94) higher average level of systolic blood pressure. A 1-SD increment in classical monocytes (CD14CD16) was associated with 2.01 mmHG (95% CI 0.79-3.24) lower average systolic blood pressure. There were no associations of CD4 T helper cell subsets with average systolic blood pressure.

CONCLUSION: These findings suggest that the innate immune system plays a role in levels of SBP whereas there were no associations with adaptive immune cells.

1 aDelaney, Joseph, A C1 aOlson, Nels, C1 aSitlani, Colleen, M1 aFohner, Alison, E1 aHuber, Sally, A1 aLanday, Alan, L1 aHeckbert, Susan, R1 aTracy, Russell, P1 aPsaty, Bruce, M1 aFeinstein, Matt1 aDoyle, Margaret, F uhttps://chs-nhlbi.org/node/865802657nas a2200289 4500008004100000022001400041245010000055210006900155260001600224520177800240100001702018700002402035700002302059700002202082700002102104700002002125700002002145700001902165700002202184700002002206700002302226700002002249700002202269700001702291700002302308856003602331 2022 eng d a2055-582200aAssociation of immune cell subsets with incident heart failure in two population-based cohorts.0 aAssociation of immune cell subsets with incident heart failure i c2022 Sep 123 a

AIMS: Circulating inflammatory markers are associated with incident heart failure (HF), but prospective data on associations of immune cell subsets with incident HF are lacking. We determined the associations of immune cell subsets with incident HF as well as HF subtypes [with reduced ejection fraction (HFrEF) and preserved ejection fraction (HFpEF)].

METHODS AND RESULTS: Peripheral blood immune cell subsets were measured in adults from the Multi-Ethnic Study of Atherosclerosis (MESA) and Cardiovascular Health Study (CHS). Cox proportional hazard models adjusted for demographics, HF risk factors, and cytomegalovirus serostatus were used to evaluate the association of the immune cell subsets with incident HF. The average age of the MESA cohort at the time of immune cell measurements was 63.0 ± 10.4 years with 51% women, and in the CHS cohort, it was 79.6 ± 4.4 years with 62% women. In the meta-analysis of CHS and MESA, a higher proportion of CD4+ T helper (Th) 1 cells (per one standard deviation) was associated with a lower risk of incident HF [hazard ratio (HR) 0.91, (95% CI 0.83-0.99), P = 0.03]. Specifically, higher proportion of CD4+ Th1 cells was significantly associated with a lower risk of HFrEF [HR 0.73, (95% CI 0.62-0.85), <0.001] after correction for multiple testing. No association was observed with HFpEF. No other cell subsets were associated with incident HF.

CONCLUSIONS: We observed that higher proportions of CD4+ Th1 cells were associated with a lower risk of incident HFrEF in two distinct population-based cohorts, with similar effect sizes in both cohorts demonstrating replicability. Although unexpected, the consistency of this finding across cohorts merits further investigation.

1 aSinha, Arjun1 aSitlani, Colleen, M1 aDoyle, Margaret, F1 aFohner, Alison, E1 aBůzková, Petra1 aFloyd, James, S1 aHuber, Sally, A1 aOlson, Nels, C1 aNjoroge, Joyce, N1 aKizer, Jorge, R1 aDelaney, Joseph, A1 aShah, Sanjiv, S1 aTracy, Russell, P1 aPsaty, Bruce1 aFeinstein, Matthew uhttps://chs-nhlbi.org/node/917500763nas a2200217 4500008004100000022001400041245014700055210006900202260001600271100002000287700002400307700002300331700002600354700001900380700002300399700002000422700002200442700002000464700002500484856003600509 2022 eng d a1556-387100aImmune cell subpopulations as risk factors for atrial fibrillation: The Cardiovascular Health Study and Multi-Ethnic Study of Atherosclerosis.0 aImmune cell subpopulations as risk factors for atrial fibrillati c2022 Oct 181 aFloyd, James, S1 aSitlani, Colleen, M1 aDoyle, Margaret, F1 aFeinstein, Matthew, J1 aOlson, Nels, C1 aHeckbert, Susan, R1 aHuber, Sally, A1 aTracy, Russell, P1 aPsaty, Bruce, M1 aDelaney, Joseph, A C uhttps://chs-nhlbi.org/node/924703449nas a2200517 4500008004100000022001400041245015400055210006900209260001200278300001000290490000800300520194800308653002002256653003102276653003102307653001402338653001102352653002202363653001102385653001402396653001702410653001802427653002002445653002602465653000902491653001402500653001102514653002502525100002602550700002102576700001902597700002302616700002402639700002202663700002002685700001702705700001702722700002102739700001702760700002502777700002702802700002202829700002002851700002402871856003602895 2022 eng d a1879-148400aMonocyte subsets, T cell activation profiles, and stroke in men and women: The Multi-Ethnic Study of Atherosclerosis and Cardiovascular Health Study.0 aMonocyte subsets T cell activation profiles and stroke in men an c2022 06 a18-250 v3513 a

BACKGROUND AND AIMS: Despite mechanistic data implicating unresolving inflammation in stroke pathogenesis, data regarding circulating immune cell phenotypes - key determinants of inflammation propagation versus resolution - and incident stroke are lacking. Therefore, we aimed to comprehensively define associations of circulating immune phenotypes and activation profiles with incident stroke.

METHODS: We investigated circulating leukocyte phenotypes and activation profiles with incident adjudicated stroke in 2104 diverse adults from the Multi-Ethnic Study of Atherosclerosis (MESA) followed over a median of 16.6 years. Cryopreserved cells from the MESA baseline examination were thawed and myeloid and lymphoid lineage cell subsets were measured using polychromatic flow cytometry and intracellular cytokine activation staining. We analyzed multivariable-adjusted associations of cell phenotypes, as a proportion of parent cell subsets, with incident stroke (overall) and ischemic stroke using Cox regression models.

RESULTS: We observed associations of intermediate monocytes, early-activated CD4 T cells, and both CD4 and CD8 T cells producing interleukin-4 after cytokine stimulation (T and T, respectively) with higher risk for incident stroke; effect sizes ranged from 35% to 62% relative increases in risk for stroke. Meanwhile, differentiated and memory T cell phenotypes were associated with lower risk for incident stroke. In sex-stratified analyses, positive and negative associations were especially strong among men but null among women.

CONCLUSIONS: Circulating IL-4 producing T cells and intermediate monocytes were significantly associated with incident stroke over nearly two decades of follow-up. These associations were stronger among men and not among women. Further translational studies are warranted to define more precise targets for prognosis and intervention.

10aAtherosclerosis10aCD4-Positive T-Lymphocytes10aCD8-Positive T-Lymphocytes10aCytokines10aFemale10aFollow-Up Studies10aHumans10aIncidence10aInflammation10aInterleukin-410aIschemic Stroke10aLymphocyte Activation10aMale10aMonocytes10aStroke10aT-Lymphocyte Subsets1 aFeinstein, Matthew, J1 aBůzková, Petra1 aOlson, Nels, C1 aDoyle, Margaret, F1 aSitlani, Colleen, M1 aFohner, Alison, E1 aHuber, Sally, A1 aFloyd, James1 aSinha, Arjun1 aThorp, Edward, B1 aLanday, Alan1 aFreiberg, Matthew, S1 aLongstreth, William, T1 aTracy, Russell, P1 aPsaty, Bruce, M1 aDelaney, Joseph, Ac uhttps://chs-nhlbi.org/node/909002665nas a2200313 4500008004100000022001400041245010000055210006900155260001600224520170900240100002001949700002101969700002501990700002002015700002402035700002202059700001502081700002102096700002402117700003002141700002002171700002202191700001902213700002002232700002302252700002002275700002002295856003602315 2023 eng d a1432-082700aAssociation of Immune Cell Subsets with Incident Hip Fracture: The Cardiovascular Health Study.0 aAssociation of Immune Cell Subsets with Incident Hip Fracture Th c2023 Aug 313 a

In this study, we aimed to evaluate the association of innate and adaptive immune cell subsets in peripheral blood mononuclear cells (PBMCs) with hip fracture. To conduct this study, we used data from the Cardiovascular Health Study (CHS), a U.S. multicenter observational cohort of community-dwelling men and women aged ≥ 65 years. Twenty-five immune cell phenotypes were measured by flow cytometry from cryopreserved PBMCs of CHS participants collected in 1998-1999. The natural killer (NK), γδ T, T helper 17 (Th17), and differentiated/senescent CD4CD28 T cell subsets were pre-specified as primary subsets of interest. Hip fracture incidence was assessed prospectively by review of hospitalization records. Multivariable Cox hazard models evaluated associations of immune cell phenotypes with incident hip fracture in sex-stratified and combined analyses. Among 1928 persons, 259 hip fractures occurred over a median 9.7 years of follow-up. In women, NK cells were inversely associated with hip fracture [hazard ratio (HR) 0.77, 95% confidence interval (CI) 0.60-0.99 per one standard deviation higher value] and Th17 cells were positively associated with hip fracture [HR 1.18, 95% CI 1.01-1.39]. In men, γδ T cells were inversely associated with hip fracture [HR 0.60, 95% CI 0.37-0.98]. None of the measured immune cell phenotypes were significantly associated with hip fracture incidence in combined analyses. In this large prospective cohort of older adults, potentially important sex differences in the associations of immune cell phenotypes and hip fracture were identified. However, immune cell phenotypes had no association with hip fracture in analyses combining men and women.

1 aElam, Rachel, E1 aBůzková, Petra1 aDelaney, Joseph, A C1 aFink, Howard, A1 aBarzilay, Joshua, I1 aCarbone, Laura, D1 aSaha, Rick1 aRobbins, John, A1 aMukamal, Kenneth, J1 aValderrábano, Rodrigo, J1 aPsaty, Bruce, M1 aTracy, Russell, P1 aOlson, Nels, C1 aHuber, Sally, A1 aDoyle, Margaret, F1 aLanday, Alan, L1 aCauley, Jane, A uhttps://chs-nhlbi.org/node/947802516nas a2200325 4500008004100000022001400041245014100055210006900196260001300265300000900278490000600287520149900293653001801792653003101810653002401841653002201865653002801887653002301915100001901938700002301957700002101980700002002001700002002021700002402041700002202065700002002087700002402107700002302131856003602154 2023 eng d a2398-923800aCirculating differentiated and senescent lymphocyte subsets and incident diabetes risk in older adults: The Cardiovascular Health Study.0 aCirculating differentiated and senescent lymphocyte subsets and  c2023 Jan ae3840 v63 a

INTRODUCTION: Cellular senescence is a feature of aging implicated in the pathophysiology of diabetes mellitus (DM). Whether senescent lymphocytes are associated with the future occurrence of DM is uncertain.

METHODS: We used cryopreserved peripheral blood mononuclear cells collected from 1860 Cardiovascular Health Study participants (average age 80.2 years) and flow cytometry immunophenotyping to evaluate the longitudinal relationships of naive (CD45RA ), memory (CD45RO ), senescent (CD28 ), and T effector memory RA (TEMRA) (CD28 CD57 CD45RA ) CD4 and CD8 T cells, and memory B cells (CD19 CD27 ), with the risk of incident DM. In exploratory analyses we evaluated the relationships of 13 additional innate lymphocyte and CD4 and CD8 subsets with incident DM risk.

RESULTS: Over a median follow-up time of 8.9 years, 155 cases of incident DM occurred. In Cox models adjusted for demographic variables (age, sex, race, study site and flow cytometry analytical batch) or diabetes risk factors (demographic variables plus education, body mass index, smoking status, alcohol use, systolic blood pressure, hypertension medication use and physical activity), no significant associations were observed for any CD4 , CD8 or CD19 cell phenotypes with incident DM.

CONCLUSIONS: These results suggest the frequencies of naive, memory and senescent T cells and memory B cells are not strongly associated with incident DM risk in older adults.

10aCD28 Antigens10aCD8-Positive T-Lymphocytes10aCellular Senescence10aDiabetes Mellitus10aLeukocytes, Mononuclear10aLymphocyte Subsets1 aOlson, Nels, C1 aDoyle, Margaret, F1 aBůzková, Petra1 aHuber, Sally, A1 ade Boer, Ian, H1 aSitlani, Colleen, M1 aTracy, Russell, P1 aPsaty, Bruce, M1 aMukamal, Kenneth, J1 aDelaney, Joseph, A uhttps://chs-nhlbi.org/node/9240