Patterns of IgA Autoantibody Generation, Inflammatory Responses and Extracellular Matrix Metabolism in Patients with Alcohol Use Disorder

Recent data have emphasized the role of inflammation and intestinal immunoglobulin A (IgA) responses in the pathogenesis of alcoholic liver disease (ALD). In order to further explore such associations, we compared IgA titers against antigens targeted to ethanol metabolites and tissue transglutaminase with pro- and anti-inflammatory mediators of inflammation, markers of liver status, transferrin protein desialylation and extracellular matrix metabolism in alcohol-dependent patients with or without liver disease and in healthy controls. Serum IgAs against protein adducts with acetaldehyde (HbAch-IgA), the first metabolite of ethanol, and tissue transglutaminase (tTG-IgA), desialylated transferrin (CDT), pro- and anti-inflammatory cytokines, markers of liver status (GT, ALP) and extracellular matrix metabolism (PIIINP, PINP, hyaluronic acid, ICTP and CTx) were measured in alcohol-dependent patients with (n = 83) or without (n = 105) liver disease and 88 healthy controls representing either moderate drinkers or abstainers. In ALD patients, both tTG-IgA and HbAch-IgA titers were significantly higher than those in the alcoholics without liver disease (p < 0.0005 for tTG-IgA, p = 0.006 for Hb-Ach-IgA) or in healthy controls (p < 0.0005 for both comparisons). The HbAch-IgA levels in the alcoholics without liver disease also exceeded those found in healthy controls (p = 0.0008). In ROC analyses, anti-tTG-antibodies showed an excellent discriminative value in differentiating between ALD patients and healthy controls (AUC = 0.95, p < 0.0005). Significant correlations emerged between tTG-IgAs and HbAch-IgAs (rs = 0.462, p < 0.0005), CDT (rs = 0.413, p < 0.0001), GT (rs = 0.487, p < 0.0001), alkaline phosphatase (rs = 0.466, p < 0.0001), serum markers of fibrogenesis: PIIINP (rs = 0.634, p < 0.0001), hyaluronic acid (rs = 0.575, p < 0.0001), ICTP (rs = 0.482, p < 0.0001), pro-inflammatory cytokines IL-6 (rs = 0.581, p < 0.0001), IL-8 (rs = 0.535, p < 0.0001) and TNF-α (rs = 0.591, p < 0.0001), whereas significant inverse correlations were observed with serum TGF-β (rs = −0.366, p < 0.0001) and CTx, a marker of collagen degradation (rs = −0.495, p < 0.0001). The data indicate that the induction of IgA immune responses toward ethanol metabolites and tissue transglutaminaseis a characteristic feature of patients with AUD and coincides with the activation of inflammation, extracellular matrix remodeling and the generation of aberrantly glycosylated proteins. These processes appear to work in concert in the sequence of events leading from heavy drinking to ALD.Peer reviewe

Blood Cell Responses Following Heavy Alcohol Consumption Coincide with Changes in Acute Phase Reactants of Inflammation, Indices of Hemolysis and Immune Responses to Ethanol Metabolites

Aberrations in blood cells are common among heavy alcohol drinkers. In order to shed further light on such responses, we compared blood cell status with markers of hemolysis, mediators of inflammation and immune responses to ethanol metabolites in alcohol-dependent patients at the time of admission for detoxification and after abstinence. Blood cell counts, indices of hemolysis (LDH, haptoglobin, bilirubin), calprotectin (a marker of neutrophil activation), suPAR, CD163, pro- and anti-inflammatory cytokines and autoantibodies against protein adducts with acetaldehyde, the first metabolite of ethanol, were measured from alcohol-dependent patients (73 men, 26 women, mean age 43.8 ± 10.4 years) at baseline and after 8 ± 1 days of abstinence. The assessments also included information on the quantities of alcohol drinking and assays for biomarkers of alcohol consumption (CDT), liver function (AST, ALT, ALP, GGT) and acute phase reactants of inflammation. At baseline, the patients showed elevated values of CDT and biomarkers of liver status, which decreased significantly during abstinence. A significant decrease also occurred in LDH, bilirubin, CD163 and IgA and IgM antibodies against acetaldehyde adducts, whereas a significant increase was noted in blood leukocytes, platelets, MCV and suPAR levels. The changes in blood leukocytes correlated with those in serum calprotectin (p < 0.001), haptoglobin (p < 0.001), IL-6 (p < 0.02) and suPAR (p < 0.02). The changes in MCV correlated with those in LDH (p < 0.02), MCH (p < 0.01), bilirubin (p < 0.001) and anti-adduct IgG (p < 0.01). The data indicates that ethanol-induced changes in blood leukocytes are related with acute phase reactants of inflammation and release of neutrophil calprotectin. The studies also highlight the role of hemolysis and immune responses to ethanol metabolites underlying erythrocyte abnormalities in alcohol abusers.publishedVersionPeer reviewe

Separate and Combined Effects of Lifestyle Risk Factors on Biomarkers of Liver Function, Inflammation and Lipid Status

Elintapoihin liittyvät terveysongelmat yleistyvät nyky-yhteiskunnissa. Lukuisat elintapojen riskitekijät – kuten haitallinen alkoholinkäyttö, tupakointi, ylipaino, fyysinen passiivisuus ja puutteellinen ruokavalio – voivat aiheuttaa kroonisia sairauksia ja elin- iän lyhentymistä. Sen sijaan terveellisten elintapojen omaksumisen on osoitettu liittyvän pidentyneeseen eliniänodotteeseen. Tässä tutkimuksessa selvitettiin erilaisten elintapatekijöiden yhteyttä maksan toimintaa kuvaaviin maksaentsyymiaktiivisuuksiin, tulehdustilojen biomarkkeriin sekä seerumin lipidiprofiiliin käyttäen laajaa kansallista väestötutkimusaineistoa (FINRISKI). Tutkimusaineisto on kerätty kuudelta eri alueelta Suomessa vuosina 1997, 2002 ja 2007. Tutkimuksen otos, 22 327 perustervettä henkilöä iältään 25–74 vuotta, poimittiin satunnaistettuna otoksena ikä- ja sukupuoliryhmittäin. Tiedot terveydentilasta, alkoholinkäytöstä, tupakoinnista, fyysisestä aktiivisuudesta ja kahvinkulutuksesta kerättiin haastatteluin ja kyselylomakkein. Fyysisiä mittauksia käyttäen kerättiin tiedot painosta ja pituudesta sekä vyötärönympäryksestä. Tutkimukseen osallistujat raportoivat alkoholinkäyttönsä viimeisimmän 12 kuukauden ajalta, ja käyttömäärien mukaan heidät jaettiin absolutisteihin sekä Maailman terveysjärjestön WHO äskettäin määrittelemiin alkoholinkäytön riskikategorioihin; vähäisen, kohtalaisen, korkean sekä erittäin korkean riskin kategoriaan. Lisäksi osallistujat ryhmiteltiin sen mukaan, miten usein juominen oli ollut humalahakuista. Seerumin maksaentsyymit (gamma-glutamyylitransferaasi GT ja alaniiniaminotransferaasi ALAT), C-reaktiivinen proteiini (CRP) ja lipidiprofiili määritettiin vakiintuneilla kliiniskemiallisilla menetelmillä. Eri elintapatekijöiden (alkoholinkäyttö, tupakointi, fyysinen passiivisuus sekä ylipaino ja lihavuus) riskipisteet määritettiin asteikolla 0–8. Riskipisteiden mukaan osallistujat jaoteltiin ryhmiin ja näin pystyttiin arvioimaan eri elintapatekijöiden yhteisvaikutuksia. Alkoholinkäytön mukainen WHO:n riskikategoria oli melko lineaarisesti yhteydessä kohonneisiin GT-, ALAT- ja CRP-arvoihin. Alkoholinkäyttö vaikutti myös merkitsevästi poikkeavien lipidiarvojen esiintymiseen. Nämä havainnot säilyivät merkitsevinä GT-, ALAT- ja lipidiarvojen osalta, vaikka aineisto vakioitiin iällä, vyötärönympärysmitalla, liikunnan määrällä, tupakoinnilla ja kahvin kulutuksella. Humalahakuisen juomisen useus oli merkitsevästi yhteydessä GT-aktiivisuuksiin sekä miehillä (p < 0.0005) että naisilla (p < 0.0005) ja ALAT- aktiivisuuksiin miehillä (p < 0.0005). Alkoholin kokonaiskulutuksen mukaan vähäisen riskin kategoriaan kuuluvilla osallistujilla, jotka joivat humalahakuisesti useammin kuin kerran kuukaudessa, oli selvästi korkeammat GT- (p < 0.0005) ja ALAT-aktiivisuudet (p < 0.0005) kuin saman riskikategorian alkoholinkäyttäjillä, joiden juominen ei ollut humalahakuista. Epäsuotuisien elintapariskitekijöiden kokonaismäärän sekä seerumin GT-, ALAT-, CRP-, kolesteroli-, HDL-, LDL- ja triglyseridiarvojen välillä havaittiin yhdenmukaisia annos-vastesuhteita. Verrattuna niihin, joilla ei ollut riskitekijöitä, kaikkien tutkimuksen kohteena olleiden biomarkkereiden osalta viite- tai tavoitearvoista poikkeavien arvojen monimuuttujakorjatut ristitulosuhteet (OR) olivat merkitsevästi korkeammat niillä, joiden riskipisteiden summa oli kaksi tai enemmän. Tarkasteltaessa ryhmää, jossa oli eniten riskitekijöitä, selvimmät ristitulosuhteiden nousut havaittiin miesten seerumin GT-aktiivisuuksissa: OR 26.6 (12.4–57.0), ALAT-aktiivisuuksissa: OR 40.3 (5.3–307.8), CRP-arvoissa: OR 16.2 (7.8–33.7) ja triglyseridiarvoissa: OR 14.4 (8.6–24.0). Rasvamaksaan viittaavan rasvamaksaindeksin eli FLI-arvon ≥ 60 esiintyminen lisääntyi miehillä 2.4 %:sta 81.9 %:iin, kun kokonaisriskipisteet nousivat 0 pisteestä 7–8 pisteeseen (p < 0.0005) ja naisilla vastaavasti 0 %:sta 73.5 %:iin (p < 0.0005). Merkittävimmät yksittäiset vaikutukset rasvamaksaan viittaavan FLI-arvon (≥ 60) todennäköisyyteen havaittiin fyysisellä passiivisuudella (p < 0.0005 molemmilla sukupuolilla) ja alkoholinkulutuksella, erityisesti miehillä (p < 0.0005). Tutkimus osoittaa, että laboratoriokokeiden systemaattinen käyttö potilaiden seurannassa saattaa parantaa elintapoihin ja käyttäytymiseen liittyvien terveysriskien arviointia. Tutkimuksen tulokset myös korostavat humalahakuisen alkoholinkäytön mahdollisia haitallisia seurauksia maksan toimintaan jopa niillä, joiden alkoholin kokonaiskulutus on vähäisen riskin kulutustasoa. Useiden epäsuotuisien elintapariskitekijöiden samanaikaiseen esiintymiseen osoitettiin liittyvän selviä poikkeavuuksia maksan toimintaa, elimistön tulehdustilaa ja lipidiprofiilia kuvaavissa laboratoriotutkimuksissa sekä suuri maksan rasvoittumisen todennäköisyys. Laboratoriotutkimusten käyttö voi olla hyödyllistä myös interventioissa, joiden päämääränä on ylläpitää terveellisiä elintapoja.Health problems associated with lifestyle are becoming increasingly common in modern societies. The main lifestyle risk factors which contribute to the incidence of chronic diseases and premature death include alcohol drinking, cigarette smoking, excess body weight, physical inactivity and poor diet. On the other hand, a lack of lifestyle-related risk factors has been shown to be associated with prolonged life expectancy. The present work explores the associations between lifestyle risk factors and biomarkers of liver function, inflammation and lipid status in a large population- based sample (the National FINRISK Study). The material had been collected from six geographical areas in Finland during the years 1997, 2002 and 2007 and provided an age and gender-stratified random sample which included 22,327 apparently healthy individuals aged 25–74 years. Data on health status, alcohol consumption, smoking, physical activity and coffee drinking were collected from structured interviews and questionnaires, and, weight, height and waist circumference were ascertained by means of physical measurements. Self-reported alcohol consumption data for the past 12 months were used to classify the participants into subgroups of abstainers and World Health Organization (WHO) risk drinking categories representing low, moderate, high and very high risk drinkers. The participants were also classified into subgroups according to their frequencies of binge drinking. Serum liver enzymes (gamma-glutamyltransferase (GGT), alanine aminotransferase (ALT), C-reactive protein (CRP) and lipid profiles were measured using standard laboratory procedures. Risk scores for the lifestyle factors (alcohol consumption, cigarette smoking, physical inactivity and excess body weight) were established on a 0–8 scale and used to classify the population into lifestyle-related risk categories, which also allowed estimation of the joint effects of the various lifestyle factors. The WHO risk drinking category was fairly well linearly related to the occurrence of elevated GGT, ALT and CRP values, and alcohol drinking was also a significant determinant of serum lipid abnormalities. Significantly higher odds for abnormal GGT, ALT and lipid profiles were found in the alcohol drinkers after adjustment for age, waist circumference, physical activity, smoking and coffee intake, while the frequency of binge-type drinking showed a significant association with GGT levels in both men (p < 0.0005) and women (p < 0.0005) and with ALT in men (p < 0.0005). Even among the individuals with low risk total alcohol consumption, higher GGT (p < 0.0005) and ALT (p < 0.0005) activities were observed in those with binge drinking episodes more than once a month than in those with no such episodes. Distinct dose-response associations were found between the total number of lifestyle-related risk factors and serum ALT, GGT, CRP, cholesterol, high-density lipoprotein, low-density lipoprotein and triglycerides (p < 0.0005 for a linear trend in all comparisons). When compared with the subjects without any risk factors, the multivariable-adjusted odds ratios for abnormalities in all biomarkers were significantly higher in those with a risk score of two or more. The most notable increases in ORs in the subjects with high numbers of risk factors were observed among men with respect to serum GGT: 26.6 (12.4–57.0), ALT: 40.3 (5.3–307.8), CRP: 16.2 (7.8–33.7) and serum triglycerides: 14.4 (8.6–24.0). The occurrence of a fatty liver index (FLI) ≥ 60 indicating the presence of fatty liver, increased from 2.4% in men with zero risk factors to 81.9% in those with a risk score of 7–8 (p < 0.0005 for a linear trend) and from 0% to 73.5% in women (p < 0.0005). The most striking impacts on the likelihood of FLI ≥ 60 were observed for physical inactivity (p < 0.0005 for both genders) and alcohol consumption (p < 0.0005 for men). The data indicate that systematic use of laboratory tests may improve the assessment of health risks related to lifestyle and behaviour. These results also emphasize the adverse effects of binge-type alcohol drinking on hepatic function even in individuals with low-risk overall alcohol consumption. Combinations of several unfavourable lifestyle factors are associated with distinct abnormalities in laboratory tests for liver function, inflammation and lipid status and a high likelihood of hepatic steatosis. The use of biomarkers could also benefit the assessment of interventions aimed at maintaining a healthy lifestyle

Liver enzymes in alcohol consumers with or without binge drinking

Abstract Background: While alcohol use is linked with a wide variety of health problems, the question of whether differences in drinking patterns could yield different outcomes has remained unclear. Patients and methods: We measured liver enzymes (ALT, GGT) from alcohol consumers with or without binge drinking from a population-based sample in Finland, where binge-type drinking is common. Data on alcohol use, diet, body weight, lifestyle (smoking, coffee consumption, physical activity), and health status were collected from 19225 subjects (9492 men, 9733 women), aged 25–74 years. The participants were subsequently classified to subgroups, both according to the frequencies of binge drinking and the amounts of regular alcohol intake (low-, medium-, and high-risk drinking). Results: The quantity of regular alcohol use was roughly linearly related with GGT and ALT activities. ANOVA analyses of the trends according to the frequency of binge drinking showed a significant GGT increase in both men (p &lt; 0.0005) and women (p &lt; 0.0005), and a significant increase of ALT in men (p &lt; 0.0005). In those with low-risk overall consumption, markedly higher GGT (p &lt; 0.0005) and ALT (p &lt; 0.0005) occurred in those with binge drinking more than once a month, compared with those with no such occasions. Binge drinking occurring ≤1/month also resulted in higher GGT (p &lt; 0.0005) and ALT (p &lt; 0.05) activities. Conclusions: These results emphasize possible adverse consequences of binge drinking on hepatic function even in those with low-risk overall consumption. The pattern of drinking should be more systematically implicated in clinical recommendations for drinking reduction

Comparison of serum calprotectin, a marker of neutrophil activation, and other mediators of inflammation in response to alcohol consumption

Abstract Aims: Previous studies have indicated that heavy alcohol intake stimulates inflammation and impairs the body's ability to regulate inflammation. The aim of this study was to compare changes in neutrophil calprotectin and a wide spectrum of other inflammatory mediators in response to heavy alcohol drinking. Methods: Serum calprotectin (a marker of neutrophil activation), suPAR, CD163, and pro- (IL-6, IL-8, TNF-α) and anti-inflammatory (IL-10, TGF-β) cytokines were measured from 61 alcohol-dependent subjects (46 men, 15 women, mean age 43.6 ± 11.0 years) at the time of admission for detoxification and after 8 ± 2 days of abstinence. These biomarkers were also measured from age- and sex-matched healthy controls representing abstainers or light drinkers. The clinical assessments included detailed clinical interviews on the amounts and patterns of alcohol consumption and assays for biomarkers of alcohol consumption (GGT, CDT, MCV, GGT-CDT) and liver function (AST, ALT). Results: The subjects with alcohol use disorder showed significantly higher concentrations of serum calprotectin (p &lt; 0.0005), suPAR (p &lt; 0.01), CD163 (p &lt; 0.01), IL-6 (p &lt; 0.0005), IL-8 (p &lt; 0.0005), TNF-α (p &lt; 0.001), and IL-10 (p &lt; 0.0005) than healthy controls. These inflammatory mediators, except for CD163, remained elevated after the 8 ± 2-day period of supervised abstinence, which resulted in significant decreases in the biomarkers of alcohol consumption and indices of liver status. The AUC (0.855) for serum calprotectin in differentiating between the heavy drinkers and healthy controls was equal or equivalent with those of the conventional biomarkers of alcohol consumption (GGT:0.835 or CDT:0.803). Conclusions: The data indicate that neutrophil calprotectin is released in response to heavy alcohol intake in a sensitive manner and may be associated with perpetuation of inflammation in patients with alcohol use disorder. Serum calprotectin may also prove to be a useful biomarker for inflammatory activity in alcohol-consuming patients

Impacts of unfavourable lifestyle factors on biomarkers of liver function, inflammation and lipid status

Abstract Background: Adopting a healthy lifestyle is associated with prolonged life expectancy. The main modifiable lifestyle-related risk factors are hazardous alcohol drinking, smoking, excess body weight and lack of physical activity. Our aim was to estimate the impact of unfavourable lifestyle factors on abnormalities in laboratory tests reflecting liver status, inflammation and lipid metabolism in a population-based cross-sectional study. Methods: The study included 22,273 participants (10,561 men, 11,712 women) aged 25–74 years from the National FINRISK Study. Data on alcohol use, smoking, body weight, and physical activity were recorded from structured interviews. The risk scores for the various life style factors were established on a 0–8 scale and used to stratify the population in classes to allow estimates of their joint effects. Serum liver enzymes (GGT, ALT), C-reactive protein (CRP) and lipid profiles were measured using standard laboratory techniques. Results: Consistent dose-response relationships were observed between the number of unfavourable risk factors and serum levels of GGT, ALT, CRP, cholesterol, HDL, LDL and triglycerides (p &lt; 0.0005 for linear trend in all comparisons). When compared with those with zero risk factors, the multivariable-adjusted odds ratios (ORs) for abnormalities in all biomarkers were significantly higher in those with a sum of risk score two or more. The most striking increases in ORs in the group with the highest numbers of risk factors were observed among men in serum GGT: 26.6 (12.4–57.0), ALT: 40.3 (5.3–307.8), CRP: 16.2 (7.8–33.7) and serum triglycerides: 14.4 (8.6–24.0). Conclusions: The data support the view that the presence of unfavourable life style risk factors is associated with distinct abnormalities in laboratory tests for liver function, inflammation and lipid status. Such biomarkers may prove to be of value in the assessment of interventions aimed at reducing unfavourable risk factors and in helping individuals in long-term maintenance of lifestyle modifications