Anti-annexin V autoantibodies and vascular abnormalities in systemic sclerosis: a longitudinal study

Abstract Background: Annexins are a group of conserved proteins which exert several regulatory functions on various cellular activities. Increased frequency and levels of antibodies against annexin V have already been observed in several autoimmune diseases including systemic sclerosis (SSc), but their role as a vascular biomarker is unknown. The aim of this study was to determine the serum levels and the dynamical behavior of anti-annexin V antibodies over a 24 months follow-up in patients with SSc. Methods: In this bicentric cross-sectional study, 70 patients with SSc were consecutively selected from March 2016 to April 2017. Demographic and clinical features, including the presence of active DUs, were collected. Serum anti-annexin V IgG and IgM antibodies were measured at baseline and after 6, 12 and 24 months of follow-up. Videocapillaroscopy was performed in all patients. Results: Among the 70 SSc patients included anti-annexin V IgG was found in 11 patients (15.7%) (range of 15.88-39.48 U/mL) and anti-annexin V IgM in 10 patients (14.3%) (range of 14.16-22.69 U/mL) at baseline. During follow-up, the number of patients who were positive for anti-annexin V IgG and IgM remained stable over 24 months. Among the patients with positive anti-annexin V IgG at baseline the frequency of patients with necrosis or amputation of extremities, forced vital capacity less than 70% and pulmonary arterial hypertension (PAH) was significantly higher than in patients with negative anti-annexin V IgG antibodies. Patients with anti-annexin V IgG had also a higher Raynaud's Condition Score and a higher Health Assessment Questionnaire Disability Index (HAQ-DI) than patients without these antibodies at baseline. Patients with positive anti-annexin V IgM at baseline presented a higher frequency of PAH, compared to those with negative anti-annexin V IgM at baseline. Conclusions: Anti-annexin V antibodies are stable and do not change their positivity during a 24 month follow-up in SSc patients. Anti-annexin V IgG was associated with more severe interstitial lung involvement and digital microangiopathy, and patients with anti-annexin V IgG or IgM had a higher occurrence of PAH indicating an association of these biomarker with more severe disease.(AU)

Serum Annexin V and Anti-Annexin V levels and their relationship with metabolic parameters in patients with type 2 diabetes

SUMMARY BACKGROUND We investigated the serum annexin V and anti-annexin V levels and their relationship with metabolic parameters in patients recently diagnosed type 2 diabetic. METHODS A total of 143 patients recently diagnosed type 2 diabetes and 133 control subjects were included in the study. Body mass index (BMI), hs-CRP, HOMA-IR, carotid intima-media thickness, and serum levels of annexin V and anti-annexin V were investigated. RESULTS HOMA-IR, serum hs-CRP, and carotid intima-media thickness were found to be statistically significant. The Pearson correlation analysis revealed a statistically significant positive relationship between the carotid intima-media thickness and the annexin V level (r=0.29, p=0.006*). A statistically significant positive relationship was also detected between the Annexin V level and level of serum hs-CRP (r=0.29 p=0.006*). CONCLUSION A positive relationship was observed between the carotid intima-media thickness and annexin V at the end of our investigation. In this regard, we also believe that serum levels of annexin V may be increased for cardiovascular protection in the elevation of carotid intima-media thickness.

RESUMO OBJETIVO Investigar os níveis séricos de anexina V e antianexina V e sua relação com os parâmetros metabólicos em pacientes diabéticos tipo 2 recém-diagnosticados. MÉTODOS Foram incluídos no estudo 143 pacientes e 133 controles com diabetes tipo 2 recém-diagnosticado. O índice de massa corporal (IMC), PCR-as, Homa-IR, espessura íntima média carotídea e níveis séricos de anexina V e antianexina V foram investigados. RESULTADOS O Homa-IR, a PCR-s do soro e a espessura média da carótida foram estatisticamente significantes. A análise de correlação de Pearson revelou uma relação positiva estatisticamente significante entre a espessura média da carótida e anexina V (r=0,29; p=0,006 *). Foi também detectada uma relação positiva estatisticamente significativa entre o nível de anexina V e o nível sérico de PCR-as (r=0,29, p=0,006*). CONCLUSÃO Também foi observada uma relação positiva entre a espessura média da carótida e anexina V no final da nossa investigação. A esse respeito, também pensamos que os níveis séricos de anexina V podem ser aumentados para proteção cardiovascular na elevação da espessura média da carótida.

Annexin V and adhesion molecules expressions on erythrocytes in sickle cell anemia patients treated with hydroxyurea

To evaluate hydroxyurea [HU] induced changes in the expression of phosphatidylserine as well as various cell surface adhesion molecules in red cells of sickle cell anemia patients. The red cell membrane expression of CD36, CD49d. CD71 and annexin V were tested using flow cytometry and results were correlated with percent of HbF, as measured by 1-IPLC, as well as other hematological variables. The present study included 60 patients with sickle cell anemia [SCA], 40 of them underwent treatment with HU for a period of 12-18 months and the other 20 patients were newly diagnosed as [SCA], beside 20 healthy subjects as controls. Hydroxyurea caused a significant increase in Hb concentration, MCV values and HbF percentage. HU effectively decreased total teaeocyc count, total platelet counts. reticulocytes percentage and expression of CD36, CD71, CD49d and annexin V from pretreatment values. However, these values continued to be significantly higher in treated patients than those of controls. A significant positive correlation was found between Hb concentration and percent of HbF and also between reticulocyte counts and CD49d expression. A significant negative correlation was found between Hb concentration and percent of reticulocytes, CD36, CD49d, Annexin and CD71 expressions. A significant negative correlation was also found between reticulocyte percent and percent of HbF, CD36, Annexin and CD7I expressions. Treatment of sickle cell anemia with hydroxyurea causes a significant increase in HbF together with a significant reduction of CD36, CD7I and CD49d adhesion molecules and annexinV expressions on the surface of erythrocytes. These findings could have clinical relevance, as inhibition of adhesion receptor expression or activity may explain part of the beneficial effects of HU

Placental histomorphology in unexplained foetal loss with thrombophilia.

BACKGROUND & OBJECTIVES: Acquired and genetic thrombotic conditions, both organ and non organ specific, are associated with increased foetal wastage. This study was carried out to examine the placenta from women with abnormal pregnancies and a history of unexplained foetal loss, and to associate with maternal thrombophilia status. METHODS: Placentas from eight women with history of unexplained foetal loss were analyzed for histopathological characteristics. All the women were simultaneously screened for the common acquired and genetic thrombophilia markers i.e., lupus anticoagulants ( LA), IgG / IgM antibodies for anticardiolipin (ACA), beta2 glycoprotein 1 (beta2GPI) and annexin V, protein C (PC), protein S (PS), antithrombin III (AT III), factor V Leiden ( FVL) mutation, prothrombin (PT) gene G20210A, methylene tetrahydrofolate reductase (MTHFR) C 677T, endothelial protein C receptor (EPCR) 23 bp insertion and plasminogen activator inhibitor ( PAI-1 4G/5G) polymorphisms RESULTS: Six of eight women were positive for one or more thrombophilia markers. The placenta in all the cases except one, showed the characteristic features of infarct fibrin deposition and calcification. Among two women who were negative for thrombophilia, one showed clear evidence of thrombus in the placental sections while the other did not show any characteristic infarcts in the placental sections. INTERPRETATION & CONCLUSION: Our findings showed that the histopathological examination of the placentas confirmed thrombophilia as the aetiological cause of thrombosis in 6 of the 8 women. The presence of thrombus in a negative thrombophilia woman suggests yet unidentified thrombophilia markers or probably non-haemostatic factors causing thrombosis.

Keratinocyte and lymphocyte apoptosis: relation to disease outcome in systemic lupus erythematosus patients with and without cutaneous manifestations

To investigate the relation of keratinocyte and lymphocyte apoptosis and macrophage function to disease activity and severity in SLE patients with and without cutaneous manifestations. Fifty SLE patients [25 with cutaneous manifestations [group I], 25 without cutaneous manifestations [group II]] and 20 normal controls [group III] were studied. SLEDAI score was used to assess lupus activity. Peripheral lymphocyte apoptosis by Annexin V, macrophage function by serum neopterin and immunohistochemical detection of apoptotic cells in the skin by p.53 were done. Renal biopsy was done in indicated cases. Mean SLEDAI score was significantly higher in group I than II [18.6 +/- 6, 8.8 +/- 2.7 respectively, p<0.001]. The mean percentage of peripheral apoptotic lymphocytes was significantly higher in group I compared to group II and III [55.3 +/- 21.4, 25.6 +/- 8. 7 and 19.4 +/- 3.2 respectively, p<0.001] and so was the serum neopterin level [27.5 +/- 7.3, 14.9 +/- 2.7, 9.4 +/- 1.1 respectively, p<0.001]. The mean number of P53+ve keratinocytes of group I was significantly higher than group II and III [20.6 +/- 5.4, 1.6+0.5, 1.7 +/- 0.4 respectively, p<0.001]. A higher percentage of class IV and V glomerulonephritis was found in group I [47%, 26%, respectively] compared to group II [11% both] [p<0.001]. The mean number of p53+ve keratinocyte showed a significant positive correlation to SLEDAI score, percentage of peripheral apoptotic lymphocytes and serum neopterin [p<0.001]. Accumulation of apoptotic keratinocytes and lymphocytes in SLE seems to be crucial in the pathogenesis of skin lesions and in triggering systemic disease activity and organ damage

Annexin V as index of apoptosis in aorta of STZ induced diabetic rats

Diabetes mellitus [DM] is a chronic disease associated with hyperglycemia and the production of reactive oxidative intermediates and a disturbed antioxidant defense mechanism. Apoptosis or programmed cell death is a fundamental component of tissue differentiation and development, it also plays a central role in DM. The highly regulated mechanism of apoptosis involves an externalization process of phosphatidylserine [Ps]. Annexin V binds with high affinity to Ps-exposing apoptotic cell and can inhibit thereby the procoagulant and proinflammatory activities of the dying cell. Heat shock proteins have been shown to protect organism in vitro and in vivo against oxidative stress which plays a role in apoptosis in DM. HSP72 inhibits mitochondria! cytochrome release and subsequent caspase activation that leads to apoptosis. Fifty male albino rats weighing 170-200gm were used in this study. They were divided in to 5 equal groups, each of 10 rats. Group I: Control group. Group II: Diabetic group. Type 2 diabetes mellitus was induced by intraperitoneal injection of streptozotocin at a dose of 40mg/kg body weight. Group III: Diabetic group, treated with 1 unit insulin injected subcutaneously daily. Group IV: Diabetic group, receiving vit. E 600mg/kg B.Wt injected intramuscularly, 3 times/week, All groups were sacrified one month after the beginning of the experiment. Blood samples were obtained from retro-orbital vein for assessment of glucose and malondialdehyde [MDA], then Animals were sacrificed and aortic tissues were removed,PCR for Annexin V and for HsP72 were done. Compared to control group, fasting serum glucose is significantly higher in the diabetic group [group II] [p<0.05] and it decreased significantly with administration of insulin [group III]. MDA is significantly higher in the diabetic group II compared with control one [p<0.05]. It decreased significantly with administration of insulin [group III] or vitamin E [group IV] in comparison to diabetic group [group II]. Administration of vitamin E and insulin [group V] leads to significant reduction in MDA back to control level. The expression of Annexin V is significantly higher in the diabetic group [group II] compared to control group [p<0.05]. Administration of insulin [group III] or vitamin E [group IV] decreases it significantly. The expression of HSP72 is significantly lower in the diabetic group compared to control one [p<0.05]. It increased significantly with administration of insulin [group III] or vitamin E [group IV]. Serum MDA level rise in STZ induced diabetic rats as a marker of oxidative stress and administration of vitamin E was found to normalize this level, in addition a significant rise in expression of Annexin V as a marker of apoptosis in aorta of STZ induced diabetic rats with decreased expression of HSP72 which may be involved in cellular protection against oxidative stress in DM

Enhanced apoptosis of peripheral blood mononuclear cells from chronic hcv patients is associated with reduced caspase 3 activity

Background: Infection with hepatitis C virus [HCV] is a major cause of chronic liver disease throughout the world. Down-regulation of the immune response plays a major role in HCV persistence. Recent investigation suggest that apoptosis of peripheral blood mononuclear cells [PBMCs] contributes to such down-regulation

Objective: The current study investigates apoptotic changes in PBMCs and their relation to caspase-3 and -8 activities in patients with chronic HCV infection

Methods: Apoptosis were investigated by measuring annexin-V binding using flowcytometry and DNA fragmentation using agarose gel electrophoresis, and caspases-3 and -8 specific activities were also measured in 43 chronic HCV patients and 10 normal control subjects

Results: A significantly higher percent of annexin-V positive PBMCs was found in chronic HCV patients than controls [p<0.0001]. DNA fragmentation was detected in PBMCs from 20/43 patients [46.5%] but not from controls. There was no statistically significant difference between HCV-PCR positive and negative patients as regards the degree of PBMCs apoptosis. Caspase-3 activity was significantly lower in patients than controls [p=0.001], was significantly lower in HCV-PCR positive than controls [p=0.001] and was significantly higher in patients with PBMCs DNA fragmentation [p=0.005]. On the other hand, caspase-8 activity was comparable in both patients and control groups. However, patients with PBMCs DNA fragmentation showed statistically significant higher activity than those without [p=0.023]. There was a statistically significant direct correlation between caspase-3 and caspase-8 activities in the patients groups [r=0.56,p<0.0001]

Conclusion: Chronic HCV infection is associated with PBMCs apoptosis irrespective of the presence of viremia. However, this apoptosis is independent on activation of either caspase-3 or caspase-8