Associations of Genes for Killer Cell Immunoglobulin-like Receptors and Their Human Leukocyte Antigen-A/B/C Ligands with Abdominal Aortic Aneurysm.

Abdominal aortic aneurysm (AAA) is an immune-mediated disease with a genetic component. The multifactorial pathophysiology is not clear and there is still no pharmacotherapy to slow the growth of aneurysms. The signal integration of cell-surface KIRs (killer cell immunoglobulin-like receptors) with HLA (ligands, human leukocyte class I antigen molecules) modulates the activity of natural killer immune cells. The genetic diversity of the KIR/HLA system is associated with the risk of immune disorders. This study was a multivariate analysis of the association between genetic variants of KIRs, HLA ligands, clinical data and AAA formation. Genotyping was performed by single polymerase chain reaction with sequence-specific primers using commercial assays. Patients with HLA-A-Bw4 have a larger aneurysm by an average of 4 mm (p = 0.008). We observed a relationship between aneurysm diameter and BMI in patients with AAA and co-existing CAD; its shape was determined by the presence of HLA-A-Bw4. There was also a nearly 10% difference in KIR3DL1 allele frequency between the study and control groups. High expression of the cell surface receptor KIR3DL1 may protect, to some extent, against AAA. The presence of HLA-A-Bw4 may affect the rate of aneurysm growth and represents a potential regional pathogenetic risk of autoimmune injury to the aneurysmal aorta.

Polymorphisms of Antigen-Presenting Machinery Genes in Non-Small Cell Lung Cancer: Different Impact on Disease Risk and Clinical Parameters in Smokers and Never-Smokers.

Lung cancer is strongly associated with cigarette smoking; nevertheless some never-smokers develop cancer. Immune eradication of cancer cells is dependent on polymorphisms of HLA class I molecules and antigen-processing machinery (APM) components. We have already published highly significant associations of single nucleotide polymorphisms (SNPs) of the ERAP1 gene with non-small cell lung cancer (NSCLC) in Chinese, but not in Polish populations. However, the smoking status of participants was not known in the previous study. Here, we compared the distribution of APM polymorphic variants in larger cohorts of Polish patients with NSCLC and controls, stratified according to their smoking status. We found significant but opposite associations in never-smokers and in smokers of all tested SNPs (rs26653, rs2287987, rs30187, and rs27044) but one (rs26618) in ERAP1. No significant associations were seen in other genes. Haplotype analysis indicated that the distribution of many ERAP1/2 haplotypes is opposite, depending on smoking status. Additionally, haplotypic combination of low activity ERAP1 and the lack of an active form of ERAP2 seems to favor the disease in never-smokers. We also revealed interesting associations of some APM polymorphisms with: age at diagnosis (ERAP1 rs26653), disease stage (ERAP1 rs27044, PSMB9 rs17587), overall survival (ERAP1 rs30187), and response to chemotherapy (ERAP1 rs27044). The results presented here may suggest the important role for ERAP1 in the anti-cancer response, which is different in smokers versus never-smokers, depending to some extent on the presence of ERAP2, and affecting NSCLC clinical course.

Wharton's jelly-derived mesenchymal stem cells in the treatment of four patients with alopecia areata.

BACKGROUND: Alopecia areata (AA) is the second most common cause of non-scarring alopecia. Little is known on the etiopathogenesis of AA. It is considered an autoimmune disease, with T lymphocytes and antibodies directed against hair follicle structures. Topical and systemic therapies are used for the treatment of AA, but none of the therapies used to date have a permanent therapeutic effect. OBJECTIVES: To evaluate the efficacy and safety of AA treatment through a single intradermal injection of a suspension of allogeneic MSCs extracted from Wharton's jelly (WJ-MSCs) into the alopecia foci. MATERIAL AND METHODS: The study involved 4 AA patients who underwent experimental therapy with a suspension of WJ-MSCs. The AA intensity was measured using the SALT score. This measure was performed 3 times during treatment: 1st measure (SALT0) prior to treatment; 2nd measure (SALT12) 12 weeks after the treatment; and 3rd measure (SALT24) 24 weeks after the treatment. Furthermore, during each follow-up visit (6, 12, 18, and 24 weeks after the administration of WJ-MSCs) the patient's general condition (physical examination) and local condition were assessed, their mood was evaluated, and a photo of the scalp was taken. RESULTS: Hair regrowth was observed in all patients by an average of 67% at the sites where the cell suspension was administered. In all cases, we observed greater dynamics of hair regrowth in the first 3 months after the treatment, with an average increase of 52.2%, compared to the following 3 months, with an average of 32%. CONCLUSIONS: The results of the applied intradermal injections of an allogeneic WJ-MSC suspension were positive with hair growth observed in all participants and the therapy was found to be safe, with no side effects.

Evaluation of plasma levels of tissue factor and tissue factor pathway inhibitor in patients with psoriasis.

INTRODUCTION: Psoriasis is a chronic, recurrent, inflammatory skin disorder with systemic involvement. It has recently been established that psoriasis is associated with an increased cardiovascular risk. Chronic skin-specific inflammation may promote atherosclerosis. Myocardial infarction or stroke can also be a result of underlying haemostasis disorders. Disorders in fibrinolysis and thrombosis in patients with psoriasis have been observed by many authors. AIM: This study points to the key role played by the tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in the extrinsic pathway of blood coagulation and the potential influence of microvascular disorders in inflamed psoriatic skin on TF and TFPI activity. MATERIAL AND METHODS: The study included 47 patients with active psoriasis vulgaris, hospitalized in the Dermatological Ward of the Regional Specialist Hospital, Research and Development Centre in Wroclaw, as well as 18 people from the control group. RESULTS: There were significant differences in the blood concentrations of TF and TFPI in patients with psoriasis when compared to the control group. A low TFPI concentration in psoriatic patients may indicate an increased risk of atherosclerosis. Interpretation of a decreased level of TF in patients with psoriasis is difficult because it seems to be at odds with observations among patients with other atherosclerosis risk factors such as hypertension, hyperlipidaemia, diabetes or smoking. CONCLUSIONS: It appears that further studies are necessary to explain this problem, perhaps to include an evaluation of TF levels in psoriatic skin.

Single Nucleotide Polymorphisms of the ERAP1 Gene and Risk of NSCLC: A Comparison of Genetically Distant Populations, Chinese and Caucasian.

An effective cytotoxic immune response to neoplastic cells requires efficient presentation of antigenic peptides to T lymphocytes by HLA class I (HLA-I) molecules. The HLA-I-bound peptide repertoire depends on antigen-processing machinery molecules. Aminopeptidase residing in endoplasmic reticulum 1 (ERAP1) trims peptides to the optimal length for HLA-I binding. Single nucleotide polymorphisms (SNPs) in the ERAP1 gene result in changes in aminopeptidase activity and specificity. This may affect susceptibility to cancer. However, non-small cell lung carcinoma (NSCLC) has not been studied in this respect. We compared genotype and haplotype frequencies of four coding, nonsynonymous ERAP1 SNPs, rs26653G > C, rs26618T > C, rs30187C > T, and rs27044C > G, in NSCLC occurring in two genetically distant populations, Chinese and Poles. We found associations of all four SNPs with NSCLC in Chinese but not in Poles. The differences in ERAP1-NSCLC associations might be explained by highly significant differences in SNP genotype frequencies between Chinese and Poles (except for rs26618). In accordance with this, the most frequent ERAP1 haplotypes were distributed differently in cases versus controls in Chinese, but not in Poles. Our findings add to the differences between Orientals and Caucasians in genetics of disease susceptibility.

[Selected endothelial hemostatic markers in patients with peripheral arterial disease after endovascular revascularization and restenosis formation]. / Wybrane sródblonkowe markery hemostazy u pacjentów z miazdzyca tetnic konczyn dolnych po wewnatrznaczyniowej rewaskularyzacji i po powstaniu restenoz.

Surgical and endovascular revascularization of ischemic legs in patients with peripheral arterial disease (PAD) can damage the arterial wall (endothelial and smooth muscle cells). Hemostatic factors released during endothelial dysfunction can lead to restenosis. 1. Determination of selected endothelial hemostatic factors in PAD patients and a reference group. 2. Prospective observation of new restenosis appearance in PAD patients after endovascular revascularization. 3. Comparison of selected endothelial hemostatic factors between non-restenotic and restenotic PAD patients. 150 PAD patients after endovascular revascularization - 90 men and 60 women, aged 44-88 (mean 65.5) years - were examined. During one-year observation after the revascularization procedures in 38 PAD patients restenosis occurred, when blood samples were also collected. The reference group consisted of 53 healthy persons - 44 men and 9 women, aged 20-56 years. Blood was drawn in the morning into 3.2% sodium citrate at a ratio of 9:1. Tissue factor (TF), tissue factor pathway inhibitor (TFPI), thrombomodulin (TM), von Willebrand factor (vWF) and tissue plasminogen activator (t-PA) were measured in plasma with commercial tests using the enzyme immunoassay. In the plasma of PAD patients after revascularization, the concentrations of TF and vWF were significantly higher, TM lower, TFPI and t-PA similar compared to the reference group. Six months after revascularization the level of TF had increased and vWF had significantly decreased. The endothelial hemostatic factors before and after restenosis did not significantly differ except TF, which after restenosis was higher. Increased TF and vWF levels in PAD patients indicate arterial endothelial cell damage, by atherosclerotic and revascularization processes. In PAD patients with restenosis compared to these patients before restenosis the determined endothelial hemostatic factors, except TF level, did not significantly differ. Perhaps TF participates in restenosis formation.

Correlation of HIWI and HILI Expression with Cancer Stem Cell Markers in Colorectal Cancer.

BACKGROUND/AIM: Cancer stem cells (CSCs) constitute a sub-population of tumor cells that possess stem cell properties, such as self-renewal and the ability of differentiation. The presence of CSCs is associated with metastatic potential, treatment resistance and poor patient prognosis. Recently, aberrant expression of P-element induced wimpy testis proteins-PIWI (HIWI and HILI) has been identified in various types of tumors. The aim of the study was to evaluate the clinical significance of the HIWI and HILI expression and its relationship with cancer stem cells markers in 72 patients with colorectal carcinoma (CRC). MATERIALS AND METHODS: The expression level of HIWI and HILI and cancer stem cells markers in paired cancerous and non-cancerous tissues was measured by real-time reverse transcription-polymerase chain reaction (RT-PCR) assay. Immunohistochemistry was performed to confirm the observed changes on mRNA level and detect tissue localization of PIWI proteins. RESULTS: Significantly higher mRNA levels of HIWI and decreased HILI mRNA were measured in colorectal cancer tissues compared to corresponding non-cancerous samples. The changes in HIWI mRNA level in cancer tissues were correlated with OCT4 expression. Positive correlations between HILI level and SOX2 were also observed in cancerous tissues. CONCLUSION: Our results indicate a reciprocal regulation between HIWI, HILI and some CSCs markers in colorectal cancer.

Tissue factor and other hemostatic parameters in patients with advanced peripheral artery disease after endovascular revascularization - search for hemostatic factors which indicate restenosis.

BACKGROUND: In patients with peripheral artery disease (PAD) a hypercoagulable state and thromboembolic complications occur. Revascularization procedures increase this state, sometimes leading to restenosis. Restenosis following balloon angioplasty (PTA)and stent implantation is ≥ 50% of artery stenosis. OBJECTIVES: To determine the concentration of tissue factor (TF), tissue factor pathway inhibitor (TFPI), thrombin-antithrombin (TAT) complexes, fibrinogen and D-dimers in the blood of patients with PAD after peripheral endovascular revascularization of the lower legs and in PAD patients with restenosis. MATERIAL AND METHODS: The study included 150 patients with PAD, 90 men and 60 women, aged 44-88 (mean 65.5) years, after successful peripheral angioplasty (PTA) and/or with stenting. During the 6 months after the revascularization procedures, restenosis occurred in 27 patients. The reference group consisted of 53 healthy persons (44 men and 9 women, aged 20-56 years). Blood was drawn in the morning into 3.2% natrium citrate at a ratio of 9 : 1. The concentration of TF, TFPI, TAT complexes and D-dimers were measured in plasma with commercial tests using an enzyme immunoassay. Fibrinogen was determined with coagulometer. RESULTS: In the plasma of patients with PAD after endovascular revascularization, the concentrations of TF, TAT complexes, fibrinogen and D-dimers were significantly higher compared to the reference group. During the six months of observation, 27 patients developed restenosis. The results of hemostatic factors in patients with restenosis were compared with the same patients before restenosis and the group of 123 PAD patients after endovascular revascularization. TF and fibrinogen levels in the 27 patients with restenosis were significantly higher than in the group of PAD patients before restenosis. CONCLUSIONS: Statistically significantly higher levels of tissue factor (TF) and fibrinogen in PAD patients with new restenosis, compared to those without restenosis after endovascular revascularization, indicate they can participate in the formation of restenosis.

Two new cases of KIR3DP1, KIR2DL4-negative genotypes, one of which is also lacking KIR3DL2.

The killer immunoglobulin-like receptor (KIR) genes KIR2DL4, KIR3DL2, and KIR3DP1 are present in virtually all humans. KIR2DL4 encodes a receptor present on uterine and decidual natural killer (NK) cells and some peripheral blood NK cells. Its only known ligand is the human leukocyte antigen-G molecule expressed on extravillous trophoblasts, and on tissues in some diseases. KIR3DL2 binds HLA-A*03 and HLA-A*11 as well as HLA-B*27 dimers, and microbial CpG DNA. KIR3DP1 is a pseudogene. During our immunogenetic studies we found two individuals, one from Lower Silesia district in Poland, and another from Western Ukraine, who were reproducibly negative for KIR2DL4 and KIR3DP1 genes, using three different PCR systems. Both individuals displayed very similar genotypes, possessing only KIR3DL3, KIR2DL3, KIR2DP1, KIR2DS1, and probably a rare variant of KIR2DL1. The Pole had also KIR3DL2, which the Ukrainian was apparently lacking. The Lower Silesia has been populated after the Second World War by a remarkable percentage with displaced people from Western Ukraine, which might contribute to genetic similarity of the two individuals described here.

Activity of thrombin-activatable fibrinolysis inhibitor in the plasma of patients with abdominal aortic aneurysm.

Patients with abdominal aortic aneurysm (AAA) experience impaired balance between fibrinolysis and coagulation, manifested by increased prothrombotic tendency and intensified inflammatory processes. The aim of this study was to evaluate the TAFI activity level (thrombin activatable fibrinolysis inhibitor) in the plasma of AAA patients. Plasma levels of PAI-1 (plasminogen activator inhibitor type 1), urokinase-type plasminogen activator and uPAR (urokinase-type plasminogen activator receptor) were measured as markers of fibrinolytic activity. The study showed that the activity of the thrombin-activatable fibrinolysis inhibitor in the plasma of AAA patients was significantly lower than in the plasma of the control individuals (64.6 ±â€Š10.1 vs. 54.2 ±â€Š10.9%, P < 0.0001). TAFI activity positively correlated with the white blood cell count (r = 0.486, P < 0.005). The uPAR concentration in the AAA patients was statistically significantly higher than in the control group and positively correlated with TAFI activity (r = 0.409, P = 0.02). The levels of PAI-1 and D-dimers (fibrin fragments) were significantly higher in patients with AAA than in the control group (44.3 ±â€Š17.5 vs. 21.7 ±â€Š8.7 ng/ml and 1869.6 ±â€Š1490.1 vs. 181.5 ±â€Š188.6 ng/ml, respectively). Lowered activity of the fibrinolysis inhibitor TAFI may heighten the blood fibrinolytic potential in AAA patients and contribute to the development of comorbidities. Therefore, TAFI participation in AAA pathogenesis cannot be excluded.

[The role of proteomics in prognosis and treatment of cardiovascular diseases]. / Rola proteomiki w prognozowaniu i leczeniu chorób ukladu sercowo-naczyniowego.

Despite advances in medicine and biological sciences the cardiovascular diseases (CVD) are still a major cause of morbidity and mortality in societies around the world. It is therefore necessary to improve the treatment methods used so far, to look for new therapeutic targets and specific, clinically useful biomarkers. Classic research related to the circulatory system are primarily concerned with histopathological descriptions of pathophysiological processes, and mapping the structure and expression of genes involved in the development of cardiovascular diseases. This information, however, does not fully explain the molecular basis of the CVD occurrence. That gap between traditional research in the pathophysiology field and the latest knowledge at the DNA level is effectively filled in by proteomics, the science dealing with the structure and function of the human proteome. This article presents main directions of the proteomics development in explaining pathomechanisms of formation and diagnosis of cardiovascular diseases and atherosclerosis accounting for the latest integrated research methods including techniques for protein and peptide fractionation, mass spectrometry and protein arrays.

Selected clotting factors in blood of patients with abdominal aortic aneurysms.

BACKGROUND: Tissue factor (TF), tissue factor pathway inhibitor (TFPI) and vascular endothelial growth factor A (VEGF-A) present in vascular structures take part in blood coagulation and in organ revascularisation. The concentration of thrombin-antithrombin complexes (TAT) in blood of patients with abdominal aortic aneurysms (AAA) reflects thrombin-generation. AIM: To determine the concentration of TF, TFPI, VEGF-A and TAT complexes in blood of patients with AAA and to consider if these factors after clot formation can play a role in the pathogenesis of abdominal aortic aneurysms. METHODS: Forty eight patients (43 men and 5 women) in the age of 59-80 (mean 72) years with AAA were examined. The blood was drawn in the morning to 3.2% natrium citrate in proportion 9:1. The concentration of TF, TFPI, VEGF-A and TAT complexes were measured in plasma with commercial kits using enzyme immunoassay. RESULTS: In plasma of patients with AAA the mean concentration of TF was elevated almost twice and TAT complexes were three times higher compared with controls. But the mean levels of TFPI and VEGF-A were similar as in control group. CONCLUSIONS: Increased concentrations of TF and TAT complexes indicate on high thrombin-generation, hypercoagulability and formation in abdominal aortic aneurysm of intraluminal thrombus, which can induce proteolytic processes in aortic wall.