Assessment of donor specific IgG Anti-HLA subclasses before and after kidney transplantation in Upper Egypt: A Single Center Study.

Donor specific antibodies (DSAs) are known as the leading cause of antibody mediated rejection (AMR), graft loss in kidney transplant (KT) recipients. DSAs characteristics, as immunoglobulin (Ig) classes, subclasses, and strength, are important to assess the immunological risk, early prediction of AMR, and therefor proper management. This longitudinal, case control study included 32 KT recipients at Assiut University Urology Hospital and 10 age and sex matched normal subjects as the control group. Total IgG, its subclasses and anti-human leukocyte antigen (anti-HLA) panel reactive antibody (PRA) were detected pre-transplantation (pre-TX), at 6-12- and 24-36-months post-TX. Rejection occurred in 4 recipients, 3 of them had high total IgG, IgG1 and/or IgG3. IgG2 and IgG4 were normal in all recipients. There were preformed anti-DSAs antibodies in 3/32 recipients (9.4%). Of these, two recipients became negative with no rejection occurred. The third recipient had high post-TX mean fluorescence intensity (MFI) and AMR occurred. The pre-TX PRA was negative in 29/32 recipients (90.6%). The PRA was negative in 8/29 recipients (27.6%) and the remaining 21/29 recipients (72.4%) developed de novo DSAs post-TX (MFI < 3000->10000). Rejection occurred with both low and high MFI. In 11 recipients, anti-HLA class I and II were not different between pre-TX, 3-6- and 24-36 months post-TX with no rejection occurred. The frequency and median levels of total IgG, IgG1 and IgG3 were increased in all recipients 24-36 months post-TX when compared with their levels pre-TX and 6-12 months post-TX in the 11 recipients and with the control group. The graft survival time significantly decreased in recipients with positive post-TX class I PRA. In conclusion, preformed DSAs may persist post-TX or turn negative. De novo DSAs developed post-TX even in non-sensitized recipients. Serum total IgG, IgG1 and IgG3 frequency increase 2-3 years post-TX.

CXCR4 as a prognostic marker in Egyptian chronic lymphocytic leukemia patients.

Chronic lymphocytic leukemia (CLL) has variable clinical presentations, and molecular and biological prognostic markers. The C-X-C chemokine receptor 4 (CXCR4) and its ligand stromal cell-derived factor-1 (SDF-1) play an important role in trafficking of lymphocytes and monocytes. The aim was to study lymphocyte expression of CXCR4 and its prognostic value in CLL. A case control study was carried out on 30 newly diagnosed CLL cases and 30 healthy controls. Fludarabine, cyclophosphamide, and rituximab (FCR) was the standard treatment. Flowcytometric measurement of CXCR4 expression on lymphocytes was done. CXCR4 was significantly higher in patients than controls (81.67 ± 17.95 vs. 11.78 ± 2.78; P< 0.001). CXCR4 was significantly higher (P<0.001) in high risk CLL (93.63 ± 6.78) vs. intermediate risk (82.50 ± 7.13) and low risk (75.84 ± 12.23). CXCR4 was significantly higher (P<0.001) in non-responders (91.63 ± 6.98) vs. partial responders (83.11 ± 5.55) and complete responders (70.11 ± 4.44). CXCR4 was significantly lower in survivors vs. non-survivors (80.89 ± 5.09 vs. 85.43 ± 5.51; P< 0.001. CXCR4 had significant positive correlation with WBCs (r=0.45, P=0.01) and lymphocytes (r=0.40, P=0.01) measured at diagnosis. In conclusions, expression of CXCR4 in newly diagnosed CLL is significantly high. CXCR4 increased expression is associated with poor prognosis and resistance to the therapy.so it can be used as prognostic tool.

Evaluation of Serum Protein 14-3-3η (Eta) as a Novel Biomarker for Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is one of the most common systemic autoimmune diseases. New markers are needed for early diagnosis of RA as seronegativity in both early and established RA remains a major limitation of both anticitrullinated protein antibodies (ACPA) and rheumatoid factor (RF). The 14-3-3η protein may represent a novel biomarker for the detection of RA. We evaluated the diagnostic performance of serum 14-3-3η protein in early and established cases of rheumatoid arthritis and we compared the diagnostic accuracy of it with those of the well-known RA markers (e.g. RF and ACPA). Sera from 50 patients with RA (20 early and 30 established) based on the 2010 ACR / EULAR Rheumatoid Arthritis Classification Criteria, 15 patients with non-RA arthritis as diseases control group (8 patients with OA and 7 patients with SLE) and 14 healthy controls were enrolled in the study. Serum RF was determined by latex, ACPA and 14-3-3η protein were determined by ELISA. Serum 14-3-3η protein levels in patients with RA were significantly higher (P=0.001*) as compared to healthy individuals. For serum 14-3-3η diagnostic accuracy in RA; Receiver operating characteristic curves (ROC) analysis comparing patient with RA with healthy control showed AUC (0.916) at optimum cutoff of > 2.5ng/mL, and a sensitivity of 100%, a specificity of 78.57%, a PPV of 94.3, and an NPV of 100. No significant difference in 14-3-3η protein serum levels was found between early and established RA groups. It was positive in 100% of early and established RA patients who were seronegative for RF and ACPA. It is concluded that, 14-3-3η protein could improve the sensitivity of RA diagnosis and cover the shortage of detection of RF and ACPA in RA patients.

Glypican-3 and Melanoma Antigen Genes 1 and 3 as Tumor Markers for Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is the commonest liver cancer; its incidence and prevalence are continuously increased. Glypican3 (GPC3), melanoma antigen-1, 3 genes (MAGE1 and 3) are tumor markers used in HCC. We evaluated their role in HCC detection and assessed their relation to tumor parameters. Three groups, HCC group, liver cirrhosis group and a control group were studied. AFP, GPC3, and MAGE1 and 3 mRNA were determined in all study subjects. Tissue GPC3 was examined in patients with HCC only. Serum AFP and GPC3 were elevated in HCC group compared to other groups (P < 0.000 and P < 0.001, respectively). AFP at cutoff 44.4ng/ml and GPC3 at cutoff 5.6µg/L resulted in 81% and 90.1% sensitivity, 73.3% and 92.6% specificity, respectively. The combined measurement of both increased the sensitivity and the specificity to 100% and 93.3%, respectively. GPC 3 was detected in tissues of 81.0% of the cases. MAGE-1 and MAGE-3 genes expression were detected in 61.9% and 52.4%, respectively in HCC cases but not in other groups. GPC3, MAGE1and 3 were increased with advanced tumor stage, size, and nodule numbers. We concluded that GPC3 is a promising diagnostic marker for HCC, and MAGE 1 and 3 could be helpful in early detection of extrahepatic metastasis of HCC.

Diagnostic Role of RASSF1A and p16INK4a Promoter Gene Hypermethylation in Serum DNA of Lung Cancer Patients: Clinicopathological Significance.

The most common inactivation mechanism of tumor suppressor genes, RASSF1A and p16INK4a, in lung cancer is hypermethylation. We detected the methylation status of RASSF1A and p16INK4a in serum of lung cancer patients using methylation-specific PCR and analyzed their clinicopathological significance. Each of RASSF1A and p16INK4a hypermethylation was detected in 31.1% cancer patients but not in benign lung lesion patients. Hypermethylation was preferentially observed in small cell lung cancer (SCLC) for RASSF1A (50%), but not for p16INK4a. In non-small cell lung cancer (NSCLC), RASSF1A and p16INK4a hypermethylation were found in 27% and 37.8% respectively. Hypermethylation of RASSF1A was not correlated with clinicopathological character. While, p16INK4a hypermethylation was associated with age >60 years, smoking and squamous cell carcinoma (SCC) (P = 0.033), but not with gender and pathological stages of NSCLC. Sensitivity and specificity of each gene were 31.1% and 100% respectively and the sensitivity improved with evaluation of a combination of the two genes (55.6%). These findings suggest that serum RASSF1A and p16INK4a hypermethylation are promising diagnostic method for detection of lung cancer. As regard the clinicopathological characteristics, p16INK4a hypermethylation may provide a more specific approach than RASSF1A hypermethylation.

Evaluation of transforming growth factor alpha and vascular endothelial growth factor in diagnosis of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most frequent malignant tumors. It is important to detect disease and recurrence at its earlier period. We aimed to evaluate the usefulness of TGF-alpha and VEGF in diagnosis of HCC patients. Thirty patients with liver cirrhosis, 30 patients with confirmed HCC and 20 healthy volunteers were subjected to abdominal ultrasonography, and alpha-fetoprotein, TGF-alpha and VEGF were assessed. Serum level of AFP was significantly higher in HCC than cirrhotic patients and controls and in cirrhosis patients than controls. The level of TGF-alpha was significantly increased in HCC and cirrhosis groups than in control group with no difference between cirrhosis and HCC groups. Serum VEGF was higher in HCC than in cirrhosis group and in both groups than in control group. Sensitivity and specificity of makers in diagnosis of HCC were 63%, 90% respectively for AFP using a cutoff value of 19.96 ng/ml; 60% and 92% for VEGF at cut off 268 and 73% and 84 % for TGF-alpha using a cutoff value of 13.95 pg/ml. VEGF may be useful serum marker for detection of HCC in addition to traditional markers.

Mannose binding lectin gene polymorphism and preclinical carotid atherosclerosis in patients with systemic lupus erythematosus.

Patients with systemic lupus erythematosus (SLE) have increased risk of atherosclerosis and CVD that cannot be explained by traditional risk factors. Previous studies indicated that mannose binding lectin (MBL) may modify the development of atherosclerosis. This study was designed to investigate association of MBL gene polymorphism with occurrence of preclinical atherosclerosis in SLE. The study included 46 patients with SLE and 17 age and sex matched controls. MBL2 genotypes were assessed in patients and controls by the PCR-RFLP method and intima-media thickness of the common carotid artery (cclMT) was determined by means of ultrasonography. Also, serological markers were measured and the disease activity index (SLEDAI) was estimated. SLE patients had higher frequency of MBL A/B + B/B genotypes (47.8%) than controls (29.4%). ccIMT was higher in patients having A/B, B/B, A/B+B/B genotypes when compared with wild genotype (A/A). Patients with A/B+B/B genotypes showed high serum level of LDL, TG, ESRI, CRP and SLEDAI score, but low level of HDL, C3, and C4 compared to wild genotype. ccIMT of mutant SLE subgroup correlated well with SLE risk factors for atherosclerosis. In conclusion, mutant genotypes of MBL may be atherogenic as SLE patients had a higher IMT, which correlated significantly with SLE risk factors for atherosclerosis.

Anti-C1q antibodies, sCD40L, TWEAK and CD4/CD8 ratio in systemic lupus erythematosus and their relations to disease activity and renal involvement.

Due to the unpredictable nature of lupus nephritis (LN), it would be clinically valuable to discover a reliable biomarker for disease activity and progression. The aim of this study is to evaluate the role of anti-C1q antibodies, sCD40L and TWEAK in systemic lupus erythematosus (SLE) and their relation to disease activity and kidney involvement. This study included 47 patients with SLE, 28 with LN and 19 without LN, as well as, 20 healthy subjects as controls. All subjects underwent complete history, examination and estimation of disease activity index (SLEDAI) and renal SLEDAI. The following investigations were done for all subjects: anti-C1q antibodies, sCD40L, TWEAK and CD4/CD8 ratio, in addition to complete blood picture, ESR, kidney function tests, ANA, anti-ds DNA antibodies and C3, C4. Anti-C1q antibodies, sCD40L and TWEAK and anti-dsDNA were significantly higher in SLE patients than controls (P < 0.001 for each), while C3, C4 and CD4/CD8 ratio were significantly lower (P < 0.001, 0.05 and 0.001 respectively). In LN patients, anti-C1q antibodies, sCD40L and TWEAK were significantly higher than non LN patients (P < 0.001 for each). Anti-C1q antibodies, sCD40L and TWEAK correlated with traditional disease activity parameters (C3, C4, anti-dsDNA and SLEDAI) as well as rSLEDAI. Levels of serum TWEAK correlated with the development of LN in patients with SLE. We concluded that anti-C1q antibodies, sCD40L and TWEAK may be used as serum biomarkers for the assessment of disease activity and development of LN.

Proinflammatory cytokines, soluble Fas receptor, nitric oxide and angiotensin converting enzyme in congestive heart failure.

We measured serum interleukin-2 receptor (sIL-2R), tumor necrosis factor-a (TNF-a), Fas receptor (sFas), nitric oxide (NO), and angiotensin converting enzyme (ACE) activity in 45 patients with congestive heart failure (CHF) of different etiologies. The relatioship between these bioindices and the severity of heart failure was analysed. Patients were classified according to the etiology of heart failure into: 15 patients with rheumatic valvular heart disease (RHD), 17 with ischemic heart disease (IHD) and 13 with idiopathic dilated cardiomyopathy (DCM). Patients were further classified according to severity of CHF following the New York Heart Association classification (NYHA) into: NYHA class II (n= 7), NYHA class III (n=20) and NYHA class IV (n=18). Eighteen healthy subjects were included as controls. Serum sIL-2R, TNF-alpha and sFas levels were determined by ELISA while serum NO and ACE levels were measured by colorimetric methods. Doppler Echocardiography was performed for all participants. Levels of sIL-2R, TNF-alpha, sFas, NO, and ACE were significantly higher in CHF patients than controls. Levels of the bioindices varied according to the CHF etiology. TNF-a level was the only one that had significant differences among different subgroups (RHD, IHD and DCM). The levels of sIL-2R, TNF-alpha, NO and sFas in patients with NYHA class IV were significantly higher than class II or III. Moreover, sIL-2R, TNF-alpha and NO levels were significantly higher in patients with diastolic dysfunction than patients with normal diastolic function. A significant positive correlations were found between sFas and both TNF-alpha and sIL-2R and between TNF-alpha and both NO and diastolic function. In addition, significant positive correlations were found between TNF-alpha and sIL-2R in both IHD and RHD patients and between sIL-2R and both ACE in IHD patients and diastolic function in DCM patients. It is concluded that a relationship exists between immune system activation, apoptosis and renin- angiotensin system in CHF and this may play a significant role in the pathophysiology and prognosis of the disease.

Thyroid disorders and autoantibodies in systemic lupus erythematosus and rheumatoid arthritis patients.

To determine the patterns of thyroid dysfunction and autoantibodies associated with SLE and RA patients, twenty patients with SLE and another group of twenty with RA were studied. The results were compared with those of twenty apparently healthy age- and sex- matched controls. All patients were subjected to complete history taking, thorough clinical examination and joint examination. All patients and controls were subjected to the following investigations: T3, T4, TSH, antithyroglobulin antibodies (ATGAb) and thyroid peroxidase antibodies (TPOAb). Also, complete blood picture, ESR, RF, ANA, CRP and LE cells were done. This study revealed that thyroid disorders were significantly increased in SLE patients (50%) when compared to RA (15%) (P<0.05). In SLE group, 20% had euthyroid sick syndrome, 20% had hypothyroidism (10% subclinical and 10% biochemical), and 10% had hyperthyroidism (5% subclinical and 5% biochemical). However, in RA, 10% had hypothyroidism (subclinical) and 5% had subclinical hyperthyroidism. TPOAb was found in 15% of SLE and 5% of RA patients and 10% of controls, but the titres were higher in SLE and RA patients. Also, ATGAb was found in 5% of SLE, 30% of RA patients and 10% of controls, but the titres were higher in SLE and RA patients. It is concluded that thyroid abnormalities are more implicated with euthyroid sick syndrome and hypothyroidism (subclinical and overt) than hyperthyroidism in SLE patients. SLE and RA were associated with antithyroid antibodies (TPOAb in SLE and ATGAb in RA). Performance of thyroid function tests in patients with SLE, in particular and RA as a part of the biochemical and immunological profiles, may help in early detection of associated thyroid disorders.