HLA-DQB1* alleles and genetic susceptibility to type 1 diabetes mellitus.

AIM: To determine human leukocyte antigen (HLA)-DQB1 allele association with susceptibility to type 1 diabetes (T1D) and to clinical and laboratory findings. METHODS: This study was conducted on 85 unrelated Egyptian children with T1D recruited consecutively from the Pediatric Diabetes Endocrinology outpatients Clinic; Mansoura University Children's Hospital, Egypt. Patient mean follow up period was 2.5 years. Patients were subdivided according to level of HbA1c (optimal/suboptimal control < 8.5% and poor control ≥ 8.5%). The control group consisted of 113 unrelated age- and sex-matched healthy subjects without T1D or other autoimmune diseases. Genomic DNA extraction was done for all subjects using a DNA isolation kit. HLA-Class II-DQB1 allele typing was carried out with a polymerase chain reaction-sequence-specific oligonucleotide probe using a INNO-LiPA HLA-DQB1 update kit. RESULTS: Significant differences were detected between Egyptian patients with T1D and control groups in the frequencies of DQB1*02 [44.4% vs 18.6%, corrected P value (Pc) < 0.001] and DQB1*03 (41.2% vs 24.4%, Pc < 0.001). Significant differences were also observed between control groups and T1D patients in the frequencies of DQB1*05 (14.6% vs 7.2%, P = 0.029) and DQB1*06 (34.1% vs 7.2%, P < 0.001). However, after correction for multiple comparisons, the significance was retained for HLA-DQB1*06 (Pc < 0.001) but lost for HLA-DQB1*05. HLA-DQB1*0201, *0202, *030201 were positively associated with T1D (Pc = 0.014, Pc < 0.001, and Pc < 0.001 respectively), while HLA-DQB1*060101 was negatively associated (Pc < 0.001) with the condition. Although the HLA-DQB1 alleles 030101 and 050101 were significantly higher in controls (P = 0.016, P = 0.025 respectively), both of them lost statistical significance after correction of P value. The frequency of the HLA-DQB1 genotypes 02/02, 02/03, and 03/03 was higher in T1D patients, and the frequency of the genotypes 03/06, 05/06, and 06/06 was higher in controls, these differences being statistically significant before correction. After correction, the genotypes 02/02, 02/03 in T1D, and the genotypes 03/06, 06/06 in controls were still significant (Pc = 0.01, Pc < 0.001, Pc < 0.001, and Pc = 0.04, respectively). Non-significant associations were found between the frequency HLA-DQB1 alleles and genotypes in T1D in relation to the grade of diabetic control, Microalbuminuria, age, gender, age of presentation, weight, height, frequency of diabetic ketoacidosis (P = 0.42), serum cholesterol, and fasting and post-prandial level of C-peptide (P = 0.83, P = 0.9, respectively). CONCLUSION: The Current work suggests that HLA-DQB1 alleles *030201, *0202, *0201, and genotypes 02/03, 02/02 may be susceptibility risk factors for development of T1D in Egyptian children, while the HLA-DQB1*060101 allele, and 03/06, 06/06 genotypes may be protective factors. HLA-DQB1 alleles and genotypes do not contribute to microalbuminuria or grade of diabetic control.

Cytokeratin 20 and vascular endothelial growth factor as molecular markers in Egyptian patients with colorectal cancer.

PURPOSE: To investigate the expression of cytokeratin 20 (CK20) and vascular endothelial growth factor (VEGF) in the peripheral blood of colorectal cancer (CRC) patients, and correlate the findings with the pathologic data of the patients. METHODS: This study was carried out on 50 subjects, 40 patients with histologically confirmed colorectal carcinoma undergoing elective surgery and 10 healthy individuals matched for age and sex. Total RNA extraction followed by real time quantitative RT-PCR and real time TaqMan quantitative assay for peripheral blood expression of CK20 and VEGF was done for both patients and controls. RESULTS: (1) Statistically significant high levels of CK20,VEGF, CEA (p = 0.000 each) and CA19-9 (p = 0.002) in CRC patients when compared with controls; (2) Statistically significant increase in the expression of CK20 in advancing CRC stage C (p = 0.001) and with LN metastasis (p = 0.000); (3) Statistically significant increase in the expression of VEGF in advancing CRC stage C (p = 0.002), pathologic grade (p = 0.038), and with LN metastasis (p = 0.004); and (4) statistically positive correlation between CK20 and VEGF expressions, and also between these markers and CEA level. CONCLUSION: CK20 and VEGF expressions in peripheral blood of CRC patients are promising molecular markers for CRC progression and metastasis.

Vascular endothelial growth factor, p53, and the H-ras oncogene in Egyptian patients with bladder cancer.

AIM: To evaluate the relationship between vascular endothelial growth factor (VEGF), p53, and the H-ras oncogene and different clinicopathological parameters in Egyptian patients with Schistosoma-associated transitional cell carcinoma of the bladder. METHODS: The study included 50 patients with transitional cell carcinoma for whom radical cystectomy and urinary diversions were carried out. VEGF and p53 protein expressions were evaluated with an immunohistochemical staining method, and H-ras oncogene mutations were analyzed with a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. RESULTS: High grade tumors revealed higher p53 immunostaining than low grade tumors (P = 0.016). p53 and VEGF protein expressions, as well as H-ras oncogene mutations, had an insignificant impact on patient outcomes (P = 0.962, P = 0.791, and P = 967, respectively). Cancer extension to regional lymph nodes was associated with poor outcomes (P = 0.008). CONCLUSION: VEGF, p53 and the H-ras oncogene have no relation to patient survival and outcome in Schistosoma-associated transitional cell carcinoma.

HLA-class II alleles in Egyptian patients with hepatocellular carcinoma.

UNLABELLED: Hepatocellular carcinoma (HCC) is linked to environmental, dietary, and life style factors. Its incidence and distribution vary widely among ethnic groups, sex, and geographic regions. HBV and HCV Infection, liver cirrhosis, male gender, and old age are important risk factors of HCC. Variability in outcome following exposure, and the clustering of HCC within families raise the possibility that genetic factors are also involved in susceptibility to HCC. The Major Histocompatibility Complex (MHC) plays a key role in anti-virus and tumor defense. HLA polymorphism is implicated in conferring genetic susceptibility to a large number of immune-mediated diseases, including some cancers. The association between HLA class II antigen and HCC in different ethnic populations that has been reported is controversial. Therefore, the aim of this work was to study the association between HLA class II-DRB1 and DQB1 polymorphism and HCC in Egyptian patients and to investigate their role as risk factors for the development of HCC. METHODS: HLA-class II (DRB1 and DQB1) typing was done by SSP for 100 subjects; 50 patients suffering from HCC (45 males and 5 females) with age range 40-64 years (51.16 years (y) +/- 6.16); and 50 normal healthy control subjects. RESULTS: 1. A significantly increased frequency of DRB1*04, and DQB1 *02 in HCC patients versus control group (p = 0.016, and 0.032, respectively) was found; 2. A significantly decreased frequency of DQB1*06 (p = 0.032) was found; 3. A significantly increased frequency of DRB1*07 (odds ratio (OR) = 4.929) was found; and 4. A significantly decreased frequency of DRB1*15 (OR = 0.316) was seen. In conclusion, while some alleles are significantly associated with HCC (DRB1*04, DQB1*02) and others are not associated (DQB1*06); therefore, it can be concluded that the DRB1*04 and DQB1*02 alleles might be risk factors for the occurrence of HCC (OR = 4.373 and 3.807, respectively), and DQB1*06 may be a protective allele (OR = 0.259).

Expression of HLA-G in cancer bladder.

This study comprised 42 patients with cancer bladder, who underwent radical cystectomy. The aim of work is to determine if HLA-G is expressed on tumor cells, derived from cancer bladder. We studied HLA-G-mRNA expression using RT-PCR and HLA-G cell surface expression by immunohistochemistry (IHC) staining technique. HLA-G was expressed in 28.6 % of cancer cases as determined by PCR and on 16.7% of cases determined by IHC staining. The sensitivity, specificity and accuracy of IHC were 58.3%, 96% and 81.7% respectively as compared to PCR results. There was a highly significant increase in the expression of HLA-G on cancer bladder cases with metastatic prostate infiltration (P= 0.021). It is concluded that HLA-G is ectopically expressed on cancer bladder malignant cells both at molecular and protein levels. However, it is not significantly associated with histologic type, tumor grade, stage, T category, schistosomiasis and lymph node involvement.

Proinflammatory cytokines (IL-12 and IL-18) in immune rheumatic diseases: relation with disease activity and autoantibodies production.

Interleukin-18 (IL-18) and its inducer IL-12 have multiple biological activities that are important in generating Th1 responses and inflammatory tissue damage. We investigated serum concentration of the novel proinflammatory Th1 cytokine; IL-18, and its inducer IL-12 in patients with immune rheumatic diseases. Group I comprised32 patients of systemic lupus erythmatosus (SLE), Group II comprised 36 patients of rheumatoid arthritis (RA). Group III comprised 9 patients (2 patients of Behcet, 2 patients of Dermatomyositis, 2 patients of Sicca syndrome, one patient of Scleroderma, and 2 patients of Mixed connective tissue disease). Group IV is a control group consists of 21 sex and age matched healthy subjects and correlated their levels with autoantibody concentration (ANA and ds-DNA), clinical grades and SLE disease activity index (SLEDAI). Serum IL-18, IL-12, ANA and ds-DNA were measured by enzyme immuno sorbent assay. IL-18, IL-12 and ANA were significantly higher in the three studied groups than in the control group (IL-18; P < 0.001 in the three groups, IL-12; P = 0.019, P = 0.002, and P = 0.006, and ANA; P < 0.001, P = 0.002,and P = 0.006, respectively).ds-DNA was significantly higher in SLE patients than in control group (P < 0.001). There were significant positive correlations between; A) levels of IL-18,and both ANA and ds-DNA in SLE patient (r = 0.41,P = 0.001, r = 0.58 and P=0.001 respectively); and B) IL-18 and ANA in both RA and group III patients (r = 0.32, P = 0.005, r = 0.61and P = 0.022 respectively). Also, there were significant positive correlation between the levels of IL-18 and clinical grades of the three groups (r = 0.60,P = 0.001, r = 0.79,P = 0.001, r = 0.78 and P= 0.001 respectively). In SLE patients , IL-18 concentration shows significant positive correlation with SLEDAI score (r = 0.76, P = 0.001). In conclusion, the elevation of proinflammatory cytokines (IL-18 and IL-12 ) may trigger the inflammatory process in immune rheumatic diseases and IL-18 is correlated with disease activity