In the peripheral blood mononuclear cells (PBMCs) of HCV infected patients the expression of STAT1 and IRF-1 is downregulated while that of caspase-3 upregulated.

Several peripheral blood mononuclear cell (PBMC) defects have been linked with hepatitis C virus (HCV) infection, including alterations in cytokine secretion and increased cell death. This study was performed to investigate the expression levels of signal transducer and activator of transcription 1 (STAT1), interferon regulatory factor 1 (IRF-1), and caspase 3 in PBMCs of patients infected with HCV. STAT1, IRF-1, and caspase 3 expression levels were compared in PBMCs from 19 untreated (naïve) HCV+ patients, 8 treated (sustained responder [SR]) HCV patients, and 20 HCV- healthy controls. Moreover, PBMCs from naïve HCV+ patients and SR-HCV patients were also evaluated for HCV RNA expression. The expression levels of STAT-1 and IRF-1 were significantly downregulated in PBMCs from naïve HCV+ patients (P  Keywords: PBMC; hepatitis C virus; STAT1; IRF-1; caspase-3.

Cytokine gene polymorphisms of TNFα, IL-6, IL-10, TGFß and IFNγ in the Saudi population.

Studies have demonstrated associations between cytokine gene polymorphisms and ethnicity. In the present work the authors examine polymorphisms in the genes encoding interleukin-6 (IL-6), IL-10, interferon-gamma (IFNgamma), tumour necrosis factor-alpha (TNFalpha) and transforming growth factor-beta (TGFbeta1) using the polymerase chain reaction sequence-specific primer (PCR-SSP) method in 150 healthy unrelated Saudis, and results compared with those from other studied populations. The genotype distributions were consistent with the Hardy-Weinberg equilibrium, and the genotype frequencies observed among Saudis showed both similarity and difference to other populations. The most notable difference was in the distribution of IL-6, where the Saudi population showed a lower CG genotype frequency compared with White American (22% vs. 39.2%, P = 0.004), Italian (22% vs. 50%, P < 0.0001) and Brazilian (22% vs. 40.8%, P < 0.0001) populations. The study population also showed a higher frequency of the IL-6 GG genotype compared with White Americans (72% vs. 45.1%, P < 0.0001), Italians (72% vs. 41%, P < 0.0001) and Brazilians (72% vs. 49.3%, P < 0.0001). These results may have significant clinical relevance to the understanding of prevalent diseases in Saudi Arabia.

Human leucocyte antigens: their association with end-stage renal disease in Saudi patients awaiting transplantation.

Most patients with chronic renal failure develop end-stage renal disease (ESRD) that requires renal transplantation. This study investigates the possible associations between human leucocyte antigen (HLA) Class I and Class II molecules with ESRD. Genotyping data (HLA) obtained between 2005 and 2009 on 235 unrelated Saudi patients (147 males, 88 females; mean age: 58 +/- 7 years) with ESRD awaiting renal transplantation were assessed retrospectively at the King Khalid University Hospital. Data were compared with the results on 60 normal, healthy, unrelated Saudi individuals (37 males and 23 females; mean age: 51 +/- 5 years). HLA Class I and Class II antigens were detected by lymphocytotoxicity and a polymerase chain reaction (PCR) method using DNA sequence-specific primers. Although present in small numbers, HLA Cw2 was found in significantly fewer patients (n = 11; 4.68%) compared to normal subjects (n = 9; 15%) and was found to confer protection against ESRD (P = 0.005; relative risk [RR]: 3.594, 95% confidence interval [CI]: 1.415-9.126). Among the HLA Class II antigens, HLA DQB1*03(8) was detected more frequently in the patient group (n = 65; 27.6%) than in the normal controls (n = 9; 15%) and was positively associated with risk of ESRD (P = 0.04; RR: 0.462, 95% CI: 0.215-0.991). No significant differences were observed between the two groups in respect of HLA-A2, HLA-B50(21), HLA-B51(5) and HLA-Cw7 (HLA Class I), and HLA-DRB1*04, HLA-DRB1*07 and HLA-DQB1*02 (HLA Class II). Occurrence of the most frequent HLA alleles was no different between the ESRD group and the controls. The protective role of HLA-Cw2 and the marginal susceptibility associated with HLA-DQBI*03(8) for ESRD requires further investigation.

Clinical and therapeutic features of brucellosis: An 11-year study at a tertiary care hospital in Riyadh, Saudi Arabia.

To study the varying presentations, risk factors, and treatment outcomes among patients with physician-diagnosed brucellosis. This retrospective analysis evaluated all cases of brucellosis reported at King Khalid University Hospital during 2003-2013. Data were retrieved from patient records and a laboratory information system. Descriptive statistics were generated to summarize the study variables. Fisher's exact test or Pearson's chi-square test was used to compare categorical variables. Out of 163 patients identified with brucellosis, 76.7% of patients were culture positive. Fever was the most frequent symptom (85.9%), followed by arthralgia (46.6%). The most common clinical signs was splenomegaly (12.9%), followed by hepatomegaly (11.0%). Laboratory investigations revealed lymphocytosis and anemia in 66.3% and 55.2% of the patients, respectively. Approximately half of the patients (47.8%) had high erythrocyte sedimentation rates, and 56.4% had neutrophilia. Raw milk consumption and direct contact with animals were reported by 45.4% and 16.0% of patients, respectively. Treatment failure and relapse were observed in 8 (5.7%) cases. All treatment failures and relapses occurred among children <= 10 years of age or adults > 45 years old (11.0% vs. 0%; p = 0.006). Our findings demonstrate that raw milk consumption can be a substantial factor in brucellosis prevention in Saudi Arabia. Laboratory findings, along with the observed pattern in clinical signs and symptoms, can potentially mean underdiagnosis of mild cases. Age was the only factor associated with unfavorable treatment outcomes.

A new method for measuring clustering in suspension between accessory cells and T lymphocytes.

Specifying the molecular basis and clinical significance of cluster formation between antigen-presenting cells and T lymphocytes will be important in many areas of immunology. In this paper we describe a novel and reproducible technique for measuring cluster formation in suspension between purified human blood monocytes and purified autologous T lymphocytes, and its application to determining the effects of recall antigens and mitogen. Blood monocytes and T lymphocytes from eight normal subjects were separately prelabelled with two different carbocyanine dyes prior to co-culture in suspension with or without antigen (PPD, SKSD) or mitogen (PHA). At 24 h the co-cultures were examined for cluster formation by ultraviolet microscopy and flow cytometry. Control experiments showed that the carbocyanine dyes were non-toxic in vitro, that cell labelling was stable for culture periods up to 120 h, and that the two dyes did not leak from cell to cell. By this technique we measured the proportion of monocytes clustering one or more T lymphocytes in the presence and absence of recall antigen or PHA. There was a close correlation between visual and flow cytometric measurement of monocyte: T lymphocyte clustering (p < 0.001) as well as a close relationship between the ability of the two recall antigens to increase the extent of clustering above baseline (p < 0.001). Antigen-increased cluster formation did not correlate with baseline clustering, unlike PHA-increased clustering, which was related to baseline levels (p = 0.02), suggesting the operation of distinct mechanisms. The method is applicable to measuring cell-cell associations in suspension during extended periods of culture, as well as for the study of agents which might modify intercellular adhesion processes.

A method of preparing blood leucocytes for flow cytometry which prevents upregulation of leucocyte integrins.

LeuCAM (CD11/CD18) cell-surface antigens are easily upregulated on cell manipulation ex vivo. A procedure for preparing leucocytes, in which human blood is immediately treated ex vivo with buffered formaldehyde and then the erythrocytes and platelets are removed by lysis and differential centrifugation, has been successfully applied to the analysis of LeuCAM antigen expression by flow cytometry. We show that the increased expression of monocyte CD11/CD18, which occurs when mononuclear leucocytes are separated by a standard Lymphoprep density gradient separation, can be avoided if cells are fixed immediately. Following this fixation polymorphs are unable to upregulate CD11/CD18 in response to fMLP stimulation in vitro. The technique produces lymphocyte, polymorph and monocyte populations that can be clearly defined on the basis of forward scatter and side scatter, and preserves the expression of various surface antigens; the percentages of gated lymphocytes expressing CD3, CD4, and CD8 were similar to those obtained using a commercial fixing and lysis solution. The processing does not render cells permeable to antibodies, as evidenced by our failure to stain cells with antibodies to intracellular antigens. We believed the method to be useful for measuring CD11/CD18 expression on blood leucocytes from normal or pathological specimens and to have application to the measurement of other cells surface antigens which may also be upregulated by the separation procedures.

Garlic and onion sensitization among Saudi patients screened for food allergy: a hospital based study.

BACKGROUND: Detection of specific IgE antibodies against food materials indicates allergic sensitization. Some very widely consumed foods materials such as garlic and onion have rarely been investigated for their allergenic potential. OBJECTIVES: To assess the presence of garlic and onion specific IgE antibodies in patients investigated for food allergy. METHODS: Radioallergosorbent test (RAST) results of 108 patients with clinical suspicion of food allergy who were specifically screened for garlic and onion specific IgE antibodies along with other food allergens were analyzed retrospectively at King Khalid University Hospital between January 2008 and April 2009. This group of patients included 73 males and 35 females with mean age 27+13.2 years. Estimation of garlic and onion specific IgE antibodies was performed by radioallergosorbent test (RAST) using Pharmacia ImmunoCAP 250 analyzer. RESULTS: Out of the 108 patients 15 (13.8%) had garlic and onion specific IgE antibodies in their sera. Garlic specific IgE antibodies with the RAST scores between one to four were present in 14 and onion specific IgE were detected in 13 patients. For garlic specific IgEs majority of patients (08) had RAST score of one (0.35-0.69 kU/L) and for onion specific IgE antibodies seven patients had RAST score of two (0.70-3.49 kU/L). Among these patients 12 (80%) were found to have coexisting specific IgE antibodies against garlic and onion. CONCLUSION: The presence of garlic and onion specific IgE antibodies in a sizeable number of patients indicate sensitization and allergenic potential of these food materials.

Increased CD11/CD18 expression on peripheral blood leucocytes of patients with sarcoidosis.

Sarcoidosis is a multisystem disease of unknown etiology characterized by non-caseating granulomata, formed mainly from macrophages surrounded by lymphocytes and plasma cells. Using a novel method for the preparation of blood leucocytes for flow cytometry, we report increased expression of LeuCAMs (CD11/CD18) on peripheral blood leucocytes of 11 Caucasian and 10 Afro-Caribbean patients with sarcoidosis compared with age-, sex- and race-matched controls. Whilst the percentages of the cells expressing CD11/CD18 were no different, the density, expressed as mean fluorescence intensity (MFI), was greater for all leucocytes in sarcoids than in normal individuals. The expression of intercellular adhesion molecule-1 (ICAM-1), a ligand for LFA-1 which is expressed on all leucocytes, was not significantly different from normal, whereas HLA-DR was expressed more intensely on sarcoid monocytes (P less than 0.01) and blood lymphocytes (P less than 0.005) than control cells. Our findings are consistent with leucocyte activation although we were unable to confirm reports of elevated tumour necrosis factor-alpha (TNF-alpha) in the patients' plasma using an ELISA. Increased expression of adhesion molecules on peripheral blood leucocytes may play a role in the cellular extravasation, aggregation, and granuloma formation seen in sarcoidosis.

Circulating adhesion molecules in sarcoidosis.

Sarcoidosis is a disease of unknown etiology characterized by non-caseating granulomata together with a number of systemic abnormalities. We have recently shown these include increased expression of the integrins CD11/CD18 on peripheral blood leucocytes. Here we have measured serum levels of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1), E-selectin and vascular cell adhesion molecule-1 (VCAM-1) in 23 patients and 14 normal controls using antigen capture sandwich ELISAs. Median circulating E-selectin levels in the patients were nearly three times those of the controls (P < 0.0001, Mann-Whitney U-test), whilst ICAM-1 but not VCAM-1 levels were only slightly elevated. These results show that endothelial cell activation and shedding of E-selectin into the circulation are additional features of the pathology of sarcoidosis.

Increased prevalence of asthma in Saudi Arabia.

BACKGROUND: Bronchial asthma is among the most common chronic illnesses of childhood. A number of reports in the recent past suggest that the prevalence of asthma is increasing globally. OBJECTIVE: To investigate the changing prevalence of asthma in the Kingdom of Saudi Arabia. SUBJECTS AND METHODS: Two populations of schoolchildren between the ages of 8 and 16 years were studied using an internationally designed protocol in 1986 and 1995. The questionnaire used in these studies was very similar to the one used in the International Study of Allergy and Asthma in Childhood. A total of 2,123 school-children in 1986 (Jeddah and Riyadh) and 1,008 schoolchildren in 1995 (Hail and Gizan) were enrolled in the surveys. These cross-sectional studies of randomly selected schoolchildren were statistically analyzed using ANOVA and a Z test. RESULTS: The comparison of data between Riyadh versus Hail (inland desert dry environment) and Jeddah versus Gizan (coastal humid environment) revealed that the prevalence of asthma in the similar populations increased significantly from 8% in 1986 to 23% in 1995 (P < .0001). Likewise, the prevalence of allergic rhinitis also increased from 20% to 25% (P < .003) since 1986. However, no significant change in the prevalence of eczema (from 12% to 13%) was noted between 1986 and 1995. CONCLUSIONS: The study indicates that there was a significant increase in the prevalence of bronchial asthma and, to a lesser extent, in the prevalence of allergic rhinitis in the Kingdom of Saudi Arabia during this 9-year period. The study also revealed increased exposure to environmental factors such as tobacco smoke and indoor animals in Saudi houses. It seems that the continuing changes in contemporary life may well have contributed to the increased prevalence of asthma in the country.

Normal and sarcoid alveolar macrophages differ in their ability to present antigen and to cluster with autologous lymphocytes.

Human bronchoalveolar macrophages from normal individuals function poorly as accessory cells for the presentation of common recall antigens. In sarcoidosis, alveolar macrophages (AM) are reported to be effective accessory cells for the presentation of such antigens. In this study normal and sarcoid AM were compared with blood monocytes for their ability to act as accessory cells in presenting tuberculin purified protein derivative (PPD) and streptokinase-streptodornase (SKSD) to autologous T lymphocytes, or to form spontaneous, antigen- or mitogen-induced clusters with the T cells. When compared to autologous monocytes, normal AM failed to present the two recall antigens effectively. Likewise normal AM formed very few clusters with T lymphocytes when compared to monocytes, even in the presence of antigens or the mitogen phytohaemagglutinin (PHA). In contrast, sarcoid AM presented both antigens as effectively, and were equally effective as monocytes in forming clusters with T lymphocytes, spontaneously and in further response to antigen or mitogen. The results suggest that in sarcoidosis enhanced accessory cell function and enhanced cluster formation may be related features of bronchoalveolar macrophage populations.

Inhibition of leucocyte adhesion molecule upregulation by tumor necrosis factor alpha: a novel mechanism of action of sulphasalazine.

The effects of the cytokine tumour necrosis factor alpha and the calcium ionophore A23187 upon CD11a, CD11b, CD11c and CD18 leucocyte membrane expression was analysed in whole blood using monoclonal antibodies and flow cytometry. Both agents significantly increased the density of CD11b/CD18 membrane expression on monocytes and granulocytes, but had no effects on adhesion molecule expression on lymphocytes. The effects of sulphasalazine, 5-aminosalicylic acid (5-ASA) and sulphapyridine upon adhesion molecule upregulation were then examined; 10(-3) and 10(-4) M sulphasalazine and 5-ASA significantly reduced tumour necrosis factor alpha induced CD11b/CD18 upregulation on monocytes and granulocytes but had no effects upon A23187 mediated upregulation. Sulphapyridine was inactive. These results suggest that sulphasalazine and 5-ASA may interfere with mechanisms of leucocyte recruitment in inflammatory bowel disease.

Alveolar macrophage accessory cell function in bronchial asthma.

The capacity of peripheral blood monocytes and alveolar macrophages (AM) obtained by bronchoalveolar lavage (BAL) to present recall antigens, namely, tuberculin purified protein derivative (PPD) or streptokinase-streptodornase (SKSD), to highly purified autologous T-cells has been studied in 11 asthmatic and 11 healthy, nonatopic normal subjects. In the asthmatic group, AM accessory cell function was variable, and most subjects were unable to present either recall antigen as effectively as blood monocytes, although one asthmatic subject demonstrated larger proliferative responses than blood monocytes for both antigens. AM accessory cell activity was not antigen-specific, and there was a correlation between accessory cell efficacy for the two antigens (r = 0.92; confidence interval, 0.53 to 0.98). Furthermore, a correlation existed between the percentage lymphocyte count in the BAL fluid and the ratio of macrophage to monocyte antigen-presenting capability for both PPD (r = 0.92; 95% confidence interval, 0.83 to 0.99) and SKSD (r = 0.90; 95% confidence interval, 0.45 to 0.98). In the normal subjects, AM were also unable to act effectively as accessory cells for the presentation of PPD and SKSD in the majority of subjects. No correlation existed between the percentage lymphocytes in BAL fluid and the ratio of AM to monocyte accessory cell function. These results suggest an association between AM accessory function and the presence of BAL lymphocytes in bronchial asthma.