Iranian Journal of Allergy, Asthma and Immunology
Volume:1 Issue: 2, Jun 2002

The use of immunoglobulin Y (IgY) instead of polyclonal antibodies prepared in mammals has been reported recently. These antibodies are found to have better properties in terms of specificity and ease of large scale production. In this study, proteins from egg yolk were extracted with acid and chloroform. The result of cellulose acetate electrophoresis at pH 9 showed some extra bands which were further purified with sodium sulfate precipitation (20%). The purified band was observed in the y globulin region. SDS-PAGE eletrophoresis of the product showed impurities which were removed by T-gel chromatography. Following that only H-chainof immunoglobulin Y with a molecular weight of about 66 KD was observed. Using the above procedure, a yield of around 40.7 mg IgY / egg was obtained.

Congenital lobar emphysema is a rare cause of respiratory distress during infancy which is cured by surgery. We are reporting three cases of congenital lobar emphysema with different presentations of the disease. Chest x-ray was the basis of diagnosis but confirmatory lung computerized tomography was also used. The emphysematous lobe was left upper lobe in two infants and right upper and middle lobes in the third one. Resection of the affected lobes was performed with good results.

We have reviewed the medical records of 40 patients with disseminated (Bacille Calmette-Guerin) BCG from 1996 to 1999 in the Immunology Department of the Children Hospital, Medical Center, Tehran university. These patients are divided in to 3 groups:1. Patients who had disseminated (Bacille Calmette-Guerin) BCG after vaccination and their diagnosis was chronic granulomatous disease.2. Patients who had disseminated (Bacille Calmette-Guerin) BCG and were diagnosed as having cell mediated immunodeficiency.3. Patients whose Nitroblue Tetrazulium and CMI were around normal, but they could not kill intracellular mycobacterium, because of confirmed deficiency of interferon-gam ma receptor and IL12 receptor.Interferon-gamma receptor or CD''119 was checked in 6 patients. In one patient interferon-gamma receptor deficiency was confirmed by flowcytometric analysis. In other patients, this marker was around normal, but presumably they had IL12 receptor deficiency, which we were unable to detect in our laboratory. In some patients this marker should be checked after preparation of more laboratory facilities.

Interleukin-4 (IL-4) is produced by T-helper cells type 2 (TH2) and induces IgE synthesis. T-helper cells type 1 (TH1) produce interferon-gamma (IFN-gamma) which suppresses TH2 and reduces IL-4 induced IgE production. In this: ;tudy, we demonstrated that the levels of specific IgE in the serum of atopic children (n=20) were elevated while IL-4 production was increased and IFN-gamma secretion was reduced, compared to those of control group. Inter!eukin-4-induced IgE synthesis by peripheral blood mononuclear cells of atopic children in vitro was blocked in the presence of IFN-gamma. In addition, levels of soluble CD23-which is specifically induced by IL-4 were significantly elevated in our atopic patients. The data indicate that enhanced production of IL-4 and lowered IFN-gamma secretion by T-helper cells correlate with the elevated specific IgE levels in the serum of atopic children.

Anti-neutrophil cytoplasmic autoantibodies have been detected in patients with some autoimmune and vascular diseases such as Wegner''s granulomatosis, polyarthritis nodosa and systemic lupus erythematosus. Indirect immunofluorescence technique has been employed to detect these autoantibodies. By this method, two general patterns of antineutrophil cytoplasmic antibodies were seen: a cytoplasmic (C-ANCA) and a perinuclear form (P-ANCA). These antibodies have also been observed in uveitis. In this study, the presence of antineutrophil cytoplasmic antibodies in 25 patients with uveitis and its relationship with anatomical location of the autoimmune reaction was evaluated. According to the results, antineutrophil cytoplasmic antibodies were detected in 16% (4 out of 25) of the patients, all of which were C-ANCA type. The results also showed that there was no significant correlation between the presence of antineutrophil cytoplasmic antibodies and anatomical location of the disease (p=0.65).

This study was undertaken to assess vitamin C status in allergic children. Twenty-six allergic patients and 46 apparently healthy controls aged 7-16 years of both sexes were introduced. All patients were diagnosed being allergic based on their histories, physical examinations and laboratory findings. Blood samples were obtained between 09.00-11.00 to determine total serum immunoglobulin E, histamine, plasma vitamin C and complete blood cell count. Stool examinations and urinealysis were also done. Although total serum immunoglobulin E levels were significantly higher in patients than in controls, the serum histamine and plasma vitamin C levels showed no significant difference between the two groups. Surprisingly, patients with allergic dermatitis and food allergy had significantly lower plasma vitamin C levels than patients with asthma and rhinitis. Also, female patients tended to have higher plasma vitamin C but lower total serum immunoglobulin E levels compared to the male patients but none of these differences were significant. Healthy boys, on the other hand, had significantly higher plasma vitamin C than healthy girls. Our findings did not confirm the previous reports which showed decreased plasma vitamin C levels in allergic patients. We concluded that the plasma vitamin C levels in different allergies might be infuenced by such factors, such as sex, type and the stage of allergic disease, besides those affecting intake.

Immunoglobulin E is one of the five classes of immonoglobulins that plays an important role in allergic diseases. Production of monoclonal antibodies by a single clonotype against different epitopes of immunoglobulin E has high priority in development of diagnostic kits.In this study, an attempt was made to produce monoclonal antibodies against human immunoglobulin E. Balb/c mice were immunized with semipurified immunoglobulin E and spleen cells fused with SP2.0 mouse myeloma eel! line in the presence of polyethylene glycol. Supernatant of hybridoma cells was screened for detection of antibody by enzyme linked immonosorbent assay method. Cloning of selective high absorbance wells were done with limiting dilution method. The suitable clone (monoclone) was selected by enzyme linked immunosorbent assay and confirmed by immunoblot. The subclass of the chosen monoclonal antibodies was determined and the clones freezed and kept in liquid nitrogen.During this study three successful fusions were carried out, which resulted in development of 156 clones with high production of anti-IgE. Fourteen clones with the highest titres were selected for cloning. After limiting dilution more than 100 monoclonal antibodies were produced and the suitable (me (GJ0F7), i.e.; the clone which displayed the high absorbance in reaction with purified immunoglobulin E and the lowest cross-reactivity with immunoglobulin M, immunoglobulin G and immoglobulin A was chosen. In immunoblotting, presence of high density band in reaction with immunoglobulin E was confirmed. The suitable mab was shown to be IgG 1 subclass with kappa light chain. It seems that, this mab could be successfully used in diagnostic kits.

Shwaehman syndrome, next to cystic fibrosis, is the second cause of congenita! exocrine pancreatic insufficiency in children. It appears as steatorrhea, recurrent infections and hematologic abnormalities such as neutropenia, skeletal dysplasia and short stature. In this study, we reviewed 3 patients'' histories. All of them showed cellular chemotaxic defect. One of them had been affected by nephrotic syndrome and finally succumbed to septic shock. This patient suffered from skeletal disorders.

Adenylyl cyclase is a membrane-bound enzyme that catalyzes the conversion of ATP to cAMP. The inhibition of adenylyl cyclase was carried out by measuring the ability of the macrophage chemotactic protein-1 to inhibit the torskolin-induced enzyme activity. Adenylyl cyclase activity in the presenc of macrophage chemotactic protein-1 was decreased compared to that in controls [2.11+/-0.15 (mean +/- SD.) vs. 6.83 +/- 0.45, activity (/nnol cAMP/mg protein/min).

The adhesion molecules are involved in adhesion of leukocytes to endothelial cells and other immune cells. Not only adhesion molecules have membrane form, but they also have soluble form. In this paper, we attempted to investigate the concen tration of soluble adhesion molecules in the type 1 diabetic patients (n = 40) and healthy subjects (n = 10). The results indicated that there was an increase in the concentration of soluble adhesion molecules: sICAM-1, sVCAM-1 and sELAM-1, in the sera of diabetic patients rather than the healthy ones (p = 0/00). Increased concentration of sVCAM-1 with the insulin injection dose (p = 0.01) and sIC AM-1 with duration of disease (p = 0.05) indicated significant reversel linear con-elation in the patients'' groups. Also, sELAM-1 with sVCAM-1 and sICAM-1 showed significant direct linear correlation in the same group (p ~ 0.01). Adhesion mol ecules were determined by the sandwich ELIS A principle. One of the most important factors in the development of atherosclerosis is the ad hesion of monocytes to endothelial cells. Tlierefore, in this study, the percentage of expression of membrane form of VCAM-1, not ICAM-1, was increased on 10000 monocytes of diabetic patients type-1 (n = 20) in comparison with normal subjects (n = 20, p ~ 0.05). The expression tensive of membrane form of CAMs on the surface of monocytes was performed by flowcytometry technique. We concluded that the increase in sICAM-1, sVCAM-1 specially sELAM-1 and also mVCAM-1 indicated endothelial activation, stimulatability of leukocyte cells and increased interaction to endothelial cells. Although, scientists are not aware about the role of elevated CAMs concentration, it can be suggested that the in creasing level of sICAM-1, sVCAM-1, sELAM-1 and mVCAM may reflect cel lular expression, taction of CAMs in autoimmune disease and also provide a po tential therapeutic target for human IDDM.