Effect of Inonotus Obliques Extracts on Proliferation and Caspase-3 Activity in Human Castro-Intestinal Cancer Cell Lines

We studied the effects of hot water extract of Inonotus obliquos mushroom on the proliferation and apoptosis of the human colon adenocarcinoma, HT-29 and the human stomach adenocarcinoma, SNU-484 cell. Cells were maintained with Dulbecco's modified Eagle medium/Ham's F-12 nutrient mixture supplemented with 10% fetal bovine serum at 37 degrees C in a humidified CO2. For the cell proliferation experiments, cells were seeded in 35 mm dishes, and were treated with the various concentrations of the extract for the different time course. Apoptosis was measured by caspase-3 activity. When we incubated HT-29 cells for 24, 48, 72, and 96 hours after treatments, the cell proliferation was more suppressed with more treatment time. In case of the human stomach cancer cell, SNU484, the extract significantly decreased the cell number. Thus, the treatment of 1.5 mg/ml extract decreased almost half of the cell number. Caspase-3 activity in HT-29 was increased by the treatment of mushroom extracts. In SNU484, caspase-3 activity tended to increase in proportion to the amounts of the extracts and the treatment of Inonotus obliquos affected the activity a lot. Therefore, Inonotus obliquos is suggested for the prevention of gastro-intestinal cancer and strongly recommended for the treatment of stomach cancer.

Comparison of Skin Prick Test Results between Crude Allergen Extracts from Foods and Commercial Allergen Extracts in Atopic Dermatitis by Double-Blind Placebo- Controlled Food Challenge for Milk, Egg, and Soybean

Skin Prick Test (SPT's) are performed to identify the causes of allergy. However, low diagnostic accuracy is a limitation to SPT, for which many possible causes have been suggested. The protein composition and allergenicity of crude allergen extracts from foods and commercial allergen extracts for SPT were analyzed. Clinical significances of SPT using crude allergen extracts from foods were compared with those using commercial allergen extracts. A total of 292 atopic dermatitis patients were involved in this study. Crude allergen extracts were prepared from milk, egg white, egg yolk, and soybean. The protein composition of food allergen extracts and commercial allergen extracts of milk, whole egg, white, egg yolk, and soybean were compared by SDS-PAGE. The allergenicity was tested by the immunoblotting method using immune sera. SPTs were performed using crude and commercial allergen. Double-blind placebo- controlled food challenge (DBPCFC) was performed to verify the SPT results and to compare the clinical significance of crude and commercial allergen extracts. Protein composition differed markedly between crude and commercial allergen extracts. By immunoblotting, crude and commercial allergen extracts showed different allergenicity. The SPT results using crude and commercial allergen extracts showed significant differences. The prevalence of milk, egg and soybean allergy was over 35% in atopic dermatitis. The accuracy of SPT using crude allergen extracts from foods was significantly higher than that using commercial allergen extracts. In the case of soybeans, the result of SPT using commercial allergen extract was clinically insignificant for the prediction of soybean allergy. The source of allergen extract was very important for the appropriate SPT in food allergy. The accuracy of SPT might be improved using the appropriate allergen source for food allergy.

Clinical application of histamine prick test for food challenge in atopic dermatitis

Determining positive food challenges are not easy as there is an absence of simple and objective tests. Histamine, an essential mediator for allergic reactions, is involved in the pathogenesis of atopic dermatitis (AD) and food challenges can change histamine levels. The significances of a prick test with histamine (histamine prick test, HPT) relating to the interpretation of food challenges in AD were evaluated. A total of 467 AD patients participated in this study. Skin prick tests, identification of specific IgE and open food challenge were conducted for the identification of food allergy. Elimination diet was performed with HPT. HPTs were conducted before and after food challenges. The wheal sizes by HPT were significantly decreased after an elimination diet. The relative changes of wheal sizes significantly correlated with those of clinical severity scores in AD patients (p<0.001). The wheal sizes in HPT were increased with a positive provocation in open food challenges. In conclusion, HPT may be a simple and objective test to interprete the results of food challenges in patients with AD. The exact mechanisms of the changes in skin reactivity by HPT need further investigation.

Plasma Interferon(IFN)-gamma, Interleukin(IL)-4, IL-5 and IL-10 Concentrations in Atopic Dermatitis / 소아알레르기및호흡기학회지

PURPOSE: There is reduced IFN-gamma production with increased IL-4 production in atopic dermatitis. IgE production is known to increase from an imbalance of IFN-gamma and IL-4 production. IgE overproduction is regarded as a major problem in the pathogenesis of atopic dermatitis. In this study we evaluated the significances of plasma IFN-gamma, IL-4, IL-5 and IL-10 concentrations in atopic dermatitis. Also the correlation between IL-4 and IgE levels as well as IFN-gamma and IgE levels were tested. METHODS: One hundred and five(105) atopic dermatitis patients who fulfilled the criteria of Hanifin and Rajka were tested as an atopic dermatitis patient group. Forty(40) normal controls who have not had any personal or family history of allergic diseases were tested as a normal control group. Routine hematologic tests, plasma IgE levels and total eosinophil counts were tested in both groups. Also plasma IFN-gamma, IL-4, IL-5 and IL-10 concentrations were measured using high-sensitive IL-4, IL-5, IL-10 and IFN-gamma ELISA kits in both groups. RESULTS: There was no noticeable difference in WBC counts between the atopic dermatitis group and the normal control group. In comparison, eosinophil percents in WBC and total eosinophil counts were significantly high in the atopic dermatitis group. Plasma IgE levels were also markedly elevated in the atopic dermatitis group. Plasm IFN-gamma levels were significantly low in the atopic dermatitis group(0.58+/-2.12 pg/ml) as compared with normal control group(5.20+/-2.60pg/ml)(P<0.01). IL-4 and IL-5 were not detected in normal controls. But in the atopic dermatitis group plasma IL-4 concentration was 1.00+/-2.05pg/ml and IL-5 was 2.18+/-1.96pg/ml. Plasma IL-10 concentration was significantly low in the atopic dermatitis group(2.36+/-3.38 pg/ml) as compared with the normal control group(9.78+/-4.52pg/ml)(P<0.01). CONCLUSIONS: Plasma IFN-gamma, IL-4, IL-5 and IL-10 levels were clinically significant in atopic dermatitis. However, plasma IL-10 levels of the atopic dermatitis was lower as compared to that of the normal subject. There was no significant correlation among plasma IFN-gamma, IL-4 levels, and blood IgE levels.

Decrease of Plasma Th2 Cytokines by Interferon (IFN)-gamma Therapy in Atopic Dermatitis

PURPOSE: Atopic dermatitis is characterized by reduced IFN-gamma production and increased IL-4 production. As a result, IgE production increases in atopic dermatitis. In the previous studies, it was reported that recombinant IFN-gamma therapy is effective in treatment of severe atopic dermatitis. In this study, changes of plasma IFN-gamma, IL-4, IL-5 and IL-10 concentration by IFN-gamma therapy were studied in atopic dermatitis. Changes of plasma IgE levels and eosinophil counts were also investigated in the present report. METHODS: Sixty-five atopic dermatitis patients were studied. Diagnostic criteria for atopic dermatitis were those used by Hanifin and Rajka. Patients received 2x106 units/m2 IFN-gamma by subcutaneous injection eighteen times for six weeks. The following investigations were performed : complete blood cell count, total IgE, eosinophil percentage and total eosinophil count in addition to plasma IFN-gamma, IL-4, IL-5 and IL-10 concentration. RESULTS: Plasma concentrations of IL-4, IL-5 and IL-10 decreased by IFN-gamma therapy in atopic dermatitis. However, plasma IFN-gamma concentration was not changed. No significant correlations among the changes of IgE, eosinophil counts and plasma cytokine concentrations were detected. CONCLUSION: Plasma concentrations of Th2 cytokine such as IL-4, IL-5 and IL-10 decreased by IFN-gamma therapy. This study suggests that Th2 cytokines might not be produced simulaneously. and that changes of Th2 cytokines might not affect the quantitiative changes of IgE and of eosinophil count.

Decrease of Plasma Th2 Cytokines by Interferon (IFN)-gamma Therapy in Atopic Dermatitis

PURPOSE: Atopic dermatitis is characterized by reduced IFN-gamma production and increased IL-4 production. As a result, IgE production increases in atopic dermatitis. In the previous studies, it was reported that recombinant IFN-gamma therapy is effective in treatment of severe atopic dermatitis. In this study, changes of plasma IFN-gamma, IL-4, IL-5 and IL-10 concentration by IFN-gamma therapy were studied in atopic dermatitis. Changes of plasma IgE levels and eosinophil counts were also investigated in the present report. METHODS: Sixty-five atopic dermatitis patients were studied. Diagnostic criteria for atopic dermatitis were those used by Hanifin and Rajka. Patients received 2x106 units/m2 IFN-gamma by subcutaneous injection eighteen times for six weeks. The following investigations were performed : complete blood cell count, total IgE, eosinophil percentage and total eosinophil count in addition to plasma IFN-gamma, IL-4, IL-5 and IL-10 concentration. RESULTS: Plasma concentrations of IL-4, IL-5 and IL-10 decreased by IFN-gamma therapy in atopic dermatitis. However, plasma IFN-gamma concentration was not changed. No significant correlations among the changes of IgE, eosinophil counts and plasma cytokine concentrations were detected. CONCLUSION: Plasma concentrations of Th2 cytokine such as IL-4, IL-5 and IL-10 decreased by IFN-gamma therapy. This study suggests that Th2 cytokines might not be produced simulaneously. and that changes of Th2 cytokines might not affect the quantitiative changes of IgE and of eosinophil count.

Thymopentin Therapy in Atopic Dermatitis

PURPOSE: Atopic dermatitis is a chronic relapsing inflammatory skin disease that results from allergic reaction. Steroid therapy has become a major therapeutic modality for treatment of atopic dermatitis. Several immunomodulatory therapies have been tried for atopic dermatitis. In this study thymopentin therapy was performed and its clinical effects and laboratory results were evaluated. METHODS: Atopic dermatitis with typical clinical symptoms were included in this study. Twelve patients were treated with subcutaneous injection of thymopentin ( (Imupentin ) at a dose of 1 mg/kg, three times per week for eight weeks. Clinical scores were measured by Hanifin scoring system before and at the end of the treatment. General hematologic laboratory tests such as hemoglobin, hematocrit, WBC count, neutrophil percentage, lymphocytes percentage and eosinophil percentage as well as blood IgE level were conducted at the same time. RESULTS: The clinical severity of atopic dermatitis patients was markedly improved by thymopentin therapy. The clinical severity score improved by 85.5%. Three patients showed complete remission of clinical status at the end of thymopentin therapy. Hemoglobin, hematocrit, WBC counts, neutrophil percentage, and lymphocyte percentage were not affected by the thymopentin therapy. Eosinophil percentage and blood IgE level were significantly reduced statistically. CONCLUSION: Thymopentin therapy is an effective immunomodulatory therapeutic modality in atopic dermatitis. Thymopentin decreased eosinophil percentage and blood IgE levels without other hematologic changes.

CD5 mRNA Overexpression in Lymphocytes of Allergic Patients / 대한면역학회지

PURPOSE: The CD5 molecules are pan-T cell antigens and are found on a minor subpopulation of B cells. CD5 antigens are involed in an intracellular signal transduction as well as in an intercellular signal transduction between CDS+ T cell/CD72+ B cell by CD5/CD72 interaction. CD5 antigens are known to be participated in classic immune reactions and in this study CDS mRNA expressions by lymphocytes were examined in allergic patients controls, acute febrile infectious disease controls and normal controls to elucidate the possibility of CDS involvement in allergic immune reactions. METHODS: Fifteen allergic patients, ten patients of acute febrile infectious disease patients and ten normal controls were studied. Venous blood was drawn and mononuclear cells were separated. T cells and B cells were separated using immunomagnetic beads. Total RNA was extracted and RT-PCR (reverse transcriptase - polymerase chain reaction) was done to detect CDS antigen mRNA expression. RESULTS: 1) CDS mRNA overexpressions were detected in allergic patient controls as compared to that in acute febrile infectious controls. CDS mRNA was not detected in normal controls. Semiquantitative CD5 mRNA expressions were measured as relative expressions of CD5 to GAPDH. Relative quantities of CD5 mRNA expressions were 90.656.24% in allergic patient controls and 23.76+3.58% in acute febrile infectious patients. CONCLUSIONS: CDS mRNA overexpression is a characteristic phenomenon in allergic immune reactions. From these result, CD5/CD72 pathway might be the preference immune mechanism in allergic immune reaction and the further study for the exact mechanism of CDS involvement in allergic immune reactions may be necessary

Increased serum interleukin-10 level in Kawasaki disease

Serum IL-10 level in Kawasaki disease(KD) was tested. In the KD patients' sera during the acute phase, the levels of IL-10 were markedly elevated (122.0 +/- 39.1 pg/ml) compared to 3.7 +/- 1.7 pg/ml in the control subjects (p< 0.001). The serum IL-10 levels remained elevated in the subacute phase (16. 7 +/- 9.7 pg/ml, p< 0.001) and were restored to the normal levels(7.9 +/- 3.9 pg/ml) during the convalescent phase. In the patients with acute febrile disease, the serum IL-10 level increased significantly (34.4 +/- 14.1 pg/ml, p< 0.001) compared to that of the age-matched control subjects, but were not as high as in acute phase of KD(p< 0.005). This increase in serum IL-10 levels in KD may contribute to the up-regulation of humoral immunity and to the down-regulation of acute inflammation due to an increase in proinflammatory cytokines.

Decreased CD5+ B cells during the acute phase of Kawasaki disease

We investigated the changes of CD5+ B cells in the peripheral blood of 20 Kawasaki disease (KD) patients. The percentage of CD5+ B cells in the total lymphocytes and in the total B cells significantly decreased during the acute phase of KD(p< 0.01), compared to that in the age-matched normal control subjects. After intravenous immunoglobulin(IVIG) treatment, the percentage of CD5+ B cells increased, but was still lower than that in the normal controls(p< 0.01). During the convalescent phase of the disease, the percentage of CD5+ B cells was restored to the normal levels. The levels of CD5+ B cell percentage in the total B cells of the patients with acute febrile disease showed similar levels to age-matched normal controls. The decreased CD5+ B cells in the patients with KD provides an additional abnormal immunological finding during the acute phase of the disease.

Effects of intravenous immune globulin on the peripheral lymphocyte phenotypes in Kawasaki disease

The effect of intravenous immune globulin (IVIG) on the lymphocyte phenotypes in acute Kawasaki disease (KD) was studied in a random trial of IVIG-and-aspirin versus aspirin-alone. Before therapy, patients in each treatment group had an increased percentage of B cells, and a decreased percentage of T cells, CD4 T cells, CD8 T cells and CD5+ B cells. There was no significant difference in immunologic parameters between the two groups measured before therapy. Patients treated with IVIG-and-aspirin had by the fourth day developed a highly-significant increase in T cells, CD4 T cells and CD8 T cells and a decrease in B cells. Despite the decrease of B cells, there were significant increases in CD5+ B cells in both treatment groups. However, the degree of increase in the IVIG-and-aspirin treated group was significantly more noticeable than that in the aspirin-alone treated group. These findings indicate that treatment with IVIG restores the T- and B- cell abnormalities, especially CD5+ B-cell abnormalities found in patients with acute KD.

Effects of intravenous immune globulin on the peripheral lymphocyte phenotypes in Kawasaki disease

The effect of intravenous immune globulin (IVIG) on the lymphocyte phenotypes in acute Kawasaki disease (KD) was studied in a random trial of IVIG-and-aspirin versus aspirin-alone. Before therapy, patients in each treatment group had an increased percentage of B cells, and a decreased percentage of T cells, CD4 T cells, CD8 T cells and CD5+ B cells. There was no significant difference in immunologic parameters between the two groups measured before therapy. Patients treated with IVIG-and-aspirin had by the fourth day developed a highly-significant increase in T cells, CD4 T cells and CD8 T cells and a decrease in B cells. Despite the decrease of B cells, there were significant increases in CD5+ B cells in both treatment groups. However, the degree of increase in the IVIG-and-aspirin treated group was significantly more noticeable than that in the aspirin-alone treated group. These findings indicate that treatment with IVIG restores the T- and B- cell abnormalities, especially CD5+ B-cell abnormalities found in patients with acute KD.

In vitro immunization-IV: antibody response of mouse splenocytes grown in a mixture of conditioned media, thymocytes, and bone marrow cells

No abstract available.

The effect of conditioned media cultured from the thymocytes and the sphenocytes on the regulation of antibody isotypes; in vitro immunization-ill

No abstract available.

Antibody Response of Mouse Splenocytes Using Mixture of Supernatants of Thymocytes, Adherent and Non-adherent Splenocytes: In-vitro Immunization-II

We adapted one of the in-vitro immunization methods to induce antibody responses and confirmed the success of the immunization by enzyme-linked immunosorbent assay (ELISA) without hybridization. We have previously reported several methods of in-vitro immunization using different conditioned media. Here we introduce another method of in-vitro immunization using a mixture of three types of supernatant (thymocytes, and adherent and non-adherent splenocytes of mouse). Splenocytes were immunized in-vitro by a recombinant human growth hormone (rhGH) with the above conditioned media, and the results by ELISA showed a much higher optical density than the other in-vitro immunization methods that we had previously reported. Humoral responses as a result of in-vitro immunization to soluble antigens were easily confirmed by ELISA using the above-conditioned media. This finding indicates that any other conditions thought to be critical by other researches may not be essential for in-vitro immunization.