Satoru Tsuruta

Correlation of house dust mite-specific lymphocyte proliferation with IL-5 production, eosinophilia, and the severity of symptoms in infants with atopic dermatitis.

Because house dust mite (HDM)-specific IgE antibody (IgE-RAST) is usually not detectable in infants with atopic dermatitis (AD), HDM has not been regarded as the cause of infantile AD. The level of HDM-specific lymphocyte proliferation (expressed as stimulation index measured by flow cytometry [SIF]), however, was found to be markedly elevated in AD infants. This suggests that the sensitization of T cells to HDM extract occurs even in infancy. Moreover, the level of HDM-SIF is correlated closely with the severity of infantile AD, suggesting that HDM is a major cause of this disease not only in adults and children but also in infants. Although the level of HDM-SIF did not correlate with the level of HDM-specific IgE-RAST in infants with AD, it did intimately correlate with the absolute number of peripheral blood eosinophils. Because T lymphocytes are known to secrete some cytokines, such as IL-5, that enhance the proliferation of eosinophils, we measured the IL-5 production by peripheral blood mononuclear cells in infants with AD on stimulation with HDM extract. The amount of IL-5 production is significantly higher in infants with AD, as well as in children with AD, than that found in nonatopic control subjects. Moreover, the level of IL-5 production is correlated closely with the level of HDM-SIF in infants with AD. Taken together, these results suggest that HDMs play an important role in the development of infantile AD by inducing IL-5 production from HDM-specific T lymphocytes.

IL-4 production by PBMCs on stimulation with mite allergen is correlated with the level of serum IgE antibody against mite in children with bronchial asthma.

BACKGROUND Although IL-4, IL-13, and IFN-gamma are known to affect IgE synthesis, it remains unclear which one plays the most important role in in vivo IgE synthesis in atopic patients. OBJECTIVE The aim of this study is to clarify the difference in importance among these cytokines in up-regulation of IgE synthesis in atopic patients. METHODS We measured IL-4, IL-13, and IFN-gamma production by PBMCs on stimulation with house dust mite (HDM) in 23 children, 3 to 15 years old, with bronchial asthma (BA) and analyzed the correlation with HDM-specific IgE antibody levels expressed as HDM IgE radioallergosorbent test (RAST) results. RESULTS The production of IL-4 and IL-13 by PBMCs on stimulation with HDM was significantly higher in children with BA than in nonatopic control subjects (IL-4, 752.9 +/- 365.9 vs 312.3 +/- 230.0 fg/mL, P <.001; IL-13, 21.9 pg/ml [<12.0-77.6] vs <12.0, P <.01). IL-4 production showed a close positive correlation with HDM IgE RAST (r = 0.71, P <.001), which was distinctly stronger than that between IL-13 production and HDM IgE RAST (r = 0.46, P <.05). IFN-gamma production was neither different between children with BA and nonatopic control subjects (7. 24 [1.54-33.90] pg/mL vs 11.2 [1.66-75.9] pg/mL) nor correlated with HDM IgE RAST levels. Essentially the same result was obtained by stimulation of PBMCs with a purified HDM major allergen Der f 1. CONCLUSION IL-4 is likely to be the most important cytokine in up-regulation of in vivo IgE synthesis against HDM in children with BA.

Unique profile of IL-4 and IFN-γ production by peripheral blood mononuclear cells in infants with atopic dermatitis

Although the level of house dust mite (HDM)-specific lymphocyte proliferation is high in infants with atopic dermatitis (AD), the level of HDM-specific IgE antibody (HDM-IgE-RAST) is usually very low or negative. To elucidate the cause of the deficient HDM-specific IgE antibody formation in infants with AD, we examined the profile of IL-4 and IFN-gamma production by HDM-stimulated PBMCs. The amount of IL-4 production was higher in infants with AD and in children with AD (3 to 15 years) than in the nonatopic control subjects. Although the amount of IFN-gamma production in children with AD was lower than that found in nonatopic children, it was higher in infants with AD than in nonatopic infants. This result suggests that HDM-specific helper T lymphocytes in infants with AD have not yet differentiated into TH2 but rather stayed at the stage of TH0. The level of IgE-RAST for egg white (EW) is already elevated in infants with AD. The amount of IL-4 produced by EW-stimulated PBMCs was comparable to that produced by HDM-stimulated PBMCs in infants with AD. However, the amount of IFN-gamma produced by EW-stimulated PBMCs was distinctly lower than that produced by HDM-stimulated PBMCs in infants with AD. Although there was no correlation between the amount of IL-4 production by HDM-stimulated PBMCs and the level of HDM-IgE-RAST in infants with AD, the amount of IL-4 production by EW-stimulated PBMCs was closely correlated with the level of EW-IgE-RAST. These results suggest that it is not the lack of IL-4 but rather a relative increase in IFN-gamma production by HDM-specific helper T lymphocytes that causes the deficiency of HDM-specific IgE-antibody synthesis in infants with AD.

Development of the Capacity of Peripheral Blood Mononuclear Cells to Produce IL-4, IL-5 and IFN-γ upon Stimulation with House Dust Mite in Children with Atopic Dermatitis

Background: Imbalances of IL-4 and IFN-γ production are widely known to increase IgE synthesis in allergic individuals, and IL-5 is known to play a critical role in the pathogenesis of various allergic diseases. However, little is known about how Th cells specific to house dust mite (HDM) develop the capacity to produce these cytokines in children with atopic dermatitis (AD). Objective: This study aims to clarify when HDM-specific Th cells develop the capacity to produce IL-4, IL-5 and IFN-γ in children with AD. Methods: The production of IL-4, IL-5 and IFN-γ by peripheral blood mononuclear cells (PBMCs) upon stimulation with HDM was measured in 91 children with AD and 37 control subjects. The changes in the cytokine production with age were analyzed transectionally and longitudinally. Results: IL-4 production by HDM-stimulated PBMCs in children with AD was already increased before age 1. Thereafter, it continuously rose until reaching a near-maximum level at age 2. Growth-related changes in the production of IL-5 resembled those in the production of IL-4 and peaked at age 1. The production of these cytokines was very low in control subjects up to age 2 and then gradually increased, albeit never above the levels measured in AD children. The production of IFN-γ in children with AD reached a near-maximum level during the first year of life and diminished after age 3. Although IFN-γ production in controls was lower than that in AD children during infancy, it continuously rose even after age 3 and surpassed the levels measured in AD children. The level of serum HDM-specific IgE antibody began to increase after age 1 following the rise of IL-4 production. Essentially the same relationship among IL-4, IFN-γ and IgE synthesis was seen in a longitudinal study of 6 AD infants, in whom HDM-specific IgE was initially negative but later turned positive. Conclusions: These findings suggest that the capacity of HDM-specific Th cells to produce IL-4, IL-5 and IFN-γ develops and effectively matures during the first 3 years of life in children with AD.

Differences in cytokine production by peripheral blood mononuclear cells (PBMC) between patients with atopic dermatitis and bronchial asthma

It is widely accepted that type 2 helper T (Th2) lymphocytes play a crucial role in the pathogenesis of atopic dermatitis (AD) as well as bronchial asthma (BA). We measured the amounts of IL‐5 and interferon‐gamma (IFN‐γ) produced by PBMC upon stimulation with house dust mite (HDM) or Candida albicans (CA) in 17 children (3–15 years) with AD, and compared these values with those of 16 children with BA. Although IL‐5 production by PBMC upon stimulation with HDM in patients with AD was significantly higher than that in 13 non‐atopic controls (geometric mean = 23.4 pg/ml versus 5.9 pg/ml, P < 0.05), it was significantly lower than that in patients with BA (177.8 pg/ml, P < 0.001). The amount of IL‐5 produced by PBMC upon stimulation with CA was also significantly lower in patients with AD than in those with BA (7.2 pg/ml versus 100.0 pg/ml, P < 0.001). The production of IFN‐γ by PBMC stimulated with HDM or CA was also significantly lower in patients with AD than in those with BA (HDM 4.3 pg/ml versus 12.6 pg/ml, P < 0.05; CA 6.5 pg/ml versus 60.3 pg/ml, P < 0.001). Consequently the ratio of IL‐5 to IFN‐γ production was high not only in patients with BA but also in those with AD. These findings suggest that there are some differences in the regulation of in vivo cytokine production between patients with AD and those with BA, although a Th2‐dominant profile is common to both.

Successful human leukocyte antigen one antigen-mismatched related bone marrow transplantation in a 6-year-old boy with leukocyte adhesion deficiency syndrome

inherited immunodeficiency characterized by repetitious severe infections and delayed wound healing because of defective adherence and migration of leukocytes. This disease is caused by inappropriate membrane expression of the leukocyte-associated integrins LFA-1 (CD11a–CD18), Mac-1 (CD11b–CD18) and p150,95 (CD11c–CD18).1–4 The severe phenotype of this disease demonstrates the complete absence of expression of these molecules. This leads to a profound dysfunction, mainly of granulocytes, which are unable to migrate towards sites of infection and ingest and kill microorganisms efficiently. Allogenic stem cell transplantation has been proven to be the most effective, as well as curative, therapy for the disease.5 There are no suitably matched donors for the majority of patients and, thus, human leukcocyte antigen (HLA) nonidentical stem cell transplantation has been performed. However, non-identical related transplants have many problems, such as engraftment failure and a high incidence of acute and chronic graft-versus-host disease (GVHD).6,7 In the present study, we report on a patient with LAD who has been successfully treated by HLA mismatched bone marrow transplantation. This is the first stem cell transplantation performed in Japan. Case report

IFN-γ Plays a Dominant Role in Upregulation of Candida-Specific IgE Synthesis in Patients with Atopic Dermatitis

Background: Although Candida albicans (CA) is known to induce Th1 clones that suppress IgE synthesis, serum IgE antibody against CA is often increased in atopic patients. This study aims to elucidate the mechanism of IgE synthesis against CA in atopic patients. Methods: We measured the production of IL-4 and IFN-γ by peripheral blood mononuclear cells (PBMCs) from atopic patients upon stimulation with CA and examined the correlation with the level of serum IgE antibody against CA. Results: The level of serum CA-specific IgE antibody (CA-IgE) was significantly higher in patients with atopic dermatitis (AD) than in patients with bronchial asthma (BA) (geometric mean = 3.6 vs. 0.27 UA/ml, p < 0.02) (UA = unit allergen), while there was no difference in the level of house dust mite-specific IgE antibody between them (67.6 vs. 87.1 UA/ml). Although IL-4 production by PBMCs upon stimulation with CA in patients with AD was not significantly different from that in patients with BA (mean = 359.1 vs. 515.3 fg/ml), IFN-γ production was significantly lower in the former than in the latter group (8.1 vs. 56.2 pg/ml, p < 0.001). Consequently, the ratio of IL-4/IFN-γ production was apparently higher in patients with AD than in those with BA, which corresponds to the difference between them in the level of serum CA-IgE. A significant negative correlation was seen in patients with AD between IFN-γ production by CA-stimulated PBMCs and the level of serum CA-IgE (p < 0.05). Conclusions: IgE synthesis against CA in atopic patients may be precipitated not by enhancing IL-4 production, but by reducing IFN-γ secretion.

Infantile hemangioendothelioma of the thymus with massive pleural effusion and Kasabach‐Merritt syndrome: Histopathological, flow cytometrical analysis of the tumor

Infantile hemangioendothelioma of the thymus is a rare disease. We describe a patient who developed a large anterior mediastinal mass, severe thrombocytopenia and massive pleural effusion at 1 month of age. Glucocorticosteroid and irradiation therapy had no effect on either the tumor size or clinical symptoms and the tumor was resected subtotally. Three months after the subtotal resection, the remaining tumor had almost disappeared and the symptoms had resolved. The patient has now been well for 1 year after surgery without evidence of recurrence. The tumor tissue was characterized by prominent vascular endothelial proliferation intermixed with a normal thymic structure, producing a picture consistent with that of an infantile hemangioendothelioma in the thymus, lmmunohistochemically, the tumor cells showed positive staining for vimentin, factor VIII and CD34. The DNA stemline and proliferative activity were examined by flow cytometry, which revealed a diploid stemline with a low growth fraction. DNA content and cell cycle analyses of the tumor tissue may be useful for predicting the biological behavior of the tumor.

Anaphylaxis caused by casein used in artificially marbled beef: A case report

Naturally marbled beef is popular in Japan because its tenderness and juiciness practically allow theflavor to flow into themouth.1 The marbled texture is a characteristic of “Wagyu” beef from a Japanese cattle breed acclaimed for its tender meat and derived from native Asian cattle. Therefore, Wagyu is usually very expensive in Japan. In some restaurants, meat injected with beef fat (“artificially marbled beef”) may be served as a low priced substitute for genuine marbled beef. Artificially marbled beef sometimes contains sodium caseinate (casein) as a food additive. We report on a patient with cows milk allergy who showed an anaphylactic reaction following the ingestion of casein-containing artificially marbled beef. A 2-year-old boy with a history of adverse reactions to cows milk, hens egg and wheat was served a beef dish intended to be free of these ingredients that his family had pre-ordered at a hotel restaurant. Within minutes of taking one bite of beef, periorbital edema and wheezing appeared, then the child lost consciousness. He was taken to the emergency room where he was treated with intramuscular adrenaline, intravenous antihistamine and corticosteroids. His symptoms resolved within 2 h and he was discharged with warnings about the possibility of delayed reactions. Two weeks later, his parents consulted our hospital searching for the cause of their sons anaphylactic reaction. We asked them for comprehensive details of his food intake: he had not eaten anything other than a beef steak among several meals and had never experienced any allergic reactions to beef itself. Therefore, we surmised that the beefmay have been processed.We contacted the hotel restaurant and found that they had used artificially marbled beef which contained casein as one of the food additives. We obtained samples of the same beef as consumed by the patient from the hotel restaurant and performed prick-to-prick tests. We used histamine and saline as positive and negative controls, respectively and accepted mean wheal diameter > 3 mm as positive. The mean wheal diameters were as follows: histamine(7.5 mm), heated sample(15 mm), unheated sample(11.5 mm) and saline(0 mm). His serumwas analyzed using the CAP system (Pharmacia, Uppsala, Sweden) for total IgE (133 IU/ml) and for specific IgE to cows milk (41.9 UA/ml) and casein (47.6 UA/ml). We analyzed the milk protein content in the artificially marbled beef using an enzyme-linked immunosorbent assay (FASTKITTM; Nipponham, Tokyo, Japan). The content of 1.193 mg/ml was sufficient to cause generalized reactions in cows milk-allergic patients. From these results, we