Search

A patient with acute lymphocytic leukaemia (ALL) is described in whom the cells originated in the pre-B cell lineage but showed an aberrant expression of T4 antigen. A B-cell origin of the leukaemic cells was indicated by reactivity for several monoclonal antibodies specific for the B-cell lineage and by cytoplasmic staining with FITC-labelled anti human mu chains.

OBJECTIVES—To investigate the expression and regulation of CD80, CD86, and CD28 costimulatory molecules in sialoadenitis and interstitial nephritis in patients with Sjogren's syndrome (SS). METHODS—Expression of CD80, CD86, and CD28 molecules was studied by immunohistochemical staining of lip biopsy specimens obtained from patients who had sialoadenitis associated with SS, and renal biopsy specimens obtained from patients who had interstitial nephritis associated with SS. To elucidate the mechanism of de novo expression of CD80 and CD86 antigens, their induction by cytokines in human salivary duct cell line (HSG) and renal cortical epithelial cells (HRCE) by cell enzyme linked immunosorbent assay (ELISA) was quantitatively investigated. RESULTS—In patients with severe sialoadenitis, CD80 and CD86 were strongly expressed on ductal epithelial cells. In contrast, these antigens were not found in the minor salivary glands of normal subjects or of patients with mild sialoadenitis. Some infiltrating cells expressed CD28. In patients who had interstitial nephritis associated with SS, some tubular epithelial cells expressed CD86 but not the CD80 antigen. Unstimulated HSG cells did not express CD80 or CD86. Interferon γ (IFNγ) consistently up regulated levels of CD80 and CD86. In contrast, tumour necrosis factor α (TNFα), interleukin 1β (IL1β), IL2, and IL4 had no effect on either CD80 or CD86 levels. Unstimulated HRCE did not express CD80 or CD86. IFNγ consistently up regulated CD86 expression. No CD80 expression was found on tubular cells. TNFα, IL1β, IL2, and IL4 had no discernible effects. CONCLUSIONS—Salivary ductal cells in patients with SS can express CD80 and CD86 costimulatory molecules in response to IFNγ. Tubular epithelial cells in patients who have interstitial nephritis associated with SS express only CD86 molecules. In patients with SS, salivary ductal cells and tubular epithelial cells may activate infiltrating CD28 positive T lymphocytes by presenting antigens to T cells, potentially leading to tissue destruction.

The purpose of this study was to determine whether interferon-gamma (IFN-gamma) induced CD69 expression by eosinophil precursors. Eosinophil precursors were induced from CD34+ cord blood cells using recombinant human interleukin-3 (IL-3) and interleukin-5 (IL-5). On day 14 of culture, cells constitutively expressed CD69 and the IFN-gamma receptor (IFN-gammaR). Stimulation with IFN-gamma for 24 h did not affect IFN-gammaR expression by the cells. On the other hand, IFN-gamma significantly upregulated CD69 expression by the precursors after 24 h of incubation. A specific JAK2 inhibitor (AG-490) caused a concentration-dependent suppression of IFN-gamma-induced CD69 expression by the precursors. In conclusion, these results indicate that IFN-gamma induces CD69 expression by eosinophil precursors via the activation of JAK2.

SUMMARYThe purpose of this study was to determine whether the JAK pathway is involved in eosinophil activation and survival through IFN-γ receptor signalling in human peripheral eosinophils. Eosinophils were purified from the blood of six atopic disease patients by anti-CD16 magnetic bead-negative selection. IFN-γ significantly up-regulated survival and CD69 expression in 24–48 h cultured eosinophils. Further, IFN-γ induced tyrosine phosphorylation of JAK2 in eosinophils, as indicated by Western blot analysis. Finally, the specific JAK2 inhibitor AG-490 inhibited the tyrosine phosphorylation of JAK2, IFN-γ-induced survival and CD69 expression in eosinophils. In conclusion, these results indicate that IFN-γ induces eosinophil survival and CD69 expression through the activation of JAK2 in peripheral eosinophils, suggesting that JAK2 may play a significant role in eosinophil regulation by IFN-γ–IFN-γR interaction.

We produced computer-assisted instruction (CAI) software for bronchial asthma patients (asthma educational system with computer-assisted instruction; ASTCAI) to assist in self-management and avoid asthmatic attacks and death. ASTCAI is a question-and-answer program operating in a multimedia environment, and was evaluated from questionnaires which 33 patients were asked. Thirty-two patients could perform ASTCAI without any assistance. The responses of 31 patients (94%) indicated that they had no difficulty with manipulation, and 29 patients (88%) stated that the program was beneficial to control of their asthma. Elderly patients (over 65) required more time than younger adults. Emergency visits or admissions of at least 1 year after the first CAI trial decreased in eight out of 26 patients, while only two patients deteriorated compared to the previous year. Our results show that CAI is feasible for most patients, and through active self-learning CAI can improve motivation for self-management as well as supplement the physician's instructions.

Background: Human umbilical vein endothelial cells (HUVEC) are an important source of hematopoietic cytokines, and interleukin-3 (IL-3)- and IL-5-induced eosinophil differentiation from CD34+ cells has been observed. To show the supportive effects of endothelial cells on eosinophil differentiation, we examined the effects of cocultured HUVEC on IL-3 and IL-5-induced eosinophil differentiation from human umbilical cord blood CD34+ cells. Methods: CD34+ cells were obtained from the heparinized umbilical vein blood of 10 volunteers using a CD34-conjugated magnetic bead positive direct selection procedure. With HUVEC in Transwell, CD34+ cells were then cultured for 14–28 days. In neutralizing experiments on HUVEC-derived cytokines, antibodies to both stem cell factor (SCF) and granulocyte/macrophage colony-stimulating factor (GM-CSF) were added to the cell cultures. Results: Cocultured HUVEC upregulated IL-3 and IL-5-induced eosinophil differentiation from CD34+ cells on day 28 of culture by 75.0%. The eosinophilopoietic effect of HUVEC was significantly only when the cells were present in the culture from day 15 to day 28. Addition of anti-SCF antibody or anti-GM-CSF monoclonal antibody to the culture significantly suppressed HUVEC-combined IL-3- and IL-5-induced eosinophil differentiation on day 28 of culture by 49.2 and 55.0%, respectively. Conclusions: These results indicate that several cytokines including GM-CSF and SCF from HUVEC promote IL-3- and IL-5-induced eosinophil differentiation from CD34+ cells.

Background: We reported on the constitutive interferon–γ receptor (IFN–γR) expression on eosinophils. But signal transduction through IFN–γR on eosinophils remains to be elucidated. In this study, we examined the involvement of the Jak/Stat pathway in the signaling of eosinophils after IFN–γR conjugation by the ligand binding. Methods: Purified peripheral eosinophils were stimulated with IFN–γ at 37°C for 1–60 min. Tyrosine phosphorylation of IFN–γR, Jak1, Jak2, and Stat1α was examined by immunoblotting. Gel–shift assay was also examined to show the formation of Stat1α–DNA complexes. Results: We show that binding of IFN–γ to human eosinophils initiated a series of events that resulted in the rapid tyrosine phosphorylation of not only the IFN–γRα chain but also Jak1, Jak2, and Stat1α. In addition, IFN–γ enhanced the DNA–binding activity of Stat1α. Conclusion: These data indicate that IFN–γ affects eosinophils through its specific receptor and utilizes the Jak/Stat pathway as its mode of signaling.

It has been unclear whether or not the interferon-gamma receptor (IFN-gammaR) on human peripheral eosinophils is completely functional. Accordingly, we examined the expression of IFN-gammaR alpha- and beta-chains and the association of the two chains after binding of IFN-gamma to the receptor.Peripheral blood eosinophils were obtained from 8 patients (bronchial asthma, n=6, and hypereosinophilic syndrome, n=2), and expression of the alpha- and beta-chain was investigated by flow cytometry with specific antibodies. Expression of mRNA for the alpha- and beta-subunits was also investigated by the reverse transcriptase-polymerase chain reaction.Eosinophils from all the patients were positive for both the alpha- and beta-chains by flow cytometry. In addition, mRNA for both chains was detected by the polymerase chain reaction. Furthermore, coprecipitation of the alpha- and beta-chains was only found after eosinophil stimulation with IFN-gamma.Human peripheral eosinophils apparently express both the alpha- and beta-chains of the IFN-gammaR, and the two chains may only form the receptor complex after ligand binding.

In order to determine regulatory mechanisms of eosinophil apoptosis, we examined the effect of recombinant IL-5 and interferon-gamma (IFN-γ) on eosinophil apoptosis and bcl-2 expression. rhIL-5 (2.5 ng/ml) significantly inhibited eosinophil apoptosis in 96 h in vitro culture compared with medium only-cultured eosinophils (89.4 ± 3.6% versus 31.3 ± 12.2% (mean ± s.d.); n = 7, P

It has been shown that CD4+ lymphocytes and eosinophils pathologically infiltrate the dermis in skin eruptions due to hypersensitivity to the administration of certain drugs associated with peripheral eosinophilia. However, the mechanisms involved in drug related skin eruptions with eosinophilia are largely unknown. There are several methods of diagnosing drug hypersensitivity, but no single in vitro method is available for the detection of the sensitizing drug. In order to elicit the mechanism of drug related skin eruption with eosinophilia and to establish effective methods for diagnosing drug hypersensitivity, we studied in vitro IL-5 production by peripheral mononuclear cells (PBMC) from patients with drug related skin eruptions (n = 8). Significant increase in IL-5 from exposure to drug alone (n = 3) and drug with PHA (n = 3) were noted, whereas PBMC from normal subjects (n = 7) exhibited no such stimulation. Six of the eight cases showed a significant decrease in the number of peripheral blood eosinophils when the skin eruptions healed compared with those at the onset. Histopathological features revealed mild to moderate infiltration of eosinophils. Immunohistopathologically, infiltration of CD4+ and CD8+ lymphocytes and CD4+ cells were predominant within the dermis of drug related skin eruption sites in all cases. This study suggested that the IL-5 production of sensitized mononuclear cells might be a critical mediator in drug hypersensitivity with eosinophilia and an important diagnostic marker.

A questionnaire on the treatment of asthma was sent to 586 physicians. They consisted of specialists authorized by the Japanese Society of Allergology and councillors of the society who were treating patients with bronchial asthma. Of the total of 306 (52%) respondents, 241 replied to questions relating to adult asthma and 129 to questions relating to childhood asthma (including duplicate replies). For acute treatment, methods most commonly selected by physicians were, in increasing order of popularity, for adults: parenteral aminophylline, oxygen inhalation, parenteral steroids and parenteral adrenaline; for schoolchildren (6–16 years): parenteral aminophylline, inhaled β-stimulant, oxygen inhalation, inhaled β-stimulant + disodium cromoglycate (DSCG), parenteral steroids; and for infants (≤ 5 years): inhaled β-stimulant, parenteral aminophylline, oxygen inhalation, inhaled β-stimulant + DSCG and parenteral steroids. For maintenance treatment, methods most commonly selected by physicians were, in increasing order of popularity, for adults: oral administration of sustained-release theophylline, inhaled steroids and DSCG inhalation; for schoolchildren (6–16 years): DSCG inhalation, oral administration of sustained-release theophylline, oral administration of antiallergic agent preparation and (β-stimulant + DSCG inhalant; and for infants: DSCG inhalation, oral administration of sustained-release theophylline, oral administration of antiallergicagent preparation and β-stimulant + DSCG inhalant. The questionnaire results clearly showed that different drugs were selected for the treatment of asthma in adults, schoolchildren and infants.

The expression of high-affinity IgE receptor (FcERI) on alveolar macrophages was examined in order to determine the association of alveolar macrophages with IgE-dependent inflammation. Immunostai

To clarify the mechanism by which visceral adipose tissue developed in growth hormone (GH) deficient patients, and was reduced by GH supplementation, the effects of GH on the triglyceride metabolism in the visceral and subcutaneous adipose tissues of the rats were compared.Visceral and subcutaneous adipose tissues were obtained from omentum and abdominal wall. [14C] deoxyglucose uptake and the triglyceride synthetic activity from [14C] oleic acid, and heparin-releasable lipoprotein lipase activity were not changed by the addition of GH (10ng/mL) in both visceral and subcutaneous adipose tissues, but the lipolysis by GH (10ng/mL) was much more remarkable in the visceral adipose tissue than in subcutaneous adipose tissue, whereas adrenaline-induced lipolysis occurred almost equally in both tissues.These results suggest that the preponderance of visceral adipose tissue in GH deficiency might partly be due to the lack of GH-preferential lipolysis in visceral adipose tissue.

A questionnaire on asthma management was sent to 586 physicians, consisting of specialists authorized by the Japanese Society of Allergology and councillors of the Society, who were treating patients with bronchial asthma. Of the total of 306 (52%) respondents in November 1993, 241 replied to questions relating to adult asthma and 129 to questions relating to childhood asthma (including duplicate replies). Responses to the questionnaire on the diagnosis, patient education and management of asthma indicated that a reduced number of patients with severe asthma were seen in 1993 in both Pediatric and Internal Medicine Departments compared with 5 years before, despite the increase in total number of asthma patients in Japan. Specifiic IgE radioallergosorbent test (RAST) measurements were frequently performed instead of skin testing for diagnosis, and eosinophil count and bronchodilator response served as an adjunct to the diagnosis. Patients were frequently asked detailed questions about aspirin-induced asthma, which accounted for 8.8, 2.2 and 1.5% of patients with asthma in the adult, schoolchildren (6–16 years) and infant (≤ 5 years) groups, respectively. In achieving ‘control of asthma’, first priority was given to coping with the symptoms in children aged 5 years or less and to enabling routine daily life activities in patients 6 years of age or older. Usefulness of peak flow measurements was widely recognized and a detailed plan for allergen avoidance (house dust) was often given to patients.

We report a case of dilated cardiomyopathy with hyperthyroidism. A 28-year-old man was admitted because of congestive heart failure and atrial fibrillation, and was newly diagnosed as having hyperthyroidism. Despite administration of antithyroid medication, he developed recurrent congestive heart failure. An echocardiogram revealed a moderately dilated left ventricle with diffuse hypokinesis. Though his thyroid function normalized, the patient's cardiac dysfunction did not improve. Beta-blocker therapy was begun with subsequent improvement in clinical symptoms. This suggests that beta-blocker treatment may be effective in patients with atrial fibrillation associated with cardiomyopathy and hyperthyroidism.

現在発売されているいわゆる抗アレルギー薬・抗喘息薬は,ケミカルメディエータ遊離抑制薬とケミカルメディエータ拮抗薬に分けられ,便宜的に抗ヒスタミン薬作用をもつ抗アレルギー薬と抗ヒスタミン作用をもたない抗アレルギー薬に分類した.この薬物は,慢性に経過するアレルギー性気管支喘息で,軽症,中等症の患者に適応がある.急性発作に効果は期待できない.少なくとも4週間以上使用してその効果を評価する.

OBJECTIVE To analyze the clonality of T cell receptor (TCR) V beta 2- and V beta 13-positive T cells, which are predominantly expressed in the lips of patients with Sjogren's syndrome (SS). METHODS The junctional sequences of complementary DNA clones encoding TCR V beta 2 and V beta 13 genes were determined by the polymerase chain reaction. Forty-one V beta 2 and 45 V beta 13 clones established from the lips of 3 SS patients were sequenced. RESULTS The V beta 2/J beta 2.3 pair was enriched in 2 of the 3 patients (44% and 46% of the clones, respectively), and the V beta 13/J beta 2.1 sequence was dominant in 2 of the 3 (23% and 45%). These pairs were not used preferentially in peripheral blood lymphocytes from the same patients. CONCLUSION Infiltrating V beta 2- and V beta 13-positive T cells from the lips of all 3 patients with SS were polyclonal, but the junctional usage of cells from 2 lip samples was restricted, compared with cells from peripheral blood. This suggests that not all expanded cells from the lips of SS patients are stimulated by superantigens.

Previous studies suggested that the eosinophil recruitment into the site of cutaneous late-phase reaction (LPR) was dependent on IgE antibody and mast cells. In this study, we determined the role of CD4+ T cells and CDS+, T cells in causing antigen-induced eosinophil recruitment of LPR in mouse skin. Eosinophil infiltration into the subcutaneous tissue of ovalbumin (OVA)- sensitized BALB/c mice was biphasic, reaching the first peak at 6 h after the subcutaneous challenge with OVA and the second peak at 24 to 48 h. The in vivo depletion of CD4+ T cells by pretreatment with anti-L3T4 monoclonal antibody (mAb) significantly decreased the second peak (at 24 h and 48 h), but not the first peak (at 6 h), of OVA-induced eosinophil infiltration into the skin of OVA-sensitized mice. However, the depletion of CD8+ T cells by pretreatment with anti-Lyt-2 mAb had no significant effect on either the first peak or second peak of OVA-induced cutaneous eosinophilia. Pretreatment with anti-murine interleukin-5 (IL-5) mAb also decreased the second peak, but not the first peak, of OVA- induced cutaneous eosinophilia. In contrast to the inhibitory effects of depletion of CD4+ T cells and of anti-IL-5 mAb on the second peak of antigen-induced cutaneous eosinophilia, disodium cromoglycate and a selective antagonist for platelet activating factor (PAF) CV-6209 decreased the first peak of OVA-induced cutaneous eosinophilia in the mouse. These results indicate that CD4+ T cells, but not CD8+ T cells, cause the second peak of antigen-induced eosinophil recruitment of cutaneous LPR and that IL-5 mediates this eosinophil recruitment. In contrast, the first peak of antigen-induced eosinophil recruitment of cutaneous LPR is mediated by mast cells and PAF.

The objective of this study was to clarify the specificity of anticardiolipin antibodies (aCL). Eighteen monoclonal hybridoma aCL from systemic lupus erythematosus (SLE)-prone MRL/Mp-lpr/lpr mice were established, and the reactivity of monoclonal aCL to phospholipids, DNA, nuclei of human epithelial cells, platelets, vascular endothelial cells, heparin, protein C and thrombomodulin was examined. All the 18 monoclonal aCL reacted with phosphatidylserine and some showed reactivity to phosphatidylinositol and phosphatidylcholine. Six of 16 monoclonal aCL were demonstrated to have the property of lupus anticoagulant. Monoclonal aCL were classified into three categories, in terms of DNA-binding specificity. Ten of 18 aCL had characteristics of antinuclear antibodies. Six of 11 aCL reacted with platelets. Three of 18 aCL were bound to vascular endothelial cells and to heparin. No monoclonal aCL reacted with protein C or thrombomodulin. Therefore, the conclusion was made that monoclonal aCL from SLE mice showed a polyspecific nature.

OBJECTIVE To analyze the T cell receptor V beta gene on double-negative (DN) alpha/beta T cells, which are increased in number, on peripheral blood lymphocytes (PBL) from patients with systemic sclerosis (SSc). METHODS The DN alpha/beta T cells were sorted by flow cytometry from PBL obtained from 3 patients with SSc. The V beta repertoire was analyzed by polymerase chain reaction. RESULTS Only 1 or 2 V beta genes (V beta 5/7, 5, or 17) were predominantly expressed on DN alpha/beta T cells from these 3 patients. CONCLUSION The V beta repertoire on DN alpha/beta T cells in PBL from patients with SSc is rather restricted.

SUMMARY Previous studies have shown that substance P induces granulocyte infiltration in mouse skin, which is mediated through mast cell degranulation. However, it is not yet known whether the direct effect of substance P on vascular endothclial cells is involved in the granulocyte infiltration in the skin. To solve this issue, we used the N-terminal peptide substance P1–9 (SP1–9), which is active for mast cells but inactive for vascular endolhelial cells, and the c-terminal peptide SP6–11, which is active for vascular endothelial cells but inactive for mast cells, since substance P activates both mast cells and vascular endothelial cells. The subcutaneous administration of substance P (10−7−10−5 m) caused granulocyte (neutrophil and eosinophil) infiltration in the skin of BALB/c mice 6 h after the injection. SP1–9 (10−5–10−4 m) also caused granulocyte infiltration of mouse skin which was associated with mast cell degranulation. In contrast. SP6–11 (10−7-10−4 M), which was found to increase the vascular permeability of endothelial cells in mouse skin, induced no significant granulocyte infiltration nor mast cell degranulation. However, SP6–11 (10−5 10−4 m) enhanced SP1–9 -induced granulocyte infiltration in the skin without any significant increase in mast cell degranulation. We conclude that substance P causes granulocyte infiltration in mouse skin through both mast cell degranulation induced by the N-lerminal peptide of substance P and the activation of vascular endothelial cells induced by the c-terminal peptide of substance P.

Infiltrating T cells around salivary glands in the lips of Sjogren's syndrome (SjS) patients are crucial in the pathogenesis of this disease. To analyze the nature of infiltrating T cells, their T cell receptor repertoire was examined with quantitative polymerase chain reaction. The repertoire of V beta transcripts in lips of SjS was not restricted; however, the V beta 2 and V beta 13 genes were predominantly expressed on the T cells of lip specimens in six and four of seven lips, respectively. Predominance of these genes was specific in lips because no predominant V beta transcripts were found in lips from healthy subjects and PBLs from SjS patients. These results indicated that the V beta 2- and V beta 13-positive T cells expanded specifically and preferentially in SjS lips, thereby suggesting the possible role in triggering the autoimmunity of this disease.

血清抗カルジオリピン抗体と急性心筋梗塞発症の関係を全身性エリテマトーデス患者176例を対象に検討した.抗カルジオリピン抗体陽性者では59例中5例に急性心筋梗塞が発症したのに対し陰性者では117例中1例のみしか発症せず,両者の間に有意な差が認められた(p

This study analyzed T-lymphocyte-receptor V beta genes of infiltrating lymphocytes into the pancreases of 12- to 16-wk-old NOD mice with severe insulitis and 5-wk-old mice with mild insulitis by the quantitative polymerase chain reaction method. The V beta transcripts on infiltrating T lymphocytes into pancreases with severe insulitis in older NOD mice were diverse. In contrast, the V beta 11 gene transcript was predominantly expressed on T lymphocytes in the pancreas in younger NOD mice with mild insulitis, suggesting the possible role of V beta 11+ T lymphocytes in triggering insulitis in this species.

We have previously shown that neutral endopeptidase (NEP; EC 3.4.24.11) regulates neuropeptide-induced responses. Recently, Pierart et al. reported that NEP degraded purified interleukin-1 (IL-1) using thymocyte proliferation assay [1]. Since IL-1 is an important cytokine in the immune response and inflammation, we have assessed whether NEP hydrolyzes recombinant human IL-1β using three assay systems (bioassay, immunoassay, and HPLC analysis). NEP on the NALM-6 cells (both intact cells and the solubilized plasma membrane fraction) efficiently hydrolyzed Met 5 -enkephalin and substance P. However, NEP did not significantly decrease the amount of rhIL-1β assessed by the growth inhibitory activity of a human melanoma, by the immunoassay, or by the direct analysis on HPLC. Therefore, we conclude that NEP does not significantly hydrolyze rhIL-1β. Our results suggest that, in contrast to the regulatory role of NEP in neuropeptide-induced responses, NEP is not a regulatory enzyme for IL-1-induced responses.

We investigated the distribution of neutral endopeptidase (NEP; EC 3.4.24.11) activity, a possible regulatory enzyme for neuropeptide-induced leukocyte activations, in each cell type of human blood leukocytes. The NEP activity assessed by an NEP inhibitor phosphoramidon-sensitive Met5-enkephalin degrading activity was present in neutrophils and the common acute lymphoblastic leukemia antigen (CALLA)-positive leukemic cells (59 pmol/min/106 cells and 62 pmol/min/105 cells, respectively); however, the NEP activity was virtually absent in lymphocytes, monocytes, eosinophils, basophils, CALLA-negative leukemic cells, or a promyelocytic cell line HL-60. The enzymatic activity was characterized as NEP on the basis of the values of kinetic parameters (Km = 61 μM, Kcat = 1,692 min-1, and Kcat/Km = 28 min-1/μM-1) and the values of IC50 of two NEP inhibitors phosphoramidon and thiorphan (7.4 nM and 8.4 nM, respectively). The distribution of NEP detected immunocytochemically using anti-NEP monoclonal antibodies was also found to be parallel with the distribution of NEP activity among peripheral blood leukocytes.

皮膚筋炎・多発性筋炎の患者では他の膠原病に比して急性間質性肺炎がとくに多数みられた.自験例の71例の皮膚筋炎のうち, 18例に急性問質性肺炎がみられ,このうち10例が治療に抵抗し予後不良であった.こうした予後不良群は男性に多く,皮膚症状を有し, CPKなどの筋原性酵素の上昇がなく,炎症反応に乏しい症例で,筋炎に対するステロイド剤の投与後に発症することが多かった.予後不良群では剖検5例のうち4例はUIPであったが, 1例はサイトメガロウイルス性肺炎であり,また抗ウイルス抗体価の変動がみられた症例もあり,皮膚筋炎・多発性筋炎に伴う急性間質性肺疾患の原因の一つとしてウイルスの関与の可能性が考えられた.胸部X線写真からの予後判定は難しかった.また特定の自己抗体と急性間質性肺炎の予後には相関がなかった.

A novel immunosuppressive compound, FK-506, isolated from Streptomyces has potent immunosuppressive activities. To investigate the effect of FK-506 on the course of diabetes in nonobese diabetic (NOD) mice, we gave this drug to these animals, from the age of 8 weeks, intraperitoneally, in doses of 0.1 mg (2.5 mg/kg/day) or 0.01 mg (0.25 mg/kg/day) three times a week. Overt diabetes were observed in 75.5% of control mice by the age of 20 weeks. In contrast, no diabetes occurred in mice given 0.1 mg of FK-506. Sixteen percent of mice treated with 0.01 mg of the drug became diabetic. Administration of this drug prevented the progression of insulitis in NOD mice. The mice given 0.1 mg of FK-506 lost weight, but this was reversible.

We have produced hematopoietic chimeric mice from an embryonic stem (ES) cell line earring Ly-1 cDNA under the control of IgH promoter and enhancer. Various amounts of serum IgM (5–86% of total IgM) in chimeric mice were of ES origin and 30–60% of IgM-positive B cells from the chimeric mice analyzed were reconstituted from ES cells. Using these chimeric mice, the expression of the Ly-1 transgene on lymphoid tissues was examined by polymerase chain reaction assay with primers specific for the transgene, and by cell sorter analysis. Transcription of the Ly-1 transgene was detected in spleen cells, thymocytes and lymph node cells; however, the expression of the Ly-1 molecule was observed only on lipopolysaccharide (LPS)-stimulated splenic IgM-positive B cells but not on resting splenic B cells. There was no significantly increased expression of Ly-1 on splenic T cells and thymocytes. Thus, our findings demonstrate that conventional splenic B cells could express the Ly-1 transgene on their surface in vivo after LPS stimulation. Also discussed is the ES-derived chimeric hematopoietic system.

To determine the mechanism by which substance P (SP) activates human neutrophils, we examined the potency of SP for inducing chemotaxis, lysozyme release, and increase in cytosolic free Ca2+ concentration ( [Ca2+] i) in human blood neutrohils. We also examined the effects of EGTA and of a formyl-Met-Leu-Phe (FMLP) antagonist on the responses. SP (10-6 to 10-4 M) induced chemotaxis, lysozyme release, and the increase in [Ca2+] i of the neutrophils dose-dependently. Preincubation with EGTA (10 mM) decreased the SP-induced increase in [Ca2+] i by 90%, whereas EGTA decreased the FMLP-induced increase in [Ca2+] i only by 37%, suggesting that the activations of human neutrophils by SP are dependent on the influx of extracellular Ca2+. An FMLP antagonist, boc-Phe-Leu-Phe-Leu-Phe, inhibited the FMLP-induced chemotaxis and increase in [Ca2+] i, but it did not inhibit the SP-induced respones.We suggest that SP induces the activations of human neutrophils by the different mechanism from that of FMLP.

We investigated the effects of F-1322 (N-[2-[4-(benzhydryloxy)piperidino]ethyl]-3-hydroxy-5-(3-pyridylmethoxy)-2-naphthamide), a new compound that inhibits both thromboxane A2 synthetase and 5-lipoxygenase and that functions as a histamine antagonist, on the Ascaris antigen-induced late asthmatic response and pulmonary eosinophilia in guinea pigs. Oral administration of F-1322 (10–100 mg/kg) inhibited the antigen-induced late asthmatic response in a dose-dependent manner. Histological analysis revealed that F-1322 prevented the accumulation of eosinophils in the airways and this was paralleled by a decrease in the number of eosinophils and lymphocytes recovered in bronchoalveolar lavage fluid. F-1322 (0.1–10 μM) inhibited eotaxin-induced chemotaxis and actin polymerization of eosinophils in vitro in a concentration-dependent manner, while oral administration of F-1322 dose-dependently suppressed the migration of eosinophils into the airways in vivo in response to infusion of interleukin 5 and eotaxin in combination. F-1322 may, thus, improve the late asthmatic response in this model, in part, by preventing the accumulation of eosinophils in the airways. The pharmacological profile of F-1322 indicates that this drug is likely to be useful in the treatment of allergic diseases such as asthma.

In order to determine the inflammatory mechanisms of skin lesions in patients with drug hypersentivity, we examined eosinophil activation and interleukin-5 (IL-5) production in infiltrating lymphocytes. First, we showed that the number of peripheral eosinophils and the level of serum IL-5 at the eruption-active stage were both significantly higher than those in healed skin eruptions. Histological and immunohistological examination revealed that CD4+ T cells and eosinophils significantly more densely infiltrated drug eruptions than control skin lesions. The infiltrating eosinophils were also shown to be activated by immunostaining using anti-secreted formed eosinophilic cationic protein monoclonal antibody. The expression of mRNA for IL-5 in the infiltrating mononuclear cells at drug eruptions was shown by in situ hybridization. These results suggest that infiltrating CD4+ T cells might regulate both peripheral and tissue eosinophils and facilitate allergic inflammation at drug eruptions by means of IL-5 production.

We recently confirmed that pemirolast potassium, an antiallergic agent, markedly inhibits migration and proliferation of vascular smooth muscle cells. It has also been reported that pemirolast inhibits intimal hyperplasia in animal experiments.To elucidate the preventive effects of pemirolast on restenosis after percutaneous transluminal coronary angioplasty (PTCA), 227 patients were enrolled in this prospective, randomized trial. A total of 205 patients who were compatible with the protocol were analyzed (pemirolast group, 104 patients with 140 lesions; control group, 101 patients with 133 lesions). Patients in the pemirolast group received 20 mg/d of pemirolast from 1 week before PTCA until the time of follow-up angiography (4 months after PTCA). Angiographic restenosis was defined as diameter stenosis/=50% at follow-up. Restenosis rates were significantly lower in the pemirolast group than in the control group (24.0% vs 46.5% of patients, 18.6% vs 35.3% of lesions, P.01, respectively). During 8 months of follow-up, there were no coronary events (death, myocardial infarction, coronary artery bypass surgery, or repeated PTCA) in 81.7% of the pemirolast group and in 63.4% of the control group (P =.013).This study suggested that pemirolast would be useful in the clinical setting to prevent restenosis after PTCA.

Background: Expression of the tumor necrosis factor-α receptor by human eosinophils has not been determined. We examined the surface expression and mRNA for tumor necrosis factor-α receptors I and II (TNF-αRI and TNF-αRII) on peripheral eosinophils. Methods: Eosinophils were obtained from 17 asthma patients and 3 healthy volunteers. Flow cytometry was used to examine receptor expression, and the reverse transcriptase-polymerase chain reaction was used to examine receptor mRNA expression. Results: Flow cytometry revealed that 16 out of 20 subjects had eosinophils expressing TNF-αRI and 18 subjects had eosinophils expressing TNF-αRII. All eosinophils expressed at least one receptor. The mRNA for TNF-αRII was detected in all eosinophils examined, but the signals for TNF-αRI mRNA were only found in cells expressing this receptor. Conclusions: Human peripheral eosinophils show heterogeneous expression of TNF-αRI and TNF-αRII.

In order to determine whether or not IFN-gammaR is associated with regulatory mechanisms on human eosinophil function, we examined the expression of functional IFN-gammaR on human peripheral eosinophils. In this study, peripheral blood eosinophils were obtained from seven normal controls and 12 patients (bronchial asthma, n = 9, and hypereosinophilic syndrome (HES), n = 3), and the purity of eosinophils was 97.11 +/- 2.31%, n = 19. We first showed that anti-IFN-gammaR alpha-chain MoAb reacted with all tested eosinophils of both normal controls and patients by flow cytometry analysis. We also showed expression of mRNA for the alpha-chain of IFN-gammaR in all purified eosinophils of six individuals. Further, to characterize IFN-gammaR on eosinophils, we did binding experiments with 125I-IFN-gamma on purified peripheral eosinophils. The linear Scatchard plot indicated a single type of high-affinity binding sites (dissociation constant (Kd) = 3.89-4.95 x 10(-10) M, numbers of binding sites = 183-233/cell, n = 3). To determine whether IFN-gammaR on eosinophils is functional, we examined surface eosinophilic cationic protein (ECP) and CD69 induction after IFN-gammaR ligation with recombinant human IFN-gamma (rhIFN-gamma) on eosinophils by flow cytometry. rhIFN-gamma stimulation significantly induced both ECP and CD69 expression on the 2-18 h-cultured eosinophils in a dose-dependent manner. Further, the effects of rhIFN-gamma stimulation were significantly blocked by both a neutralizing anti-IFN-gamma MoAb and a blocking anti-IFN-gammaR MoAb. These results suggest that human peripheral eosinophils express functional IFN-gammaR.

A decrease in smooth muscle cells is observed in advanced atherosclerotic lesion. To understand this mechanism, we selected oxysterols as candidates for toxic lipid, and examined their cytotoxicity on human cultured vascular smooth muscle cells, together with the manner of cell death. In the presence of 7-ketocholesterol or 7 beta-hydroxycholesterol (50 mumol/L), the percentage of detached cells increased significantly with dose dependency, and an increase in detached cell number and DNA nick detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling study (TUNEL) preceded an increase in lactate dehydrogenase released into the medium. DNA extracted from smooth muscle cells incubated with 7-ketocholesterol or 7 beta-hydroxycholesterol showed a laddering pattern on agarose electrophoresis. In the presence of 7-ketocholesterol or 7 beta-hydroxycholesterol, fragmented DNA quantified by the quantitative sandwich enzyme immunoassay was significantly increased. From these results, it is proposed that 7-ketocholesterol and 7 beta-hydroxycholesterol are toxic to smooth muscle cells, and that this cytotoxicity is mediated by apoptosis.