抗原および抗ヒトIgEに対する好塩基球の反応性. 1.好塩基球数の変化

Blood basophils in subjects with bronchial asthma migrate from blood stream into local allergic reaction sites after inhalation of antigen into airways. The phenomena can be observed by changes in number of basophils in the peripheral blood. The peripheral basophil count is at a normal level in non-attack stage of asthmatics as in healthy subjects. The basophil count increases in pre-attack stages, and decreases during attack stages. These findings suggest that number of basophils changes in close relation to asthma cycle, and that by observing number of basophils in the peripheral blood, it is possible to detect near future attacks. The decrease in number of basophils and histamine release by stimulation with antigen and anti-IgE can be observed in vitro in whole blood and basophils separated by density gradient centrifugation and by counterflow centrifugation elutriation. The decreased number of basophils in the peripheral blood represents migration of the cells into local allergic reaction sites.気管支喘息では,気道への抗原吸入後,好塩基球は末梢血よりアレルギー反応局所へと遊走する｡この現象は,末梢血中の好塩基球数の変動により観察することができる｡気管支喘息患者の好塩基球数は,非発作時には健常人とほぼ同じで正常範囲内にあるが,発作前段階で増加し,発作出現とともに減少する傾向を示す｡これらの所見は,好塩基球が喘息発作と密接な関連を持ちながら変動すること,そして,これらの変動を観察することにより,近い将来の発作を予知することが可能であることを示唆している｡抗原あるいは抗ヒトIgE刺激による好塩基球数の減少およびヒスタミン遊離は,全血法あるいは濃度勾配法やcounterflow centrifugation elutriation法により分離された好塩基球を用いてin vitroで観察することができる｡末梢血中の好塩基球数の減少は,好塩基球がアレルギー反応局所へと遊走したことを示している

Component-Resolved in Vitro Diagnosis in Peach-Allergic Patients

BACKGROUND: The in vitro diagnosis of pollen-related food allergy presents low specifi city and reproducibility with many conventional extracts. This can be improved using natural purifi ed allergens, recombinant purifi ed allergens, or both. OBJECTIVE: We compared specifi c immunoglobulin (Ig) E determination (sIgE), the basophil activation test (BAT), the histamine release test (HRT), and the cellular allergen stimulation test (CAST) using natural and recombinant allergens in the diagnosis of peach allergy. METHODS: Thirty-two peach allergic patients were studied. Skin prick tests were performed with commercial peach and extract with Mal d 1, nPru p 3, and profi lin (nPho d 2). sIgE, BAT, CAST, and HRT were determined using rPru p 3, rMal d 3, rBet v 1, rMal d 1, and rMal d 4. RESULTS: Agreement between the techniques was good with all the allergens, except HRT with rMal d 1 and rMal d 4. With rPru p 3, sIgE, CAST, BAT, and HRT showed sensitivity values of 88%, 81%, 72%, and 69% and specifi city values of 100%, 93%, 97%, and 83%, respectively. In patients with systemic symptoms or contact urticaria, the values were 100%, 85%, 81%, and 81%. In patients with oral allergy syndrome, sensitivity to profi lins or homologues of Bet v 1 was detected in 100% of the cases by all the techniques, except by HRT with rMal d 1, which detected 66% of the cases. CONCLUSIONS: The use of single allergens in the in vitro diagnosis of peach allergy by specifi c IgE determination, BAT, and CAST offers high specifi city and sensitivity, with better results than the HRT

Complete blood count: absolute or relative values?

Background: For many years, relative values based on 100 quantified cells have been used to assess blood counts in the field of hematology. However, modern blood counting machines have recently made it possible to determine absolute counts. Thus, the current study assessed whether the determination of relative values, based on 100 cells counted, or the determination of absolute values is more accurate in hematology. Methods: To calculate the errors of absolute counts and of quotients, we used two independent methods to determine the errors. For the error calculation, we first performed a Gaussian error calculation. Second we identified the errors using daily control checks and examined the high limit of the actual errors (precision) on the Sysmex XE5000 hematological analyzer. Results: Our findings indicated that the accuracy of the relative values was always much higher compared to the absolute values. Conclusion: This finding can be explained by “combined errors” which affect absolute cell counts and which are directed for all cell counts of one run into the same direction. These types of errors are reduced by quotient formation as shown here for the basophils. The accuracy of the absolute values obtained from the hematology machines of the latest generation was acceptable due to the very high number of cells quantified

Biocompatibility and tissue regenerating capacity of crosslinked dermal sheep collagen

The biocompatibility and tissue regenerating capacity of four crosslinked dermal sheep collagens (DSC) was studied. In vitro, the four DSC versions were found to be noncytotoxic or very low in cytoxicity. After subcutaneous implantation in rats, hexamethylenediisocyanatecrcrosslinked DSC (HDSC) seldom induced an increased infiltration of neutrophils or macrophages, as compared with normal wound healing; whereas new formation of collagen was observed. DSC crosslinked with glutaraldehyde (GDSC) followed by reaction with NaBH4 shortly after implantation showed an increased infiltration of neutrophils with a deviant morphology. Furthermore, a high incidence of calcification was observed, which may explain the minor ingrowth of giant cells and fibroblasts, and the poor formation of new rat collagen. Acyl azide-crosslinked DSC (AaDSC) first induced an increased infiltration of macrophages, and then of giant cells, both with high lipid formation. AaDSC degraded at least twice as slowly as HDSC and GDSC, finally leaving a matrix of newly formed rat collagen. Samples crosslinked with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide (ENDSC) induced the same mild cellular reaction as HDSC; whereas, similar to AaDSC, the degradation rate was slow and an optimal rat collagen matrix was formed. Of the crosslinked DSC samples, ENDSC seems most promising for tissue regeneration

Contact urticaria to giraffe hair

Background: Immediate-type hypersensitivity to animal proteins is a common problem in people occupationally exposed to animals. Methods: A 19-year-old female working as a voluntary zookeeper in her off-time suffered from hives on her forearms following contact to the fur of a giraffe. For diagnostic evaluation, skin prick tests, assessment of specific serum IgE antibodies, and basophil activation tests were performed. Results: Skin prick tests with a standard series of common aeroallergens were positive for various pollens. Prick testing with native materials was positive for extracts of hair from two different giraffe subspecies in the patient, but not in control subjects. By CAP-FEIA, no specific serum IgE antibodies to dander of a large variety of animals were found in the patient. In the basophil activation test, expression of the activation marker CD63 was induced by extract of giraffe hair on the cells from the patient, but not on those from unaffected controls. Conclusions: This patient suffers from an `exotic' immediate-type contact allergy to giraffe hair. Copyright (C) 2005 S. Karger AG, Basel

Efecto del aceite esencial de Minthostachys verticillata sobre la respuesta inmune de pacientes alérgicos a ácaros del polvo

El aceite esencial de hojas de M. verticillata fue obtenido por destilación en corriente de vapor y analizado por cromatografía gaseosa. Células mononucleares de sangre periférica de pacientes alérgicos, fueron estimuladas con aceite esencial. La proliferación fue ensayada mediante método colorimétrico del MTT o 3,(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide. Se cuantificaron células CD4 + , CD8 + y B en cultivos de linfocitos y en sobrenadantes, IFN- γ e IL-13. Se determinó la liberación de la enzima β -hexosaminidasa de basófilos adicionados de aceite esencial, y desafiados con el alergeno. Los efectos del aceite esencial fueron comparados con los de drogas antialérgicas. Los principales constituyentes identificados fueron pulegona, mentona y limoneno. Aceite esencial incrementó los valores de células CD4 + , CD8 + y B (p<0.002), estimuló la síntesis de IFN- γ y redujo los niveles de IL-13. El aceite esencial disminuyó la liberación de β -hexosaminidasa de basófilos (p<0.0001) con mayores efectos que los de las drogas ensayadas. Aceite esencial estimuló la desviación Th1 y redujo la liberación de la enzima β -hexosaminidasa de basófilos de pacientes alérgicos.The essential oil from leaves of Minthostachys verticillata was obtained by steam distillation and analyzed by gas chromatography. Peripheral blood mononuclear cells from allergic patients were stimulated with essential oil. By application of MTT or 3,(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide colorimetric test the proliferation was assayed. In lymphocyte cultures CD4+, CD8+ and B cells were quantified, and in supernatants, IFN-γ and IL-13. The liberation of β-hexosaminidase enzyme was determined for basophils with essential oil added and challenged with allergen and its effects were compared with those of anti-allergic drugs. The main constituents identified were pulegone, menthone and limonene. Essential oil increased absolute values of CD4+, CD8+ and B cells (p<0.002), stimulated IFN-γ synthesis and reduced IL-13 levels. Essential oil diminished β-hexosaminidase liberation by basophils (p<0.0001), with effects majors to those of the drugs tested. Essential oil stimulated the Th1 deviation and reduced β-hexosaminidase enzyme liberation by basophils from allergic patientsFil: Cariddi, Laura Noelia. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Moser, Marina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Andrada, Melisa. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Demo, Mirta Susana. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Zygadlo, Julio Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Sabini, Liliana Ines. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Maldonado, Ana Maria. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentin

Cloned mouse cells with natural killer function and cloned suppressor T cells express ultrastructural and biochemical features not shared by cloned inducer T cells.

We have examined the morphology, cytochemistry, and biochemistry of mouse leukocyte subsets by analyzing cloned leukocyte populations specialized to perform different immunologic functions. Cloned cells expressing high-affinity plasma membrane receptors for IgE and mediating natural killer (NK) lysis and cloned antigen-specific suppressor T cells contained prominent osmiophilic cytoplasmic granules similar by ultrastructure to those of mouse basophils. Both clones also incorporated 35SO4 into granule-associated sulfated glycosaminoglycans, expressed a characteristic ultrastructural pattern of nonspecific esterase activity, incorporated exogenous [3H]5-hydroxytryptamine, and contained cytoplasmic deposits of particulate glycogen. By contrast, cloned inducer T cells lacked cytoplasmic granules and glycogen, incorporated neither 35SO4 nor [3H]5-hydroxytryptamine, and differed from the other clones in pattern of nonspecific esterase activity. These findings establish that certain cloned cells with NK activity and cloned suppressor T cells express morphologic and biochemical characteristics heretofore associated with basophilic granulocytes. However, these clones differ in surface glycoprotein expression and immunologic function, and the full extent of the similarities and differences among these populations and basophils remains to be determined

カンジダ特異的IgG(4)抗体と気管支喘息

Serum levels of Candida-specific IgG(4) antibodies were examined in 66 patients with bronchial asthma, relating to patient age and asthma severity. 1. The levels of Candida-specific IgG(4) antibodies were the highest in patients with 60+years of age, compared to the levels in cases with 0-39 and 40-59 years of age. 2. In cases with 40-59 years of age, the levels of Candida-specific IgG(4) were significantly higher in cases with long-term steroid therapy (severe intractable asthma) than in cases without steroid regimen. The results suggest that increased levels of Candida-specific IgG(4) were observed in relation to patient age and asthma severity.66例の気管支喘息症例を対象に，血清中カンジダ特異的lgG(4)抗体を測定し，その血中レベルと年齢および喘息の重症度との関連について検討を加えた。1．血清カンジダ特異的IgG(4)値は，0～39才，40～59才の年齢層にくらべ，60才以上の年齢層において高く，年齢によりその値が変動することが 示唆された。2．40～59才の年齢層では，血清カンジダ特異的IgG(4)は，ステロイド非使用例にくらべ，ステロイド依存性重症難治性喘息症例で有意の高値を示し，この年齢層では，カンジダ特異的IgG(4)抗体がその発症病態に関連している可能性が示唆された。以上，カンジダ特異的IgG(4)抗体の上昇は，高年齢層の症例（60才以上）では全般的に，また40～59才の年齢層で重症型喘息症例において観察されることが明らかにされた