Prolonged organ culture reduces the incidence of endothelial immune reactions.

PURPOSE The number of antigen-presenting cells decreases during organ culture of corneoscleral discs. This might result in a decrease of immune reactions with increasing duration of organ culture. To investigate this hypothesis, we performed a retrospective analysis of all penetrating keratoplasties that were consecutively performed over the last 5 years.PATIENTS AND METHODS All cases of penetrating keratoplasties (n=1006) were divided into two groups, with the division made at the median of the storage time (21 days). These two groups were compared by a Cox proportional hazards survival model regarding the incidence of endothelial immune reactions, clear graft survival, and chronic endothelial cell loss following penetrating keratoplasty considering patient's age, donor's age, and risk situation as co-variates.RESULTS We observed statistically significantly fewer endothelial immune reactions (20.1% (95% confidence interval 15.5-24.5%) after 2 years) in the group with a storage time of more than 21 days compared with the group with a storage time of <21 days (26.5% (95% confidence interval 21.6-31.2%) after 2 years). However, the duration of organ culture did not have a statistically significant effect on clear graft survival or chronic endothelial cell loss.CONCLUSION Our results demonstrate that an increased duration of organ culture leads to a lower incidence of endothelial immune reactions following penetrating keratoplasty. However, we do not recommend increased storage times in general as overall graft survival did not improve. The reason for this apparent paradox may be that the endothelial cell count decreases during storage time.

Hypersensitivity pneumonitis induced by Shiitake mushroom spores.

Hypersensitivity pneumonitis (HP) is a pulmonary granulomatosis involving an immunoallergic mechanism caused by chronic inhalation of antigens, most frequently organic substances, as well as chemicals. We report the first European case of hypersensitivity pneumonitis due to the inhalation of Shiitake mushroom spores. A 37-year-old French Caucasian man with a one-month history of persistent dry cough, shortness of breath and loss of weight was admitted to our hospital on December 2010. Anamnesis showed he was involved in mushroom production beginning in the summer of 2010. His temperature on admission was 36.6°C and he had a normal blood pressure (135/90 mmHg). Bilateral fine crackles were audible in the base of both lungs. Pulmonary function tests showed a mild restrictive pattern with decreased DLco and a PaO(2) of 65 mmHg, Chest CT scan revealed reticulo-nodular shadows, slight ground glass opacities, liner atelectasis, and subpleural opacities in both lung fields. Bronchoscopy was normal but cytological examination of BAL revealed a predominant lymphocytosis (55%). Serum precipitins to the Shiitake mushroom spores were positive (3 precipitins arcs with high intensity) and as a result we advised the patient to cease his mushroom production activities. The diagnosis of hypersensitivity pneumonitis due to inhalation of Shiitake mushroom spores was established as a result of the improvement of all of his clinical symptoms, i.e., cough, weight loss, bilateral fine crackles, mild restrictive pattern of pulmonary function, and reticulo-nodular shadows on chest CT, once exposure was eliminated. Recent interest in exotic mushrooms varieties, e.g., Shiitake, in developed countries because of their possible medicinal properties might increase the potential risk of HP among mushrooms workers. Therefore, healthcare professionals have to take this new potential respiratory disease into account.

Recombinant allergens combined with biological markers in the diagnosis of allergic bronchopulmonary aspergillosis in cystic fibrosis patients.

Allergic bronchopulmonary aspergillosis (ABPA) is a frequent complication in cystic fibrosis patients. The diagnosis remains difficult and requires a combination of clinical, radiological, biological, and mycological criteria. The aim of this study was to analyze the added value of two recombinant antigens, rAspf4 and rAspf6, associated with the detection of specific IgG; precipitins; total IgE; and Aspergillus fumigatus in sputum for the diagnosis of ABPA. In a retrospective study, we determined the specific IgE responses to these recombinants in 133 sera of 65 cystic fibrosis patients. We selected an average of five serum samples from each of the 17 patients with ABPA (13 proven and 4 probable ABPA) and from 3 patients with Aspergillus bronchitis and rhinosinusitis. One serum sample for the 45 patients without ABPA was tested. The sensitivity of specific IgE detection against rAspf4 calculated per patient (92.3%) was significantly higher (P < 0.05) than that of rAspf6 (53.8%). When rAspf4 IgE detection was associated with anti-Aspergillus IgG enzyme-linked immunosorbent assay (ELISA) and precipitin detection, the sensitivity rose to 100%. The specificities of rAspf4 and rAspf6 IgE detection were 93.7% and 91.6%, respectively. Other diagnostic criteria had slightly lower specificities (87.5% for anti-Aspergillus IgG ELISA, 89.6% for precipitins, 84.4% for total IgE, and 85.0% for positive A. fumigatus culture in sputum). In conclusion, this retrospective study showed the relevance of rAspf4 IgE detection, in combination with other biological markers (Aspergillus IgG ELISA, precipitins, and total IgE), for improving the biological diagnosis of ABPA.

Severe asthma with fungal sensitization: a case report and review of literature.

There is a substantial body of evidence supporting an association between asthma severity and fungal exposure and sensitization. Fungal allergens are a recognized risk factor for severe asthma. We describe the case of a 44-year-old asthmatic whose asthma control deteriorated after moving to a new flat with walls covered in mould. Allergic bronchopulmonary aspergillosis was excluded. Although sensitization to Candida was demonstrated by a positive Candida-specific radioallergosorbent test, the patient did not entirely satisfy the criteria for a diagnosis of allergic bronchopulmonary candidiasis. The patient's asthma control improved after engaging in a monthly washing regimen of the walls. This case further demonstrates the association between fungal sensitization and asthma severity. The term severe asthma with fungal sensitization has been recently coined to describe this phenomenon.

Case report: A rare cause of miliary nodules -- allergic bronchopulmonary aspergillosis.

Allergic bronchopulmonary aspergillosis (ABPA) is a complex immune hypersensitivity reaction to the fungus Aspergillus fumigatus, which occurs in patients with asthma and cystic fibrosis. The common radiological findings include fleeting pulmonary opacities, bronchiectasis and mucoid impaction. Uncommon radiological findings encountered in ABPA include lung masses, perihilar opacities simulating hilar adenopathy and pleural effusions. Herein, we describe a 36-year-old man who presented to the chest clinic with miliary nodules on a chest radiograph and was diagnosed as having ABPA confirmed on both serology and lung biopsy. This is the first report of ABPA presenting as randomly scattered nodules on CT.

Aspergilloma formation in cavitary sarcoidosis.

Pulmonary cavitation is rather uncommon in patients with sarcoidosis, and aspergilloma is even more uncommon in such cases. Here, we present the case of a 63-year-old female patient with cavitary lung disease who had been under treatment for TB for 9 months. A diagnosis of pulmonary sarcoidosis was established based on the fiberoptic bronchoscopy finding of noncaseating granuloma. Treatment with corticosteroids led to a dramatic improvement in symptoms. While under treatment for sarcoidosis, the patient developed an aspergilloma. She presented immediate skin test reactivity to Aspergillus fumigatus, as well as positivity for A. fumigatus serum precipitins. This is the first reported case of aspergilloma formation in a patient with cavitary sarcoidosis in India.

Aspergilloma formation in cavitary sarcoidosis / Formação de uma aspergilloma em sarcoidose com cavitação

Pulmonary cavitation is rather uncommon in patients with sarcoidosis, and aspergilloma is even more uncommon in such cases. Here, we present the case of a 63-year-old female patient with cavitary lung disease who had been under treatment for TB for 9 months. A diagnosis of pulmonary sarcoidosis was established based on the fiberoptic bronchoscopy finding of noncaseating granuloma. Treatment with corticosteroids led to a dramatic improvement in symptoms. While under treatment for sarcoidosis, the patient developed an aspergilloma. She presented immediate skin test reactivity to Aspergillus fumigatus, as well as positivity for A. fumigatus serum precipitins. This is the first reported case of aspergilloma formation in a patient with cavitary sarcoidosis in India.

A cavitação pulmonar é rara em pacientes com sarcoidose, e o aspergiloma é ainda mais raro nestes casos. Apresentamos o caso de uma paciente de 63 anos com doença pulmonar cavitária em tratamento para a TB por 9 meses. Estabeleceu-se o diagnóstico de sarcoidose pulmonar com base nos achados de granuloma não-caseoso na fibrobroncoscopia. Houve grande melhora dos sintomas com o tratamento com corticosteroides. A paciente desenvolveu um aspergiloma durante o tratamento para a sarcoidose. Houve reação imediata ao teste cutâneo para Aspergillus fumigatus, assim como resultado positivo para precipitinas de A. fumigatus no soro. Este é o primeiro caso relatado de formação de aspergiloma em um paciente com sarcoidose com cavitação na Índia.

Assessment of four serological techniques in the immunological diagnosis of farmers' lung disease.

Farmers' lung disease (FLD) is a pulmonary disease that results from repeated inhalation of antigens from mouldy hay or straw. The objective of this prospective study was to assess the reliability of four serological techniques in FLD diagnosis. Sera from 15 consecutive patients with FLD, 15 healthy control farmers and 30 urban controls were analysed using four serological techniques [electrosyneresis (ES), Ouchterlony double diffusion (DD), ELISA and Western blot (WB)] with four antigens (Absidia corymbifera, Eurotium amstelodami, Wallemia sebi and Saccharopolyspora rectivirgula). In the authors' region, ES on cellulose acetate with A. corymbifera antigen was the most relevant diagnostic tool for discriminating FLD patients from healthy exposed farmers (sensitivity 87 %, specificity 100 %). DD tests were in accordance with ES, but their discriminatory power was lower. No threshold indicating both good sensitivity and specificity could be established with ELISA. WB analysis failed to identify specific bands for FLD. This study demonstrates the efficacy of determining precipitin levels with an appropriate technique, using a panel of antigens consistent with the specific exposure of a given area.

Diagnostic value of serum precipitins to mould antigens in active hypersensitivity pneumonitis.

Serum precipitins have a controversial diagnostic value in hypersensitivity pneumonitis (HP). The present authors' objective was to assess their diagnostic value by developing scores from a panel of specific antigens tested by two techniques (electrosyneresis and double diffusion) to discriminate active HP from other interstitial lung diseases. Consecutive patients presenting with a condition for which HP was considered in the differential diagnosis were included in the study. All patients underwent the same standardised diagnostic procedure, including precipitin tests performed in routine conditions. Clinical manifestations, bronchoalveolar lavage and high-resolution computed tomography defined the presence or absence of HP. Receiver-operating characteristic curves and logistic regression were used to develop the serological scores. A total of 122 patients (including 31 cases of HP) were included in the study. Five antigens from the panel were selected for the serological scores (Absidia corymbifera, Eurotium amstelodami, Wallemia sebi, Saccharopolyspora rectivirgula and mesophilic Streptomyces sp.). Electrosyneresis was more discriminative than the double-diffusion technique. Predictive negative values varied 81-88% and predictive positive values varied 71-75% for prevalence of HP 20-35%. In conclusion, serological scores using a panel of relevant antigens may guide both biological and clinical practice in areas of high prevalence of hypersensitivity pneumonitis.

Specific IgG to Thermoactynomices vulgaris, Micropolyspora faeni and Aspergillus fumigatus in building workers exposed to esparto grass (plasterers) and in patients with esparto-induced hypersensitivity pneumonitis.

BACKGROUND: Esparto is one the most frequent causes of hypersensitivity pneumonitis in Spain. OBJECTIVE: Determination of risk factors in the working environment that could explain the sensitisation process, and assessment of the differences in specific IgG levels to Aspergillus fumigatus, Saccharopolyspora rectivirgula and Thermoactynomices vulgaris in patients with hypersensitivity pneumonitis induced by esparto, exposed healthy plasterers and control population. METHODS: Determination of precipitins and specific IgG to Aspergillusfumigatus, Saccharopolyspora rectivirgula and Thermoactynomices vulgaris in the three previously mentioned groups. Questionnaire on possible risk occupational and extra-occupational factors. RESULTS: Both healthy and exposed plasterers have higher levels of specific IgG to Aspergillus fumigatus, Saccharopolyspora rectivirgula and Thermoactynomices vulgaris than the healthy controls. The patients had higher levels of IgG than exposed healthy plasterers only to Thermoactynomices vulgaris. Precipitins were detected in only two patients. There were no occupational factors influencing on the sensitisation process. CONCLUSIONS: Specific IgG is an occupational exposure marker among plasterers, but it has not been possible to establish a cut off point that differentiates exposed subjects from affected ones. This determination has a greater sensitivity than precipitins. We did not identify occupational or extra-occupational risk factors that facilitate the sensitisation process.

Significance of precipitins and asymptomatic lymphocytic alveolitis: a 20-yr follow-up.

What is the significance of serum precipitins and lymphocytic alveolitis in a healthy subject exposed to antigens responsible for hypersensitivity pneumonitis (HP)? This study was done to evaluate the 20-yr outcome of asymptomatic dairy farmers with or without precipitins or lymphocytic alveolitis in their bronchoalveolar lavage (BAL). Twenty-seven of the initial 43 farmers were restudied. Of the missing 16, 11 refused the follow-up, three had died and two could not be located. The restudied farmers had a clinical evaluation, lung function tests and a high resolution computed tomography (HRCT). Only one subject, an asthmatic, had dyspnoea, none described present or past symptoms suggestive of HP. Of those who were no longer on the farm, none had quit because of respiratory problems. Lung function changes were as expected with aging. The only difference was that farmers with positive precipitins had a statistically significant greater decrease in their forced expiratory volume in one second than the sero-negative farmers. Four HRCTs showed signs of minimal parenchymal abnormalities; these were not more prevalent for subjects with or without precipitins or BAL lymphocytosis. Serum precipitins and asymptomatic lymphocytic alveolitis in an asymptomatic, exposed dairy farmer have no clinically meaningful long-term consequences.

Clinical diagnosis of hypersensitivity pneumonitis.

The diagnosis of hypersensitivity pneumonitis (HP) is difficult and often relies on histopathology. Our objective was to identify diagnostic criteria and to develop a clinical prediction rule for this disease. Consecutive patients presenting a condition for which HP was considered in the differential diagnosis underwent a program of simple standardized diagnostic procedures. High-resolution computed tomography scan and bronchoalveolar lavage (BAL) defined the presence or absence of HP. Patients underwent surgical lung biopsy when the computed tomography scan, BAL, and other diagnostic procedures failed to yield a diagnosis. A cohort of 400 patients (116 with HP, 284 control subjects) provided data for the rule derivation. Six significant predictors of HP were identified: (1) exposure to a known offending antigen, (2) positive precipitating antibodies to the offending antigen, (3) recurrent episodes of symptoms, (4) inspiratory crackles on physical examination, (5) symptoms occurring 4 to 8 hours after exposure, (6) and weight loss. The area under the receiver operating characteristic curve was 0.93 (95% confidence interval: 0.90-0.95). The rule retained its accuracy when validated in a separate cohort of 261 patients. The diagnosis of HP can often be made or rejected with confidence, especially in areas of high or low prevalence, respectively, without BAL or biopsy.

Use of co-immunoelectrodiffusion to detect presumed disease-associated precipitating antibodies, and time-course value of specific isotypes in bird-breeder's disease.

BACKGROUND AND OBJECTIVE: The practical value of immunological diagnosis of bird-breeder's disease (BBD) is controversial, because of difficulties in distinguishing active disease patients from simple contact subjects. The aim of this study was to determine the diagnostic and prognostic value of (a) presumed disease-associated antibodies precipitating pigeon antigens (immunoglobulin A (IgAp) and P2 component), (b) characterization of specific isotypes (IgG, IgM, and IgA), and (c) antibody kinetics after antigen eradication. METHODS: 405 subjects (775 sera) in contact with birds were studied [by means of co-immunoelectrodiffusion (Co-IED) and enzyme-linked immunofiltration (ELIFA)] with soluble extracts of pigeon droppings and squab crop milk. These patients were divided into two groups based on the final clinical evaluation of the patients' physicians, which was taken as the gold standard (positive in 90 and negative in 315 cases). RESULTS: On the basis of this gold standard, the detection of presumed disease-associated precipitating antibodies by Co-IED had a specificity of 95.5%, a sensitivity of 98.7%, an accuracy of 98%, and positive and negative predictive values of 95.5% and 98.7%, respectively. Most of the patients with a final positive diagnosis of BBD had specific IgG, IgM, and IgA antibodies by ELIFA. After antigen eradication, anti IgAp and/or P2 antibodies disappeared more rapidly than other precipitating systems. CONCLUSION: Identification by Co-IED of precipitating immune complexes IgAp and/or P2 significantly reinforces the intrinsic credibility of immunological diagnosis of BBD. Compared to these presumed disease-associated precipitating antibodies, detection and time course of specific IgM, IgA antibodies, provided no additional diagnostic value or prognostic arguments to judge disease activity after antigen eradication.

Specific immune response to occupational antigens in asymptomatic egg processing workers.

BACKGROUND: Hen's egg proteins are known sensitizers and may cause occupational respiratory diseases in exposed workers. The study was conducted in order to ascertain the prevalence of sensitization to work-related antigens in asymptomatic egg processing factory workers. METHODS: Clinical history, respiratory function, atopy and the presence of specific antibodies to environmental and occupational antigens were evaluated in 77 workers. A total of 116 unexposed and non-professionally exposed controls were also studied. The antigen concentration of the factory environment was measured. RESULTS: Specific IgE and precipitin positivity to egg components was more frequently present in workers than in the two control groups. The egg white protein concentration was higher in the egg breaking area. CONCLUSIONS: The avian proteins inhaled induced immune responses to occupational antigens in asymptomatic, professionally exposed subjects. Measures to decrease environmental antigen concentrations and thorough clinical monitoring of sensitized workers are advocated.

Symptoms and serum precipitins in workers exposed to dry sausage mould: consequences of exposure to sausage mould.

OBJECTIVE: Hypersensitivity pneumonitis due to dry sausage mould has been reported in workers who brush off the excess mould which coats dry sausage. Prevalence of symptoms and sensitization to mould among these pork-butchery workers is unknown. The aim of the study was to assess the clinical, radiographic, functional, and immunological features in exposed and non-exposed workers in semi-industrial pork butcheries. PATIENTS AND METHODS: Symptoms, and serum precipitins against mould extracts, were studied in workers in semi-industrial pork butcheries. Of 600 workers asked to participate, 123 (20.5%) were included. Fifty-nine workers, exposed to dry (raw) sausage mould and Penicillium nalgiovense were compared with 64 non-exposed subjects, for symptoms, chest X-rays, spirometry and CO-transfer measurements. Precipitating antibodies were detected by immunoelectrophoresis and electrosyneresis. RESULTS: Sneezing, cough, dyspnoea, nasal obstruction, headache, and discomfort were significantly more frequent in the exposed group at work and after work than in the control group (P < 0.05). The prevalence of precipitating antibodies for sausage mould was higher in the exposed group (37%) than in the non-exposed group (9%) (P < 0.01). The mean number of precipitating lines measured by electrosyneresis was higher in exposed workers than in non-exposed workers for mould extract (1.09 vs 0.28, P < 0.05) and for Penicillium nalgiovense (1.77 vs 0.33, P < 0.05). No specific X-ray opacity or lung function impairment was found in either group. CONCLUSIONS: Clinical symptoms and sensitization to Penicillium nalgiovense are frequent among workers exposed to mould during brushing in dry sausage plants.

Partial characterization of serum immunoglobulins of Oreochromis mossambicus.

Serum immunoglobulins of O. mossambicus were purified using chromatography methods--CM affinity gel blue chromatography followed by two step purification involving a combination of ion-exchange and gel filtration chromatography. Studies revealed that O. mossambicus produces only one class of high molecular weight macroglobulin as determined by molecular sieving by Sepharose CL 6-B. Immunoelectrophoresis of purified O. mossambicus serum against rabbit anti O. mossambicus serum gave only a single precipitin line. Further analysis of the immunoglobulin by SDS-PAGE showed that the IgM macroglobulin weighs about 900,000 Da, composed of mu-like heavy chain weighing about 90 kDa each and light chains weighing about 30 kDa each.

[The humoral reactions of bioincompatibility in the early period after the heterotopic autograft of veins into the coronary bed and the implantation of synthetic heart valves]. / Humoral'ni reaktsiï bionesumisnosti u rann'omu periodi pislia heterotopichnoi autotransplantatsiï ven u koronarne ruslo ta implantatsiï syntetychnykh klapaniv sertsia.

Humoral manifestations of bioincompatibility were studied at early stages (during several hours and days) after operations using non-peculiar to heart trans- and implants: aortocoronary bypass at 102 patients with coronarosclerosis and implantation of artificial cardiac valves at 145 patients with endocarditis. In the various postoperation stages the formation of the structurally and functionally different types of the nonclonal specific autoprecipitins to autological membranocellular components was revealed at recipients by methods of double immunodiffusion and immunoelectrophoresis. These autoprecipitins such as: early, activated and tardly, synthesized under effect of alien agent--may be useful as diagnostic and prognostic indicators of bioincompatibility and its clinical consequences at the earliest period after cardiac operations using trans- and implants.

[The manifestations of the immediate bioincompatibility reaction in patients undergoing restorative heart operations]. / Proiavy nehainoï reaktsiï bionesumisnosti u khvorykh pry vidnovnykh operatsiiakh na sertsi.

The generation in plasma the anodic autoprecipitating protein, or autoprecipitin (AAP), as the bioincompatibility reaction under influence of a number of alien components during cardiac operations with using cardiopulmonary bypass, heterotopic vascular autotransplants and artificial valve implants was studied in 257 patients. Selective formation of the AAPs in result of their inductors effect was discovered by the methods of immunodiffusion and immunoelectrophoresis in blood of patients within operation periods and early postoperation hours. Repeated and reiterated generation of the AAPs due to bioincompatibility in patients with coronarosclerosis and endocarditis, undergoing prosthetic cardiac operations may favour developing reaction of immediate hypersensitivity with complications and has prognostic value.

Precipitating Pseudomonas aeruginosa antibodies and antimicrobial therapy in cystic fibrosis patients.

Forty patients with cystic fibrosis were studied bacteriologically and serologically. Precipitating Pseudomonas aeruginosa antibodies were monitored by crossed-immunoelectrophoresis (CIE) in order to evaluate the possibility of preventing chronic colonization by Pseudomonas aeruginosa by cycles of antimicrobial therapy. Sputum or pharyngeal aspirate and serum samples from all patients were analyzed by means of spread on selective media and CIE, respectively. Significant differences in the number of precipitins were obtained: noncolonized and intermittently colonized patients had no precipitins, whereas the number of precipitins in the chronically colonized patients varied from 11 to 44. An increase in the number of precipitins could be a good marker for initiation of therapy with antimicrobial agents that are either active against Pseudomonas aeruginosa or able to inhibit the release of virulence factors.