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Although flow cytometry is increasingly used as a tool to diagnose hematologic malignancies, the reporting format of acute leukemia immunodiagnosis is still imprecise and sometimes vague, often reflecting old guidelines. Thus, the purpose of the present work was to make the reporting format for the immunological diagnosis of acute leukaemia easy and clear to understand. This work represents part of a more articulated series of technical guidelines that the Italian Society for Cytometry (GIC) is currently processing. Thirteen separate recommendations, covering all aspects of an acute leukemia cytometry report, are listed. According to our suggestions, the report must contain clear statements about: 1. demographic identification of patient; 2. identification of the hospital or division sending the sample; 3. type of specimen (bone marrow aspirate, peripheral blood, other biological fluids); 4. timing of observation (first diagnosis or follow-up); 5. diagnostic hypothesis made by the sender; 6. list of antigens and type of immunofluorescence analysis carried out; 7. absolute number of cells in the sample; 8. quality of the sample, in terms of viability; 9. general description of the gating procedure; 10. immunophenotype of blast cells; 11. description of cells surrounding blasts; 12. diagnostic conclusions; 13. definition of an antigen panel (when applicable) for the detection of minimal residual disease. As an example of a final report we present a case of acute myeloid leukaemia with t(8;21) translocation; in filling this report, we followed all the 13 points of the checklist described in the paper.

The current burden of allergic diseases, estimated by both direct and indirect costs, is very relevant. In fact the cost estimation for rhinitis amount globally to 4-10 billion dollars/year in the U.S. and to an average annual cost of 1089 euros per child/adolescent and 1543 euros per adult in Europe. The estimated annual costs in Northern America for asthma amounted to 14 billion dollars. Consequently, preventive strategies aimed at reducing the clinical severity of allergy are potentially able to reduce its costs. Among them, specific immunotherapy (SIT) joins to the preventive capacity the carryover effect once treatment is discontinued. A number of studies, mainly conducted in the US and Germany demonstrated a favourable cost-benefit balance. In the nineties, most surveys on patients with allergic rhinitis and asthma reported significant reductions of the direct and indirect costs in subjects treated with SIT compared to those treated with symptomatic drugs. This is fully confirmed in recent studies conducted in European countries: in Denmark the direct cost per patient/year of the standard care was more than halved following SIT; in Italy a study on Parietaria allergic patients demonstrated a significant difference in favor of SIT plus drug treatment for three years versus drug treatment alone, with a cost reduction starting from the 2nd year and increasing to 48% at the 3rd year, with a highly statistical significance which was maintained up to the 6th year, i.e. 3 years after stopping immunotherapy, corresponding to a net saving for each patient at the final evaluation of 623 euros per year; in France a cost/efficacy analysis comparing SIT and current symptomatic treatment in adults and children with dust mite and pollen allergy showed remarkable savings with SIT for both allergies in adults and children.

Background: three main problems hamper the identification of wheat food allergens: 1) lack of a standardized procedure for extracting all of the wheat protein fractions; 2) absence of double-blind, placebo-controlled food challenge studies that compare the allergenic profile of Osborne’s three protein fractions in subjects with real wheat allergy; 3) lack of data on the differences in IgE-binding capacity between raw and cooked wheat. Methods: sera of 16 wheat challenge positive patients and 6 with wheat anaphylaxis were used for SDS-PAGE/immunoblotting of the three Osborne’s protein fractions (albumin/globulin, gliadins, glutenins) of raw and cooked wheat. Thermal sensitivity of wheat LTP was investigated by spectroscopic approaches. IgE cross-reactivity between wheat and grass pollen was studied by blot inhibition. Results: The most important wheat allergens were the alpha-amylase/trypsin inhibitor subunits, which were present in all three protein fractions of raw and cooked wheat. Other important allergens were a 9-kd lipid-transfer-protein in the albumin/globulin fraction and several low-molecular weight glutenin subunits in the gluten fraction.........................

The efficacy and safety of sublingual immunotherapy (SLIT) are currently supported by clinical trials, metaanalysis and post-marketing surveys. Practice parameters for clinical use of SLIT are proposed here by a panel of Italian specialists, with reference to evidence based criteria. Indications to SLIT include allergic rhinoconjunctivitis, asthma, and isolated conjunctivitis (strength of recommendation: grade A). As to severity of the disease, SLIT is indicated in moderate/severe intermittent rhinitis, persistent rhinitis and mild to moderate asthma (grade D). SLIT may be safely prescribed also in children aged three to five years (grade B), and its use in subjects aged more than 60 years is not prevented when the indications and contraindication are ascertained (grade D). The choice of the allergen to be employed for SLIT should be made in accordance with the combination of clinical history and results of skin prick tests (grade D). Polysensitisation, i.e. the occurrence of multiple positive response does not exclude SLIT, which may be done with the clinically most important allergens (grade D). As to practical administration, co-seasonal, pre co-seasonal, and continuous schedules are available, being the latter recommended for perennial allergens or for pollens with particularly prolonged pollination, such as Parietaria (grade D). For pollens with relatively short pollination, such as grasses and trees (cypress, birch, alder, hazelnut, olive) the pre coseasonal and perennial schedules are preferred (grade C). The build-up phases suggested by manufacturers can be safely used (grade A), but they can be modified according to the patient’s tolerance (grade C). A duration of SLIT of 3-5 years is recommended to ensure a long-lasting clinical effect after the treatment has been terminated (grade C).

Walnut is the most common cause of allergic reactions to tree nuts, as reported by large population studies. Two major allergens of walnut have been identified up until now: a 2S albumin and a vicilin-like protein. OBJECTIVE: This study was designed to identify the walnut major allergens in the Italian population and to compare the walnut IgE-binding profile in patients with or without pollen allergy. METHODS: We selected 46 patients either with oral allergy syndrome confirmed by open oral challenge or with systemic symptoms after ingestion of walnut. These patients' sera were used for the immunoblotting of walnut extract; the identified allergens were purified by HPLC and sequenced. A peach-walnut cross-inhibition study was then performed. RESULTS: The only major allergen recognized by our study population was a 9-kd lipid transfer protein (LTP), recognized by 37 patients. Two other minor allergens of approximately 9-kd molecular weight, both belonging to the vicilin family, were recognized by 10 patients. IgE binding to walnut LTP was completely inhibited by peach LTP. CONCLUSION: In Italian patients with walnut allergy confirmed by documented history of severe systemic reactions or by open oral food challenge, the major allergen is an LTP. The sensitization to this protein seems to be secondary to the sensitization to peach LTP, which acts as the primary sensitizer. LTP and vicilins were able to sensitize patients not allergic to pollen.

Background. Hazelnut major allergens up to now identified are a 18 kd protein homologous to Bet v 1 and a 14-kd allergen homologous to Bet v 2. No studies reported hazelnut allergens as recognized by patients with positive double-blind placebo controlled food challenges (DBPCFCs) or allergic to hazelnut but not to birch. Objective. In this study we characterized the hazelnut allergens by studying the IgE reactivity of 65 patients with positive DBPCFC and of 7 patients with severe anaphylaxis to hazelnut. Methods. Hazelnut allergens were identified by SDS-PAGE and IgE immunoblotting. Further characterization was made by: a) amino-acid sequencing; b) evaluation, with EAST inhibition, of IgE binding properties of raw and roasted hazelnut, and c) assessment of cross- reactivity with different allergens by immunoblotting inhibition. Results. All the sera from the DBPCFC positive patients recognized a 18 and a 47 kd allergen; other major allergens were at a m.w. of 32 and 35 kd. The binding to the 18 kd band was inhibited by birch extract, demonstrating its homology with the birch major allergen, and was abolished in roasted hazelnut. The 47 kd allergen is a sucrose-binding protein; the 35 kd allergen is a legumin; the 32 kd allergen is a 2S albumin. The patients with severe anaphylactic reactions to hazelnut showed a specific IgE reactivity to a 9 kd allergen, totally inhibited by purified peach LTP, which was heat-stable. Conclusions The major allergen of hazelnut is a 18 kd protein, homologous to Bet v 1; the 9 kd allergen is likely a lipid transfer protein; other major allergens have a m.w. of 47, 32 and 35 kd.

Two almond IgE-binding proteins were N-terminally sequenced and identified as almond 2S albumin and conglutin ã. Localisation and conservation of IgE binding in a 6 kDa peptide obtained by endoproteinase digestion of 2S albumin was shown.