Hypersensitivity to chironomid Chironomus salinarius (non-biting midge living in the lagoon of Venice) in a child with serious skin and respiratory symptoms.

We are here describing the case of a ten-year-old child, without any allergyc family antecedent, who presented a growing skin and respiratory symptomatology in contact with swarms of non-biting midges Chironomus Salinarius in the summers 1984-1985-1986. These diptera have been greatly multiplying since the beginning of the summer 1982. The skin tests and the RAST-ELISA with common air-borne allergens resulted negative. On the contrary the prick-test with a suspension of the entire body of the Chironomus Salinarius and the radio-allergo-sorbent-test, specially prepared, resulted strongly positive. We point out that the massive reproduction of these midges, which may provoke allergy, in drainage areas and lagoons is a consequence of man interference on the natural water equilibrium.

Monoclonals and allergy. A review, with experimental contribution.

The role of monoclonal antibodies in allergy has been explored. First the status of art of monoclonal research in general is reviewed, by outlining a monoclonals identikit and the relevant technologies employed for their development. The attention is then focused on the present impact of monoclonals in the allergological field, first considering a general outline, and then the important steps of standardization of monoclonal antibodies. A comprehensive hint is made concerning the monitoring of immunotherapy, with future extrapolations on developing anti-idiotype vaccines, of which same examples can already be found in the infectious field, thus leaving the way open in allergy as well. A second section deals with the experimental contribution of the Authors, with the description of the preparation of the allergenic extracts of D.F., with details of the relevant steps (rabbit immunization; extract characterization; techniques used for monoclonal screening and characterization). The results obtained are discussed in relation to the techniques employed, weighing the reciprocal advantages and drawbacks.

Immunological agents in the respiratory tract: their role in homeostasis and immunopathogenesis of some morbid conditions of the airways.

Some immunological agents, which are expected to interfere with homeostasis and immunopathogenesis of some morbid conditions of the airways, have been reviewed. Attention was focused on some immunoglobulins (IgG, IgA, IgE) and other immunoeffectors such as alveolar macrophages, eosinophils, lymphocyte subsets, interleukins. Other substances, which prospective relation with immunity has been lately highlighted, were considered as well (fibronectin, neuropeptides). The possible pathological role of these immunoeffectors has been discussed.

Specific immunotherapy with allergoids in atopic subjects, hyporeactive to other vaccines.

Allergoids comply with the therapeutic requirement to reach high doses of the injected allergen, since the modifications of the allergens cause a loss of allergenicity, though the immunogenicity is being retained. Considering that it is possible to use a starting dose 40 times higher in comparison with retard vaccines, we thought it very expedient to treat with allergoids 18 patients allergic to parietaria, and 12 patients allergic to grass pollen, who had shown no clinical improvement after immunotherapy, lasting for at least three years with retard vaccines. The therapeutic results obtained are the following: improvement in 66% of the parietaria allergic patients after the first year, and of the 72% in the second year, of the 75% of the grass pollen allergic patients after the first year and of the 83% in the second year. They can be interpreted as due to the fact that such patients are always poorly reactive to the doses of allergens commonly present in retard vaccines. On the contrary, they do react to allergoids, both due to the high amount of allergen administered, and to the fact that polymerization brings about a stronger epitopic expression matched by a higher immunogenic activity. Moreover, the data are also indicative for a good relationship between the antibody titer of antigen-specific blocking IgG and the clinical effectiveness valued by means of symptom and medication scores.

[New findings on the immunology of bronco-82]. / Nuove acquisizioni sulla immunologia del bronco-82.

The effect of Bronco-82 treatment, a bacterial polyvalent vaccine, was determined on some of the most significant parameters of the immune response, i.e. PHA-induced blastogenesis, secretory IgA levels, IL 2 receptor expression, human T-cell activation antigen (MLR 3) and gamma-interferon production. The results obtained pointed out a high immunostimulating activity of the vaccine on the parameters considered. Some aspects of the complex immunomodulating effects of Bronco-82 are discussed.

Immunomodulators and allergy.

The role played by immunomodulation in allergy is critically reviewed. The prospective mechanisms of immunomodulation, specifically including those showing relation with allergic phenomena, are taken into consideration, examining the main interfering factors. Type I, IgE-mediated immunoreaction, in its two aspects, IgE synthesis and mediator's release, is analyzed, bearing in mind some agents involved or affecting these processes, mimicking immunomodulation. Other agents, such as Corynebacterium granulosum derivatives and glucocorticoids have been also appraised, to highlight their action in this connection.

In vivo and in vitro study of antigens and allergens in pollen extracts of Graminaceae.

The standardization of the diagnostic and immunotherapeutic products plays an even more definite role in allergology. The investigations performed in the last few years allowed to attain the purification and characterization of the allergenic extracts, both from the allergenic and antigenic viewpoint, implementing the most sophisticated techniques. The identification of the allergens in the extracts is of primary importance in the standardization and purification of the allergens themselves. The allergenic activity may be determined both in vitro and in vivo and the combination of the two techniques allows reaching a higher reliability in the assay. The Authors thought it very timely to verify the reliability of the in vivo and in vitro assays by employing four allergenic extracts of Graminaceae pollens, due to their importance in the allergological field. Phleum, Lolium, Cynodon, Anthoxanthum, were selected for the investigations. The extracts were also characterized by immunoprecipitation techniques. The results allowed to state that good results may be obtained by the in vivo assay (prick test) of allergenic extracts, with a precision comparable to that obtainable with the RAST inhibition, even using a restricted sample of allergic patients. It was ascertained that it is possible to express the allergenic activity versus a reference extract (Phleum). The three extracts (Anthoxanthum, Cynodon, Lolium), for which good antisera were obtained, were antigenically characterized, by means of CRIE and CIE, and the major allergens were identified.

Attempts at characterizing the allergens of cat saliva and fur.

In the framework of a wide program aimed at identifying various animal allergens, it was deemed interesting to study cat fur and saliva antigens. Cat fur and saliva extracts were prepared. Cat saliva antiserum was raised in rabbits with cat saliva as antigen. Immunochemical techniques such as isoelectric focusing, blotting, CIE, CRIE, fused rocket immunoelectrophoresis, line immunoelectrophoresis were implemented in order to identify the allergens and their isoelectric point, to perform the antigenic and allergenic characterization of the candidate allergen, to define their molecular weights and last, but not least, to carry out the quantitative assay towards a standard. On the whole the results obtained, besides confirming some previous results, pointed out by literature (presence of Cat 1 and albumin in the extracts) do stress the presence of other allergens, and the possibility to implement isoelectric focusing and blotting, which avoids the use of animal antisera, for the allergen detection in biological materials.

[Immunotherapy of allergic manifestations]. / Immunoterapia delle manifestazioni allergiche.

The Authors, after outlining the epidemiologic role of the allergic disorders, which places them among the pathologies covered by the wide chapter of social diseases, deal with the immunogenetic aspects and the particular conditions of the atopic habitus, atopic condition and atopic disease. The latest trends of antigen non specific immunotherapy, with the relevant possibility of eliciting a non specific suppressive mechanism of the IgE response, are considered. The antigen specific immunotherapy is then discussed under its various aspects: prophylaxis, efficacy, indications, modalities of treatment, unwanted reactions, immunological modifications, possible causes of its failure, medico-legal problems relevant to such therapy. Finally, the latest perspectives in the field of specific immunotherapy are envisaged, such as the use of allergoid vaccines, of liposomes as immunoadjuvants or the stimulation and the induction of suppressive factors of the IgE response.

[Characterization and standardization of allergen extracts]. / Caratterizzazione e standardizzazione degli estratti allergenici.

The allergenic extracts employed in the diagnostic or therapeutic fields are prepared from the different raw materials employing different extraction fluids: it has been shown that the solute contains, besides the allergens, many other substances. The analytical techniques, of which some of the most useful are described, should be implemented for the characterization and standardization of the extracts. The results obtainable, when using these techniques, for the extract characterization, are presented. Subsequently, the most important allergens identified and purified are briefly reviewed. The use of the analytical techniques described allows verifying the changes occurring in the allergenic extracts during storage. The implementation of the same allows the evaluation of the possibility, in subsequent times, of preparing, therefore standardizing, replicated allergenic extracts, similar to the former insofar as the allergenic and antigenic characteristics are concerned.

[Diagnostic use of a radioenzyme test in urticaria-like disorders and in urticaria-angioedematous syndromes]. / Impiego diagnostico di un test radioenzimatico nelle forme orticarioidi e nelle sindromi orticario-angioedematose.

The Authors report the results obtained with the histamine radioenzymatic test in the evaluation of the histamine content of granulocytes of 91 subjects, suffering from urticaria and urticaria-angioedema syndrome. The laboratory investigation was also integrated, according to the clinical implications, by other in vitro tests such as: kallikrein, RAST, PRIST, secretory IgA, precipitins assays. In urticaria-angioedema syndrome the quantitative and functional evaluation of C1-esterase inhibitor was also performed, to exclude the heredity of these pathologic forms. Basing on the results obtained, the authors expect that the granulocyte histamine radioenzymatic assay is highly reliable from the diagnostic viewpoint in the urticaria and angioedema forms.

[Immunodiagnosis and immunotherapy of allergic forms]. / Immunodiagnostica ed immunoterapia delle forme allergiche.

The Authors outline the immunological and immunopathogenic bases of atopy, pointing out the possible mechanisms involved in the impaired control capacity on IgE synthesis (hyperreactivity of NK cells, histamine-H2 receptors interaction, adenylcyclase stimulation, etc.). The in vivo and in vitro techniques employed in the allergological diagnosis are examined, with particular reference to the more recent tests (RAST, RAST Spot Test, Histamine radioenzymatic assay, in vitro IgE spontaneous synthesis, etc.). The latest findings in the allergological immunotherapy are then surveyed, including both the antigen non specific immunotherapy and the relevant possibility of inducing a non specific suppressor mechanism of the IgE response, and the antigen specific immunotherapy, which, besides the classical (aqueous, retard) vaccines, exploits the monovalent allergenic extracts, the allergoids (formaldehyde and glutaraldehyde) and the modified allergens (urea, PEG/PVA, D-GL, polysaccharides, photooxydation).

[RAST: a micromethod using whole blood (spot test)]. / RAST: impiego di un micrometodo su sangue intero (spot test).

The discovery of the role played by IgE immunoglobulins in allergopathies has allowed developing a radioimmunotechnique called RAST, that permits the detection of antireaginic antibodies, present in tiny concentrations in serum and secretions. One factor so far limiting the total use of RAST in the pediatric field, was supplied by the amount of serum required, difficult to obtain especially in the newborn. Reported herein, the Authors show the results obtained by using spot test on whole blood, in which the performance of the RAST requires a few drops of blood. The spot test has the same diagnostic validity (sensitivity, specificity, reproducibility etc.) as RAST performed on serum. The optimal ratio both as to drops of blood and as the ratio between specific and aspecific bound radioactivity, has been obtained by using 7 and 10 mm-diameter disks. Moreover these disks, once they have been dried at room temperature, turned out to be stable for several weeks. The Authors hold that the application of spot test in the pediatric and neonatal fields avoids psychic trauma of blood withdrawal sometimes difficult and allows a deeper insight in the etiological factors of atopic diseases to start a correct therapeutic treatment.

[Spot test: use of a micromethod with whole blood in allergy diagnosis]. / Spot test: impiego di un micrometodo su sangue intero nella diagnostica allergologica.

A limiting factor for the use of some in vitro techniques in the allergological diagnostic (RAST, PRIST, Precipitins) in the pediatric field, was the amount of serum needed for their carrying out, very difficult to obtain, especially in neonatology. In the present report the Authors record the results obtained by using a micromethod on the whole blood, called "Spot Test", which consists in the use of a particular base on which the blood drops are directly absorbed. The "Spot Test" showed the same sensitivity and reproducibility of the allergometric tests performed on the serum, therefore the correlation coefficients "r" for the RAST (r = 0.96), PRIST (r = 0.92) Precipitins (r = 0.89) showed to be highly significant for a close correlation between the two tests. Besides the "Spot test" turned out to be endowed, once the bases disc-blood dried with a stability at room temperature for several months (four months) without losing its sensitivity or specificity. This allows the possible sending of these bases to Specialized Centers without any particular problems of storage.

[Research on a new allergenic extract adsorbed on L-tyrosine. Pharmacological and immunotherapeutic findings]. / Ricerche su un nuovo estratto allergenico adsorbito su L-tirosina. Indagini farmacologiche ed immunoterapiche.

Antigen-specific immunotherapy, whose usefulness has been widely and objectively documented in the last few years, and whose therapeutic effectiveness depends on the administration of high doses of antigen, exploits the use of retard allergenic extracts which turn out to be more tolerated, and liable to a lower occurrence of side-effects. The Authors describe the results obtained with pollinosis vaccine adsorbed on L-tyrosine and employed on 206 patients with seasonal allergic conditions. Pharmacological experiments were performed on these vaccines, to establish their general tolerability (on mice and guinea-pigs), their local tolerability and the prolonged tolerability on rats. The results of the pharmacological experiments show that the vaccine does not cause any side-effects or toxic phenomena, that the growth curve of the animals treated is superposable to that of the controls, and that the product is perfectly absorbed without irritation or phlogosis of the peripheral tissues. As far as the clinical data are concerned a high percentage of cases (56.6%) classified as "optimal" were observed, while only a low percentage (4.54%) showed results classified as "null". Concerning the occurrence and the frequency of side-effects we should like to point out that only in one of the 206 cases tested (0.48%) was there local itching. These data are particularly significant, when compared with the side effects registered during immunotherapy with the aqueous vaccines.

[Immunomodulators]. / Immunomodulatori.

A comprehensive picture of the most well-known immunomodulators has been drawn, focussing the attention on their immunopharmacological profile and their prospective clinical use. Besides, the cell targets of the immunologically active agents are also considered. The role of cyclic nucleotides in the mechanisms of immune regulation as well as the importance of the host characteristics in this connection are carefully analyzed. Some general considerations are finally set forth on the usefulness of the immunopharmacology investigation both in defining the action of the agents under study and in suggesting their potential role in the design of new agents for immunotherapy.

[Use of staphylococcal protein A for the detection of rubella virus IgM antibodies]. / Impiego della proteina A stafilococcica per la ricerca di IgM specifiche verso il virus della rosolia.

The ability of Staphylococcus aureus protein A to bind human serum immunoglobulin of different classes and the conditions which allow a selective reduction of IgG in comparison to other immunoglobulin classes were preliminarly studied. Working under standardized conditions, a recovery of 3.5% of the initial value for IgG, of 40-50% for IgM and of 70-75% for IgA was obtained. Rubella HI antibodies, before and after staphylococcus treatment, were titred in 88 sera collected from adult women at different times from hexantema and in 210 sera of 109 subjects exposed to eventual contamination. The results were compared with the ones obtained by means of sucrose density gradient centrifugation followed by titration of rubella HI antibodies in the fractions. The tests proved that HI antibodies titration, before and after serum treatment with staphylococcal protein A, can be used, as presumptive test, for the research of rubella specific IgM antibodies. In fact, all the sera, on which the presence of rubella IgM antibodies was proved by sucrose density gradient centrifugation, were also positive after staphylococcus treatment. In 15% of sera resulting positive for HI rubella antibodies after staphylococcus treatment, usually at low titer, the presence of specific IgM antibodies was not detected by serum fractionation on sucrose density gradient ("false positive" cases). The results of serological diagnosis, carried out by using treatment with staphylococcal protein A and serum fractionation on sucrose density gradient, matched the clinical-epidemiological data. Twenty-seven women, out of 28 examined during the first three months of pregnancy and for whom serological diagnosis of a "recent" rubella virus infection was negative, had normal children at normal time. Therefore only in one case a spontaneous abortion occurred for uncertain causes. Rubella virus was isolated from the embryo of a woman who interrupted pregnancy because asymptomatic rubella virus infection was serologically diagnosed in the second month.

[Use of a new radioenzymatic test in urticaria and drug allergy]. / Impiego di un nuovo test radioenzimatico nelle forme orticariodi e nelle allergie da farmaci.

The Authors report preliminary data obtained with the histamine release test, evaluated with a radioenzymatic technique, in patients with a symptomatology highly indicative for penicillin allergy (21 cases) and in patients with urticarial forms (20 cases) or atopic dermatitis (5 cases). This technique, suitably modified, turned out very sensitive and able to detect histamine concentrations in the picogram range. The use of this test in type I immunoreactions, after Ig removal and fixation (IgE, IgG4) from basophils of atopic and normal subjects, pointed out its remarkable advantages (sensitivity, specificity, reliability) in comparison with other techniques. The results of our case-list, even if preliminary seem to be indicative for an extension of the trial to a wider number of patients both with allergies of pharmacological type and with urticarial forms or atopic dermatitis.