Intracutaneous Tests Research Articles

The allergenic and antigenic properties of spore extracts of Aspergillus fumigatus: A comparative study of spore extracts with mycelium and culture filtrate extracts

The antigenic properties of conidial preparations of Aspergillus fumigatus were compared with mycelial and culture filtrate extracts by use of sera of patients with aspergillosis and hyperimmune rabbit serum. Biologic activities were tested by intracutaneous tests. The immunologic parameters used were: precipitating properties in double-diffusion, IgG binding by ELISA, IgE binding by enzyme inhibition assay, and by enzyme allergosorbent test. It has been demonstrated that the components that are released after disintegration of spores have positive titers in all immunologic assays. The immunologic properties of these spore plasma extracts are comparable with mycelial and/or culture filtrate extracts that are in accordance with the corresponding skin reactivities. Components that are released spontaneously from spores are only weakly positive or negative in the immunologic assays and demonstrate a very low biologic activity.

Detection of immunologically sensitized isocyanate workers by RAST and intracutaneous skin tests

Antibody estimations by RAST were performed on 621 partially selected isocyanate workers whose case histories were well-documented. Two hundred forty-seven of the workers were clinically suspected to be sensitized to isocyanates since they had suffered from occupationally related recurrent asthmatic attacks, chronic obstructive lung diseases, bronchitis, rhinitis, conjunctivitis, cutaneous hypersensitivity reactions, and/or fever. RAST studies with seven different isocyanates conjugated to human serum albumin (HSA) demonstrated that 14% of these symptomatic subjects had significant levels of specific-IgE antibodies. On the other hand, only one of the 374 asymptomatic workers had such antibodies. In contrast to toluene diisocyanate (80:20% mixture of 2, 4 and 2, 6-isomers, if not otherwise mentioned)-HSA, a toluene diisocyanate-ovalbumin conjugate exhibited no antigenicity in nearly all of the cases. Intracutaneous skin testing with five different isocyanate-HSA conjugates performed in 203 workers produced immediate-type wheal-and-flare reactions in 14 of the 53 symptomatic persons but in none of the 150 asymptomatic probands. There was a good overall correlation between RAST and skin test results. Our findings provide strong evidence for IgE-mediated sensitization to isocyanates in a subgroup of the subjects with isocyanate-induced diseases. By means of RAST and/or skin testing these immunologically sensitized persons can be identified.

Induction of benzylpenicilloyl specific antibodies, including IgE, by long-term administration of benzylpenicillin.

Using the ELISA and passive haemagglutination test we investigated the humoral immune response induced by long-term administration of benzylpenicillin in patients. After the second week of the course in twenty-four out of sixty-one patients, synthesis of benzylpenicilloyl (BPO) specific IgG, IgA, IgM and/or IgE could be demonstrated. BPO-specific IgE was always found in combination with BPO-specific IgG and/or IgM. No difference in the humoral immune response was found between patients who developed clinical hypersensitivity reactions and those who did not. In a follow-up study the disappearance of BPO-specific immunoglobulins of all classes was observed. Results of the epicutaneous tests showed that a long-term course of benzylpenicillin did not evoke delayed hypersensitivity. However intracutaneous tests with benzylpenicilloyl-polylysine or benzylpenicillin showed positive immediate type reactions. From the results it can be concluded that a BPO-specific IgE response is not necessarily associated with the development of a clinical hypersensitivity reaction.

Detection and expression of DNA homologous to the tox gene in nontoxinogenic isolates of Corynebacterium diphtheriae.

Three probes have been described which can be used to detect the presence of DNA sequences homologous to the tox gene of Corynebacterium diphtheriae. Probes "A" and "B" detected sequences coding for A and B fragments of diphtheria toxin, respectively. The third "A-B" probe contained both the A and B coding sequences. The B probe was completely unambiguous in detecting only toxin-related sequences, and the A probe was only slightly less so. The efficacy of the probes was tested on a series of toxinogenic and nontoxinogenic isolates of C. diphtheriae. All isolates which were toxinogenic as characterized by the gel immunodiffusion technique gave positive reactions with the probes. Of particular interest was the finding that 14 of 43 nontoxinogenic isolates also carried DNA homologous to both the A and B probes. All 14 isolates were nontoxinogenic by the rabbit intracutaneous test as well as by the gel immunodiffusion test; however, 12 of them produced ADP-ribosylating activity, whereas two were negative. The isolates producing ADP-ribosylating activity belonged to a cohort of cultures, of which 11 were isolated in South Dakota and 1 was isolated in Montana. Genomic DNAs of all 12 appeared to be identical when restriction enzyme digest patterns were compared, and the same fragment carried the tox gene in all of them. The tox-bearing nontoxinogenic isolates from Alaska and Florida each had unique restriction patterns and did not produce ADP-ribosylating activity. A number of genomic fragments of all the tox-bearing nontoxinogenic isolates hybridized with beta converting phage DNA. The significance of these observations to the natural history of diphtheria was discussed.

Study of immunological parameters in farmer's lung.

A cross-sectional epidemiological study was carried out on 2932 farmers (response rate 92%) living in Northern Italy. A questionnaire on respiratory symptoms was supplied, chest X-ray taken and simple spirometry tests made. Thirty-nine subjects reported attacks of breathlessness associated with fever after exposure to mouldy hay; thirty-six had a clinical history of farmer's lung and X-ray and/or respiratory function changes indicative of chronic farmer's lung; 113 were classified as having bronchial asthma. These 118 subjects and a random sample of 131 non-symptomatic subjects were examined by intracutaneous skin tests and double diffusion precipitation tests with antigens associated with farmer's lung. All sera positive by precipitation were examined by immunoelectrophoresis, counterimmunoelectrophoresis, and C1q binding test, and Bovine conglutinin test in solid phase for the detection of circulating immunocomplexes. 27.8% cases of chronic farmer's lung, 4.4% of cases of bronchial asthma and 2.3% of non-symptomatic subjects showed precipitating antibodies against farmer's lung antigens. Of the precipitin-positive subjects, the majority had precipitating antibodies against Micropolyspora faeni. No differences in immunoelectrophoregrams in number and pattern of precipitation arcs for M. faeni were found between chronic farmer's lung and bronchial asthma subjects. Circulating immune complexes were present in 50% of chronic farmer's lung patients, 40% of those with bronchial asthma and 33.3% of non-symptomatic subjects. Three farmer's lung subjects (9.3%) showed immediate skin-test reactions to rural environmental allergens.

Evaluation of infusion sets by biological and chemical testing

Some of the biological and chemical tests used in toxicological evaluation of synthetic polymers were correlated. The individual tests (sperm cell motility, tissue culture inhibition, intracutaneous test, optical absorption at 220–360 nm, reducing impurities) did not mutually correspond. The final estimation of the tested device is assessed as a sum of these units. Therefore, it is necessary to evaluate the results of the individual tests in arbitrary units.

Stabilization of a diluted aqueous mite allergen preparation by addition of human serum albumin. An intracutaneous test study.

Stability of diluted Spectralgen Dermatophagoides farinae allergen preparations in phenol-saline with, and without human serum albumin (HSA) has been studied by intracutaneous tests in mite-allergic patients. The phenol-saline reconstituted mite allergen preparation lost activity to less than 1/10 of initial activity within 1 week, while mite allergen preparation reconstituted with solvent containing HSA was stable for at least 8 months after reconstitution. Human serum albumin probably prevents adsorption of allergens to glass surfaces which may be a problem for very diluted allergen preparations.

Allergic reactions to measles (rubeola) vaccine in patients hypersensitive to egg protein

We evaluated two children with allergy to egg-white protein (ovalbumin) who had generalized urticaria, angioedema, and respiratory difficulty after immunization with live rubeola vaccine. In both patients, serum IgE reactive with ovalbumin-related antigens in the vaccine was demonstrated. Subsequent evaluation of 24 children with ovalbumin allergy revealed that those who had positive ovalbumin skin tests, but no clinical reaction to egg white, were skin test negative on prick and intradermal testing with measles vaccine and were safely immunized. They had no detectable IgE directed against the rubeola vaccine, although IgE directed at ovalbumin was present. Six patients who had severe allergic hypersensitivity reactions on exposure to ovalbumin had IgE antimeasles vaccine antibody and had positive reactions after intracutaneous or intradermal testing with the vaccine. These patients were safely immunized with increasing volumes (0.05 ml increments every 20 minutes) of measles vaccine to receive the full dose. These studies suggest that children with severe allergic hypersensitivity to egg white should be screened with an intracutaneous test prior to immunization with measles vaccine; however, children who have positive skin tests but no clinical reaction to ovalbumin exposure are at minimal risk for hypersensitivity reactions to measles immunization, as previously reported.

Diagnostic significance of late cutaneous allergic responses and their correlation with radioallergosorbent test.

In order to study the clinical significance of late cutaneous allergic responses 178 intracutaneous tests were done on the forearm of forty-three allergic patients with perennial allergy and twelve patients with pollen allergy. The allergen concentration was chosen to give a moderate immediate response with a weal diameter of about 15 mm. Late cutaneous allergic responses (LCAR) were elicited in eighty-two intracutaneous tests in thirty-one patients with perennial allergy and in nine intracutaneous tests in eight patients with pollen allergy, with almost all the allergens frequently responsible for allergy in this area. Their occurrence was found to be related to the size of the immediate reaction. Almost all the patients showing LCAR had positive provocation tests and were clinically sensitive to the corresponding allergen.

Studies on bacterial hypersensitivity in man. Interrelationship between skin reactions to bacterial peptidoglycan and serum peptidoglycan antibody titers.

The nature and extent of bacterially induced allergies are difficult to define. Since peptidoglycan, the main component of the cell wall of almost all bacteria, has been available in a highly purified, chemically and immunologically well-defined form, investigation of the allergological significance of this cell component is feasible. Intracutaneous tests were carried out on 181 test subjects with five different peptidoglycan (PG) preparations from Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus Pyogenes. The results of the investigation were compared with the result of determination of serum PG antibodies and serum IgE concentrations. It was shown that test subjects with dual and later reactions to three different staphylococcal PGs displayed significantly higher PG antibody titers than test subjects with negative reactions. Such a relationship could not be found with the cutaneous reactions to streptococcal PG. The total serum IgE values were very much higher in test subjects with immediate reactions to staphylococcal PG than in test subjects with a negative reaction. Typical Arthus reaction or late granulomatous reactions were not observed. Humoral antibodies are involved at least in part in the elicitation of dual and late reactions. Thus, there are interesting parallels to allergy to fungal spores and organic dusts.

The role of penicillin in the pathogenesis of chronic urticaria.

Intracutaneous tests with penicilloyl-polylysine (PPL) and benzyl-penicillin G (PG) were performed in 245 patients suffering form chronic recurrent urticaria, including physical urticaria. Positive results were observed in fifty-nine patients (24%). Sera from fifty-seven of these fifty-nine patients were investigated for circulating anti-penicilloyl antibodies by an enzyme-linked immunosorbent assay (ELISA) and a passive haemagglutination test (HA). The results were compared to those form the ELISA and HA in a control group of thirty-five patients who had shown clinical allergic reactions to penicillin and had positive skin tests to PPL and/or PG. The in vitro tests revealed positive results in 12.3% and 37.1% respectively. In forty-three patients the course of the positive intracutaneous tests to penicillin, together with the duration of chronic urticaria, was followed over a period of time up to 3.5 years. In twenty-two out of forty-two patients with positive intracutaneous tests to penicillin, a diet free of dairy products proved to have a curative effect, compared to two out of forty control subjects with chronic urticaria and negative skin tests to penicillin. These studies indicate that penicillin has an important role in the aetiology and maintenance of chronic urticaria.

Successful renal transplantation in a patient with anaphylactic reaction to Solu-Medrol (methylprednisolone sodium succinate)

Following the intravenous infusion of Solu-Medrol ® (methylprednisolone sodium succinate), anaphylactic shock developed in a 41 year old man on two consecutive occasions, six weeks apart. Intracutaneous testing with different components of the Solu-Medrol Mix-O-Vial ® demonstrated that the patient was allergic to methylprednisolone sodium succinate but not to other ingredients. In view of the recent increase in the popularity of steroid pulse therapy, we believe that all clinicians should be aware of this unusual and potentially fatal reaction to Solu-Medrol.

Widespread cutaneous candidiasis and tinea infection masking mycosis fungoides

A 71-year-old female with a widespread double mycotic infection caused by C. Albicans and T. rubrum was discovered to be suffering from mycosis fungoides. Clinically she was found to have large, polycyclic erythematous plaques with scaly, slightly infiltrated borders, covering almost all areas of the glabrous skin, and also involving the scalp (with no hair penetration), the soles and palms, toe-webs, finger and toe nails; there was also perleche and oral thrush. Cultures yielded C. albicans from most of the skin lesions, from the scalp, mouth, finger nails and urine and stool specimens, and T. rubrum from intermingled skin specimens, from the palms and soles and toe-nails. Blood culture was negative as were intracutaneous tests with fungal antigens and tuberculin. Histological examination confirmed the fungal invasion of the horny layer and at the same time revealed an underlying pathologic picture of mycosis fungoides, the lesions having been masked by the mycotic eruption. Intensive cytostatic and antifungal therapy led to a transient improvement but shortly thereafter there was a relapse of the fungal and lymphoproliferative manifestations and the patient died in septic shock.

Induction of chronic arterial hypertension in rats by repeated transient hypertensive rises in blood pressure. Possible pathogenetic role of delayed hypersensitivity against arteries and arterioles.

Repeated transient rises in blood pressure to hypertensive levels were induced by intravenous injections of angiotensin once or twice weekly. This procedure induced chronic arterial hypertension in five of fourteen rats (36%) within 4-6 weeks of starting the injections. Intracutaneous tests using homogenized common carotid arteries and histological examination of the kidneys gave support to the hypothesis that the transient rises in blood pressure to hypertensive levels resulted in a delayed hypersensitivity (DHS) reaction against components in small arteries and arterioles. This DHS reaction seemed to be responsible for an increased permeability of arterial vessels to plasma components causing exudative thickening of the walls of small arteries and arterioles and thereby narrowing of their lumina. When the lumina of small arteries and arterioles are narrowed, peripheral resistance to blood flow increases, and arterial hypertension results.

A critique of evaluation studies of sublingual and intracutaneous provocative tests for food allergy

Food allergy and means for testing for it continue to remain “controversial”. Clinical ecology, a specialty area emphasizing allergy to foods and environmental chemicals, has been resisted by conventional medicine, one reason being that its testing methods have been found in evaluation studies to be inadequate. Evaluation studies are examined and a number of problematical aspects of some of them are noted.

Mechanism of functional reorganization of paralyzed intercostal muscles in rats with experimental botulism

Altogether 183 C57BL/6 mice were used to determine the optimum dose of phytohemagglutinin (PHA) to be equal to 3 microgram/g liniment tela, which reduced the time of wound healing by 33%. Experiments on 255 heterobred rabbits of both sexes revealed that the natural animal population could be distributed into three groups according to delayed type hypersensitivity (DTH) to PHA: highly sensitive, moderately sensitive and poorly sensitive, in a ratio of 1:1, 5:1. Microscopic, cytological and histological analyses have demonstrated that PHA exerts a selective therapeutic effect on wound healing. The reduction of the time required for wound healing by 38% and positive changes in cellular dynamics of inflammation have been observed only in the animals with a weak DTH to PHA. The PHA intracutaneous test is recommended for selection of patients being in want of pharmacological correction of wound healing.

Perenniale allergische Rhinitis ? wie sicher ist die allergologische Diagnostik?

In an analysis of 322 patients with probable non-seasonal rhinitis, 90 were discovered to have clinically proven allergy to the house dust mite Dermatophagoides farinae. The total correspondence between the skin test and RAST amounts to 64.8%, between the skin test and intranasal provocation test 60.5%, and between RAST and INT 68%. When one considers the positive results from the case history, the skin test, the RAST and the INT on the 90 ascertained allergic cases, one can say the following concerning the efficiency of these four allergological testing methods as they apply to the house dust mite D. farinae. Approximately 30% of those patients with an indicative case history and ca. 60% of those with a positive skin test are allergic. In these cases the RAST is in 50% and the intranasal provocation test is positive in 74%. The third diagnostic method to be carried out after that of the case history and intracutaneous skin test, should be the intranasal provocation test. The RAST should follow a negative provocation test.

Immune phenomena in patients with pustular bacterid.

In 20 patients with histologically confirmed pustular bacterid of immunological status was checked including lymphocyte and granulocyte components of the cellular immune system as well as HLA typing. For comparison the same investigations were carried out in 20 healthy controls. In contrast to the controls, in the group of patients the chemotactic activity of polymorphonuclear leukocytes as well as their ability to engulf either vital or heat-inactivated Candida albicans cells was found to be slightly impaired (p less than or equal to 0.01 and 0.05, respectively). Killing of C. albicans cells by polymorphonuclear leukocytes were strikingly impaired (p less than or equal to 0.001). NADPH-dependent oxidase activity, however, was found to be normal. Impairment of chemotaxis and killing activity could not be substantiated in AB pool serum, thus a serum-dependent disorder of granulocyte function has to be assumed. Lymphocyte response to phytohemagglutinin was reduced in all patients, whereas intracutaneous test reactivity to microbial antigens was normal. T and B cell counts in peripheral blood were within normal range. HLA typing revealed a significantly increased prevalence rate of HLA-B14 as compared with the control group and other patients.

Skin testing in chronic urticaria.

In 164 patients with chronic recurrent urticaria, the value of skin tests was investigated, Patch tests carried out with the standard series of the International Contact Dermatitis Research Group and with a series of penicillins revealed positive reactions in 22 (35/162) and 6.9% (11/158), respectively. IN 21.5% (33/152) positive intracutaneous tests to cilligen and/or penicillin G were observed. 35% (51!147) of the patients had one or more positive intracutaneous tests with inhalants. Scratch tests with food allergens showed positive reactions in 12.5% (12/95). There was no difference in results between patients with idiopathic urticaria or angio-oedema and those with physical urticaria. The value of these findings is discussed. Most of the patients with penicillin allergy, were recommended to be on a diet free from milk and milk products. In 50% this was successful.

A method for eliciting indurated DTH reactions to soluble protein antigens in the flank skin of mice: correlation of visual measurements with 125I-UdR uptake indices.

A new technique for eliciting specific indurated delayed type hypersensitivity (DTH) reactions to soluble protein antigens in the flank skin of mice is reported here. We show that immunization with soluble antigens emulsified in complete Freund's adjuvant emulsion allows elicitation of indurated reactions upon intracutaneous test of sensitized mice with antigen on alum precipitates. With this method it is possible to measure simultaneously the gross induration of the cutaneous DTH reactions and correlate it with the degree of mononuclear cell infiltration as quantitated by the cPM of 125I-UdR uptake in mononuclear cells at biopsied reaction sites and confirmed by histological examination.