John I. Wypych

Studies of coexisting honeybee and vespid-venom sensitivity

Honeybee and vespid venom-specific IgE were measured by RAST in randomly selected sera of 87 patients who had had anaphylactic reactions after insect stings. Overall there was a poor correlation between the titers of honeybee venom and yellow jacket or hornet venom-specific IgE. Sera from nine patients with high titers of both honeybee venom and yellow jacket venom-specific IgE were selected for RAST-inhibition studies, with these two venoms as coupling and inhibiting antigens. Three patterns of IgE-antibody specificity were detected. Four patients had unique antibody activity with no cross-reactivity between the yellow jacket and honeybee venom-specific IgE. This is probably the most common pattern in patients with dual sensitivity. Three patients reacted to a major allergen in yellow jacket venom cross-reacting with a minor allergen in honeybee venom. Their RAST-inhibition patterns demonstrated that the yellow jacket-venom RAST was inhibited by yellow jacket venom only and the honeybee-venom RAST was inhibited by both yellow jacket venom and honeybee venom. Two patients had the opposite pattern with honeybee-venom RAST inhibited by honeybee venom only and the yellow jacket RAST inhibited by both honeybee venom and yellow jacket venom. These latter patients reacted to a major allergen in honeybee venom that was cross-reacting with a minor allergen in yellow jacket venom. Studies with rabbit antisera raised to vespid and honeybee venoms demonstrated major antigens that were unique to each family that did not cross-react and several minor cross-reacting antigens.

Comparison of the allergenicity and antigenicity of Polistes venom and other vespid venoms

The crossantigenicity of Polistes venom with other vespid venoms was examined with rabbit and human antisera. Venom preparations from various Polistes species were obtained by electrical stimulation of individual insects and venom sac dissection. Rabbit antibodies were raised to the venom (P. apachus) and venom sac extract (P. exclamans). Human antisera were obtained from patients allergic to Polistes and other vespid venoms. The venom appeared to be more potent than the venom sac preparations in reactions with rabbit IgG and human IgE antibodies. Among the Polistes species, P. exclamans, P. instablis, and P. apachus venoms showed several lines of precipitation with rabbit antisera, and P. annularis and P. fuscatus venoms only one line, suggesting quantitative or qualitative antigenic differences. In RAST analysis, most sera reacted equally to all Polistes species but occasional exceptions were noted, again suggesting differences in venom allergens. P. exclamans-coupled discs gave the most consistent results. In gel diffusion experiments, there was no crossreactivity between Polistes and yellow jacket venoms and only limited crossreactivity between Polistes and hornet venoms. Patients sensitive to Polistes venom showed varying degrees of reactivity to yellow jacket and hornet venoms in RAST analysis. Patients sensitive to other vespid venoms also showed varying degrees os sensitivity to Polistes venom. Polistes venom appears to contain a genus-unique antigen (allergen). In addition, there appear to be some crossreacting antigens in Polistes and other vespid venoms but to a much lesser degree than found previously in the analysis of the relationship of yellow jacket and hornet venoms.

Clinical and immunological studies of beekeepers

Thirty‐four beekeepers were interviewed and their blood assayed for the presence of antibodies reacting with bee venom, bee venom phospholipase A (PLA), and whole bee body extract. Following a bee sting, most beekeepers experienced only minimal local tissue reaction. Their serum contained high levels of total antibodies (primarily IgG) reacting to bee venom and phospholipase A. These antibody titres correlated with the frequency of bee stings.

Local intranasal immunotherapy for grass-allergic rhinitis

In a double-blind controlled study, local intranasal immunotherapy was evaluated for the treatment of grass pollenosis. On the basis of serum grass-specific IgE levels, 50 grass-allergic patients were randomly divided into three groups and treated with either an aqueous solution of mixed-grass extract, a formaldehyde-modified mixed-grass extract (allergoid), or a histamine solution (placebo). Intranasal solutions were administered in gradually increasing doses over a preseasonal 10 wk period, adverse local reactions from the aqueous grass extract were frequent during treatment. Few adverse reactions occurred from the allergoid or histamine solutions. During the pollen season, patients receiving both grass extracts had much lower symptom/medication scores than patients receiving placebo. The severity of eye symptoms was the same in all groups. After treatment, serum grass-specific IgE rose in patients receiving aqueous and allergoid extract; no change was noted in patients receiving placebo therapy. Grass nasal secretory-specific IgA titers in each group did not change during the study. The results of this study suggest that local intranasal immunotherapy with either aqueous or allergoid grass extracts is clinically effective for the treatment of grass pollenosis. Adverse reactions associated with the aqueous extract may limit its usefulness. No correlation was present between the secretory immune response and clinical benefit.

Comparison of the allergenicity and antigenicity of yellow jacket and hornet venoms

The immunologic properties of yellow jacket and hornet venoms were compared by measuring their reaction with rabbit antisera and human IgE and IgG antibodies. Anti-hornet venom rabbit serum showed precipitin bands unique to hornet venom and several bands crossreacting with yellow jacket venom. Anti-yellow jacket venom rabbit serum reacted with yellow jacket venom but failed to react with the hornet venoms. Most sera from patients who had had allergic reactions after vespid stings reacted with yellow jacket and hornet venoms in RAST analysis. A few sera reacted with only one of the venoms. RAST inhibition studies confirmed the crossreactivity of these IgE antibodies. The IgG antibody response of 14 patients was measured after yellow jacket venom immunotherapy. All had rising titers of yellow jacket venom-specific IgG. There was also an increase in the IgG antibody response measured with hornet venom in the majority of patients. The rise was significant with yellow hornet venom (p less than 0.02) but failed to reach significance with bald-faced hornet venom (p greater than 0.05). In IgG radioimmunoassay inhibition studies using yellow jacket venom-coupled discs, yellow jacket venom was considerably more potent than hornet venom. These studies indicate major crossreactivity between yellow jacket and hornet venoms. In this group of patients, yellow jacket venom appeared to be the primary allergen.

Local intranasal immunotherapy for ragweed allergic rhinitis II. Immunologic response

Local nasal immunotherapy (LNIT) was administered in a double-blind study to 67 subjects. Twenty-three received an unmodified ragweed extract (RW), 24 received a glutaraldehyde polymer of ragweed extract (PRW), and 21 received placebo. Serum ragweed-specific IgE (S-IgE), ragweed-specific nasal secretory (NS-) IgE, secretory IgA (SIgA) and IgG, and NS-albumin were measured. RW therapy caused a significant increase in ragweed-specific S-IgE (p less than 0.005) and NS-SIgA (p less than 0.05). PRW therapy caused a significant rise in ragweed-specific NS-SIgA (p less than 0.001). NS-IgE (p less than 0.05), and NS-IgG (p less than 0.01). Ragweed-specific S-IgG was not affected by any of the treatments. There was no consistent correlation between NS-antibody levels and symptom/medication scores.

Local Intranasal Immunotherapy with High-Dose Polymerized Ragweed Extract

Thirty-one ragweed-allergic patients received preseasonal local intranasal immunotherapy (LNIT) with high doses of gluteraldehyde-polymerized ragweed extract (average total dose 544 micrograms antigen E). Minimal side effects were reported during treatment and did not interfere with the dosing schedule. During the ragweed pollen season, LNIT-treated patients had lower symptom scores for sneezing, rhinorrhea and nasal congestion than a comparable group of untreated ragweed-allergic patients. There was no difference in ragweed-induced eye symptoms between the two groups. Secretory ragweed-specific IgA and IgG rose following LNIT treatment. Absolute antibody titers and changes in titers did not correlate with clinical improvement. LNIT with the polymerized ragweed did not block the seasonal rise in serum ragweed-specific IgE. These results suggest that LNIT with high-dose polymerized ragweed extract is a safe, simple and effective form of immunotherapy.

Stinging insect allergy: Detection and clinical significance of venom IgE antibodies

Venom-specific IgE antibodies in 109 sera from patients who had had immediate systemic allergic reactions following insect stings were measured by the radioallergosorbent (RAST) procedure. The majority of sera contained IgE antibodies to either bee, yellow jacket, or hornet venoms. Some sera had positive RAST reactions with 2 or 3 venoms, but others contained single venom-specific IgE antibodies. Of 24 patients who had large local reactions, the sera of 12 contained venom IgE antibodies. The RAST procedure provides an accurate means of documenting IgE-mediated allergic sensitivity to stinging insects.

The inhibitory effect of serum factors on measurement of IgE aspergillus antibodies by RAST

The RAST value obtained in the A. fumigalus system, from sera of patients with allergic bronchopulmonary aspergillosis (ABPA) is dependent not only on the reagin titre but also on blocking factors that depress the RAST score. These factors are primarily specific for A. fumigatus and can be removed by an A. fumigatus immuno‐adsorbent. However, some blocking activity could be found in normal sera even after absorption with an A, fumigates immunoadsorbent and seems to be non‐specific. All of these blocking factors influence the specificity of the RAST, and complicate the comparison of the quantitation of IgE antibodies in different sera. The use of discs rather than cellulose particles and the use of dilute sera result in a RAST assay more proportional to the specific IgE antibodies to A. fumigates.

Local nasal immunotherapy: Efficacy of lowdose aqueous ragweed extract

In previous studies preseasonal local nasal immunotherapy (LNIT) with moderate doses of aqueous ragweed extract (mean total dose 59 micrograms of AgE and 139 micrograms of AgE) was an effective treatment for ragweed hay fever; however, local adverse reactions during therapy were common. This study evaluated the clinical and immunologic responses to LNIT by use of lower doses of aqueous ragweed extract in order to minimize these adverse reactions. Patients were administered preseasonal LNIT for 7 wk and received a mean total dose of 4.7 micrograms of AgE. During the ragweed season, symptom/medication scores (SMS) of the treated patients were equivalent to SMS of untreated patients. Serum ragweed-specific IgE and nasal secretory ragweed-specific IgA rose slightly in the treated patients but not to the extent observed in previous studies. After the ragweed season treated and untreated patients had a substantial increase in serum ragweed IgE antibody titers. No correlation could be found between antibody responses and SMS. This study indicates that LNIT with lower doses of aqueous ragweed extract is clinically ineffective.