Characteristics of Study Subjects
A total of 52 patients were enrolled in the study, of which 34 (63.6%) were male and 18 (36.4%) were female, with an age distribution of 11 (7, 24.5) years.19 patients were diagnosed with allergic rhinitis alone, and 33 patients were diagnosed with allergic rhinitis combined with asthma. Most of the patients in the rhinitis alone and rhinitis combined with asthma groups were Han ethnicity, born by natural delivery, resided in urban areas, lived in buildings < 9 floors, used mattresses, seldom observed cockroaches at home, and did not use air conditioning (Table 1).
Pollen Crude Extract and Component sIgE Sensitization Patterns
In the patients included in this study, the sensitization rates to Art v and its component Art v1 were 92.3% and 59.6%, respectively; Phl p sensitization was 76.9%, with the highest sensitization rate to its component Phl p12 (63.5%), followed by Phl p1 (25.0%); Bet v positive rate was 78.9%, with sensitization rates to its components Bet v1 and Bet v2 being 26.9% and 63.5%, respectively; Amb a and its component Abm a 1 had sensitization rates of 73.1% and 17.3%, respectively, while Amb p sensitization rate was 59.6% (Figure 1A). Further analyses showed that 64.6% of Art v positive patients exhibited Art v 1 positivity (Figure 1B); 75.0% of Phl p positive patients exhibited Phl p 12 positivity, with lower positivity rates for the other components (7.5% to 32.5%) (Figure 1C); and 75.6% of Bet v positive patients exhibited Bet v 2 positivity (Figure 1D); 23.7% of Amb a positive patients were Amb a 1 positive, whereas a comparable percentage of Amb p positive patients were Amb a 1 positive (29.0%) (Figure 1E).
Potential Connections Between Pollens
Speaman`rho correlation analysis and hierarchical clustering analysis were performed on the allergen extracts and components detected. Correlation analysis showed Bet v 2 and Phl p 12 had the highest positive correlation (r = 0.98, P < 0.05). Further a strong positive correlation (0.64 ≤ r ≤ 0.80, P < 0.05) could be observed between the allergens Art v1, Art v, Amb p, Amb a, Phl p, Phl p 12, Bet v and Bet v 12 (Figure 2A). On the other hand, hierarchical clustering analysis categorized the 15 allergens into four sensitization categories (Figure 2B). The first category included Amb p, Amb a, and Art v 1; the second category included Bet v 2, Phl p, Phl p 12, and Bet v; Art v was categorized on its own as the third category; and the fourth category comprised Phl p 6, Phl p 2, Phl p 7, Amb a 1, Phl p 5, Phl p 1, and Bet v 1.
Concomitant Sensitization to Pollens
Based on the results of cluster analysis and correlation analysis, Upset was further used to analysis the co-sensitisation between the allergens Art v and its component Art v 1, Bet v and its component Bet v 2, Amb a, Amb p, Phl p and its component Plp h 12. The highest proportion of concomitant sensitization was observed among Art v and its component Art v 1, Bet v and its component Bet v2, Amb a, Amb p, Phl p, and its component Plp h 12 (25%) (Figure 3). Moreover, there was evident cross-overlap among these pollens and components, with the proportion of any two or more simultaneous positives reaching up to 92.3%.
In addition, we divided the patients into groups according to whether the above pollen crude extract allergens were sensitizing or not, and analysed the sensitisation rates of other crude extracts and components (Table 2). The results showed that in the Art v positive group, the sensitisation rates of Phl p 12, Bet v 2, Amb a, and Amb p were as high as 64.6% to 79.2%, whereas in the Art v negative group, none of these allergens were detected positive (positivity rate of 0%), with the difference being statistically significant (all P < 0.01). Similar results were observed for the Phl p allergen, with the sensitisation rates of Phl p 12, Bet v 2, Amb a, and Amb p being significantly higher in the Phl p positive group compared to the Phl p negative group (OR: 4.15~9.00). Furthermore, in the Bet v positive samples, the positivity rates for Phl p 12 and Bet v 2 were both 75.6%, compared to 18.2% in the negative group, showing a statistically significant difference (P < 0.01). In the results we also observed a higher sensitisation rate for both Phl p and its components Phl p 12, Bet v 12 and Amb p in Amb a positive patients as well (all OR >10, P < 0.01).
SIgE Inhibition Assays Results
Based on the results of the correlation and hierarchical cluster analyses described above, there is a potential cross-reactivity between Art v, Phl p and Bet v allergens. And to test this hypothesis, we further conducted sIgE inhibition assay using crude extracts of Art v, Phl p and Bet v as inhibitors (Figure 4 and Table 3). The sIgE inhibition assay results showed that the self-inhibition rates of Art v, Phl p and Bet v were 85.60%, 79.20% and 40.00%, respectively. Art v exhibited an inhibition rate greater than 73.2% against Phl p and its component Phl p 12, as well as Bet v and its component Bet v 2, while Phl p showed inhibition rates of 79.20% to 89.87% against its component Phl p 12 and Bet v and its component Bet v 2. Whereas, Bet v had inhibition rates of 11.9 to 59.8% against Phl p and Art v, with a notably higher inhibition rate of 76.80% against Phl p 12.