Construction by artificial intelligence and immunovalidation of hypoallergenic mite allergen Der f 36 vaccine.

Background: The house dust mite (HDM) is widely recognized as the most prevalent allergen in allergic diseases. Allergen-specific immunotherapy (AIT) has been successfully implemented in clinical treatment for HDM. Hypoallergenic B-cell epitope-based vaccine designed by artificial intelligence (AI) represents a significant progression of recombinant hypoallergenic allergen derivatives. Method: The three-dimensional protein structure of Der f 36 was constructed using Alphafold2. AI-based tools were employed to predict B-cell epitopes, which were subsequently verified through IgE-reaction testing. Hypoallergenic Der f 36 was then synthesized, expressed, and purified. The reduced allergenicity was assessed by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and basophil activation test. T-cell response to hypoallergenic Der f 36 and Der f 36 was evaluated based on cytokine expression in the peripheral blood mononuclear cells (PBMCs) of patients. The immunogenicity was evaluated and compared through rabbit immunization with hypoallergenic Der f 36 and Der f 36, respectively. The inhibitory effect of the blocking IgG antibody on the specific IgE-binding activity and basophil activation of Der f 36 allergen was also examined. Results: The final selected non-allergic B-cell epitopes were 25-48, 57-67, 107-112, 142-151, and 176-184. Hypoallergenic Der f 36 showed significant reduction in IgE-binding activity. The competitive inhibition of IgE-binding to Der f 36 was investigated using the hypoallergenic Der f 36, and only 20% inhibition could be achieved, which is greatly reduced when compared with inhibition by Der f 36 (98%). The hypoallergenic Der f 36 exhibited a low basophil-stimulating ratio similar to that of the negative control, and it could induce an increasing level of IFN-γ but not Th2 cytokines IL-5 and IL-13 in PBMCs. The vaccine-specific rabbit blocking IgG antibodies could inhibit the patients' IgE binding and basophil stimulation activity of Derf 36. Conclusion: This study represents the first application of an AI strategy to facilitate the development of a B-cell epitope-based hypoallergenic Der f 36 vaccine, which may become a promising immunotherapy for HDM-allergic patients due to its reduced allergenicity and its high immunogenicity in inducing blocking of IgG.

Sputum metabolomic profiling of bronchial asthma based on quadruple time-of-flight mass spectrometry.

To improve diagnosis of asthma, we tend to confirm potential biomarkers by comparing sputum metabolome profiles between asthma patients and healthy controls, using ultra-high-performance liquid chromatography coupled to quadruple time-of-flight mass spectrometry (UHPLC-QTOF/MS). Thirty endogenous metabolites contributing to the separation of asthma patients and healthy controls were tentatively identified in positive mode, such as 1-hexadecanoyl-sn-glycerol, glycerol 1-stearate, sphingosine, Phe-Ser, Tyr-Ala and Phe-Gln, and 12 endogenous metabolites were identified in negative mode, such as cytidine 2',3'-cyclic phosphate, 1-hexadecanoyl-2-(9Z-octadecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol), 1-octadecanoyl-2-(9Z-octadecenoyl)-sn-glycero-3-phosphoserine, thymidine, gamma-L-glutamyl-L-valine and adenine. Those differential metabolites were mainly participatedin glycerophospholipid metabolism, retrograde endocannabinoid signaling and metabolic pathways in positive mode and 2-oxocarboxylic acid metabolism, biosynthesis of amino acids, phenylalanine, tyrosine and tryptophan biosynthesis, valine, leucine and isoleucine degradation and metabolic pathways in negative mode. Importantly, several metabolic pathways including glycerophospholipid metabolism, inositol phosphate metabolism, and glycolysis or gluconeogenesis were found most important. These findings suggest sputum metabolomics can be used for the early diagnosis and risk prediction of asthma.

ADRB2 polymorphisms in allergic asthma in Han Chinese children.

BACKGROUND: Single-nucleotide polymorphisms (SNPs) in the human ß2 adrenergic receptor (ADRB2) gene have been linked to pathogenesis of and treatment responses in asthma. In China, where asthma is common, little information exists regarding ADRB2 variants and their effects on asthma. The current study sought to investigate the relationship between variations in ADRB2 and childhood asthma. METHODS: Blood samples were collected from 298 pediatric asthma cases treated in our hospital from July 2013 to September 2014; 304 samples were collected from healthy children for a control group. Six loci (at base positions -47, 79, -2387, 46, 523, and 491) of ADRB2 were genotyped using the TaqMan probe assay; SHEsis was used to determine haplotypes. Allele frequencies and genotype distributions were compared between groups using chi square and t tests. RESULTS: No significant differences in genotype or allele frequencies were found between the groups for the polymorphic loci -2387 bp (T>C), 46 bp (G>A), and 523 bp (C>A) (p > 0.05). However, genotype frequencies for the ADRB2 SNPs at -47 bp (C>T) and 79 bp (G>C) differed significantly between asthma and control groups (p < 0.05). Further, linkage disequilibrium was found between these 2 SNPs (D' = 0.990, r(2) = 0.962), as well as between SNPs at 46 and 523 bp (D' = 0.985, r(2) = 0.607). Finally, of the 4 haplotypes analyzed in these samples (haplotypes III, IV, IX, and XI), the frequency of haplotype III was significantly lower in the asthma group than the control group (p < 0.05; odds ratio [OR] = 0.72; 95% confidence interval [CI], 0.48 to 0.97). CONCLUSION: Variations of ADRB2 at base positions -47 (C>T) and 79 (G>C), as well as haplotype III, may contribute to susceptibility to childhood asthma.