Association between the Interaction of Key Genes Involved in Effector T-Cell Pathways and Susceptibility to Developallergic Rhinitis: A Population-Based Case-Control Association Study

<div><p>Background</p><p>Evidence suggests that interaction between key genes mediating signaling and transcriptional networks involving effector T-cell responses may influence an individual’s susceptibility to develop allergic rhinitis(AR).</p><p>Objective</p><p>The aim of this study was todetermine whether specific interactions between key genes involved in effector T-cell pathways are associated with an individual’s susceptibility to develop AR in Han Chinese subjects.</p><p>Method</p><p>A cohort of 489 patients with AR and 421 healthy controls was enrolled from the Han Chinese population in Beijing, China. AR was established by questionnaire and clinical examination, and peripheral blood was drawn from all subjects for DNA extraction. A total of 96 single nucleotide polymorphisms (SNPs) in 26 reprehensive candidate genes involved in T helper 1 (Th1), Th2, Th17, Th9 and T regulatory cell pathways were selected from the International Haplotype Mappingdatabase for Han Chinese in Beijing (CHB) population, and IlluminaGoldenGate assay was conducted for SNP genotyping. The PLINK software package was used to perform statistical analyses.</p><p>Results</p><p>Simple SNP-phenotype association analysis using logistic regression showed SNP rs8193036 in IL17A gene, rs2569254 in IL-12 and rs1898413 in RORα weresignificantlyassociatedwith AR.Simple SNP-phenotype association analysis with genetic models demonstrated thatrs2569254 in IL-12, rs1031508 in STAT4, and rs3741809 in IL-26 were likely to be recessive, rs8193036 in IL17A allelic, rs897200in STAT4 genotypic, and rs1898413 in RORα dominant. Epistasis analyses exhibited that 83 SNPs in 23 genes were significantly interactive; of which 59 interactions/SNP pairs demonstrated OR values higher than 2 or lower than 0.5, and 12 interactions/SNP pairs OR values higher than 4 or lower than 0.25. STAT3, RORα and IL-26, involved in Th17 pathway,were the mostfrequentlyinteractive genes.</p><p>Conclusion</p><p>This study suggests that interactions between several SNPs in key genes involved in effector T-cell pathways are likely to influence an individual’s susceptibility to develop AR.</p></div

Simple SNP-phenotype association results with full model.

<p>Note: SNP, single nucleotide polymorphisms; Chr, chromosome; ALLELIC, allelic model; GENO, genotypic model; DOM, dominant model; REC, recessive model; TREND, trend model.</p><p>Simple SNP-phenotype association results with full model.</p

Additional file 1 of Replication study of susceptibility variants associated with allergic rhinitis and allergy in Han Chinese

Additional file 1. Additional tables and figures

Amplification-Free, Single-Microbead-Based Cas12a Assay for One-Step DNA Detection at the Single-Molecule Level

CRISPR/Cas-based systems are highly attractive for developing next-generation diagnostic technologies because of their intrinsic merits such as simplicity, sensitivity, and specificity. However, currently, nucleic acid amplification procedures are still needed to achieve attomolar sensitivity in most CRISPR/Cas-based assays, which causes high cost, operation difficulty, and low efficiency. Herein, we combine the CRISPR/Cas12a-based assay and a single-microbead detection platform for one-step and amplification-free detection of DNA at the single-molecule level. By modifying DNA reporters on a biomimetic membrane-coated microbead, the activated Cas12a by targets will cleave these reporters and lighten the bead within 10 min. The method allows the detection of the target down to three copies in a 5 μL sample. Furthermore, we successfully apply this method for the specific identification of viral infection, foodborne bacteria, and DNA mutation in real samples without extra nucleic acid amplification. We believe that this approach offers new insights for developing CRISPR/Cas-based DNA assays in biomedical applications

RA inhibits primitive myelopoiesis by acting downstream of <i>gata4/5/6</i>.

<p>All embryos are positioned anterior left and lateral front. Embryos were microinjected with <i>gata5</i>-MO and <i>gata6</i>-MO together (B, C, G, H) at 1–2-cell stage and then treated with 10 µM DEAB (C, H) or vehicle DMSO (B, G) immediately, or microinjected with <i>gata4</i> mRNA and <i>gata6</i> mRNA together (D, E, I, J) and then treated with 250 nM RA (E, J) or vehicle DMSO (D, I) from 10 to 11 hpf, respectively. They were then examined for expressions of myeloid markers <i>lcp1</i> (A–E) and <i>mpx</i> (F–J) at 24 hpf by whole mount <i>in situ</i> hybridization. The number shown in the lower left-hand corner of each panel is the number of embryos exhibiting the typical phenotype shown in the panel to the number of embryos totally observed.</p

Retinoic Acid Signaling Plays a Restrictive Role in Zebrafish Primitive Myelopoiesis

<div><p>Retinoic acid (RA) is known to regulate definitive myelopoiesis but its role in vertebrate primitive myelopoiesis remains unclear. Here we report that zebrafish primitive myelopoiesis is restricted by RA in a dose dependent manner mainly before 11 hpf (hours post fertilization) when anterior hemangioblasts are initiated to form. RA treatment significantly reduces expressions of anterior hemangioblast markers <em>scl</em>, <em>lmo2</em>, <em>gata2</em> and <em>etsrp</em> in the rostral end of ALPM (anterior lateral plate mesoderm) of the embryos. The result indicates that RA restricts primitive myelopoiesis by suppressing formation of anterior hemangioblasts. Analyses of ALPM formation suggest that the defective primitive myelopoiesis resulting from RA treatment before late gastrulation may be secondary to global loss of cells for ALPM fate whereas the developmental defect resulting from RA treatment during 10–11 hpf should be due to ALPM patterning shift. Overexpressions of <em>scl</em> and <em>lmo2</em> partially rescue the block of primitive myelopoiesis in the embryos treated with 250 nM RA during 10–11 hpf, suggesting RA acts upstream of <em>scl</em> to control primitive myelopoiesis. However, the RA treatment blocks the increased primitive myelopoiesis caused by overexpressing <em>gata4/6</em> whereas the abolished primitive myelopoiesis in <em>gata4/5/6</em> depleted embryos is well rescued by 4-diethylamino-benzaldehyde, a retinal dehydrogenase inhibitor, or partially rescued by knocking down <em>aldh1a2</em>, the major retinal dehydrogenase gene that is responsible for RA synthesis during early development. Consistently, overexpressing <em>gata4/6</em> inhibits <em>aldh1a2</em> expression whereas depleting <em>gata4/5/6</em> increases <em>aldh1a2</em> expression. The results reveal that RA signaling acts downstream of <em>gata4/5/6</em> to control primitive myelopoiesis. But, 4-diethylamino-benzaldehyde fails to rescue the defective primitive myelopoiesis in either <em>cloche</em> embryos or <em>lycat</em> morphants. Taken together, our results demonstrate that RA signaling restricts zebrafish primitive myelopoiesis through acting downstream of <em>gata4/5/6</em>, upstream of, or parallel to, <em>cloche</em>, and upstream of <em>scl</em>.</p> </div

RA restricts the primitive myelopoiesis mainly before 11 hpf.

<p>All embryos are positioned anterior left and lateral front. Embryos were treated with vehicle DMSO (A, H, O) or with 50 nM RA (B–G, I–N) from 3 to 5 (B, I), 5 to 7 (C, J), 7 to 9 (D, K), 9 to 11 (E, L), 11 to 13 (F, M) and 13 to 26 hpf (G, N), or with 250 nM RA form 10 to 11 (P), 11 to 13 (Q), 13 to 22 hpf (R), respectively. They were then examined for expressions of myeloid markers <i>lcp1</i> (A–G, O–R) and <i>mpx</i> (H–N) at 26 hpf (A–N) or 22 hpf (O–R) by whole mount <i>in situ</i> hybridization. The number shown in the lower left-hand corner of each panel is the number of embryos exhibiting the typical phenotype shown in the panel to the number of embryos totally observed. The typical embryos expressing <i>lcp1⁺</i> cells at 22 hpf were shown in O–R. The scatter plot (S) shows the number of <i>lcp1⁺</i> cells counted from each of the embryos at 22 hpf with different treatment (control; 10–11 hpf RA treatment; 11–13 hpf RA treatment and 13–22 hpf RA treatment).</p

Candidate SNPsin the key genes involved in effector T-cell pathways.

<p>SNP, single nucleotide polymorphis</p><p>Candidate SNPsin the key genes involved in effector T-cell pathways.</p

Demographic and clinical characteristics of the study groups.

<p>Demographic and clinical characteristics of the study groups.</p

Simple SNP-phenotype association results with logistic regression.

<p>SNP, single nucleotide polymorphism; OR, odds ratio; STAT, Chi-square statistic</p><p>Simple SNP-phenotype association results with logistic regression.</p