Department of Environmental Medical Biology, Institute of Tropical Medicine and Arthropods of Medical Importance Resource Bank, Yonsei University College of Medicine, Seoul, Korea.
Convergence Research Center for Insect Vectors, College of Life Science and Bioengineering, Incheon National University, Incheon, Korea.
PLoS One. 2021 Nov 23;16(11):e0257114. doi: 10.1371/journal.pone.0257114. eCollection 2021.
Allergens present in the feces or frass of cockroaches can cause allergic sensitization in humans. The use of fecal and frass extracts for immunotherapy has been previously investigated but has not yet been fully standardized. Here, we treated cockroaches with ampicillin to produce extracts with reduced amounts of total bacteria.
We performed targeted high-throughput sequencing of 16S rDNA to compare the microbiomes of ampicillin-treated and untreated (control) cockroaches. RNA-seq was performed to identify differentially expressed genes (DEGs) in ampicillin-treated cockroaches.
Analysis of the microbiome revealed that alpha diversity was lower in the ampicillin-treated group than in the control group. Beta diversity analysis indicated that ampicillin treatment altered bacterial composition in the microbiome of cockroaches. Quantitative polymerase chain reaction revealed that almost all bacteria were removed from ampicillin-treated cockroaches. RNA-seq analysis revealed 1,236 DEGs in ampicillin-treated cockroaches (compared to untreated cockroaches). Unlike bacterial composition, the DEGs varied between the two groups. Among major allergens, the expression of Bla g 2 decreased significantly in ampicillin-treated cockroaches (compared to untreated group).
In this study, the reduced level of allergens observed in cockroaches may be related to lower amounts of total bacteria caused by treatment with antibiotics. It is possible to make a protein extract with few bacteria for use in immunotherapy.
蟑螂粪便或粪便中的过敏原可引起人类过敏致敏。已经研究了粪便和粪便提取物用于免疫疗法,但尚未完全标准化。在这里,我们用氨苄青霉素处理蟑螂,以产生细菌总量减少的提取物。
我们对 16S rDNA 进行了靶向高通量测序,以比较氨苄青霉素处理和未处理(对照)蟑螂的微生物组。进行 RNA-seq 以鉴定氨苄青霉素处理蟑螂中的差异表达基因(DEG)。
微生物组分析表明,氨苄青霉素处理组的 alpha 多样性低于对照组。β多样性分析表明,氨苄青霉素处理改变了蟑螂微生物组中细菌的组成。定量聚合酶链反应显示,氨苄青霉素处理的蟑螂几乎去除了所有细菌。RNA-seq 分析显示氨苄青霉素处理的蟑螂中有 1236 个 DEG(与未处理的蟑螂相比)。与细菌组成不同,两组之间的 DEG 存在差异。在主要过敏原中,Bla g 2 的表达在氨苄青霉素处理的蟑螂中明显降低(与未处理组相比)。
在这项研究中,蟑螂中观察到的过敏原水平降低可能与抗生素处理引起的总细菌数量减少有关。有可能为免疫疗法制作一种细菌含量少的蛋白质提取物。