| Objective:Compare the differences of intestinal fungal microecology and fungal secreted proteins between AIDS patients and healthy people,and analyze the correlation between intestinal fungal microecology,secreted protein and immune status.To explore the changes of intestinal fungal microecology,the expression of secreted proteins and the possible role of intestinal microecology changes in the damage of intestinal barrier in patients with AIDS.Methods:Two groups of people were included,namely AIDS patients(group A)and healthy people(group B).The fecal samples of two groups of people were collected,and the fungal composition of the samples was analyzed using ITS2 high-throughput sequencing technology;RT-q PCR was used to detect the expression of intestinal fungal secretory protein gene in two groups of people:phospholipase B1(PLB1),hyphal wall protein 1(HWP1),agglutinin-like sequence 3(ALS3),secreted aspartic proteases(SAPs).At the same time,the clinical data of all subjects were collected,and the CD4~+T cell count and CD4/CD8 ratio of the two groups were recorded.The statistical software SPSS 26.0 was used to analyze the basic information of the two groups,the differences between the two groups’indicators(Mann-Whitney test,independent sample t-test,t,-test)and the correlation between the indicators(Spearman correlation analysis method);Use R language(4.0.5)and QIIME2software(2020.2)to analyze community biological information;The difference was considered statistically significant(P<0.05).Results:1.Analysis of intestinal flora composition:at the phylum level,the highest relative abundance of the two groups is Ascomycete,followed by Basidiomycetes.The ratio of ascomycetes/basidiomycetes in group A(4.33)was significantly higher than that in group B(1.54).At the level of class,order,family and genus,the species with higher relative abundance in the two groups are similar,but there are also differences,and the relative abundance of the same species is different.There was no statistical difference in the species diversity(Alpha diversity)of intestinal fungi between the two groups.There were statistical differences in the composition of intestinal fungal community(Beta diversity)between the two groups;Myrmecridium under Ascomycetes and Homophron under Basidiomycetes are enriched in Group A;Ophiosphaerella and Nigrospora under ascomycetes and Flaviporus under basidiomycetes are enriched in Group B.2.The relative abundance of intestinal Candida albicans in group A was higher than that in group B,but there was no significant difference between the two groups(Z=-1.843,P=0.065);The expression of Candida albicans pathogenic factor ALS3 in group A was higher than that in group B,and the difference between the two groups was statistically significant(t=2.304,P=0.028).3.Correlation analysis:1)No obvious correlation between fungi and ALS3 was found;2)No significant correlation was found between ALS3 and CD4~+T cells and CD4/CD8;3)Flaviporus was significantly correlated with CD4~+T cell count(r=0.555,P=0.001)and CD4/CD8(r=0.507,P=0.002),and was positively correlated;Ophiosphaerella is significantly correlated with the number of CD4+T cells(r=0.450,P=0.007).Conclusion:1.The microecology of intestinal fungi in patients with AIDS has changed significantly compared with that in the healthy group.2.The ability of intestinal Candida albicans to produce ALS3 in patients with AIDS is higher than that in healthy group.3.There is a positive correlation between the fungi enriched in the healthy group and the immune level of the host.Restoring the normal intestinal microecology is conducive to maintaining the integrity of the host intestinal barrier. |
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