Changes Of Intestinal Flora In Human Immunodeficiency Virus Infected Patients And The Effect Of Combined Antiretroviral Therapy On Them

Objective To investigate the differences of intestinal flora between human immunodeficiency virus(HIV)infected and to study healthy individuals and the effect of combined antiretroviral therapy(cART)on the gut microbiota in HIV-infected patients.Methods 1.Fifty there HIV-infected patients who were hospitalized in Hangzhou Xixi Hospital from 2018 to 2019,and 6 healthy volunteers were enroled.They were divided into healthy control group(Control group),untreated HIV infected group(Untreated group),HIV-infected group with positive blood viral load after cART(cART1 group)and HIV-infected group with negative blood viral load after cART(cART2 group).Three groups,including Control group and Untreated group,Untreated group and cART1 group,cART1 group and cART2 group were compared and analyzed.Multiple laboratory indicators and clinical data of the study subjects were also collected.2.The total DNA of the collected stool specimens was extracted.After the experimental steps of detection,PCR amplification,product purification,library preparation and quality inspection,Illumina MiSeq platform was finally used to sequence DNA fragments.3.The sequencing raw data were processed,and the processed data were analyzed by bioinformatics and statistics.Results 1.Distribution of effective data length:<200bp was 18887,accounting for 1.31%.200-300bp was 12690,accounting for 0.88%.300-400bp was 157891,accounting for 10.98%.400-500bp was 1247897,accounting for 86.82%.≥500bp was 2,accounting for 0.01%.2.Analysising of operational taxonomy unit,Control group and Untreated group have 662 common OTUs,the former has 162 OTUs uniquely,the latter has 989 OTUs uniquely.Untreated group and cART1 group have 1215 common OTUs,the former has 436 OTUs uniquely,and the latter 245 OTUs uniquely.cART1 group and cART2 group have 1099 common OTUs,the former has 361 OTUs uniquely,and the latter 327 OTUs uniquely.3.For alpha diversity analysis,it is showing that Control group>Untreated group>cART2 group>cART1 group,there were no significant differences between the comparison groups.4.In the beta diversity analysis,Control group was more differentiated from Untreated group,and there were more obvious differences between Control group and Untreated group,but Untreated group also had points closer to Control group.There was no significant difference between Untreated and cART1 groups and cART1 and cART2 groups.5.In species analysis,there are four phyla,which are Firmicutes,Proteobacteria,Bacteroidetes and Actinobacteria,and their contents are above 95%.For Control group,Bacteroidetes(52.30%)>Firmicutes(42.39%)>Proteobacteria(3.8 9%)>Actinobacteria(0.71%),Untreated group,Firmicutes(54.89%)>Bacteroidetes(23.06%)>Proteobacteria(11.89%)>Actinobacteria(8.53%).For cART1 and cART2 groups,Proteobacteria(40.01%and 39.58%)>Firmicutes(37.21%and 31.19%)>Bacteroidetes(16.45%and 24.94%)>Actinobacter-ia(5.35%and 3.49%).For Control group,Bacteroides(37.15%),Prevotella(10.61%)and Megamonas(10.56%)contains more than 5%of the genera.Untreated group contains more than 5%Escherichia(10.39%),Bacteroides(6.04%),Prevotella(9.72%),Enterobacteriaceae unclassified(13.78%)and Parabacteroides(6.43%).For cART1 group more than 5%genera are Escherichia(26.68%),Bacteroides(6.30%),Prevotella(9.04%),Enterobacteriaceae unclassified(11.84%)and Klebsiella(9.13%).More than 5%of cART2 group were Escherichia(35.47%),Bacteroides(9.83%)and Prevotella(10.72%).LEfSe analysis found that Control group had more bacteria in Bacilli,Actinomycetales,and Pseudomonadales than Untreated group,and Control group had more Bacteroidetes,Verrucomicrobia,Betaproteobacteria,Negativicutes,Clostridium,Lachnospiraceae,Veillonellaceae,Bilophila and Eggerthella.Compared with cARTl group,Untreated group had more bacteria in Firmicutes unclassified,Sphingobacteriia,Actinomycetales,Stomatobaculum,Streptococcus and Oscillibacter,and cART1 group had more bacteria in Proteobacteria.Compared with cART2 group,cART1 group contained more bacteria in Ruminococcaceae unclassified and Staphylococcaceae,and cART2 group contained more bacteria in Candidatus Saccharibacteria.Conclusion 1.HIV infection leads to a reduction in the diversity of intestinal flora species,and there is a significant difference compared with the intestinal flora of healthy people,and the species composition changes greatly.2.For HIV-infected patients with positive blood viral load after cART,the intestinal flora did not develop towards a healthy direction,but further tended to be disordered.cART,to some extent,had a negative impact on the intestinal flora and further reduced its diversity.3.The species diversity of intestinal flora in HIV-infected patients with negative blood viral load after cART increased slightly,but they were still in an unhealthy state,which was similar to HIV-infected patients with positive blood viral load after cART,and the effect of different treatment outcomes on intestinal flora was not obvious.