Susceptibility Gene Polymorphisms And Immune Response Mechanisms Of Lymphocyte Subsets In Tuberculosis

Tuberculosis（TB）is a chronic respiratory disease caused by a single pathogen.The World Health Organization（WHO）estimates that the number of deaths among HIV-negative TB patients has increased from 1.2million in 2019 to 1.3 million in 2020,and China remains a country with a high burden of TB.With the development of science,it has been recognized that TB is not only an infectious disease induced by Mycobacterium.tuberculosis（M.tb）,but also a dynamic immune balance between human immune cells and M.tb.However,not everyone infected with M.tb is effectively cleared,the vast majority in the form of latent infection,and only about 10 percent of infected people develop disease in their lifetime.In recent years,scientists have found that there are many susceptible genes to TB in the human body,and most of the susceptibility genes are related to the immune system and inflammatory response of the human body.It is precisely because of the mutation of these genes that the immune efficacy of the body to M.tb is impaired.This topic through the TB susceptibility of candidate genes in patients with pulmonary tuberculosis and to analyze the expression in normal peoples,can make a clear definition of the genes and gene of TB,the interaction between genes and environment,will not only help to elucidate genetic differences in phenotype and immunology of TB,also provides the accurate diagnosis and treatment of TB is proposed.Flow cytometry was used to analyze the number of CD3⁺T cells,CD4⁺T cells,CD8⁺T cells,NK cells,NKT cells and B cell subsets in order to understand the immune response mechanism of tuberculosis patients,and provide some ideas for reasonable immune intervention of tuberculosis patients,so as to reduce the recurrence rate and drug resistance rate of tuberculosis.（1）The relationship between susceptibility gene polymorphism and pulmonary tuberculosis.This study included 113 in Active tuberculosis（ATB）patients hospitalized in the Department of Tuberculosis Medicine of the 8^th Medical Center of Chinese PLA General Hospital and 60 medical staff as normal controls（NC）.A susceptibility study of 146 Single nucleotide polymorphisms（SNPs）in genes associated with susceptibility to TB to analyze alleles,genotypes,genetic models,linkage disequilibria and gene-to-gene interactions of the SNP locus of tuberculosis susceptibility gene in Beijing area.The results showed that the C allele frequency of rs9061（SP110）in ATB patients was significantly higher than that in normal controls（χ²=6.663,OR=0.489,P=0.0098）,CC genotype frequency was significantly higher than that in normal controls（χ²=9.876,P=0.007）,dominant model（TT+CT vs CC,χ²=9.203,OR=0.361,P=0.0024）were significantly different from normal controls.The G allele frequency of rs11556887（SP110）in ATB patients was significantly higher than that in normal controls（χ²=15.041,OR=0.203,P=0.0001）,and the GG genotype frequency was significantly higher than that in normal controls（χ²=13.792,P=0.001）.The dominant model（AA+AG vs GG,χ²=12.591,OR=0.205,P=0.0004）was significantly different from normal controls.The C allele frequency of rs4327230（SP110）in ATB patients was significantly higher than that in normal controls（χ²=9.243,OR=2.366,P=0.0024）,CC genotype frequency was significantly higher than that in normal controls（χ²=9.573,P=0.008）,dominant model（GG+CG vs CC,χ²=9.373,OR=0.359,P=0.0022）were significantly different from normal controls.The A allele frequency of rs7041（VDBP）in ATB patients was significantly higher than that in normal controls（χ²=10.641,OR=2.208,P=0.0011）,and the AA genotype frequency was significantly higher than that in normal controls（χ²=17.582,P=0.0002）.The dominant model（CC+CA vs AA,χ²=16.571,OR=0.244,P<0.0001）were significantly different from normal controls.The T allele frequency of rs4588（VDBP）in ATB patients was significantly higher than that in normal controls（χ²=3.989,OR=1.714,P=0.0458）,and the TT genotype frequency was significantly higher than that in normal controls（χ²=7.171,P=0.028）.The codominant model（TT vs GG,P=0.0073）and recessive model（GG+GT vs TT,P=0.0088）were significantly different from normal controls.The GA genotype frequency of rs3761624（TLR8）in ATB patients was significantly lower than that in normal controls（χ²=7.120,P=0.028）.The C allele frequency of rs6682925（IL23R）in ATB patients was significantly higher than that in normal controls（χ²=9.820,OR=0.469,P=0.0017）,and the TT genotype frequency was significantly lower than that in normal controls（χ²=9.342,P=0.009）.The codominant model（TT vs CC,χ²=7.582,OR=0.277,P=0.0059）and recessive model（CC+CT vs TT,χ²=8.676,OR=2.727,P=0.0032）were significantly different from normal controls.The A allele frequency of rs187238（IL-18）in ATB patients was significantly higher than that in normal controls（P=0.0002）,and the AA genotype frequency was significantly higher than that in normal controls（χ²=9.342,P=0.0002）.The haplotype AAGAAA of VDBP+IL-8+TLR2+IL-2rs3733359-rs7041-rs4588-rs4073-rs1898830-rs2069762 was significantly different between ATB patients and normal controls（χ²=5.193,P=0.02）,haplotype AAGAGC was significantly different between ATB patients and normal controls（χ²=8.626,P=0.003）,haplotype AAGTAA was significantly different between ATB patients and normal controls（χ²=4.162,P=0.04）,haplotype AAGTGA was significantly different between ATB patients and normal controls（χ²=14.492,P=0.0001）,haplotype GATAAA was significantly different between ATB patients and normal controls（χ²=4.510,P=0.03）,haplotype GCGTAA was significantly different between ATB patients and normal controls（χ²=14.048,P=0.0002）,haplotype GCGTGA was significantly different between ATB patients and normal controls（χ²=10.149,P=0.001）,haplotype AAGAGA was significantly different between ATB patients and normal controls（χ²=6.190,P=0.01）,haplotype AAGTGC was significantly different between ATB patients and normal controls（χ²=5.551,P=0.02）,haplotype GATTAC was significantly different between ATB patients and normal controls（χ²=3.920,P=0.048）,haplotype GATTGC was significantly different between ATB patients and normal controls（χ2=4.620,P=0.03）,and hamotype GCGAAA was significantly different between ATB patients and normal controls（χ²=4.955,P=0.026）.The haplotype ACCAC of ADRB2+IL-4+IL-12rs1042713-rs1042714-rs2243250-rs2243268-rs4921437 was significantly different between ATB patients and normal controls（χ²=7.929,P=0.005）,and haplotype GCCAC was significantly different between ATB patients and controls（χ²=4.500,P=0.03）.The haplotype GAATGCA of IFN-γ+PTX7rs2069718-rs2430561-rs208290-rs2393799-rs7958311-rs208294-rs3751143was significantly different between ATB patients and normal controls（P=0.039）,and hamotype GAGCGTC was significantly different between ATB patients and normal controls（χ²=7.260,P=0.007）.The haplotype TACATG of IL-23Rrs10489629-rs10889677-rs11465802-rs3762318-rs6682925-rs7518660 was significantly different between ATB patients and normal controls（P=0.028）.The interaction model of IL23R gene rs10489629 and rs11465802 had the most significant susceptibility to tuberculosis,but there was no statistical difference（P>0.05）.The interaction network between rs2243268 and rs2243274 of IL-4 gene and rs10489629 and rs11465802 of IL-23R gene was the strongest,but there was no statistical difference（P>0.05）.Therefore,the C allele at rs9061（SP110）and CC genotype increased the risk of tuberculosis,and the dominant model reduced the risk of tuberculosis.The G allele and GG genotype of rs11556887（SP110）significantly increased the risk of tuberculosis,and the dominant model is reduced risk of tuberculosis.The C allele and CC genotype of rs4327230（SP110）significantly increased the risk of tuberculosis,and the dominant model reduced the risk of tuberculosis.The A allele and AA genotype of rs7041（VDBP）significantly increased the risk of tuberculosis,and the dominant model reduced the risk of tuberculosis.The T allele and TT genotype of rs4588（VDBP）significantly increased the susceptibility to tuberculosis,recessive model was a protective model to reduce the risk of tuberculosis,and codominant model was a risk model.The GA genotype of rs3761624（TLR8）is a risk genotype for pulmonary tuberculosis.The C allele and recessive model of rs6682925（IL23R）significantly increased the risk of tuberculosis,and the TT genotype was a protective genotype that reduced the risk of tuberculosis.The A allele and AA genotype of rs187238（IL-18）significantly increase the risk of tuberculosis.The VDBP+IL-8+TLR2+IL-2rs3733359-rs7041-rs4588-rs4073-rs1898830-rs2069762 haplotype AAGAAA,haplotype AAGAGC,haplotype AAGTAA,haplotype AAGTGA,haplotype GATAA,haplotype GCGTAA,haplotype GCGTGA,haplotype AAGAGA,haplotype AAGTGC,haplotype GATTAC,haplotype GATTGC,and haplotype GCGAAA was significantly different between ATB patients and normal controls.The haplotype ACCAC and haplotype GCCAC of ADRB2+IL-4+IL-12rs1042713-rs1042714-rs2243250-rs2243268-rs4921437 were significantly different in ATB patients and controls.The haplotype GAATGCA and GAGCGTC of IFN-γ+PTX7 rs2069718-rs2430561-rs208290-rs2393799-rs7958311-rs208294-rs3751143 were significantly different between ATB patients and normal controls.The IL-23R rs10489629-rs10889677-rs11465802-rs3762318-rs6682925-rs7518660 haplotype TACATG had significant difference between ATB patients and normal controls.The interaction model of rs10489629 and rs11465802 of IL23R gene had the most significant susceptibility to tuberculosis,rs2243268 and rs2243274 of IL-4 gene,rs10489629 and rs11465802 of IL-23R gene had the strongest interaction network,But there was no statistical difference.（2）Immune response mechanism of lymphocyte subsets in ATB patients.In this study,30 First-treated etiology positive ATB（FTEP-ATB）patients,28 First-treated etiology negative ATB（FTEN-ATB）patients,26Re-treated etiology positive ATB（RTEP-ATB）patients and 29 Re-treated etiology negative ATB（RTEN-ATB）patients who were hospitalized in the Department of Tuberculosis Medicine,8^th Medical Center of PLA General Hospital,were enrolled.A total of 27 Non-tuberculous pulmonary disease（NTB-PD）patients hospitalized in Department of Respiratory and Critical Care Medicine,8^th Medical Center of PLA General Hospital.The 60normal controls were further divided into 30 IGRA-positive Latent tuberculosis infection（LTBI）patients and 30 IGRA-negative Healthy controls（HC）.The number of subsets of CD3⁺T cells,CD4⁺T cells,CD8⁺T cells,NK cells,NKT cells and B cells in each group was analyzed by flow cytometry.To evaluate the immune status of ATB patients,NTB-PD patients,LTBI patients and HC,as well as the effect of bacterial load and anti-tuberculosis treatment on the immune function of ATB patients.The results showed that the number of CD3⁺T cells and CD4⁺T cells subsets in ATB patients were significantly lower than those in LTBI patients and HC,and the number of CD8⁺T cell subsets in ATB patients were significantly lower than that in HC（P<0.05）.The number of B cell subsets in ATB patients was significantly lower than that in LTBI patients and HC（P<0.05）.The number of CD3⁺T cells,CD4⁺T cells and CD8⁺T cells in recirculated ATB patients was higher than that in newly treated ATB patients,and the number of positive patients was lower than negative patients,but there was no statistical difference（P>0.05）.Therefore,the immune response function of lymphocyte subsets in ATB patients is generally lower than that in LTBI patients and healthy persons.The higher the load of M.tb,the more obvious the inhibition of the immune response function of ATB patients.Effective anti-tuberculosis treatment can restore the immune response function of ATB patients to a certain extent.