ABSTRACT
Aim of Study: To identify the factors of molecular genetic risks during the development of infection in HIV, based on the TNFα cytokine gene polymorphism in combination with HLA DRB1/DQA1/DQB1 genes, as well as to analyse their possible association with the progress of the disease. 185 HIV infected patients and 173 individuals control group have been analysed. The DNA was extracted from peripheral blood, by using QiagenQIAamp DNA kit reagents. The quality and quantity of DNA was checked by using Qubit ® fluorometer HLA typing for HLA DRB1/DQB1/DQA1* was performed by RT-PCR with sequence-specific primers (SSO). TNFα gene G–238A and G–308A polymorphic variant incidence was determined by RT-PCR analysis. Results: We have detected TNFα gene allele 308A in 11% HIV infected patients, whereas in control group this allele have been detected only in 4% patients. Although the incidence of the TNFα gene –238A allele was twice as high in the control group (6%) as in the HIV infected patients (3%), it did not prove to be statistically valid (p = 0.253). The incidence analysis of three-locus haplotypes DRB1-DQB1-DQA1 – in TNFα position-238A/G -308A/G showed that haplotypes 01:01/05:01/01:01-TNFα-238(GA)/308(GG) and 01:01/03:02/03:01 - TNFα-238(AA)/308(GG) are more frequent in the control group in comparison to the groups of infected patients. This means that these haplotypes have a protective function, which significantly affects the progress of infection. The association of 15:01/05:01/01:01 -TNFα-238(GG)/308(GG) and 03:01/05:01/01:01- TNFα-238(GG)/308(GА) genotypes indicates a high risk of developing a fulminant infection. The genetic factors of AIDS-related complex of syndromes development are associated not only with the HLA complex class II alleles, but also with the SNP polymorphism in the promoter region of cytokine genes.
ABSTRACT
Aims: One of the ways to treat HIV/AIDS is Antiretroviral therapy (ART), which consists different schemes and includes numbers of drugs. However, in view of the HLA polymorphism it is necessary to determine the best association of ART with the HLA class II haplotypes. The purpose of the current study is to evaluate various HLA class II haplotypes with ART effectiveness in HIV-infected patients. Study Design: This study is a retrospective follow up on the association of ART with the HLA class II haplotypes of HIV/AIDS patients. From 254 Latvian HIV/AIDS infected patients - 195 men and 59 women blood was collected and HLA class II heliotypes were defined. Main ART therapy parameters were observed and compared. Place and Duration of Study: Riga Stradiņš University, the Laboratory of Clinical Immunology and Immunogenetics, Riga, Latvia Riga Stradiņš University, the Department of Infectology, Riga, Latvia Riga Eastern clinical university hospital, Infectology Centre of Latvia, Riga Methodology: The research included 254 Latvian HIV/AIDS infected patients - 195 men and 59 women. For monitoring immunological parameters were used: amount of CD4 + lymphocytes and HIV viral load, which were observed in 24-48 weeks. The efficacy criterias of ART were HIV RNA viral load <400 cop/ml - after 16-24 weeks, increase of CD4 + cell amount of 30-70 cells/μl after first 12 weeks and of 100-150 cells/μl after a year, and absence of new opportunistic infections after 12 weeks of ART treatment. For DNA extraction venous blood was used. HLA class II alleles DRB1 *, DQA1 *, DQB1 * genotyping was performed using PCR method. Results: During examining the HLA class II haplotypes the most credible association with high immunological efficacy showed haplotypes 01:01/06:02-8/01:03, 01:01/03:01/01:02, 13:01/06:02-8/01:02. After 12 weeks of treatment, CD4 + lymphocytes amount in a given group increased to 600-700 cells/μl, HIV RNA viral load decreased to 5 000 copy/ml, after 24-48 weeks of therapy - CD4 + lymphocytes amount increased to 806-900 cells/μl, and HIV viral load RNA decrease <400 copy/ml. Low immunological association of effectiveness: HLA-DRB1*/ DQB1*/DQA1* 15:01/03:01/03:01, 17:01/05:01/02:01, 17:01/03:01/05:01, 07:01/03:01/02:01, 11:01/03:01/05:01, 15:01/03:02/01:02. The frequency of HLA-DRB1 and DQB1, each allele and genotype was compared between the patients and the controls using the chisquare test. The P value and odds ratio (OR) were calculated using EPI INFO software, version 06, with 95% confidence intervals and Fisher exact correction for small numbers.[16] Conclusion: The given data shows the efficiency of ART therapy. No clinical progression of HIV (worsening of latent opportunistic infections) was observed during the ART treatment in study groups with existing haplotypes.