ABSTRACT
Aims: To find out the most frequent associations of the HLA II class loci DRB1*, DQA1*, DQB1* with the HIV/AIDS infection. Place and Duration of Study: The study took place in The Laboratory of Clinical Immunology and Immunogenetics (LCII) of Riga Stradiņš University (RSU), Riga, Latvia, Riga Eastern Clinical University Hospital, “Infectology Centre of Latvia”, between May 1991 and December 2004. Original Research Article British Journal of Medicine & Medical Research, 4(17): 4482-4500, 2014 4483 Methodology: We analysed the medical documentation of 2500 patients and included 1180 (888 men, 292 women, 185 of them in AIDS phase) HIV infected patients. Genomic DNA was extracted from the blood with phenol-chloroform extraction method. Lowresolution HLA typing for HLA- DRB1*; DQB1*; DQA1* was performed by polymerise chain reaction (PCR) with amplification with sequence-specific primers (SSP). PCR products were separated on 3% agarose, the amplified bands were visualized, and the HLA-DRB1; DQA1; DQB1 type was deduced. Results: Genetic markers of immunologic alleles upon development of HIV infection – HLA-DRB1*03(17:01); 05(11:01); 07:01; HLA-DQA1*01:01; 02:01; 03:01; 06:01; HLADQB1* 03:02; 05:01; 03:03; 03:04, as well as resistance markers connected with slow development of HIV infection – HLA-DRB1*01:01; 04:01; 06(13:01); HLA-DQA1*01:03; 04:01; 05:01; HLA-DQB1*03:01; 03:03; 04:01-2; 06:01; 06:02-8 are located in different groups of patients. High risk markers in case of HIV infection development belonging to the following groups of alleles: HLA-DRB1*03(17:01), DRB1*05(11:01), DQA1*01:01; 03:01 un DQB1*05:01; 03:02, as well as three-loci haplotypes HLA-DRB1*03(17:01)/ DQB1*05:01/ DQA1*01:01; HLA-DRB1*05(11:01)/DQB1*03:01/ DQA1 *05:01; DRB1*01:01/DQB1*03:02/ DQA1*03:01 and DRB1*01:01/DQB1*05:01/DQA1*01:01 are determined. Resistance to HIV infection development forms in the following groups of alleles: HLA-DRB1*01:01; 06(13:01), HLA-DQB1* 03:01; 06:02-8; HLA-DQA1*01:02; 01:03, as well as in haplotypes HLA- DRB1*01:01/DQB1*06:02-8/DQA1*01:02;HLADRB1* 06(13:01)/DQB1*06:02-8/DQA1*01:02; HLADRB1* 01:01/DQB1*03:01/DQA1*01:02; and HLA-DRB1*06(13:01)/DQB1*06:02-8/ DQA1*01:02 in different groups of HIV/AIDS patients. Conclusion: The prevalence of genes DRB1; DQA1; DQB1 and DRB1-DQA-DQB1 combinations in the five groups of HIV infected patients have been established. Comparative analysis was performed also in the group of healthy donors (control group). The role of the main histocompatibility complex has been established, it enables marker functions and that can be used in the additional prognostic diagnostics in case of HIV infection. The obtained results testify that upon the identification of HIV genes it is possible to understand the molecular mechanisms in case of progression of AIDS syndrome complex; this possibly can be beneficial for the determination of the clinical results of infected patients.
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.
ABSTRACT
Aims: To find out whether ongoing missense mutations in the exon 2 of DRB1*01:01 affect the operation of this protective allele in HIV patients. Place and Duration of Study: the Clinical Immunology and Immunogenetics Laboratory of Riga Stradiņš University, Riga Eastern Clinical University Hospital, “Infectology Center of Latvia”, between May 2006 and December 2011. Methodology: The study includes 200 HIV-infected patients. DNA was isolated from venous blood samples using the Qiagen QIAamp DNA kit reagents and the exon 2 nucleotide sequence of HLA was determined by the automatic sequencing – “Big Dye Terminator mix” (Applied Biosystems, USA). Statistical analysis was performed using Microsoft Excel, DOS StatCalc programs. The significance of the differences in indicators was evaluated according to reliability p£0.05. The odds ratio was calculated according to Wolf’s method. Results: We found missense: at codon 47– in 80% of cases; at codon 67– in 20% of cases; at codon 75 – in 11% of cases; at codon 82– in 10% of cases; at codon 86– in 10% of cases (p<0.05) (See Table 3). One of the HIV patients had a STOP-codon (codon 13). Besides, a balance between nucleotide transversion and transition has been observed, suggesting mutations in the exon 2 (transversion in a human genome is rare) (OR 0.05, 95% CI 0.00-0.053). Conclusion: The results of the study are not complete in order to be able to say conclusively that the existing mutations in the exon 2 of HLA-DRB1 *01:01 gene cause wrong immune response, thus the protective functions of this allele are not fulfilled. For a fuller understanding of the importance of ongoing mutations in the exon 2 in the development of HIV/AIDS, it is necessary to increase the study group.