Genetic Polymorphisms HLA Class II in SIRS and Sepsis in Children.

Aims: The aim of this research was to investigate the genetically determined predisposition to develop SIRS and sepsis by analyzing human leukocyte antigen HLA class II genes. Study Design: children defined by the criteria for SIRS of Critical Care Medicine Consensus Conference were included into the study in a prospective manner. Place and Duration of Study: Riga Stradiņš University Laboratory of Clinical Immunology and Immunogenetic, Department of Pediatrics, Rīga Stradiņš University, and Children’s Clinical University Hospital, Latvia, between January 2008 and May 2009. Methodology: Samples from children with SIRS and sepsis were collected at the Children`s Clinical University Hospital of Latvia. During the study, 65 patients with SIRS were observed. In 12 cases among SIRS patients, sepsis was confirmed. DNA was separated from lymphocytes of peripheral blood. At the same time, 100 DNA samples from healthy children were analysed. HLA typing low-resolution for HLA- DRB1; DQB1; DQA1 was performed by polymerase chain reaction (PCR) with amplification with sequence-specific primers (SSP). PCR products were separated on 3% arose, the amplified bands were visualised. The frequency of alleles and genotypes were compared between the patients and the controls using chi-square test. P-value and odds ratio were calculated using EPI INFO software version 6 with 95 % confidence intervals and Fisher exact correction for small numbers. Results: The frequency of DRB1*07:01 allele was significantly increased in patients with SIRS (OR=8.71; 95% Cl = 2.8-26.8; p<0.0001) and patients with sepsis (OR=9 .70; 95% Cl = 2.3-42.2; p<0.005). In-group SIRS was significantly increased DQB1*03:02 (OR=2.19; 95% Cl = 1.1-4.8; p<0.049) and sepsis group DQB1*03:01 (OR=2.95; 95% Cl = 1.3-7.1; p<0.013) alleles also. But the frequency of DRB1*15:01 (OR=0.50; ; 95% Cl = 0.2-0.9; p<0.038) and DQB1*06:01, (OR=0.16; 95% Cl = 0.2-1.3; p<0.042) alleles was lower in all SIRS patients than in control group. In the distribution of HLA DRB1*/DQA1*/DQB1*, such haplotypes as DRB1*07:01/DQA1*02:01/DQB1*03:02, (OR=8.08; 95% Cl = 0.9-74.2; p<0.049); DRB1*07:01/DQA1*02:01/DQB1*05:01, (OR=8.08; 95% Cl = 0.9-74.2; p<0.049) and DRB1*04:01/DQA1*03:01/DQB1*02:01-2 (OR=5.10; 95% Cl = 0.9-27.3; p<0.049) in SIRS group were frequently detected. The haplotypes DRB1*07:01/DQA1*02:01/DQB1*03:01, (OR=33; 95% Cl = 0.9-27.3; p<0.004); DRB1*07:01/DQA1*03:01/DQB1*03:01; (OR=19.80; 95% Cl = 0.9-27.3; p<0.029) and DRB1*04:01/DQA1*02:01/DQB1*03:01, (OR=19.80; 95% Cl = 0.9-27.3; p<0.03) were frequently found in septic group patients. Conclusion: Our pilot data shows that the susceptibility markers HLA-DRB1*07:01/ DQA1*02:01/ DQB1*03:01, in sepsis patients are partly consistent with literature data. The number of patients in the study is relatively small. In order to increase statistical significance of results and to prove current findings, it is important to continue the study.

Prepae B-lymphoblastoid cell lines of HLA novel allele B ~*5610 in a family / 中国输血杂志

Objective To prepare B-lymphoblastoid cell lines of HLA novel allele B*5610 in a family for further study and identification . Method Isolate mononuclear cells under aseptic conditions from the peripheral blood. After infection with Epstein-Barr virus, the cells were cultured in 20% FBS, 2?g/ml CsA RPMI 1640. Results Immortalized B-lymphoblastoid cell lines of five B *5610 carriers in a family were achieved, and the new genes were inherited stably. Conclusion Our work is important for storing and breeding the precious material of biomedicine because the B *5610 genes in the immortalized B-lymphoblastoid cell lines were inherited stably.