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Amela Šahović

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Objectives This study aims to investigate the low-resolution human leukocyte antigen (HLA)-B locus polymorphisms between unrelated healthy individuals and patients with diagnosis of seronegative spondyloarthropathies and determine risky and protective allelic groups and genotypes. Patients and methods The study included 104 healthy control individuals (52 males, 52 females; median age 43 years; range 2 to 76 years) and 96 patients (43 males, 53 females; median age 28.5 years; range 2 to 67 years) diagnosed with: ankylosing spondylitis (AS) (n=19), reactive arthritis (n=19), psoriatic arthritis (n=28) and undifferentiated spondyloarthropathies (n=30). Genomic deoxyribonucleic acid was extracted from peripheral blood to detect allelic groups of HLA class I and II. Single-specific-primer polymerase chain reaction was used for HLA genotyping and visualization of products after their separation on 1.5% agarose gel for horizontal gel electrophoresis. Results Significantly increased frequency was found for HLA-A*02 and HLA-B*27 allelic variants in all groups of patients. The increased frequency of the HLA-B*35 allelic group in the control group represents the protective gene variant for the occurrence of AS. The predisposing genotype (HLA-B*27/B*44 and B*27/B*51) for the onset of disease was only found in AS patients. Conclusion This study shows the strong association of HLA-B*27 antigen with spondyloarthropathies, which is considered a risk variant of the gene for the onset of disease. Protective and risky allelic variants and genotypes are rare and their detection as well as increased frequency are possible if larger numbers of patients are involved.

Objectives This study aims to analyze human leukocyte antigen A (HLA-A), human leukocyte antigen B (HLA-B), human leukocyte antigen C (HLA-C), HLA-DRB1*, HLA-DRB3*, HLA-DRB4*, HLA-DRB5*, HLA-DQB1* loci expression in patients with rheumatoid arthritis (RA) in the Federation of Bosnia and Herzegovina. Patients and methods Deoxyribonucleic acid was isolated from peripheral blood of 48 RA patients (22 males, 26 females; mean age 36 years; range 2 to 63 years) and 104 healthy control individuals (52 males, 52 females; mean age 43 years; range 2 to 76 years). Deoxyribonucleic acid samples were analyzed using polymerase chain reaction-sequence-specific primers and sequence specific oligonucleotides methods. Results The most frequent allelic groups in RA patients were HLA-DRB1*01 (odds ratio=2.795; 95% confidence interval: 1.441-5.421; p=0.004) and HLA-DRB1*04 (odds ratio=2.573; 95% confidence interval: 1.214-5.453; p=0.023). Among RA patients, the most frequent genotype for the allelic group HLA-DRB1*, in the light of the common epitopes theory, was observed for DRB1*01/DRB1*13. This genotype indicates an increased incidence and relative risk (odds ratio=11.09). Conclusion The most common genotype in our RA patients was DRB1*01/DRB1*13, which showed increased frequency and a high relative risk. This genotype variant may be considered a predisposing factor for the development of RA.

AIM: The research was conducted by genotyping two Human Leukocyte Antigen (HLA) gene classes. The main objective of this research was to investigate distribution and frequency of the allelic groups, genotypes and haplotypes in the gene loci of HLA class I (HLA-A*, -B*, -C*) and HLA class II (HLA-DRB1*, -DQB1*) in patients included in the program of cadaveric renal transplantation. MATERIAL AND METHODS: Our study covered 186 blood samples of patients who are registered on the list for cadaveric renal transplantation in Federation of Bosnia and Herzegovina and included 59 control, healthy unrelated individuals. For the HLA typing, we have used three different methods: micro lymphocyte cytotoxicity test (MLCT), Polymerase Chain Reaction (PCR) – Sequence Specific Primers (SSP) and PCR – Sequence-Specific Oligonucleotides (SSO) or Luminex technology. All patients and cadaveric donors were tested using the three methods because the system is polymorphic. RESULTS: Analysis of the results of genotyping HLA class I gene loci identified dominant HLA-A*02, HLA-B*35, HLA-C*07 allelic groups. Analysis of the HLA class II gene loci genotyping showed that HLA-DRB1*11 and HLA-DQB1*03 loci had the highest incidence in HLA class II. CONCLUSION: Based on our results and previous research, there were no observed differences between allelic frequencies and genotypes of healthy people and people with ESRD. Differences between allelic groups occurred, but they were not statistically significant, except HLA-C*01 (p = 0.020).

A. Sofo-Hafizović, A. Dizdarevic-Rekic, Lejla Ibričević-Balić, Lejla Burazerovic, M. Vila, EminaSuljovic-Hadzimesic, Marina Skuric-Tomic, Elma Ćatović-Baralija, Gorana Ahmetovic et al.

Entezam, A. Šahović, Michael, Colvin, J. Hilton, Makio, Ogawa

We have studied the effects of 4-hydroperoxycyclophosphamide (4- HC)on murine hemopoietlc progenitors. We found dose-dependent killing and differential sensItivItIes of colony forming cells with burst forming units-erythrocyte being most sensitive and colony-forming units-granu locyte/erythrocyte/macrophage/megakaryocytemost resistant. We also tested the effects of 4-HC on more primitive murine progenitors which wereIdentIfIable in our assaysystemwhenthe addItiOnof interleukin-3 was delayed until Day 7. We found that the senslth'itiesofthe progenitors for blast cell coloniesare similar to thoseof colony-formingunits that late-np pairing blast cell colonies were particularly resistant to 4-HC. mechanismof sensitivities of sensitIvItIesof murine afterbriefinCUbatiOnwithdiethylaminobenzaldehyde,aninhib itor ofaldehyde dehydrogenase. Progenitors for granuincyte/amcrophage colonies, granulocyte/erythrocyte/macrophage/megakaryocyte differential sensitivItIesof progenitorsdisappearedfollowing We also tested the sensitIvities ofthe progenitors to phenylketophosphainide, an analogue of 4-HC which is resistant to inactivation by aldehyde dehydrogenase. Various colony-forming units exhibited a similar dose response to this compound. These data indicate that intracellular levels of aldehyde dehydrogenase might play an Important role in differential sensitivities of murine

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