Factors influencing overall survival and GvHD development after allogeneic hematopoietic stem cell transplantation - single centre experience.

BACKGROUNDS: Allogeneic hematopoietic stem cell transplantation (alloHSCT) is a substantial therapeutic procedure for the treatment of a wide spectrum of severe diseases. Despite advancements in treatment and supportive care, alloHSCT still carries a considerable mortality risk, primarily caused by graft-versus-host disease (GvHD). Our retrospective analysis aimed to identify the factors influencing overall survival and GvHD development in HLA-identical sibling alloHSCT. We have analyzed patients' and donors' age, AB0 compatibility, recipient-donor gender match, stem cell source, time from the diagnosis to alloHSCT, conditioning regimen type, GvHD prophylaxis, and relapse. PATIENTS AND METHODS: Our study included 96 patients (54 male, 42 female) who underwent HLA-identical sibling alloHSCT. The median follow-up was 64.5 months (range 1-218 months), and the median age of both recipients and donors was 34 years. Malignant hematological diseases were the most common indications for alloHSCT. RESULTS: GvHD and its complications accounted for the highest number of deaths (N = 24; 46.2%), followed by relapse (N = 18; 34.6%). Acute GvHD developed in 30 patients (31.3%), while chronic GvHD occurred in 25 patients (26.0%), resulting in a total of 45 patients (46.9%) experiencing GvHD. Male recipients with female donors had significantly worse overall survival compared to other patients (P = 0.01; HR = 2.33). Overall survival was better in patients transplanted within 1 year from the diagnosis compared to those transplanted after 1 year (P = 0.03; HR = 1.93). No factor reached statistical significance regarding the impact on acute GvHD, chronic GvHD, or overall GvHD. CONCLUSION: We confirmed that sex mismatch, specifically in the case of a female donor and a male recipient, significantly negatively affects overall survival after alloHSCT. Additionally, overall survival is significantly shorter when the interval between the diagnosis and alloHSCT exceeds one year.

Analysis of HLA-G gene polymorphisms in Slovak women with pre-eclampsia.

OBJECTIVES: To identify possible association between the selected HLA-G gene polymorphisms and risk of pre-eclampsia. BACKGROUND: Pre-eclampsia is a serious multisystem disorder that affects women during pregnancy. Despite many research studies, the pathology of pre-eclampsia is not fully understood. Human leukocyte antigen G (HLA-G) belongs to the molecules that induce fetal acceptance by the maternal immune system. HLA-G expression was found to be impaired in the women suffering from pre-eclampsia suggesting its involvement in the development of pre-eclampsia. METHODS: 123 women with pre-eclampsia and 102 women with normotensive pregnancy were included in the study. HLA-G gene polymorphisms affecting its expression was determined, namely the HLA-G 14 bp insertion/deletion polymorphism in the 3'UTR and HLA-G 1597ΔC polymorphism tagging the HLA-G*01:05N null allele. Genotyping was performed by PCR and PCR-RFLP. RESULTS: No statistically significant differences in either allele or genotype frequencies between pre-eclampsia cases and control group have been observed (p > 0.05). CONCLUSION: Genetic predisposition of HLA-G to pre-eclampsia in Slovak women was examined for the first time. No association between analysed HLA-G gene polymorphisms and susceptibility to pre-eclampsia was observed. Further investigations are needed to determine the role of immunosuppressive molecule HLA-G in pre-eclampsia development (Tab. 5, Fig. 2, Ref. 37).

VLA4 Gene Polymorphism and Susceptibility to Multiple Sclerosis in Slovaks.

Multiple sclerosis (MS) is an inflammatory autoimmune disease occurring in genetically sensitive individuals. As migration of immune cells into the CNS is facilitated by the Very Late Antigen 4 (VLA-4) integrin molecule, the VLA4 gene may be considered as a plausible candidate genetic risk factor for susceptibility to MS. Therefore, the objective of our study was to investigate the association between two genetic polymorphisms located in the VLA4 gene and the risk of multiple sclerosis. One hundred seventeen MS patients and 165 control subjects from Slovakia were genotyped for VLA4 gene SNP polymorphisms at positions 269 (C/A) and 3061 (A/G). The same study cohorts were also genotyped for the rs3135388 polymorphism tagging the HLA-DRB1*15:01 allele, which is a known genetic factor associated with susceptibility to develop MS in many populations. Our findings show for the first time that the rs3135388 polymorphism is a strong risk factor for MS in the Slovak population. Investigation of the VLA4 gene polymorphisms revealed a significantly higher frequency of the 3061AG genotype in MS patients compared to the controls (P ≤ 0.05). We suggest that the 3061AG polymorphic variant is an independent genetic risk factor for MS development in our population as it was significantly associated with this disease. The association was also confirmed after applying multivariate logistic-regression analysis adjusted for gender, age and HLA-DRB1*15:01 positivity as possible influencing factors.

High susceptibility to pemphigus vulgaris due to HLA-DRB1*14:54 in the Slovak population.

The current work describes an association between pemphigus vulgaris (PV) and class II HLA alleles in the Slovak population, the first such study in Slovakia on the 'high-resolution level'. This work takes into account the new HLA allele nomenclature, officially adopted in 2010. In particular, we have focused on the associations between PV and DRB1*14:54 and DRB1*14:01. This case-control study was performed in a cohort of 43 PV Caucasian patients and 113 Caucasian control subjects from Slovakia. HLA typing was performed using PCR-SSP (polymerase chain reaction with sequence-specific primers). We found significantly positive associations between PV and the HLA alleles DRB1*04:02, DRB1*04:04, DRB1*14:54, DRB1*14:04, DRB1*14:05, DQB1*03:02 and DQB1*05:03. In contrast, HLA-DQB1*06, DRB1*07 and DRB1*13 were negatively associated with PV. Importantly, 93% of PV patients possessed at least one of two HLA haplotypes, DRB1*04-DQB1*03 or HLA-DRB1*14-DQB1*05. We confirmed the previously reported associations between HLA class II alleles and PV and described a new association between PV and DRB1*14:54. This allele was first described in 2005, and there has been only one report of its association with PV to date.

Characterization of MICA gene polymorphism of HLA complex in the Slovak population.

BACKGROUND: The function of the MHC class I polypeptide-related sequence A (MICA) gene, which belongs to the MHC class I chain-related genes, is to trigger cytolysis of target cells mediated by NKG2D receptor recognition in NK (Natural Killer) cells and CD8 T-lymphocytes. The MICA gene has a high degree of polymorphism, especially observed in exons 2-5. MICA allelic diversity has been reported in association with some autoimmune diseases such Behcet's disease, psoriasis and diabetes, as well as with organ rejections. AIM: The aim of this study was to analyse MICA gene polymorphism in the Slovak population, to establish frequencies of MICA alleles and to compare the results with those found in other Western Eurasian populations. No such study has been performed previously in the Slovak population. SUBJECTS AND METHODS: This study examined DNA samples from 124 unrelated Slovak individuals (51 women and 73 men with an average age of 40.3 years) using direct sequencing of MICA exons 2-5. Allele and genotype frequencies were calculated by direct counting and statistical analysis was carried out using Arlequin software. RESULTS: This study identified 15 out of 71 MICA alleles. The most frequent allele was MICA(*)008 (37.1%) followed by alleles MICA(*)002 (16.5%) and MICA(*)009 (11.3%). The rarest alleles were MICA(*)027, MICA(*)006 (both 0.8%) and MICA(*)057 (0.4%), respectively. The most frequent genotypes were 008/008 and 008/002, both with a frequency of 13.7%. Exon 5 microsatellite polymorphism screening revealed five MICA alleles, namely A4, A5, A5.1, A6 and A9. The most frequent was allele A5.1 (37.1%) and the rarest A5 (8.1%). Finally it was found that haplotype MICA*008 A5.1 was the most frequent (37.1%). CONCLUSION: A comparison of these results with those reported in the literature revealed similarity in MICA polymorphism to that found in other Western Eurasian populations. The data will be useful for further association studies on MICA polymorphism and its function.

Associations between HLA class II alleles and type 1 diabetes mellitus in the Slovak population.

OBJECTIVES: Several associations between HLA complex and diabetes mellitus type IA were found in various groups of patients of Caucasoid population. This study was therefore prompted to be conducted in Slovak population, since any such has not yet been performed in Slovak population. METHODS: Patients suffering from DM-1A originated from all regions of Slovakia. Their age ranged from 1 to 42 years; but the criterion for including the subject to the study was the definition of diagnosis in older patients before their age of 15 (Table 1). The diagnosis was set up according to internationally accepted criteria. A total of 460 patients was typed for HLA-DQB1 alleles, among them 97 also for HLA-DQA1 and 146 for HLA-DRB1 alleles. HLA-typing was performed by a PCR-SSP method. Control group consisted of 196 (DQA), 143 (DQB1) and 130 (DRB1) unrelated blood donors aged 19-55 years old irrespective of their age or sex. The data obtained were expressed in a 2 x 2 contingency table and statistical significance was calculated by the Fisher exact test. RESULTS: Among 11 HLA-DQB1 alleles tested DOB1*0302 was the most frequent in DM-1A patients (30.33% vs. 5.59% in healthy subjects (HS), followed by DQB1*0201 (22.93% vs. 12.94%, respectively). In contrast, the frequency rate of DQB1*0301 (10.66% vs. 24.48%), DOB1*0602 (2.17% vs. 10.14%) and DQB1*0603 (2.5% vs. 8.39 %) were decreased in DM-1A patients. Out of 14 DQA1 alleles the highest occurrence rate showed DQA1*0301 (30.93% vs. 17.09) and DQA1*0501 (34.02% vs. 25.76%), while DQA1*0102 (8.76% vs. 16.58%) and DQA1*0201 (6.18 % vs. 13.51%7), respectively, were found to be the least frequent. Among 13 HLA-DRB1 alleles tested, the most common occurrence rates showed DRB1*03 (26.37% vs. 9.62%) and DRB1*04 (7.19% vs. 14.23%), while the least frequent alleles were DRB 1*15 (2.74% vs. 12.31%), DRB1*07 (7.19% vs. 14.23%), and DRB1*11 (2.74% vs. 20.38%). The alleles DQB1*0302 and DQA1*0301, respectively, were present in the same individual in all DRB1*04 positive patients, suggesting that they belong to the haplotype. Similar situation was observed with the alleles DQB1*0201, DQA1*0501, and DRB*0301, respectively, forming the second HLA haplotype so characteristic for DM1A.

HLA class II allele frequencies in type 1A diabetes mellitus Slovak patients.

BACKGROUND: Diabetes mellitus type 1A (DM-1A) is an autoimmune disease in which the immune response is directed to pancreatic islet cells. DM-1A occurs in genetically predisposed individuals. Among type 1A diabetes associated genes, those of the HLA region have the greatest effect. OBJECTIVES: The aim of our study was to obtain a comprehensive survey of the HLA-DRB1 and HLA-DQB1 allele frequencies in Slovak patients suffering from DM-1A. METHODS: HLA class II genotyping was performed on genomic DNA by the PCR-SSP method according to the 12th Workshop protocol. RESULTS: Our report gives the first presentation of the distribution of HLA-DRB1 alleles (including complete DRB1*04 subtypes) and that of HLA-DQB1 alleles in the Slovak diabetic patients diagnosed at 0-18 years of age. Susceptibility is significantly associated with the alleles DQB1*0302 (OR = 7.8), DRB1*04 (OR = 4.9), DRB1*0301 (OR = 4.2) and DQB1*02 (OR = 2.2), whereas the alleles DQB*0602 (OR = 0.05), DRB1*11 (OR = 0.2), DRB1*15 (OR = 0.2) and DQB1*0301 (OR = 0.3) were found to be protective. CONCLUSIONS: Our results, consistent with other studies, show increased frequencies of known positively associated HLA class II alleles in our type 1A diabetes mellitus patients compared to the general (nondiabetic) population. The protective effect of previously reported alleles was confirmed as well. Results of our population-based study serve in clinical practice for the identification of subjects at risk of developing DM-1A among the first-degree relatives (Tab. 2, Ref. 12).

[Latent autoimmune (type 1) diabetes mellitus in patients originally classified as type 2. Divergence of etiologic markers]. / Latentný autoimunitný (typ-1) diabetes mellitus u pacientov pôvodne klasifikovaných ako typ-2. Divergencia etiologických markerov.

BACKGROUND: To assess the prevalence of markers of autoimmune insulitis (AII) in patients classified originally as having Type-2 diabetes mellitus (Type-2 DM). 386 patients subdivided according to the BMI, C-peptide and type of treatment. METHODS AND RESULTS: Age, BMI, C-peptide, Glutamic acid decarboxylase autoantibodies (GADA), HLA-DR/,-DQ alleles. Prevalence of GADA varied from < 5% in obese patients with normal/increased C-peptide to > 30% in non-obese patients with low C-peptide. In majority of GADA positive patients, the Type-1 DM high-risk HLA-DRB1*, HLA-DQB1* alleles have been found. Among them HLA-DRB1*0302 and HLA-DRB1*0201 were more frequent than HLA-DRB1*040x and HL:A-DQB1*0302. CONCLUSIONS: Significant fraction of patients classified initially as Type-2 DM may have in fact Type-1 DM. Such patients can be recognized on the basis of assessment of serological (GADA) and immuno-genetical (HLA-DR/,-DQ alleles) markers. In some patients clinical, metabolic, immune, and immunogenetic markers may disagree. This divergence stresses multifactorial genesis of diabetes. Moreover, it can also suggest that both autoimmune insulitis and insulin resistance may coexist in parallel.

[Association of type I diabetes mellitus with HLA alleles]. / Asociácia diabetes mellitus I. typu s HLA-alelami.

Insulin-dependent diabetes mellitus is a chronic autoimmune disease characterised by a loss of tolerance towards own antigene structures beta-pancreatic cells. The destruction of cells subsequently leads to the loss of insulin production. There are more factors which trigger the autoimmune response in susceptible individuals, however, they are only partially known so far. One of the predisposing factors is the genotype, while the main role is ascribed to genes of the main histocompatible complex (HLA). Out of extensive genetic and epidemiological studies, the Caucasoid population is known to have a significant association of insulin-dependent diabetes mellitus with the increased frequencies of haplotypes HLA-DRB1*04-DQA1*0301-DQB1*0302 and DRB1*0301-DQA1*0501-DQB1*0201.

[Latent autoimmune (type I) diabetes mellitus in adults. Part. II. Association of HLA antigens, status of cellular immunity and occurrence of other autoimmune diseases]. / Latentný autoimunitný (Typ-1) diabetes mellitus u dospelých. II. Cast'. Asociácia s HLA antigénmi, stav bunkovej imunity a výskyt iných autoimunitných ochorení.

AIM OF STUDY: To assess some immunological and immunogenetic aspects in patients with latent autoimmune (Type-1) diabetes mellitus (DM) of adults (LADA). SUBJECTS: 24 patients with LADA, 11 patients with Type-2 DM and 20 healthy volunteers (Pilot study). PARAMETERS TESTED: HLA-DRB1* and HLA-DQB1* alleles, parameters of cellular immunity (CD4+, CD8+, CD3/HLA-DR+, CD8/HLA-DR+, CD45RA+[CD4], CD16+CD56), CD19+, IL-4, INF-gamma and organ specific (OSA) autoantibodies (against thyroid gland, gastric parietal cells, tubuli, basal membranes of glomerulus, AMA and ABBA). RESULTS AND DISCUSSION: Type-1 DM HLA-DRB1* and HLA-DQB1* risk alleles have been found in a majority of patients with LADA. The most frequent were HLA-DRB1*0301 and DQB1*0201. Assessement of parameters of cellular immunity and cytokine profiles (IL-4 a INF-gamma) in peripheral blood did not reveal any contribution to a differentiation between Type-1 and Type-2 DM). We confirmed increased occurence of OSA in patients with LADA, what stress importance of routine screening for OSA in patients with LADA.

HLA-DRB1, -DQB1 and -DPB1 polymorphism in the Slovak population.

HLA-DRB1, -DQB1 and -DPB1 allele frequencies were investigated in a sample of the Slovak population by PCR-SSP and PCR-RFLP methods. The most frequent DRB1 alleles were DRB1*1101-5 (0.2038), DRB1*0701-2 (0.1423), and DRB1*1501-2 (0.1231). The most rare alleles found were DRB1*0901 (0.0038), and DRB1*1201 (0.015). The most common DQB1 alleles were DQB1*0301 (0.2448), DQB1*0201 (0.2098), and DQB1*0501 (0.1119), respectively. The alleles with the least occurrence rate were DQB1*0601 (0.0035) and DQB1*0401 (0.007). The most common DPB1 alleles found were DPB1*0401 (0.4329), DPB1*0402 (0.2089), and DPB1*0201 (0.1438), respectively. The least frequent alleles were DPB1*0601, *1101, and *1501 (0.0034). Allele frequencies found in our study were compared to those in Czech, Austrian, and German populations. No statistically significant differences were observed.

[Frequency of the HLA-DPB1 allele in the Slovak population]. / Frekvencia HLA-DPB1 alel v Slovenskej populácii.

The polymorphism at the HLA-DPB1 locus was established in 146 unrelated persons. The polymerase chain reaction (PCR) in combination with restriction fragment length polymorphism (RFLP) technique was used. After PCR amplification of the second exon of the HLA-DPB1 locus, the PCR products were digested with seven allele specific endonucleases. The alleles DPB1*0401 (43.29%), DPB1*0402 (20.89%), DPB1*0201 (14.39%) and DPB1*0301 (9.25%) were the most common among 15 DPB1 alleles detected in the tested group. The least frequent alleles were DPB1 *0202, *0601, *1101 and *1501 (0.34%). The comparison with allele frequencies in Austrian and German populations showed no statistically significant differences. (Tab. 2, Ref. 18.)

[Frequency of loci of HLA-DRB1* and -DQB1* alleles in the Slovak population]. / Frekvencia alel lokusov HLA-DRB1* a -DQB1* v Slovenskej populácii.

Results on HLA-DRB1* and HLA-DQB1* allele frequencies in the Slovak population by PCR-SSP method are presented. HLA-DRB1* alleles were determined in 130 and HLA-DQB1* alleles in 143 healthy unrelated individuals. The highest frequency was observed for the alleles HLA-DRB1*1101-13 (0.203), HLA-DRB1*0701 (0.142), HLA-DQB1*0301 (0.244), and HLA-DQB1*0201 (0.209). The least frequent alleles were HLA-DRB1*1402-6-9, HLA-DRB1*0901 (both 0.0038), HLA-DQB1*0401 (0.007), and HLA-DQB1*0601 (0.0035). The results obtained by DNA-typing were compared with those calculated from the serological study. No statistically significant differencies were found. The allele frequencies obtained in our study were also compared with those of the Czech and Austrian populations. No statistically significant differencies were observed. (Fig. 2, Tab. 3, Ref. 13.)

The role of HLA class II antigens in the induction of cytotoxic T lymphocytes.

Investigation of pairs of unrelated persons mismatched for a particular HLA-DQB1 or -DPB1 gene on the induction of cytotoxic T lymphocytes (CTL) revealed that HLA-DQ and HLA-DP antigens provided a slight proliferative stimulus which was, however, sufficient for the generation of CTL. Monomorphic anti-DQ and anti-DP monoclonal antibodies abrogated the induction of cytotoxic response. The results indicate that the HLA-DQ and HLA-DP antigens play a similar role to HLA-DR specificities in clinical bone marrow transplantation.