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P102
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P102
Abstracts / Human Immunology 75 (2014) 50–141
GESTATIONAL DIABETES MELLITUS TRANSCRIPTOME ANALYSIS REVEALS INDUCTION OF GENES RELATED TO THE MAJOR HISTOCOMPATIBILITY COMPLEX. Eduardo A.
Donadi, Nathalia B. Cezar, Adriane F. Evangelista, Danilo J. Xavier, Amanda F. Assis, Thais C. Arns, Maria Cristina Foss-Freitas, Milton C. Foss, Elza T. Sakamoto-Hojo, Geraldo A. Passos. FMRP-USP, Ribeirao Preto, Brazil. Gestational diabetes mellitus (GDM) is the most common metabolic disorder observed during pregnancy, and has been defined as an abnormal glucose metabolism first diagnosed during gestation. GDM women usually present postpartum diabetes, insulin resistance, metabolic syndrome, hypertension and dyslipidemia. Although few genetic markers have been described for GDM, approximately 10% of GDM patients present high probability of progression to type 1 diabetes (T1D). In a previous meta-analysis study, evaluating the transcript profiles of patients with the major types of diabetes, we reported that transcription profile of T1D patients was closer to GDM than to type 2 diabetes. Additionally, it has already been reported association of some HLA alleles with GDM, suggesting a possible immunological feature for GDM. In this study, we compared the transcription profiling of PBMCs of 18 GDM patients and 10 healthy pregnant women. Total RNA samples were hybridized to AgilentÒ 4 44 K oligo microarrays encompassing the whole human functional genome. Differentially expressed mRNAs were obtained by Rank Products analysis, which classifies transcripts according to their fold change values. The hierarchical clustering of mRNAs and samples were performed using the Cluster program, dendrograms and spatial representations of mRNAs were constructed using the TreeView software. A total of 731 differentially and significantly expressed mRNAs were observed when GDM patients were compared to controls. Molecular functions and biological pathways, as analyzed by the DAVID database, identified 130 biological processes and nine signaling pathways (P < 0.05). Several induced and biologically relevant transcripts were associated with the HLA system, including HLA-DRB6, DQB1, DQB2, DOA and DQA2. The modulation of these transcripts in GDM patients reinforces the hypothesis of a deregulation of HLA class II genes in GDM patients, deserving further attention.
P103
HIGH RESOLUTION HLA ALLELE FREQUENCIES OF STEM CELL DONORS IN MEXICO. GENETIC DIVERSITY AND ITS RELEVANCE TO IMPROVE UNRELATED DONOR SEARCHES.
Hilario Flores-A 1,2, Carmen Alaez 1,2, Diego Sanchez 2, Andrea Munguia 1, Araceli Rodriguez 1, David Garcia 1, Clara Gorodezky 1,2. 1 InDRE, Secretary of Health, Mexico City, D.F., Mexico; 2 Fundación Comparte Vida, A.C., Mexico City, D.F., Mexico. Aim: DNA-based registry typing is commonly done by SSOP and only a few registries employ sequencing at the time of recruitment, allowing to understand allele and haplotype frequencies and evaluating the size and composition of the registry, to assess the likelihood of finding matched donors. We explored the impact of the increasing number of HLA alleles in the genetic diversity of Mexicans, which will in turn impact the possibility of finding a donor for patients with a similar ethnic background. Methods: Healthy Mexicans enrolled at the DONORMO-The Mexican Unrelated Bone Marrow Donor Registry, born in different geographic areas of Mexico were HLA typed at the allele level, for A⁄, B⁄, C⁄, DRB1⁄, DQB1⁄ loci. DNA samples were sequenced in an ABI Prism 310 or in a 3500 sequencer. The SPSS 12 software was used for analysis. For A locus, 127 individuals were typed; 523 for B, 81 for C; 952 were SBT for DRB1 and 3343 for DQB1. Results: A total of 273 common and well documented alleles were detected; 91 were unique observations; 40 at A⁄ locus; 26 at B⁄; 8 at C⁄; 14 at DRB1⁄ & 3 at DQB1⁄. 65 different A⁄ alleles are present, of which 32 are A2 different variants: A⁄02:01; A2:06; A2:05; (13.4–5.5%) prevail. The prevalent B⁄ alleles were 49:01; 35:12; 35:17; 15:15; 14:02; 14:01;39:06; 39:05; 40:05; 40:08; 50:01; 52:01 51:02; 53:01; (>1.5%). Only 10 C⁄ alleles are frequent: 04:01, 07:02; 01:02; 03:04; 07:01; 16:01, 03:06; 5:01; 08:01; 08:02; (>2.5%). 16 frequent DRB1⁄ alleles are present: 04:07; 08:02; 14:06; 01:02; 04:02; 04:11; 04:04; 19:01; 14:02; 07:01: 03:01; 16:02; 01:03; 04:04; 10:01; 15:03; 15:01, 15:03 (1.8%). Unique observations were found at DQB1⁄ for 06:11; 03:08; 02:04. Mediterranean, Amerindian, Spanish, Caucasian, Semitic and African influences are clear. Conclusion: The molecular variation shown, and a one-step typing strategy using SBT since donor recruitment, is highly relevant to optimizing donor search algorithms in HSCT, for identification of potential matches, and in understanding the genetic background of this highly heterogeneous country. The data are also very useful for applications that depend on allele prevalence information such as: resolution of ambiguity during HLA typing, research and clinical practice, and the management of the general data, in the face of an ever growing list of recognized allele sequences.
P102
Abstracts / Human Immunology 75 (2014) 50–141
GESTATIONAL DIABETES MELLITUS TRANSCRIPTOME ANALYSIS REVEALS INDUCTION OF GENES RELATED TO THE MAJOR HISTOCOMPATIBILITY COMPLEX. Eduardo A.
Donadi, Nathalia B. Cezar, Adriane F. Evangelista, Danilo J. Xavier, Amanda F. Assis, Thais C. Arns, Maria Cristina Foss-Freitas, Milton C. Foss, Elza T. Sakamoto-Hojo, Geraldo A. Passos. FMRP-USP, Ribeirao Preto, Brazil. Gestational diabetes mellitus (GDM) is the most common metabolic disorder observed during pregnancy, and has been defined as an abnormal glucose metabolism first diagnosed during gestation. GDM women usually present postpartum diabetes, insulin resistance, metabolic syndrome, hypertension and dyslipidemia. Although few genetic markers have been described for GDM, approximately 10% of GDM patients present high probability of progression to type 1 diabetes (T1D). In a previous meta-analysis study, evaluating the transcript profiles of patients with the major types of diabetes, we reported that transcription profile of T1D patients was closer to GDM than to type 2 diabetes. Additionally, it has already been reported association of some HLA alleles with GDM, suggesting a possible immunological feature for GDM. In this study, we compared the transcription profiling of PBMCs of 18 GDM patients and 10 healthy pregnant women. Total RNA samples were hybridized to AgilentÒ 4 44 K oligo microarrays encompassing the whole human functional genome. Differentially expressed mRNAs were obtained by Rank Products analysis, which classifies transcripts according to their fold change values. The hierarchical clustering of mRNAs and samples were performed using the Cluster program, dendrograms and spatial representations of mRNAs were constructed using the TreeView software. A total of 731 differentially and significantly expressed mRNAs were observed when GDM patients were compared to controls. Molecular functions and biological pathways, as analyzed by the DAVID database, identified 130 biological processes and nine signaling pathways (P < 0.05). Several induced and biologically relevant transcripts were associated with the HLA system, including HLA-DRB6, DQB1, DQB2, DOA and DQA2. The modulation of these transcripts in GDM patients reinforces the hypothesis of a deregulation of HLA class II genes in GDM patients, deserving further attention.
P103
HIGH RESOLUTION HLA ALLELE FREQUENCIES OF STEM CELL DONORS IN MEXICO. GENETIC DIVERSITY AND ITS RELEVANCE TO IMPROVE UNRELATED DONOR SEARCHES.
Hilario Flores-A 1,2, Carmen Alaez 1,2, Diego Sanchez 2, Andrea Munguia 1, Araceli Rodriguez 1, David Garcia 1, Clara Gorodezky 1,2. 1 InDRE, Secretary of Health, Mexico City, D.F., Mexico; 2 Fundación Comparte Vida, A.C., Mexico City, D.F., Mexico. Aim: DNA-based registry typing is commonly done by SSOP and only a few registries employ sequencing at the time of recruitment, allowing to understand allele and haplotype frequencies and evaluating the size and composition of the registry, to assess the likelihood of finding matched donors. We explored the impact of the increasing number of HLA alleles in the genetic diversity of Mexicans, which will in turn impact the possibility of finding a donor for patients with a similar ethnic background. Methods: Healthy Mexicans enrolled at the DONORMO-The Mexican Unrelated Bone Marrow Donor Registry, born in different geographic areas of Mexico were HLA typed at the allele level, for A⁄, B⁄, C⁄, DRB1⁄, DQB1⁄ loci. DNA samples were sequenced in an ABI Prism 310 or in a 3500 sequencer. The SPSS 12 software was used for analysis. For A locus, 127 individuals were typed; 523 for B, 81 for C; 952 were SBT for DRB1 and 3343 for DQB1. Results: A total of 273 common and well documented alleles were detected; 91 were unique observations; 40 at A⁄ locus; 26 at B⁄; 8 at C⁄; 14 at DRB1⁄ & 3 at DQB1⁄. 65 different A⁄ alleles are present, of which 32 are A2 different variants: A⁄02:01; A2:06; A2:05; (13.4–5.5%) prevail. The prevalent B⁄ alleles were 49:01; 35:12; 35:17; 15:15; 14:02; 14:01;39:06; 39:05; 40:05; 40:08; 50:01; 52:01 51:02; 53:01; (>1.5%). Only 10 C⁄ alleles are frequent: 04:01, 07:02; 01:02; 03:04; 07:01; 16:01, 03:06; 5:01; 08:01; 08:02; (>2.5%). 16 frequent DRB1⁄ alleles are present: 04:07; 08:02; 14:06; 01:02; 04:02; 04:11; 04:04; 19:01; 14:02; 07:01: 03:01; 16:02; 01:03; 04:04; 10:01; 15:03; 15:01, 15:03 (1.8%). Unique observations were found at DQB1⁄ for 06:11; 03:08; 02:04. Mediterranean, Amerindian, Spanish, Caucasian, Semitic and African influences are clear. Conclusion: The molecular variation shown, and a one-step typing strategy using SBT since donor recruitment, is highly relevant to optimizing donor search algorithms in HSCT, for identification of potential matches, and in understanding the genetic background of this highly heterogeneous country. The data are also very useful for applications that depend on allele prevalence information such as: resolution of ambiguity during HLA typing, research and clinical practice, and the management of the general data, in the face of an ever growing list of recognized allele sequences.