Mahsa Rahmani; Hamid Reza Khorasani; Monireh Golpour; Ali Shabestani Monfared; Hosein Nattaj; Saeeid Abedian; Amrollah Mostafazadeh
Abstract
Background: The human leukocyte antigen (HLA) matching between organ donor and recipient is an acceptable strategy in clinical transplantation since 1964. However, in bone marrow transplantation, finding matched donors is often problematic. Thus new method for down regulation of HLA can be an alternative ...
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Background: The human leukocyte antigen (HLA) matching between organ donor and recipient is an acceptable strategy in clinical transplantation since 1964. However, in bone marrow transplantation, finding matched donors is often problematic. Thus new method for down regulation of HLA can be an alternative strategy to solve this problem. Objective: To examine the effect of serum starvation on HLA class I expression in human peripheral blood mononuclear cells (PBMCs). Methods: PBMCs were cultured in RPMI-1640 supplemented with 10% FBS (non-starved cells) as well as in medium only (starved cells) for 16, 24, 48, 72, 96h under standard cell culture conditions. The pattern of cell death and HLA class I expression was determined by flowcytometry. Antigenicity of the starved PBMCs was evaluated in a one-way mixed lymphocyte culture by MTT assay. Results: Mean fluorescence intensity (MFI) of different indicated starved PBMCs gradually decreased and this reduction was stable after 96h of re-feeding with medium containing FBS. Under serum starvation condition, PBMCs showed apoptotic cell death pattern. There was a linear correlation between percentages of cells, which exhibited the late apoptosis death pattern and serum starvation period (r=0.88, p<0.01). Surprisingly, the starved PBMCs lost their stimulatory property in mixed culture with allo-reactive lymphocyte. Conclusions: Membrane HLA class I expression could be stably reduced in 96h starved human PBMCs culture condition, decreasing their allo-reactivity while their viability rate is enough for possible clinical application.
Shirin Farjadian; Nasrin Kiyanimanesh; Abbas Abbaszadegan; Mehrdad Lotfazar
Abstract
Background: Papillon-Lefevre Syndrome (PLS) is a rare autosomal recessive disorder characterized by diffused palmoplantar keratoderma and severe periodontitis. Increased susceptibility to infections due to impairment of the immune system is considered to be involved in pathoetiology of this disease. ...
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Background: Papillon-Lefevre Syndrome (PLS) is a rare autosomal recessive disorder characterized by diffused palmoplantar keratoderma and severe periodontitis. Increased susceptibility to infections due to impairment of the immune system is considered to be involved in pathoetiology of this disease. Objective: According to the crucial function of HLA molecules in immune responses and association between certain HLA class I alleles and some periodontal or skin diseases, this study was designed to evaluate the relation of HLA class I genes and PLS. Method: HLA class I genes were typed by PCR-SSP (Polymerase Chain Reaction with Sequence Specific Primers) method in eight Iranian PLS patients and 89 healthy controls. Results: The results showed no sig-nificant difference between the patients and controls. Moreover, identical haplotypes or genotypes were also observed among PLS patients and their healthy siblings. Conclu-sion: It seems that further genes are involved in genetic susceptibility to PLS. However the results of this study showed no significant association between HLA class I genes and PLS, molecular analyses of killer immunoglobulin-like receptors (KIRs) and MHC class I chain-related gene A and B (MICA/B) in PLS may clear many obscure points about the genetic factors involved in these diseases.
Andisheh Ghashghaie Mansour; Seyyed Hamidollah Ghaffari; Kamran Ali-moghadam; Ardeshir Ghavamzadeh
Abstract
Background: HLA compatibility between transplant donor and recipient is one of the major determinants of transplant outcome. Objective: To determine HLA class I by PCR- Sequence-Specific Oligonucleotide Probe (PCR-SSOP) in cord blood donors. Methods: Genomic DNA of 142 cord blood samples registered ...
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Background: HLA compatibility between transplant donor and recipient is one of the major determinants of transplant outcome. Objective: To determine HLA class I by PCR- Sequence-Specific Oligonucleotide Probe (PCR-SSOP) in cord blood donors. Methods: Genomic DNA of 142 cord blood samples registered at the Cord Blood Bank of Iran at Hematology, Oncology, and Bone Marrow Transplantation Research Center, was prepared and HLA class I was determined by the PCR-SSOP. Results: A total of 284 HLA-A alleles was identified of which A*02 and A*24 were the most common. Among 284 HLA-B and HLA-C alleles, B*35, B*51, Cw*4 and Cw*12 were the most frequent alleles in the studied population. Conclusion: Amplification of HLA loci with PCR-SSOP has proved to be a reliable method for HLA-A, -B and -C genotyping.
