HLA genes constitute a highly polymorphic multigene system. In the present study, HLA-B oligonucleotide chips were manufactured by using a set of sequence-specific oligonucleotide probes derived from polymorphic regions in exon 2 and exon 3 of HLA-B gene spotted by microarrayer onto the aldehyde modified glass slides. In addition, the sequenced HLA-B gene clones used as standard samples were amplified from exon 2 and exon 3 by PCR. Together with the correct hybridization and wash conditions, the PCR products were bound with the array probes on the chip, and the hybridization patterns were transformed to HLA-B genotypes. The results showed that the genotypes of standard samples by the HLA-B oligonucleotide chips were completely identical with the sequenced clones. In conclusion, the oligonucleotide chip method presented here for HLA-B genotyping is a rapid, accurate, sensitive and attractive high throughput biochemical way.
Genotype ; HLA-B Antigens ; classification ; genetics ; Humans ; Oligonucleotide Array Sequence Analysis ; Sequence Analysis, DNA

This study was purposed to investigate the nucleotide sequences of a novel HLA-B*15:124 allele and its molecular mechanism. The genomic DNA from whole blood was extracted by using commercial DNA extraction kit. The sequences of exon 2, 3 and 4 of HLA-B locus in the proband were amplified by PCR with group-specific primers, the PCR products were purified by enzymes digestion, then exon 2 to 4 of HLA-B locus for both orientations was sequenced. The results showed that 2 HLA-B alleles of proband were gained after amplification and sequencing of group-specific primers, among them one was a B*40:03, another was a novel allele. After BLAST analysis, the novel allele showed nucleotides different from HLA-B*15:52 in exon 3 at nucleotide position 427 A > T and 440 G > T which resulted in amino acid change from Thr to Ser at codon 143 and Trp to Leu at conon 147. It is concluded that a novel HLA-B allele has two different nucleotides. This HLA-B allele is identified and has been officially named B*15:124 by the WHO Nomenclature Committee.
Alleles ; Base Sequence ; Exons ; Female ; HLA-B Antigens ; classification ; genetics ; Humans ; Sequence Analysis, DNA

OBJECTIVETo identify a novel HLA allele in Chinese population.

METHODSA new HLA-B allele was initially detected by an usual PCR-SSP and PCR-SSOP in routine typing HLA allele. Sequence-based typing (SBT) was used to identify and analysis the difference between the new allele and HLA-B 4409 allele.

RESULTSThe HLA-B exon 3 nucleotide sequence of the novel allele was different from all other known alleles. The allele had 3 nucleotides replaced of the closest matching B 4409 allele at nt538(G>C), nt539(A>T) and nt540 (C>G) in exon 3, resulting in an amino acid change from D(GAC) to L(CTG) at codon 180.

CONCLUSIONA novel HLA allele was confirmed by the SBT and it was officially designated as HLA-B 4446 by WHO Nomenclature Committee for Factors of the HLA System in September,2005.

Alleles ; Base Sequence ; HLA-B Antigens ; classification ; genetics ; Humans ; Male ; Molecular Sequence Data ; Sequence Analysis, DNA

Fifty-two Korean patients with Behcet's syndome were typed for HLA antigens. 52 apparently healthy Korean subjects were used as controls; 42 for HLA-A, B, C and all 52 for HLA-DR typing. HLA-B5 and DRw8 presented significantly high frequencies in all patiens. According to Shimizu's classification, HLA-B5 and DRw8 were significantly increased in the complete type; B5 in the incomplete type; DR3 in the suspected-possible type. According to Lehner's classification, HLA-DR3 was significantly increased in the neurological type; B5 in the ocular type; B5 in the ocular type. We confirmed the association of HLA-B5 with the severity of Beh et's syndrome. A relation might exist between DRw8, DR3 and Behcet's syndrome.
Adolescent ; Adult ; Behcet Syndrome/*immunology ; Female ; HLA Antigens/*analysis/classification ; Human ; Korea ; Male ; Middle Age ; Support, Non-U.S. Gov't

To know the prognosis of Behcet's disease with HLA-Bw 51 antigen, comparison studies of 24 cases of Behcet's disease were investigated. The sex distribution was 14 males (58%) and 10 females (42%). The patients were grouped into three clinical types: the complete type with 11 cases (45%), the incomplete type with eight cases (33%) and the suspect type with five cases (22%), according to the criteria established by the Behcet's Disease Research Committee in Japan (1982). They were also divided into three ocular types according to the location of the inflammation: the anterior segment type, the fundus type and the mixed type. They were divided into 10%, 10% and 60% in HLA-Bw 51 negative group and 14%, 22% and 64% in HLA-Bw 51 positive group, respectively. Skin lesions observed in 30% of the HLA-Bw 51 negative group and 70% of the HLA-Bw 51 positive group, which was statistically significant (p < 0.01). The other general symptoms and the visual acuity between the two groups were not statistically significant (p > 0.1).
Adolescent ; Adult ; Behcet Syndrome/classification/*complications/immunology ; Eye Diseases/*etiology/immunology ; Female ; HLA-B Antigens/*analysis ; HLA-B51 Antigen ; Humans ; Male ; Middle Aged ; Prognosis ; Visual Acuity

The study was aimed to investigate the immunological characteristics and prognosis of acute myeloid leukemia (AML) M(1) and to find the main points in immunology to differentiate AML M(1) from M(2), and M(1) from ALL (proB, preB, T). Immunophenotyping was performed in 41 AML M(1) patients by three-color flow cytometry analysis using CD45/SSC gating, meanwhile the cytogenetic analysis was performed in 17 patients. 51 newly diagnosed AML M(2) patients and 58 newly diagnosed ALL patients were used as control at the same time. The results showed that the positive rate of CD33 in M(1) was 100%, which was high in sensitivity, but low in specificity; the positive rate of CD11b, CD15, MPO, CD117 in M(1) were significantly lower than that in M(2) (p < 0.05); the positive rate of T-lineage antigen in Ly + AML M(1) was higher than that in M(2) (p < 0.05); compared with ALL ProB, M(1) had high expression of HLA-DR, simultaneously myeloid antigen CD13, CD15, CD33, CD117, MPO and T-lineage antigen CD4, CD7 were all highly expressed (p < 0.05); compared with ALL PreB, M(1) had high expression of HLA-DR, CD34, meanwhile myeloid antigen CD13, CD15, CD33, CD117, MPO and T-lineage antigen CD4, CD5 were all highly expressed (p < 0.05); as compared with T-ALL, the early-phase antigen HLA-DR, CD34, myeloid antigen CD13, CD15, CD33, CD117, MPO of M(1) were all significantly highly expressed (p < 0.05). In M(1), the complete remission (CR) rate in patients with CD7 positive had no statistical difference from that in patients with CD7 negative (p > 0.05); the CR rate of patients with CD34 positive had no statistical difference from that of patients with CD34 negative (p > 0.05); CR rate in M(1) was lower than that in M(2) (p < 0.05), time to reach CR was longer, the incidence of hyperleukocytic acute leukemia was higher (p < 0.05), CR rate in hyperleukocytic acute leukemia was lower (p < 0.05). It is concluded that the myeloid antigen CD33, CD13 in M(1) are highly expressed, early-phase antigen HLA-DR in M(1) is also highly expressed, but the myeloid antigen CD11b, CD15, MPO, CD117 in M(1) are lowly expressed, T-lineage antigen CD4, CD7 in M(1) are highly expressed in the meantime. There is no definite characteristic marker in immunology to differentiate M(1) from M(2), but as the positive rate of CD11b, CD15, MPO, CD117 in M(1) are significantly lower than that of M(2), CD11b, CD15, MPO, CD117 can be used as reference indicators to differentiate M(1) from M(2). AML M(1), ALL ProB, ALL PreB and T-ALL, which are difficult to differentiate in morphology can be well seperated through the analysis of immunological phenotype. CD117 is mainly expressed in AML, which is useful for the differentiation diagnosis between AML and ALL. The prognosis of M(1) is worse than that of M(2).
Adolescent ; Adult ; Antigens, CD ; analysis ; Antigens, CD7 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; CD13 Antigens ; analysis ; CD4 Antigens ; analysis ; Diagnosis, Differential ; Female ; HLA-DR Antigens ; analysis ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; classification ; diagnosis ; immunology ; Male ; Middle Aged ; Prognosis ; Sialic Acid Binding Ig-like Lectin 3 ; Young Adult

BACKGROUND: The findings from morphologic, immunologic, and cytogenetic studies have been used for diagnosis of acute leukemia. Although research regarding the relationship between each of these methods has grown significantly, the results of research in this area is still conflicting. Therefore, a comparative analysis of immunophenotyping results with the results of morphologic and cytogenetic analysis of 339 cases of acute leukemia was performed in this study. METHODS: 339 cases were chosen from the pool of patients with acute leukemia admitted to the Chonnam National University Hospital from July 1994 to December 2000 and comparative analysis was performed on the results of immunophenotyping using flow cytometry (Coulter EPICS XL Flow Cytometer) and morphological findings. Further comparative analysis was performed on 185 cases that also had available data from cytogenetic analysis. RESULTS: Based on the FAB classification for the 339 cases with acute leukemia, 63.7% (216/339) was AML, 35.4% (120/339) was acute lymphoid leukemia (ALL), and 0.9% (3/339) was unclassified acute leukemia. In the AML cases, the CD13 and CD33 expression percentages were 90.0% (188/209) and 68.1% (141/207), respectively. HLA-DR expression percentages, with the exception of M3 and M3v, comprised 72.4% (118/163) and HLA-DR was expressed at 5.1% (2/39) in M3 and M3v. 85.6% (77/90) of B lineage ALL were CD10-positive. Ly+ AML were 18.6% (40/215) and My+ ALL were 35.5% (43/121). Among the cases with available cytogenetic analysis results, 56.3% (76/135) AML and 42.9% (18/42) ALL had evidence of chromosomal abnormalities. CONCLUSIONS: Immunophenotyping results are generally consistent with the findings of morphological classifications and are useful in diagnosis of some subtypes of acute leukemia cases. However, many cases showed expression of unusual markers contrary to morphological diagnosis. Specific cytogenetic findings, t(8;21)(q22;q22), were observed, especially in CD19 and CD56 positive AML.
Chromosome Aberrations ; Classification ; Cytogenetic Analysis ; Cytogenetics* ; Diagnosis ; Flow Cytometry ; HLA-DR Antigens ; Humans ; Immunophenotyping ; Jeollanam-do ; Leukemia* ; Precursor Cell Lymphoblastic Leukemia-Lymphoma

BACKGROUND: The combination of Philadelphia chromosome (Ph) and monosomy 7(-7) was rarely observed in acute lymphoblastic leukemia (ALL). With the results from immunophenotyplc and molecular analysis, Philadelphia chromosome positive ALL with monosomy 7[Ph(+)/-7] has been considered that it may be derived from neoplastic transformation at the pluripotent stem cell level. We compared the clini-cal, laboratory, and hematological findings between 5 cases of Ph(+)/-7 and 5 cases of Ph(+) without monosomy 7 [Ph (+) /N7]. METHODS: During the period from January, 1995 to December, 1996, total 72 cases of ALL were confirmed among 259 cases of hematologic malignancy with bone marrow cytogenetic analysis. Among 72 ALL cases, 5 cases of Ph(+)/-7(monosomy 7 or 7q abnormalities) were compared with Ph only or Ph without monosomy 7(ph(+)/N7] on the hematological, immunophenotypic, other laboratory, clinical findings and event ree survival (EFS) The karyotyping of the bone marrow specimens was analysed byshort-term unsynchronized culture methods such as overnight colcemid treatment and 24 hours incubation following ethidium bromide treatment. RESULTS: The mean age of Ph(+)/-7 was 30.6+/-12.8 years, and it was significantly different from that of Ph(+)/N7 (p=0.009), Four cases of Ph(+)/-7 were classified as ALL L2 subtype, and 2 cases revealed CNS involvements. Immunophenotyping was positive in CD10, CDl9, CD2O, CD22 and HLA-DR. But one case revealed e-B-lymphoid lineage with positivity in CD34, CDl3, and CD33. The response to chemotherapy and EFS was very poor in Ph(+)/-7 group, and the mean EFS was 3.2+/-1.9 months(p=0.014). All of cases showed induction on failure in chemotherapy, relapsed with bone marrow, CNS and extramedullary involvements, and expired due to sepsis. CONCLUSIONS: Ph(+)/-7 ALL had very Poor clinical course with being resistant to chemotherapy and unfavorable prognosis, revealed L2 subtype by FAB classification, and was slightly older in ages compared with Ph(+)/N7 ALL.
Bone Marrow ; Classification ; Cytogenetic Analysis ; Demecolcine ; Drug Therapy ; Ethidium ; Hematologic Neoplasms ; HLA-DR Antigens ; Hydrogen-Ion Concentration ; Immunophenotyping ; Karyotyping ; Monosomy* ; Philadelphia Chromosome* ; Pluripotent Stem Cells ; Precursor Cell Lymphoblastic Leukemia-Lymphoma* ; Prognosis ; Sepsis

This study was to clarify whether Behcet's disease (BD) could be classified into the spondyloarthropathy (SpA) complex. It was undertaken on 58 patients with BD (BD group), 56 patients with SpA (SpA group), and 3 patients who concurrently satisfied the criteria for BD and SpA (BDSpA group). The clinical parameters and known susceptible HLA antigens were compared between BD group and SpA group. In addition, 3 patients in BDSpA group were reviewed. The prevalence of definitive sacroiliitis (SI) in BD group and SpA group was 46.4% and 5.2%, respectively. However, none had a definitive SI in healthy controls. Enthesitis was observed in 3.4% of BD group and in 50% of SpA group. The patterns of eye involvement were different between these two groups. HLA-B27 was negative in all 49 patients of BD group, whereas it was positive in 67.9% of SpA group. The prevalence of HLA-B51 was 51.7% in BD group, and that in SpA group was 21.4%. One patient in BDSpA group was considered to have concurrent BD and ankylosing spondylitis (AS). Another patient was closer to AS, and the third to BD. Conclusively, it seems that BD could not be classified into the SpA complex.
Adult ; Behcet Syndrome/*classification/immunology/pathology ; Eye/pathology ; Female ; HLA-B Antigens/analysis/immunology ; HLA-B27 Antigen/analysis/immunology ; Humans ; Lumbar Vertebrae/pathology/radiography ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pelvis ; Radioactive Tracers ; Sacroiliac Joint/pathology/radiography ; *Spondylarthritis/immunology/pathology ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo study the characteristics of the virology background of HLA-A2 restricted HBcAg(18-27) epitope mutations in HBV infected patients in China.

METHOD30 HBV sequences with different genotypes from Genbank were analyzed by bioinformatics and the mismatched primers were designed for constructing a PCR-RFLP method to screen HBcAg(18-27)V/I in China. The distributions of HBcAg(18-27)V/I of 160 samples with HBV genotype B/C infection from 8 areas in China were screened and analyzed by PCR-RFLP and sequencing. The affinity of HBcAg(18-27)V/I to HLA-A0201 was analyzed through referencing the bioinformatics websites.

RESULTSWe successfully constructed a PCR-RFLP method for screening HBcAg(18-27)V/I from genotype B/C, and only 3 samples with HBcAg(18-27)V sequence were found in the 160 samples (3/160, 1.88%). The affinity of HBcAg(18-27)I to HLA-A 0201 was lower than the one of HBcAg(18-27)V through bioinformatic analysis (HLA ligand score was 123 vs 156, and the SYFPEITHI score was 22 vs 24).

CONCLUSIONThe last amino acid of most HBcAg(18-27) sequences of epidemic HBV strains in China is isoleucine, and not valine. Therefore HBcAg(18-27) sequence background in different HBV genotypes should be thoroughly considered when using it as a reference or control in immunological research about HBV.

Adult ; China ; epidemiology ; Computational Biology ; DNA, Viral ; genetics ; Epitopes, T-Lymphocyte ; immunology ; Female ; Genotype ; HLA-A Antigens ; immunology ; Hepatitis B Core Antigens ; genetics ; immunology ; Hepatitis B virus ; classification ; immunology ; Hepatitis B, Chronic ; epidemiology ; immunology ; virology ; Humans ; Male ; Mutation ; Sequence Analysis, DNA ; T-Lymphocytes, Cytotoxic ; immunology