Objective: The Human Leukocyte Antigen (HLA) Class II is the major histocompatibility complex surface glycoproteins of humans responsible for presenting exogenous antigenic peptides which help direct specificity of immune response. In immune-cell therapy, the HLA allelic variants are of particular importance as they determine the successful activation of target cells that results to a desired therapeutic response. However, HLA Class II exhibits high polymorphism and has variable distribution in population, constituting these so-called allelic variants. Specifically, the HLA Class II DRB1 is considered the predominant locus among Filipinos. This research aimed to identify the presence of HLA Class II DRB1 allelic variants in the stem cell samples of ten (10) Filipino cancer patients by reverse transcription polymerase chain reaction (RT-PCR) amplification. Method: This study employed a PCR-based HLA Class II typing to identify the HLA Class II DRB1 allelic variant in Filipino cancer patients. Design of forward and reverse primers for HLA Class II DRB1, optimization of PCR conditions for amplifying HLA Class II DRB1, and identification of HLA Class II DRB1 allelic variants from samples by sequencing and database comparison were conducted. Results: PCR optimization showed that optimum annealing temperature for HLA DRB1 was 58.8°C with 1 mM MgCl2. PCR amplification of HLA DRB1 from ten anonymized cancer patient samples and DNA sequencing revealed that Patients 1, 2, 5, 8, 9, and 10 harbor HLA DRB1 allelic variants, particularly, the HLA DRB1*04:06:01, HLA DRB1*12:01:01, HLA DRB1*0813, HLA DRB1*04:05:01, HLA DRB1*09:01:02, and HLA DRB1*16:02:01, allelic variants, respectively. Conclusion Using the designed primers and optimized RT-PCR protocol, HLA information derived from six out of ten patient samples can be used for further applications in developing personalized or generic antigenic peptides such as dendritic cell cancer vaccine.
HLA Antigens

No abstract available.
HLA-DR Antigens*

No abstract available.
HLA Antigens* ; Humans

No abstract available.
HLA-D Antigens* ; Humans*

No abstract available.
HLA-DR Antigens* ; Keratinocytes*

Ankylosing spondylitis has been shown to be highly associated with HLA-B27 in Caucasian patients. This is also present in other ethnic groups. This study was conducted to determine the frequency distribution of HLA antigens in Filipinos and determine the association of ankylosing spondylitis. Twenty patients satisfying the criteria for ankylosing spondylitis and 192 unrelated controls were HLA-A and B typed. Blood from these subjects were typed using NIH lymphocyte microcyto-toxicity method. Of the unrelated controls, the frequencies of HLA A9 (w24), B40, A1, B5, Bw22 were increased and B13, B18, Bw35 were decreased. B27 had a frequency of 5.2%. A very significant high frequency of B27 (90%) was found in patients with ankylosing spondylitis with a very high relative risk of 163. A11 had a frequency of 55% with a relative risk of 3.37 which was not significant while B18 had an 18% frequency and a relative risk of 10.5 which was significant. This study reaffirms the high degree of association of ankylosing spondylitis with HLA B27 and suggests than B18 may be an additional genetic marker for this disease.
HLA Antigens ; Spondylitis, Ankylosing

BACKGROUND: HLA proficiency survey in Korea started in 1996 and the results of the survey were last reported in 1999. In this report, we summarized the results of the survey performed in recent 2 years. METHODS: A total of four proficiency surveys were performed, in which 54-59 laboratories participated. Each survey included 3 tests for HLA class I (serology and DNA) and class II (DNA) typing and 6 for HLA crossmatch test (3 cells x 2 sera). RESULTS: Overall concordance of serologic typing was 99.5% (436/438) for HLA-A, 95.7% (419/438) for HLA-B, and 94.8% (199/210) for HLA-C. The antigens assigned incorrectly by more than 5% of the participating laboratories were B54 (10.3%), B55 (10.3%), B27 (5.4%), Cw6 (22.9%), and C-blank (5.7%). Overall concordance rates of DNA typing were 99.7% (393/394) for HLA-A, 99.8% (415/416) for HLA-B, 100% (156/156) for HLA-C. Correct assignment of HLA-DRB1 and -DQB1 was reportred by 99.7% (98.1-100%) and 99.2% (88.9-100%) for generic and 99.2% and 98.1% (80-100%) for allelic level, respectively. Most laboratories (93.5-97.9%) were using sensitive methods of crossmatch such as T-long, T-AHG, and flowcytometry. The proportion of laboratories evaluated as unacceptable was on the average 3.1% of total laboratories. CONCLUSIONS: The rate of correct identification of HLA antigens was higher this time than in the previous survey in 1999. The rate of unacceptable crossmatch was also low enough to be satisfactory. It is thought that the proficiency survey has contributed to the high quality of HLA tests in the participating laboratories and should be continued to maintain the proficiency in Korea.
DNA Fingerprinting ; Histocompatibility Testing* ; HLA Antigens ; HLA-A Antigens ; HLA-B Antigens ; HLA-C Antigens ; HLA-DRB1 Chains ; Korea

Experimental study for extracting DNA and determining HLA of I and II class by PCR-SSO was carried out. The percentage of allele frequency was estimated in comparison with that was reported in 1991 conference. Result showed no difference in locus A and DR12. Locus A demonstrated the highest percentage with some differences of 2-4%. In Locus DR12 with the highest percentage of differences of 3%. Highest difference was estimated in locus B. In this study, B15 has 25.7% (in comparing with 0%). B62 had 0% (in comparing with 10.1%), B75 6.7% (in comparing with 0%), B7 12.2% (in comparing with 0%)
DNA ; HLA Antigens ; Gene Frequency

To determine the possible participation of genetic factors in the pathogenesis of Takayasu's arteritis and to investigate an association between HLA antigens and the disease, we performed HLA typing in twenty two patients confirmed by clinical findings and aortography, and in fifty normal Koreans as controls. HLA-A, B,C and DR antigens were tested by standard microlymphocytotoxicity method with HLA antisera, which were supplied by UCLA Tissue Typing Laboratory. The results were as follows: 1) Frequent antigens of HLA-A locus in patients were A 2(54.5%), Aw 33(31.8%), Aw 24(27.2%) and A26(13.6%) in decreasing order, and Aw 33 was more frequent in patients than in normal controls(18.0%)(relative risk: 2.1). 2) Frequent antigens of HLA-B locus in patients were Bw61(31.8%), Bw44(31.8%), Bw62(22.7%) and Bw52(13.6%) in decreasing order, and Bw61 was more frequent in patients than in normal controls(10%)(relative risk : 4.2). 3) Frequent antigens of HLA-C locus in patients were Cw3(54.5%), Cw6(50.0%) and Cw1(22.7%) in decreasing order. 4) Frequent antigens of HLA-DR locus in patients were DR6Y(36.4%), DR2(31.8%), DRw9(27.2%), DR4(27.2%) and DR28(22.7%) in decreasinng order. In MT system MT 3 was more frequent in patients(54.5%) than in normal controls(31.6%)(relative risk : 2. 6). However, the difference of HLA antigen frequencies between patients and normal controls was not statistically significant, and the association of specific HLA antigens with Takayasu's arteritis requires further studies to be confirmed.
Aortography ; Histocompatibility Testing ; HLA Antigens* ; HLA-A Antigens ; HLA-B Antigens ; HLA-C Antigens ; HLA-DR Antigens ; Humans ; Immune Sera ; Takayasu Arteritis*

BACKGROUND: HLA proficiency survey was started in 1996 in Korea, and the results of the 1996-1998 surveys were reported previously. Here, we report the results of the surveys performed in recent three years (2000-2002). METHODS: Six surveys were carried out with the participation of 52-54 laboratories. For each survey, 3 peripheral blood samples and 2 sera were distributed for 3 HLA class I serology, 3 HLA class I DNA, 3 HLA class II DNA, 6 HLA crossmatch, and 3 PRA tests. RESULTS: Overall consensus of serologic typing was similar to the results of the previous survey: HLA-A 93.5%, HLA-B 88.3%, and HLA-A, B 82.7%. There were an increasing number of the laboratories that were using DNA typing for HLA-DR (51 laboratories, 94%) and HLA-A and B (26 laboratories, 48%). Overall consensus of DNA typing was very high: HLA-A 100%, HLA-B 99.1%, HLAC 97.9%, HLA-DRB1 low/high resolution 99.2/99.0%, HLA-DQB1 low/high resolution 99.3/97.5%. HLA crossmatch (T cells) was reported by 44-49 laboratories, and the use of sensitive methods was increased: AHG 33 laboratories and flow cytometry 7 laboratories. For incompatible (positive) crossmatches, 4.9% (0-14.3%) of cytotoxicity tests and 7.1% (0-16.7%) of flow tests were reported as negative. PRA was reported by 5 laboratories only. CONCLUSIONS: The use of DNA tests for HLA typing and AHG or flow cytometry methods for HLA crossmatch tests has much increased compared to the previous report. A continuous survey program would play an important role in the standardization and maintenance of laboratory proficiency in histocompatibility testing in Korea.
Consensus ; DNA ; DNA Fingerprinting ; Flow Cytometry ; Histocompatibility Testing* ; HLA-A Antigens ; HLA-B Antigens ; HLA-DR Antigens ; HLA-DRB1 Chains ; Korea