한국 von Willebrand병 환자의 von Willebrand 인자 유전자 돌연변이 조사

BACKGROUND: We intended to find the mutations of von Willebrand factor (VWF) gene as the most important contributing factor of von Willebrand disease (VWD) in Korean patients. METHODS: In 40 known vWD patients mutations of vWF gene were sought by direct sequencing of PCR products targeting exons 18, 19, 20, 26, 28 and 52 frequently implicated as the locations of mutation. For factors other than VWF gene contributing to VWD phenotype, we tested ABO blood group and measured ADAMTS13 activity in VWD patients. RESULTS: Twenty-seven cases (67.5%) were type 1 vWD, 3 cases (7.5%) type 3, and 5 cases (12.5%) type 2A. Three cases were type 2A or 2B (7.5%) and 2 cases were suspected to be type 2N (5.0%). Among them six candidate missense mutations were found: V1279I, R1306W, R1308C, and V1316M were previously reported in type 2B and type 1 vWD, and C858W and T1477I were novel findings. All patients were heterozygotes. Blood group O was overly represented in VWD patients, while ADAMTS13 activity of the patients was not significantly different from that of normal control. CONCLUSIONS: Mutation of VWF gene detected by genetic studies can significantly improve the diagnostic accuracy, especially in subtype assignment of VWD. Two novel mutations, C858W and T1477I associated with VWD were found and expected to contribute to the elucidation of its pathophysiology.ope

Annual Report on External Quality Assesment in Hematology in Korea (2003)

Four trials of external quality assessment in diagnostic hematology were performed in 2003 with about 430 participating laboratories in Korea. We performed quality assessment for white blood cell count, hemoglobin, red blood cell count, platelet count, white cell differential count, red blood cell morphology and coagulation test. The response rate was more than 95%. The performance of quality assessment appeared to be gradually improved year by year.ope

A Case of Heparin Induced Thrombocytopenia with Circulating Anti-PF4/Heparin Antibody

Heparin-induced thrombocytopenia (HIT) is usually caused by anti-platelet factor 4 (PF4)/heparin antibodies, leading to intravascular platelet activation. Circulating anti-PF4/heaprin antibody (IgG, IgA and IgM) was detected by ELISA (Asserachrom HPIA, Diagnostia Stago, Asnieres, France) in a 61-year-old man with coronary artery disease and dyspnea on exercise. He had undergone a coronoary angiography using 2,000 unit of heparin before a procedure. On admission, laboratory testing revealed a platelet count of 296×109/L and aPTT (activated partial thromboplastin time) of 38.1 sec. The fall in the platelet count was progressive, resulting in 42% and 53% of platelet counts on the 6th and 12th days after intravenous heparin administration, respectively. He was discharged after coronary artery bypass graft. On discharge, platelet count was normalized to be 212×109/L. On the 7th day after dischage, anti-PF4/heaprin antibody was detected by ELISA (Asserachrom HPIA, Diagnostia Stago, Asnieres, France).ope

A novel fibrinogen variant (fibrinogen Seoul II; AαGln328Pro) characterized by impaired fibrin α-chain cross-linking

We report a novel fibrinogen variant (fibrinogen Seoul II), which has a heterozygous point mutation from CAA to CCA leading to AαGln328Pro. The mutation site is among several glutamine residues that serve as α-chain cross-linking acceptor sites. Fibrinogen Seoul II was found in a 51-year-old male patient and his family in Seoul, Korea. The patient was diagnosed with myocardial infarction at age 43. Eight years later he was admitted to the emergency room due to recurrence of the disease, where he expired under treatment with tissue plasminogen activator (t-PA). Fibrin polymerization curves, made using purified fibrinogen from the patient's relatives, showed a decreased final turbidity, suggesting Seoul II fibrin clots are composed of thinner fibers. This supposition was verified using scanning electron microscopy. Alpha-polymer formation by the mutant fibrinogen upon thrombin treatment in the presence of factor XIII and calcium was distinctly impaired. This result confirms that the residue Aα328 plays a pivotal role in α-chain cross-linking.ope

Annual Report on External Quality Assessment in Diagnostic Genetics in Korea (2003)

The importance of quality control for dramatically growing genetic tests continues to be emphasized with increasing clinical demands. Diagnostic genetics subcommitee of KSQACP performed two trials for cytogenetic study in 2003. Cytogenetic surveys were performed by 33 laboratories and answered correctly in most laboratories except some problems in nomenclature and analysis for FISH and complex cytogenetic abnormalities in neoplasia. The molecular genetic test surveys include M. tuberculosis, HBV, HPV, leukemia/lymphoma, ApoE genotyping, Duchenne muscular dystrophy, myoclonic epilepsy and ragged red muscle fibers, and spinal and bulbar muscular atrophy. HPV, myoclonic epilepsy and ragged red muscle fibers, and spinal and bulbar muscular atrophy were the first challenge of the genetic survey. Molecular genetic survey showed excellent results in most participants, however, HPV tests should be improved by quality control in a few laboratories. External quality assessment program for cytogenetic analysis could be helpful to give participants many chances of continuous education and of interesting case materials.ope

Evaluation of the Abbott Cell-Dyn Sapphire Hematology Analyzer

Background :The performance of Cell-Dyn Sapphire (Abbott Diagnostic, USA) was compared to the Bayer Advia 2120 (Bayer Diagnostics, USA), Sysmex XE-2100 (Sysmex Corporation, Japan), and reference microscopy. Method :Three hundred samples for routine CBC and WBC differentials were randomly chosen for a comparison analysis. The Cell-Dyn Sapphire system was evaluated according to the linearity, imprecision, inter-instrument correlations, and white blood cell differential. Result :The CBC parameters (WBC, RBC, hemoglobin and platelet) showed a significant linearity with correlation coefficients greater than 0.99 (P Conclusion :These results demonstrate that the Cell-Dyn Sapphire has a good linearity, an acceptable reproducibility, a minimal carryover, and a comparable performance with the sysmex XE-2100 and Advia 2120.ope

A Patient with IgA Monoclonal Gammopathy Presenting as Myelomatous Pleural Effusion with Axillary Node Involvement

Multiple myeloma is a cancer of plasma cells that produce monoclonal immunoglobulin, and the neoplastic plasma cells typically accumulate in the bone marrow with occasional involvement of other organs. Pleural effusion that is associated with multiple myeloma has been infrequently reported (＜6%) and myelomatous pleural effusion is extremely rare (＜1%). A 73-year-old woman was admitted to the department of dermatology for skin lesions on both arms and both ankles. A chest radiograph taken on admission showed a nodular lesion in the right upper lung and pleural effusion. Analysis of the pleural fluid revealed many atypical plasma cells, elevated levels of IgA (27.95g/L) and lambda light chain (9.16g/L), and monoclonal IgA-lambda paraprotein on immunofixation. The serum concentrations of IgA were elevated (33.08g/L) while the concentrations of IgG and IgM were decreased. Bone marrow aspirate smears contained increased levels of immature-appearing atypical plasma cells. This is only the third case of myelomatous pleural effusion that has been reported in Korea.ope

Relationship between Myelodysplastic Syndrome and Epstein-Barr Virus or Human Parvovirus Bl9 Infection

Background :The mechanisms responsible for the disturbed hematopoiesis in myelodysplastic syndrome (MDS) include the expansion of abnormal clones, defects in cellular differentiation and he perturbation in the production of hematopoietic regulatory factors. Recently, viral infection such as immunodeficiency virus is known to induce myelodysplasia. And viral infection evokes the production of several cytokines. Therefore, abnormal production of cyrokine may be a potential candidate for the pathogenesis of MDS after viral infections such as Epstein-Barr virus (EBV) and human parvovirus B19. Method :We investigated bone marrow aspiration slides from 17 patients with MDS referred for the bone marrow study, over a period from January, 1992 to April, 1996. To clarify the contribution of EBV and human parvovirus B19 infections to the pathogenesis of MDS, DNA-PCR for EBV and human parvovirus Bl9 was used. Result :The EBV and human parvovirus B19-PCR results were all negative in 17 patients with MDS. Conclusion :EBV and human parvovirus Bl9 infections may not be associated with the major pathogenesis of MDSope

Studies on E & EAC rosette formation and surface immunoglobulin of human peripheral lymphocytes

의학과/박사[한글] 인체의 말초혈액 임파구는 면역기능의 특성에 따라 흉선에서 유래된 T세포와 bursa 혹은 이에 해당하는 조직에서 유래한 B세포로 대별 할 수 있다(Roitt등, 1969). T세포는 면양 적혈구와 결합하여 E로��erythrocyte rosette)을 형성하는 특성이 있는데 이것은 흔 히 T세포의 marker로 이용된다(Brain등, 1970; Coombs등, 1970; Jondal등, 1972). B세포는 면역 형광법으로 세포 표면에 있는 면역글로부린(immunoglobulin)을 직접 인지할 수 있거나(Grey등, 1971; Fu등, 1974) EAC(erythrocyte-antibody-complement) 로��형성을 이용하여 보체에 대한 수용체를 증명 할 수도 있다(Bianco등, 1770: Mendes등, 1973). 이러한 surface marker들은 임파구증식성 종양의 기원을 구별하는데 도움이 된다(Wilson 및 Nossal, 1971). 임파구 증식성 종양중에서도 만성 임파구성 백혈병은 B세포 기원임이 밝혀졌으나(Foulis등, 1973; Piessens등, 1973) 급성 임파구성 백혈병의 경우에는 아직도 그 기원이 분명치 않다. 이에 저자는 급성 백혈병 환자를 대상으로 Iymphoblast의 surface marker를 관찰하므로써 그 기원의 이해를 돕고저 하였으며 아울러 정상 한국인의 말초혈액 임파구의 E 및 EAC로��형성율과 세포표면 면역글로부린(sIg) 양성율을 설정하는 동시에 그 실험방법에 따르는 차이를 검토하고저 본 연구를 시행하였다. 실험재료로는 건강인 65명과 1980년 3월∼8월 사이에 연세의료원에 내원한 급성 백혈병 환자 25예로부터 말초혈액을 채취하여 실험하였다. 임파구 및 blast를 포함한 단핵구세포(mononuclear cell)의 분리는 Ficoll-Mypaque방법을 이용하였다(Kim등, 1977). E로��형성율은 Holm등(1975)의 방법, EAC로��형성율은 Mendes등 (1973)의 방법에 의하였다. sIg양성율은 Davey 및 Gottlieb(1974)의 방법을 따라 직접 면역형광법을 이용하였다. 그 실험성적은 다음과 같다. 1. 건강성인(20∼45세): 57명에서의 E 및 EAc로��형성율은 각각 평균 65.3±11.O% 및 19.2±6.9%이었고 남녀의 유의한 차이는 없었다(P>0.05) 소아(4∼15세) 15명을 대상으로 실험한 결과 E 및 EAC로��형성율은 각각 평균 61.6±10.2% 및 17.8±7.37이었고 성인과 비교할때 유의한 차이는 없었다(P>0.05). 2. 건강성인(17∼35세) 22명에서의 sIg양성율은 평균 14.7±6.17이었다. 이중 10명을 대상으로 monovalent antisera를 사용하여 실험한 결과, IgG, IgM, IgD 및 IgA 양성 임파구는 각각 평균 6.7%, 7.9%, 4.2% 및 1.3%이었다. 3. B세포의 주된 marker인 EAC로��형성율과 sIg양성율 사이엔 어느 정도 직선적인 순상관 관계가 성립되나 반드시 일치하지는 않았다(γ=+ 0.59). 4. E로��형성율과 sIg양성율은 실험방법에 따라 차이가 있는데 E로��형성율은 4℃, 18시간조건에서 가장 높았고 sIg양성율은 30분보다는 18시간 방치시 더욱 높았다. 5. 급성 임파구성 백혈병 환자 14예중 T marker를 갖는 예가 5, T와 B marker를 모두 갖는 예가 1, T와 B marker를 모두 갖지 않는 예가 8이었다. 이상의 결과를 종합하면 정상 한국인의 말초혈액 임파구의 E 및 EAC로��형성율은 외국인과 비슷하며 실험방법에 따라 차이가 있음을 알았다. 또한 형태학적으로는 같은 급성 임파구성 백혈병 환자라도 면역학적 특성은 서로 다른 이질적인 성격을 보이며 T와 B marker를 갖는 예보다는 아무 marker도 갖지 않는 예가 더욱 많음을 알 수 있었다. 이러한 면역학적 분류를 더욱 많은 예에서 시도하여 보면 임상적 가치가 있을 것으로 사료된다. [영문] Lymphocytes derived from human bone marrow (B lymphocytes) express on their surface immunoglobulin(sIg) determinants (Pernis et al, 1971), and receptor for complement(Bianco et al,1970). Thymus derived lymphocytes (T lymphocytes) lack the above surface markers but bind steep red blood cells in a rosette formation (Jondalet al, 1972). Determination of lymphocyte subpopulations usually in connection with functional characterization is valuable in diagnosis of human primary or secondary immunodeficiency states(Gajl et al,1973). Moreover these recognizable surface markers have facilitated the classification of lymphoproliferative diseases for cell origin and stage of differentiation. Chronic lymphocytic leukemia(CLL) represents a neoplasm of B cell origin in most instances(Ross et al, 1973). In contrast, no homogeneous pattern has s? far emerged from the study of the surface properties of blast cells from patients with acute lymphoblastic leukemia (ALL). The present investigation is attemped to determine the surface markers of lymphoblasts in ALL and is also studied in normal subjects for reference value in Korea. In addition, the author reviewed some technical aspects of determination of surface markers. Mononuclear cells from peripheral blood were isolated by Ficoll-Hypaque technic (Kim et al, 1975). T and B lymphocytes were identified by three surface markers: cells forming rosettes with sheep red blood cells(E rosette) as a measure of T lymphocytes(Holm et al, 1975) : surface immunoglobulin by direct immunofluorescence (Davey and Gottlieb, 1974) and lymphocytes with receptor for complement observed by the EAC rosette method using human RBC treated with rabbit serum and guinea pig complement (Menses et al, 1973) as that of B lymphocytes. The results are summarized as follows: 1. The mean value of I and EAC rosette formation of lymphocytes from peripheral blood in 50 normal subjects was 65.3±11.0% and 19.2±6.9%, respectively. There was no statistically significant difference between females and males, and between adults and children. 2. The mean value of surface immunoglobulin positive lymphocytes from peripheral blood of 22 normal adults was 14.7±6.1%. The percentage of lymphocytes reacted with monospecific antiserum specific for IgM or IgG were higher than for IgD or IgA. 3. The two surface markers for B lymphocytes, EAC rosette formation and positive surface immunoglobulin were correlated proportionately to some degree(r=+0.59). 4. T7e determination of E rosette formation and sly on lymphocytes varied in different condition. The percentage of lymphocytes forming I rosettes in overnight incubation was higher in 4℃ than in room temperature or 37℃. The percentage of positive sIg lymphocytes incubated at 4℃ or 37℃ was higher in overnight condition than in short time (30 min) incubation. 5. Of 14 patients with acute lymphoblastic leukemia, 5 had T marker, 8 had neither T nor B(Null), and 1 had both(Double). These results indicate that ALL represents a heterogeneous proliferation of lymphocytes and suggest that ALL can originate in uncommitted cells that have net differentiated, or that lymphocytes cell markers may he lost or retained during malignant transformation. Finally, there is some hope that this immunological classification of acute lymphoblastic leukemia will prove to be of practical value and is feasible as a routine pretreatment investigationrestrictio

Changes of platelet parameters determined by the Bayer ADVIA™ 120 with EDTA sample age

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