BACKGROUND: The recent expansion of knowledge about various ABO alleles has led to the need for a comprehensive measure to cover the numerous polymorphisms dispersed in the ABO gene. A few studies have examined the diversity of the O allele compared to A or B subgroup alleles, resulting in antigenic changes. This study investigated the relationship between the serologic and molecular genetic characteristics of the O alleles in the Korean population. METHODS: One hundred and five samples from healthy blood group O subjects were selected randomly. The isoagglutinin titer was measured using a tube agglutination and gel microcolumn assay. The ABO alleles were analyzed by sequencing exons 6 and 7 of the ABO gene. When the origin of a heterozygous nucleotide sequence was ambiguous, it was separated into a single allele using mono-allele amplification or cloning. RESULTS: The median IgM isoagglutinin titer was eight. In contrast, the median IgG anti-A and anti-B isoagglutinin titers were 64 and 32, respectively. The IgG isoagglutinin titer showed a significant increase with age (P<0.0001). Six O alleles were observed in 105 blood group O populations by sequencing. The O01 and O02 alleles were common (0.57, 0.36). Three rare O alleles (O04, O05, and O06) and one novel non-deletional O allele were found. CONCLUSION: The distribution of isoagglutinin titers of blood group O and the genetic frequency of O alleles in this study would form the basis of the development and interpretation of ABO genotyping and serologic workup in the Korean population.
Agglutination ; Alleles ; Base Sequence ; Clone Cells ; Cloning, Organism ; Exons ; Immunoglobulin G ; Immunoglobulin M ; Molecular Biology ; Sequence Analysis

BACKGROUND: Measurement of the ABO antibody (Ab) titer is important in ABO-incompatible transplantation. However, to the best of our knowledge, no standard protocol or external survey program to measure the ABO Ab titer has been established in Korea. We investigated the current status of ABO Ab titer measurements at various laboratories in Korea and the impact of the protocol provided to reduce interlaboratory variations in the methods and results of ABO Ab titers. METHODS: The Korean external quality assessment of blood bank laboratories sent external survey samples with a questionnaire to 68 laboratories across Korea for the measurement of ABO Ab titers in May 2012. After 6 months, a second set of survey samples were sent with a standard protocol to 53 of the previously surveyed laboratories. The protocol recommended incubation at room temperature only and use of the indirect antihuman globulin method for the tube test as well as and the column agglutination test (CAT). RESULTS: Several interlaboratory variations were observed in the results, technical procedures, and methods selected for measurement. We found that 80.4% laboratories hoped to change their protocol to the provisional one. Additionally, CAT showed significantly lower variation among laboratories (P=0.006) than the tube test. CONCLUSIONS: Our study provides baseline data regarding the current status of ABO Ab titer measurement in Korea. The standard protocol and external survey were helpful to standardize the technical procedures and select methods for ABO Ab titer measurement.
ABO Blood-Group System/*immunology ; Agglutination Tests/*standards ; Antibodies/*analysis ; Humans ; Immunoglobulin G/analysis ; Immunoglobulin M/analysis ; Laboratories/*standards ; Questionnaires ; Republic of Korea ; Temperature

There are many serologic tests for syphilis. By means of the usual serologic tests, it is not possible to differentiate between patients who need therapy and those who are cured. In this paper I want to discuss the scientific developments and demonstrate the results of immunologic research in syphilis, which makes it possible to differentiate between treated and untreated cases.
Antibodies, Bacterial/analysis ; Chromatography, Gel ; Electrophoresis, Starch Gel ; Fluorescent Antibody Technique ; Hemagglutination Tests ; Human ; Immunoglobulin G/analysis ; Immunoglobulin M/analysis ; Syphilis/immunology* ; Syphilis Serodiagnosis* ; Treponema pallidum/immunology

AIMTo explore the role of humoral immunity in the pathophysiological process of freezing injury and the possible immune interference in the preventation and treatment of frostbite.

METHODSSevere experimental freezing injury model was made in Wistar rats( n = 20). The concentration of three types of immunoglobulin (IgG, IgA and IgM), two types of complement components (C3 and C4), and circulating immune complex (CIC) were measured respectively before and at 4h, 1d, 3d, and 5d after frostbite. At the same time, the tissue immune complex (TIC) in skeletal muscle and the contents of the red blood cell immune complex (RBC-IC) were also observed and then was the red blood cell immune adherence activity (RCIA).

RESULTSSerum IgG concentration decreased rapidly to the lowest level at 4 h after frostbite IgA concentration dropped to the nadir on 1 day after freezing. Decreases of both immunoglobulins were maintained during the 5 days after frostbite. The fate of both C3 and C4 were the same as those immunoglobulins. Freezing had rather less effect on IgM level. CIC concentration in serum, expressed as the percent of prefreezing increased rapidly and to the zenith on the 3 days post-freezing. By immunofluorescence microscopy, thin continuous linear pattern (IgG) was demonstrated along the SM on the first day post-freezing. Granular and nodular deposits (IgG) appeared along the SM as the time proceeded after frostbite. RBC-IC contents, expressed as the erythrocyte IC rosette rate, increased significantly and to the zenith on the 3 d post-freezing, while RCIA depressed to the nadir at the same time.

CONCLUSIONThe freezing frostbite is an immune complex related disease which have not been reported by others before.

Animals ; Antigen-Antibody Complex ; analysis ; immunology ; Frostbite ; blood ; immunology ; Immunoglobulin A ; immunology ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Immunoglobulins ; immunology ; Male ; Rats ; Rats, Wistar

PURPOSE: Kawasaki disease is a systemic vasculitis, and its etiology and pathogenesis are still not clear. Our study was undertaken to investigate the characteristics of the activation of B cells in the peripheral blood of Kawasaki disease (KD) patients and evidence of stimulation by superantigens. MATERIALS AND METHODS: Blood samples were obtained from three patients (2 males, 1 female) with KD, who were admitted to our Hospital, Seoul, Korea. The mean age was 1.2 years. Distribution of B cells was studied in the acute and subacute phases of KD patients. From the RNA of B cells, we obtained complementary DNA (cDNA) and performed polymerase chain reaction (PCR). To determine the oligoclonal expansion of immunoglobulin M (IgM) VH family, we cloned and sequenced the PCR products from each group and analyzed DNA. RESULTS: In the peripheral blood of acute phase patients, T cells were significantly decreased (p < 0.05), whereas B cells were significantly increased (p < 0.05). When the first PCR was done on the B cell chains, VH1 to VH6 were all found to be expressed. The number of micro gene clones obtained from 3 patients was 312, and they belonged to VH3, VH4 and VH5 family. M99686 germ line was most frequently used and the next most frequently used, were X92224/J, L21967 and L21964. A similar order was seen in patients. Among the clones, 20 sets of clones showed the same base sequence and this was frequent between VH2 and VH5. There was one set, which showed almost the same base sequence between different patients, and the homology was 99.5%. Twenty sets of clones that had the same base sequence showed high similarity to the germ line (94 - 100%). Among these, the clones that utilized the M99686 germ line were 4 sets which were most frequent. The 3-dimensional structure of one of these clones showed typical beta, sheet structure of immunoglobulin chains. CONCLUSION: The IgM transcripts expressed by the B cells in the peripheral blood of KD patients in the acute phase of the disease clearly showed an oligoclonal expansion, suggesting that KD is caused not by stimulation of a superantigen, but rather by a conventional antigen.
B-Lymphocytes/*metabolism ; Female ; Humans ; Immunoglobulin M/*metabolism ; Immunoglobulin Variable Region/*genetics ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome/*genetics/*immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

PURPOSE: Kawasaki disease is a systemic vasculitis, and its etiology and pathogenesis are still not clear. Our study was undertaken to investigate the characteristics of the activation of B cells in the peripheral blood of Kawasaki disease (KD) patients and evidence of stimulation by superantigens. MATERIALS AND METHODS: Blood samples were obtained from three patients (2 males, 1 female) with KD, who were admitted to our Hospital, Seoul, Korea. The mean age was 1.2 years. Distribution of B cells was studied in the acute and subacute phases of KD patients. From the RNA of B cells, we obtained complementary DNA (cDNA) and performed polymerase chain reaction (PCR). To determine the oligoclonal expansion of immunoglobulin M (IgM) VH family, we cloned and sequenced the PCR products from each group and analyzed DNA. RESULTS: In the peripheral blood of acute phase patients, T cells were significantly decreased (p < 0.05), whereas B cells were significantly increased (p < 0.05). When the first PCR was done on the B cell chains, VH1 to VH6 were all found to be expressed. The number of micro gene clones obtained from 3 patients was 312, and they belonged to VH3, VH4 and VH5 family. M99686 germ line was most frequently used and the next most frequently used, were X92224/J, L21967 and L21964. A similar order was seen in patients. Among the clones, 20 sets of clones showed the same base sequence and this was frequent between VH2 and VH5. There was one set, which showed almost the same base sequence between different patients, and the homology was 99.5%. Twenty sets of clones that had the same base sequence showed high similarity to the germ line (94 - 100%). Among these, the clones that utilized the M99686 germ line were 4 sets which were most frequent. The 3-dimensional structure of one of these clones showed typical beta, sheet structure of immunoglobulin chains. CONCLUSION: The IgM transcripts expressed by the B cells in the peripheral blood of KD patients in the acute phase of the disease clearly showed an oligoclonal expansion, suggesting that KD is caused not by stimulation of a superantigen, but rather by a conventional antigen.
B-Lymphocytes/*metabolism ; Female ; Humans ; Immunoglobulin M/*metabolism ; Immunoglobulin Variable Region/*genetics ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome/*genetics/*immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

BACKGROUND: Humoral immune response is essential for resistance against leptospirosis, and the antigens in-volved in the response might be responsible for pathoge-nesis, diagnosis or immunity. The aim of this study is to clone, overexpress and purify one of leptospiral pro-tein antigens recognized by patients' sera and elucidate it as a diagnostic antigen. METHODS: We isolated a reactive clone LIA5 from a genomic library of Leptosp ira interrogans serovar lai which expressed a 62 kDa protein co-migrated to a predominant protein of Leptospira by immunoblotting with patients' sera. The sequence of the clone with 2.7 kb leptospiral DNA was determined. The 62 kDa protein encoded by an open reading frame with 1,637 bp was overexpressed in E. coli by pT7 expression vector, and purified to homogeneity by Ni-affinity resin and further electroelution. Patients' sera were examined for IgM or IgG antibodies reacting with the recombinant antigen by strip immunoblot. RESULTS: Sequence analysis of the clone identified two genes homologous to the hsp58 and hsp 10 of Lepto-sp ira serovar cop enhageni, homologues of the E. coli heat shock protein genes groEL and groES, respectively. The Hsp58 protein was detected commonly in several leptospires by immunoblotting with both patients' sera and rabbit anti-Hsp58 antibody raised against the purified recombinant Hsp58 protein(rHsp58). The predominant class of reacting antibody against rHsp58 in in patients' sera was IgG . CONCLUSION: The results indicate that Hsp58 is a highly conserved, dominant antigen in humoral immune response to leptospirosis. Further studies are needed to test the rHSP as an immunodiagnostic antigen.
Antibodies ; Clone Cells* ; Cloning, Organism* ; Diagnosis ; DNA ; Genomic Library ; Heat-Shock Proteins* ; Hot Temperature* ; Immunity, Humoral ; Immunoblotting ; Immunoglobulin G ; Immunoglobulin M ; Leptospira* ; Leptospirosis ; Open Reading Frames ; Sequence Analysis

OBJECTIVETo observe the immunologic changes in patients who have complications after polyacrylamide hydrogel (HPG) injection.

METHODSTo compare the patients in the HPG injection group and the control group, the expressions of immunoglobulin IgG, IgM, IgA and complement C3, C4 of serum were measured by immunoturbidimetry.

RESULTSThe expressions of IgG and C3 in the injection group were lower than the control group with extremely significant difference (P < 0.01). The expressions of IgM were higher than the control group with significant difference (P < 0.05). The expression of IgA and C4 had no significant differences between the two groups. The immunologic changes had no relation to the injection doses.

CONCLUSIONSClinical application of HPG as a soft tissue filling material may cause foreign-body rejection reaction. The long-term immunologic effect of HPG needs further observation in more cases.

Acrylic Resins ; adverse effects ; Adult ; Breast ; surgery ; Complement C3 ; analysis ; Complement C4 ; analysis ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Male ; Nephelometry and Turbidimetry ; Postoperative Complications ; etiology ; immunology

OBJECTIVETo explore the potential changes in the immune function of patients with obstructive sleep apnea hypopnea syndrome (OSAHS).

METHODSWe carried out a retrospective cross-sectional study of 187 patients with established OSAHS and 20 healthy subjects (control). For all the patients, the medical history was carefully examined, and overnight sleep monitoring was carried out with detection of the humoral and cellular immunity.

RESULTSWe found a significant increase in the levels of C3 and a decrease in both the IgM level and NK cell percentage in OSAHS patients as compared to the control group (P<0.01). Correlation analysis indicated that C3 was positive correlated to AHI but inversely to the lowest pulse oxygen saturation (LSpO(2)); IgM showed a mild positively correlation to LSpO(2), and NK cells had a mild inverse correlation to AHI. The other immunological indices were not found to undergo noticeable changes or show correlations in OSAHS.

CONCLUSIONImmune function changes occur in patients with OSAHS, characterized primarily by deteriorations in the humoral and cellular immunity.

Adult ; Antibody Formation ; Complement C3 ; analysis ; Complement C4 ; analysis ; Cross-Sectional Studies ; Female ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Retrospective Studies ; Sleep Apnea, Obstructive ; blood ; immunology

OBJECTIVESTo evaluate the clinical significance of flow cytometry (FCM) to detect the cytomegalovirus (CMV) PP65 antigen in patients with CMV infection.

METHODSSamples from 35 patients without CMV infection were used as negative control. The definite diagnosis of CMV infection was based on the national criteria for CMV infection. All 136 patients with CMV infection were examined with the FCM to detect CMV PP65 antigen, real-time fluorescence quantitative-polymerase chain reaction assay (RFQ-PCR) to detect CMV-DNA and ELISA to measure the serum level of IgM antibody against CMV. The results of these 3 assays in 2 groups (isolated organ involvement and disseminated diseases) were compared and the significance of PP65 antigenemia was evaluated. A short-term follow-up was undertaken in 18 patients.

RESULTSThe percentages of PP65 positivity in blood mononuclear cells (MNC) and polymorphic nuclear leukocyte (PMNL) from 35 negative control patients were 0.21% +/- 0.09% with a range of 0 - 0.41% and 0.24% +/- 0.10% with a range of 0.12% - 0.48%, respectively, which were not significantly different (t = 0.425, P > 0.05). The 95(th) percentiles (P(95)) of PP65 in MNC and PMNL were 0.39% and 0.45%, respectively, so a cutoff value of >/= 0.50% was set. Of the 136 patients with CMV infection, 118 samples from 118 patients were positive for PP65 antigenemia with a positive rate of 86.8%, which was not statistically different from that (90.4%, chi(2) = 0.91, P > 0.05) of CMV-DNA detected by RFQ-PCR assay but it was significantly higher than that (45.6%, chi(2) = 51.50, P < 0.005) of the detection by IgM measurement. PP65 detection was correlated with urine CMV DNA amplification (chi(2) = 63.78, P < 0.01) while the different detection rates between the two assays were not statistically significant (chi(m)(2) = 1.78,P > 0.05). PP65 detection was not correlated with serum IgM measurement while the detection rates between the two were significantly different (chi(m)(2) = 52.92,P < 0.01). No significant difference was found between the detection rates of CMV infection in MNC (45/53, 84.9%) and PMNL (43/53, 81.1%) (chi(m)(2) = 0.25, P > 0.05). Higher PP65 antigenemia level was correlated with systemic CMV infection, while lower level of PP65 was either in the patients with isolated organ involvement by CMV (chi(2) = 38.51, P < 0.005) or less severe in patient's situation. PP65 antigenemia of CMV infection returned to lower level or negative in recovery stage and increased when condition of patients deteriorated.

CONCLUSIONSPP65 antigenemia detection by FCM is effective in the diagnosis of the active CMV infection. Quantitative monitoring of PP65 antigenemia is useful in the evaluation of patients with CMV infection.

Antigens, Viral ; analysis ; Cytomegalovirus ; immunology ; Cytomegalovirus Infections ; diagnosis ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Immunoglobulin M ; analysis ; Polymerase Chain Reaction