The lower expression of CD20 antigen molecules on the B cell membrane is the primary characteristic of B-chronic lymphocytic leukemia (B-CLL). In this paper, we combined laser scanning confocal microscopy (LSCM) and quantum dots labeling to detect the expression and distribution of CD20 molecules on CD20+B lymphocyte surface. Simultaneously, we investigated the morphology and ultrastructure of the B lymphocytes that belonged to the normal persons and B-CLL patients through utilizing the atomic force microscope (AFM). In addition, we measured the force spectroscopy of CD20 antigen-antibody binding using the AFM tips modified with CD20 antibody. The fluorescent images indicated that the density of CD20 of normal CD20+B lymphocytes was much higher than that of B-CLL CD20+B cells. The AFM data show that ultrastructure of B-CLL CD20+B lymphocytes became more complicated. Moreover, the single molecular force spectroscopy data show that the special force of CD20 antigen-antibody was four times bigger than the nonspecific force between the naked AFM tip and cell surface. The force map showed that CD20 molecules distributed homogeneously on the normal CD20+B lymphocytes, whereas, the CD20 molecules distributed heterogenous on B-CLL CD20+B lymphocytes. Our data provide visualized evidence for the phenomenon of low-response to rituximab therapy on clinical. Meanwhile, AFM is possible to be a powerful tool for development and screening of drugs for pharmacology use.
Antigen-Antibody Reactions ; immunology ; Antigens, CD20 ; immunology ; B-Lymphocytes ; immunology ; ultrastructure ; Binding Sites, Antibody ; Cell Membrane ; immunology ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; immunology ; Microscopy, Atomic Force ; Microscopy, Confocal ; Quantum Dots

Antibodies, Monoclonal ; pharmacology ; therapeutic use ; Antibodies, Neoplasm ; immunology ; therapeutic use ; Antibody Affinity ; Antibody Specificity ; Antigen-Antibody Reactions ; Antigens, Neoplasm ; analysis ; immunology ; Carcinoma, Hepatocellular ; immunology ; therapy ; Humans ; Liver Neoplasms ; immunology ; therapy

Immunohistochemistry (IHC) is an important auxiliary method for pathologists in routine diagnostic work as well as in basic and clinical research including exploration of biomarkers, as IHC allows confirmation of target molecule expressions in the context of microenvironment. Although there has been a considerable progress in automation and standardization of IHC, there are still many things to be considered in proper optimization and appropriate interpretation. In this review, we aim to provide possible pitfalls and useful tips for practicing pathologists and residents in pathology training. First, general procedure of IHC is summarized, followed by pitfalls and tips in each step and a summary of troubleshooting. Second, ways to an accurate interpretation of IHC are discussed, with introduction to general quantification and analysis methods. This review is not intended to provide complete information on IHC, but to be used as a basic reference for practice and publication.
Antigen-Antibody Reactions ; Automation ; Biomarkers ; Immunohistochemistry* ; Methods ; Pathology ; Publications

No abstract available.
Animals ; Antigen-Antibody Reactions* ; Guinea Pigs* ; Guinea* ; Lung* ; Mast Cells*

To overcome the present limitations of passive biochip, based on the basic principle of antigen-antibody reaction, we develop an antigen-antibody reaction model of solid-phase surface and design a novel active biochip system according to this model, which introduces the negative pressure and controlling devices to control the immunoreactions on the nitrocellulose (NC) membrane. From the computer simulation results, this is a rapid, stable, robust and practicable system, which can be used to increase the efficiency of immunoreactions and improve the reproducibility and accuracy of biochip analysis.
Antigen-Antibody Reactions ; Biosensing Techniques ; instrumentation ; Equipment Design ; Models, Biological

Febrile nonhemolytic transfusion reaction (FNHTR) is one of the most common adverse reactions to the transfusion of blood products. It is caused by antigen-antibody reactions between patient's plasma and transfused cellular components (or the reverse) or bioactive substances such as cytokines generated by leukocytes during storage of cellular blood components. Most of the antibodies involved in FNHTR are anti-HLA antibodies. However, platelet-specific antibodies and rarely granulocyte-specific antibodies may be involved in FNHTR, We found a granulocyte antibody from a 53-year-old male with FNHTR for the first time in Korea. Fever developed during transfusion of a unit of packed RBC after partial cystectomy, and subsided after the adminstration of acetaminophen. The hemolytic transfusion reaction and sepsis were excluded after investigations. Mixed passive hemagglutination test revealed that the patient had antibody against human neutrophil antigen-1b (HNA-1b), and granulocyte antigen genotyping showed granulocyte antigen mismatches between the patient (HNA-1a homozygote) and the unit of transfused RBC (HNA-1a/HNA-1b heterozygote).
Acetaminophen ; Antibodies ; Antigen-Antibody Reactions ; Blood Group Incompatibility* ; Cystectomy ; Cytokines ; Fever ; Granulocytes ; Hemagglutination Tests ; Humans ; Korea* ; Leukocytes ; Male ; Middle Aged ; Neutrophils ; Plasma ; Sepsis

BACKGROUND: Granulocyte specific antibodies are associated with several clinical conditions including febrile transfusion reaction and transfusion-related acute lung injury as well as immune neutropenias. The identification of granulocyte specific antibodies is important for the diagnosis of these disorders. However, there have been rarely confirmed clinical reports in Korea since the testing techniques are complicated and difficult to maintain. In this study, development of in-house indicator cells and renewedly establishment of the mixed passive hemagglutination assay (MPHA) as a serologic test to detect and identify granulocyte specific antibodies were conducted. METHODS: The in-house indicator cells for MPHA were made by sensitizing human Rh(D) positive O RBCs with human IgG anti-Rh(D) (DiaMed AG, Switzerland) and then combining with AHG anti-IgG (Immucor Inc., USA). To determine the optimal conditions, various combinations of anti-Rh(D) IgG sensitization strengths of indicator cells, microwell coated antigens (intact granulocyte vs. extracted granulocyte) and reaction conditions were compared. RESULTS: The best test conditions for MPHA were as follows: optimal results were obtained with the anti-Rh(D) sensitization dilutions of 1/64-1/192 and the reaction condition of 4 hours incubation at room temperature in humid chamber. Extracted granulocytes coated at the plate showed better results than intact granulocytes. HLA antigens were completely removed from extracted granulocyte antigens after acidified chloroquine treatment. CONCLUSION: Granulocyte MPHA using in-house anti-Rh(D) sensitized indicator cells was developed for the first time in Korea. The newly established MPHA would be effectively used for the diagnosis and treatment of disorders associated with granulocyte specific antigen-antibody reactions in Korea.
Acute Lung Injury ; Antibodies ; Antibodies, Anti-Idiotypic ; Antigen-Antibody Reactions ; Blood Group Incompatibility ; Chloroquine ; Granulocytes ; Hemagglutination ; HLA Antigens ; Humans ; Immunoglobulin G ; Korea ; Neutropenia ; Serologic Tests

PURPOSE: A cDNA microarray is a systematic method to identify key molecules for prognosis and for treatment response by profiling thousands of genes expressed in a single cancer. The clinical value of cDNA microarray is still being investigated in various fields. This technique could be used in detecting molecules important for cancer to develop, to monitor the effect of new cancer therapeutics, and to give a prognosis for cancer patients. We now report the results of our initial cDNA microarray data to analyze the genome pattern of colorectal cancer tissues and to evaluate the possibility of using cDNA microarrays in a clinical setting for cancer patients. METHODS: We used the general cDNA microarray technique with a 2.4 K cDNA chip provided by Macrogene company. RNA extracted from seven colorectal cancer tissues was amplified by using RT-PCR (reverse transcriptase-polymerase chain reaction), and applied to a cDNA chip to produce an antigen-antibody reaction. The results were analyzed individually and hierarchically. RESULTS: All seven tested cancer tissues were harvested from operative specimens at the Korea Cancer Center Hospital. The male-to-female ratio was 4 to 3. Five patients were TNM stage II, and two patients were stage III. Eighteen genes were upregulated in stage II patients, and 51 in stage III patients. The number of genes discriminating stage was 69, including 8 control genes, 4 ribosomal genes, 5 EST genes, 10 known non-functional genes, 23 genesof unknown function, and 19 possible cancer-related genes. A hierarchial graph showed similar patterns within a stage, which suggests that genetic patterns might affect clinical characteristics. CONCLUSIONS: Seven colorectal cancer tissues were analyzed with the cDNA microarray technique using 2.4 K cDNA chip. Authors could identify 69 genes that showed the significant change of expression. Although our reports presented the preliminary results, we think that the cDNA microarray will be able to offer an informative results to predict cancer development and progression in colorectal cancer.
Antigen-Antibody Reactions ; Colorectal Neoplasms* ; DNA, Complementary* ; Genome ; Humans ; Korea ; Oligonucleotide Array Sequence Analysis* ; Prognosis ; RNA

Acute Poststreptococcal glomerulonephritis is generally stated to be the most common cause of acute nephritis in childhood. The activity of serum complement has been studied in various diseases in an attempt to accumulate evidence of antigen-antibody reaction. In acute poststreptococcal glomerulonephritis, several investigators have reported a decrease in serum complement early in the course of disease. This paper reports on observation of serum C3 levels in 41 cases of poststreptococcal glomerulonephritis which were collected from department of pediatrics, Yonsei University Medical College, from Aug 1978 to March 1979 The results were as follows : 1. The mean value of serum C3 concetration in this group as a whole was lower than in the control group and the difference was statistically significant(p<0.001) 2. The initial reduction of serum C3 concentration did not correlate with the severity of the acute phase of the disease. 3. In those children with acute poststreptococcal glomerulonephritis, Serum C3 concentration returned to normal within 6 weeks.
Antigen-Antibody Reactions ; Child ; Complement System Proteins ; Glomerulonephritis* ; Humans ; Nephritis ; Pediatrics ; Research Personnel

Hematoxylin and eosin staining is simple and one of the most important techniques in pathological diagnosis. However, it cannot provide complete information about the disease of a patient. Immunohistochemical staining (IHC) is an important method for demonstrating the distribution of a certain molecule or antigen in tissues using specific antigen-antibody reactions. It is used in routine diagnostic work and research to explore biomarkers. In this review, I aim to provide an adequate interpretation of the results of IHC and pathological diagnosis for clinicians.
Antigen-Antibody Reactions ; Biomarkers ; Diagnosis* ; Eosine Yellowish-(YS) ; Hematoxylin ; Humans ; Methods