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

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

AB blood group is determined by A and B genes located on each chromosome which is inherited from parents. But unusual inheritance of A and B genes on the same chromosome has been reported as Cis AB. In some pedigree, the antigen-antibody reactions may occur despite of same AB phenotype, so called "Cis AB", in contradiction to the general Mendelian inheritance of blood group ABO expression. The anesthesiologists, often meet many cases of the transfusion, may anesthetise the Cis AB patients for surgery. Recently, authors experienced one case of patient with Cis AB blood group undergoing Cesarean Section.
Antigen-Antibody Reactions ; Blood Transfusion ; Cesarean Section ; Female ; Humans ; Parents ; Pedigree ; Phenotype ; Pregnancy ; Wills

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

OBJECTIVETo investigate the difference of the antigenicity of xenogenic acellular dermal matrix (ADM) implants prepared by different methods.

METHODSThe split-thickness skin sheet from swine was processed by trypsin and Triton X-100 to make xeno-ADM. Twenty-five Japanese white rabbits were divided into 5 groups, i.e. xeno-ADM(1) (conjugated with glutaraldehyde), xeno-ADM(2) (conjugated with network) and xeno-ADM(3) (no conjugation, as control), in which the ADMs were and xeno-ADM(4) (conjugated) and allo-ADM (no conjugated as control), in which the ADMs were embedded into the subcutaneous place of rabbit ear and back after that the rabbits were pre-sensitized by xeno-ADM soluble protein antigen injections. The titers of anti ADMs antibody in rabbit serum were monitored during 2 - 32 post-operative weeks and the histological changes of the embedded ADMs were observed grossly and microscopically.

RESULTSThe serum titers of anti-xeno-ADM in xeno-ADM(4) group was the highest. Whereas regardless of the sensitizing effects, the titers in all groups ranged as follows: xeno-ADM(3) > xeno-ADM(2) > xeno-ADM(1) (P < 0.05 - 0.01). About 40% serum samples in allo-ADM group exhibited positive anti-allo-ADM protein antibodies. Histologically, Evident and lasting inflammatory reaction could be found in the xeno-ADM grafting sites, which was much stronger than that in allo-ADM group. The degradation and absorption gradient of ADM was ranked as follow: xeno-ADM(3) > xeno-ADM(2) > xeno-ADM(4) > xeno-ADM(1) > Allo-ADM. Foreign body megalocytic reaction might evoke in the surrounding of conjugated ADM.

CONCLUSIONThe immunogenicity in xeno-ADM was stronger than that in allo-ADM, which could induce the host to develop immune reaction restricted by IgG. Large sheets of degenerated ADM implants could lower down the antigen-antibody reaction and ameliorate the structural destroying and degeneration absorption of ADM induced by inflammatory immune reaction.

Animals ; Antigen-Antibody Reactions ; Dermis ; immunology ; transplantation ; Male ; Rabbits ; Skin Transplantation ; Swine ; Transplantation, Heterologous

BACKGROUND: Since the first introduction of radioimmunoassay for the quantification of the thyroidstimulating hormone (TSH), more advanced analytical methods have been developed and used in laboratories. However, they are still inconvenient in that they require time-consuming procedures, special safety in handling isotopes, expensive equipment, and a highly qualified expert. METHODS: As an immunoassay system for the rapid measurement of TSH in serum, we have developed a new analytical system based on immunochromatographic assay with fluorescencelabeled anti-TSH monoclonal antibodies. The assay system is composed of a test strip housed within a cartridge and a laser-fluorescence scanner for quantification. The strip contains a sample pad, an absorption pad, and a nitrocellulose membrane where a captured antibody is immobilized and antigen-antibody reaction occurs. Fifty microL of serum was added to 50 microL of a detector solution and the mixture was loaded onto the well of the sample pad on the cartridge. After incubation for 12 min, the cartridge was quantified with the laser-fluorescence scanner. RESULTS: The calibration curve displayed linearity (R=0.95) at concentrations of 1-40 mIU/L. Intraand inter-assay imprecisions were determined to be CVs within 10%. Analytical recovery was 93.9% at 3 different concentrations and the detection limit was 0.868 mIU/L of TSH. The new assay system correlated well with an Abbott AxSYM for quantification of TSH (R=0.97, slope 0.94, N=20). CONCLUSIONS: The TSH measurement system developed in this study showed good reproducibility. However, our TSH quantification system needs some improvement to be used in the medical field because of its low analytical sensitivity. With enhanced performance in analytical sensitivity, introduction of a whole-blood type strip, and a more miniaturized fluorescence scanner, we expect the TSH analytical system to be used for point-of-care testing in the near future.
Absorption ; Antibodies, Monoclonal ; Antigen-Antibody Reactions ; Calibration ; Collodion ; Fluorescence* ; Immunoassay ; Immunochromatography* ; Isotopes ; Limit of Detection ; Membranes ; Radioimmunoassay ; Thyrotropin*

OBJECTIVE: To develop an immunoassay system for the rapid measurement of beta-hCG in serum sample, we designed an immunochromatographic assay-based one-step assay that uses two monoclonal antibodies to beta-hCG as a sandwich pair. METHODS: The assay system is composed of a test strip housed within a cartridge for sample application and a laser-fluorescence scanner for quantification. The strip contains a sample pad, an absorption pad, and a nitrocellulose membrane where a capture antibody is immobilized and antigen-antibody reaction occurs. Ten L of serum was added to 60 L of detector solution, and the mixture was loaded onto the well of the sample pad on the cartridge. After incubation for 12 min, the cartridge was scanned for quantification with the laser-fluorescence scanner. RESULTS: No cross-reactivity was observed between beta hCG antibodies and other pituitary hormones. The calibration curve displayed linearity (R2=1) at concentrations of 0-1,000 IU/L. Intra- and interassay imprecisions were determined with serum samples to be their CVs within <6% and <7%, respectively. Analytical recovery was 103-108.4% in serum samples at three different concentrations. There was high correlation between beta-hCG concentrations measured by Boditech system and those measured by Abbot AxSYM system. CONCLUSION: The new fluorescence assay system is a convenient and fast method and can be used as a good tool for detection and quantification of beta-hCG in serum samples. Furthermore, the portable system allows us to perform the tests on a sampling site in any places without bringing the samples to a laboratory.
Absorption ; Antibodies ; Antibodies, Monoclonal ; Antigen-Antibody Reactions ; Calibration ; Chorionic Gonadotropin ; Collodion ; Fluorescence* ; Immunoassay ; Immunochromatography* ; Membranes ; Pituitary Hormones