ABSTRACT
In order to develop an anti-human TNF-alpha mAb, mice were immunized with recombinant human TNF-alpha. A murine mAb, TSK114, which showed the highest binding activity for human TNF-alpha was selected and characterized. TSK114 specifically bound to human TNF-alpha without cross-reactivity with the homologous murine TNF-alpha and human TNF-beta TSK114 was found to be of IgG1 isotype with kappa light chain. The nucleotide sequences of the variable regions of TSK114 heavy and light chains were determined and analyzed for the usage of gene families for the variable (V), diversity (D), and joining (J) segments. Kinetic analysis of TSK114 binding to human TNF-alpha by surface plasmon resonance technique revealed a binding affinity (KD) of ~5.3 pM, which is about 1,000- and 100-fold higher than those of clinically relevant infliximab (Remicade) and adalimumab (Humira) mAbs, respectively. TSK114 neutralized human TNF-alpha-mediated cytotoxicity in proportion to the concentration, exhibiting about 4-fold greater efficiency than those of infliximab and adalimumab in WEHI 164 cells used as an in vitro model system. These results suggest that TSK114 has the potential to be developed into a therapeutic TNF-alpha-neutralizing antibody with picomolar affinity.
Subject(s)
Animals , Humans , Mice , Amino Acid Sequence , Antibodies, Monoclonal/chemistry , Antibody Affinity/immunology , Antibody Specificity , Base Sequence , Blotting, Western , Cell Line , Cytotoxicity, Immunologic , Enzyme-Linked Immunosorbent Assay , Immunoglobulin Variable Region/genetics , Kinetics , Mice, Inbred BALB C , Molecular Sequence Data , Neutralization Tests , Sequence Analysis, Protein , Tumor Necrosis Factor-alpha/immunologyABSTRACT
BACKGROUND: Hepatitis B virus (HBV) infection is one of the worldwide public health problem affecting about 300 million people. The envelope protein of HBV consists of three components known as preS1, preS2, and S antigen. According to the recent study, anti-HBs Ab showed effective neutralization ability against HBV from chronic hepatitis B and liver transplant patients, suggesting the possible development of therapeutic antibody. METHODS: Spleen cells immunized with S antigen of HBV were fused with myeloma cell line to obtain HBsAg specific monoclonal antibodies. High affinity antibodies against HBsAg (adr, ad and ay type) were selected by competitive ELISA method. Nucleotide sequence of the variable regions of monoclonal antibodies was analyzed by RT-PCR followed by conventional sequencing method. RESULTS: We produced 14 murine monoclonal antibodies which recognize S antigen of HBV. Two of them, A9-11 and C6-9 showed the highest affinity. The sequence analysis of A9-11 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain I (B) and light chain lambda 1, respectively. Likewise, the sequence analysis of C6-9 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain II (B) and light chain kappa 1, respectively. Neutralization assay showed that A9-11 and C6-9 effectively neutralize the HBV infection. CONCLUSION: These results suggest that A9-11 and C6-9 mouse monoclonal antibodies can be used for the development of therapeutic antibody for HBV infection.
Subject(s)
Animals , Humans , Mice , Antibodies , Antibodies, Monoclonal , Base Sequence , Cell Line , Enzyme-Linked Immunosorbent Assay , Hepatitis B Surface Antigens , Hepatitis B virus , Hepatitis B , Hepatitis B, Chronic , Hepatitis , Liver , Public Health , Sequence Analysis , SpleenABSTRACT
We experienced a case of split hand split foot(SHSF) anomaly in a male neonate who had a deficiency of the middle finger, syndactyly of the 1st and 2nd finger and a deep median cleft in both hands. He also had a deep median cleft between 2nd and 3rd toe, syndactyly of the 1st and 2nd, 3rd and 4th toe without deficiency of the middle toe in both feet. SHSF anomaly may occur either isolated or associated with other anomalies. In this case, it occurd sporadically without family history and showed an isolated type without any other specific anomalies except both posterior iris synechiae. The karyotype of patient showed 46,XY,t(7p:14q) which has not been reported yet. We reported the case with the review of the associated literatures.
Subject(s)
Humans , Infant, Newborn , Male , Fingers , Foot , Hand , Iris , Karyotype , Syndactyly , ToesABSTRACT
We have constructed several panels of MAbs which specifically recognize B-subunit of HCG (BHCG). Splenocytes from Balb/c mice immunized with B-subunit of HCG were fused with SP2/o-Ag14 myeloma cells by PEG method. Fifteen different hybridorna clones (individually named as mG10.127, mG10.61, mG9.5, mG9.18, rnG9.20, mG6.3, mG6.36, mG6.8, mG7.31, mG7.79, mG9.11, mG9.51.6, mG9.51.12, mH4.17, and mH4.4) were obtained by indirect ELISA screening and three to five successive cloning procedures. The distinct features of these MAbs were determined by specificity, western blot, isotyping, and isoelectrofocusing. All of the MAbs except mG9.20 and mG6.8 specifically bind to BHCG without cross- reaction with B-subunit of LH (BLH). In western blot analysis, all of the MAbs bind to non-denatured form of BHCG suggesting that the MAbs recognize conformation-dependent epitope of BHCG. This new panels of MAbs to BHCG should be useful for developing diagnostic reagent such as pregnancy, choriocarcinoma, Down's syndrome as well as for the fine quantitation of serum or urinary HCG.
Subject(s)
Animals , Female , Humans , Mice , Pregnancy , Antibodies, Monoclonal , Blotting, Western , Choriocarcinoma , Chorionic Gonadotropin , Clone Cells , Cloning, Organism , Down Syndrome , Enzyme-Linked Immunosorbent Assay , Mass Screening , Sensitivity and SpecificityABSTRACT
PURPOSE: This study was performed to evaluate the polymerase chain reaction (PCR) in the diagnosis of Mycoplasma pneumoniae pneumonia in comparison with the specific antibody test. METHODS: Five hundred and ten patients with pneumonia, ranging from the ages of 8 months to 15 years who were admitted in Sung-Ae and Kwangmyung Sung-Ae general hospitals from Nov. 1996 to Oct. 1997, were enrolled in this study. Specific antibody test of the serum using Serodia Myco II kit and PCR of the sputum or throat swab were performed on admission simultaneously, and follow-up antibody test was done during the convalescent stage if they showed sero-negative at first. Among the PCR positive patients, 40 were treated with erythromycin initially, and were followed with PCR on the 7th day of treatment. RESULTS: The sensitivity of PCR to the specific antibody test was 82.5%, and the specificity, 98.4%. The false positive and negative rates of PCR to the specific antibody test were 1.6% and 17.5%, respectively. Fifty-six (91.8%) of the 61 patients who showed sero-negative and PCR positive on admission were determined as sero-positive at the convalescent stage. Negative conversion of PCR at the convalescent stage was noted for 32 (80%) of 40 patients who were treated with erythromycin. CONCLUSION: The PCR was considered to be valuable due to its high sensitivity and specificity as a diagnostic method of Mycoplasma pneumoniae pneumonia. The effect of the method was more apparent than the specific antibody test in early diagnosis which is clinically important. In addition, it seems to be more useful in the appreciation of treatment and epidemiologic study than the culture method that shows low sensitivity and takes too much time.
Subject(s)
Humans , Diagnosis , Early Diagnosis , Epidemiologic Studies , Erythromycin , Follow-Up Studies , Hospitals, General , Mycoplasma pneumoniae , Mycoplasma , Pharynx , Pneumonia , Pneumonia, Mycoplasma , Polymerase Chain Reaction , Sensitivity and Specificity , SputumABSTRACT
3p partial trisomy is a rare chromosomal anomaly. We experienced a case of 3p partial trisomy in a male neonate. It was diagnosed by clinical and chromosoaml study. He had multiple anomalies such as brachycephaly, wide open fontanelle, square face, hypertelorism, mongoloid palpebral fissure, micrognathia, low set malformed ear, bilateral cleft lip and palate, double outlet right ventricle, atrial septal defect, ventricular septal defect, left ventricular hypoplasia, renal microcysts and micropenis. He was manifested intrauterine growth retardation. Peripheral blood chromosome studies showed an additional chromosomal material at the distal part of the short arm of chromosome 7. Analysis of chromosomes of family members showed that the father had normal karyotype, but the mother had reciprocal balanced translocation,46, XX, t(3;7)(p25;p22). The karyotype formula of the propositus was thus,46,XY,der(7),t(3;7)(p25;p22)mat, that is unbalanced for a duplication 3p25-->3pter, resulting from segregation of a balanced maternal translocation. Two years after patient's birth, his sister was born at 40 weeks of gestation without congenital anomalies. In the case of his sister, amniocentesis and chromosome studies had been done at 16 weeks of gestation. The result of the chromosome analysis was 46,XX,t(3;7)(p25;p22), as in her mother. We report a neonate with multiple congenital anomalies due to partial trisomy for the short arm of chromosome 3, his mother and a female sibling with t(3;7)(p25-->p22).
Subject(s)
Female , Humans , Infant, Newborn , Male , Pregnancy , Amniocentesis , Arm , Chromosomes, Human, Pair 3 , Chromosomes, Human, Pair 7 , Cleft Lip , Craniosynostoses , Double Outlet Right Ventricle , Ear , Fathers , Fetal Growth Retardation , Heart Septal Defects, Atrial , Heart Septal Defects, Ventricular , Hypertelorism , Karyotype , Mothers , Palate , Parturition , Siblings , TrisomyABSTRACT
The unilateral megalencephaly is a rare brain malformation characterized by cerebral asymmetry and cortical dysplasia caused by faulty migration of the subependymal neuroblasts. We experienced a case of unilateral megalencephaly in a two day-old male with the chief complaint of asymmetric head appearance. Large left hemisphere with agyria, pachygyria, dilatation of lateral ventricle, and the thick cortex of the ipsilateral hemisphere were showed in brain MRL, Clinical findings in this case were intractable seizure, hemiparesis, and psychomotor retardation. A review of literatures was also presented briefly.
Subject(s)
Humans , Male , Brain , Dilatation , Head , Lateral Ventricles , Lissencephaly , Malformations of Cortical Development , Paresis , SeizuresABSTRACT
We report an autopsy case of neonatal giant cell hepatitis that was presumed to be related to bacterial sepsis, endotoxemia and to the subsequent parenteral alimentation and antibiotics treatment. The patient died of candidal endocarditis and multiple brain infarcts. This female baby was born by a normal full term spontaneous delivery. Six days after delivery she developed fever and lethargy as she suffered from Cheyne-Stokes respiration with severe grunting. Blood culture grew Enterobacter and Acinetobacter. After management of the sepsis her general condition improved. On the 23rd day of admission she was found to have deep jaundice and hepatosplenomegaly. The liver became larger progressively and the edge was palpable at the umbilical level. Grade II systolic murmur was heard along the left lower sternal border. She died on the 31st day of hospitalization. Postmortem examination showed severe jaundice, hepatosplenomegaly, a large vegetation on the mitral valve and multiple petechial hemorrhages of the viscera. Microscopically the liver showed features of massive giant cell transformation, mild fibrosis and inflammatory cells, suggestive of giant cell hepatitis. Numerous yeasts and candidal pseudohyphae were seen in the cardiac vegetation, focally extending into the myocardium. There was a focus of candidal vasculitis in the bowel wall. In addition there were multiple bilateral organizing infarcts in the cerebral hemisphere as well as diffuse white matter damage associated with septicemia.