With the development of bio-technological drugs, drug immunogenicity evaluation has become key factor of clarifying safety and efficacy of these drugs. It has become the focus to establish a stable and reliable evaluation system. Due to the advantages such as continuous real-time monitoring, surface plasmon resonance (SPR) technology has been widely used in bio-technological drugs immunogenicity assessments. Our study applied this technology to detect anti-drug antibody (ADA) of a recombinant human anti-rabies monoclonal antibody NM57 in the sera of 48 volunteers admitted in phase I clinical trials. This method could satisfy the basic requirements of detection of ADA.
Antibodies, Anti-Idiotypic ; analysis ; blood ; immunology ; Antibodies, Monoclonal ; analysis ; blood ; immunology ; Antibodies, Viral ; analysis ; blood ; immunology ; Humans ; Rabies virus ; immunology ; Recombinant Proteins ; analysis ; blood ; immunology ; Surface Plasmon Resonance

BACKGROUND: Detection methods for ABO antibody (Ab) titers vary across laboratories, and the results are different depending on the method used. We aimed to compare titer values using different detection methods for the measurement of ABO Ab titers. METHODS: For ABO Ab detection, pooled group A or B red blood cells (RBCs) were reacted with each of 20 sera from blood groups A, B, or O without dithiothreitol treatment. The room-temperature (RT) incubation technique and the indirect antiglobulin test (IAT) were used in the tube test and gel card test. Flow cytometry (FCM) was performed by using anti-IgM and anti-IgG Abs. RESULTS: Regardless of the blood groups tested, the FCM assay with anti-IgM showed the highest titer compared to the tube test and gel card test with RT incubation in both. The tube test with IAT showed a higher titer than the gel card test with IAT (Gel-IAT) or FCM with anti-IgG in blood group A and B, while Gel-IAT showed the highest titer relative to the other tests, only for the anti-A Ab in blood group O. CONCLUSIONS: There were significant differences in the titers depending on the detection method used, and each method showed a different detection capacity for each ABO Ab depending on the ABO blood group tested. Therefore, caution should be exercised in interpreting ABO Ab titer results, taking into consideration the detection method used and the blood group.
ABO Blood-Group System/*immunology ; Adult ; Aged ; Agglutination Tests/instrumentation/*standards ; Antibodies/*analysis ; Antibodies, Anti-Idiotypic/analysis ; Erythrocytes/chemistry/metabolism ; Female ; *Flow Cytometry ; Humans ; Male ; Middle Aged ; Temperature

The objective of this study was to analyze the molecular genetic basis for 2 individuals with A2B phenotype of ABO subtype. The ABO group antigens on red blood cells were identified by monoclonal antibodies and the ABO antibodies in serum were detected by the standard A, B, O cells. The exon 5 to exon 7 coding region of ABO gene was amplified by polymerase chain reaction (PCR) and the PCR product was sequenced directly after the enzymes digested. The amplified product was also cloned by TOPO TA cloning sequencing kit to split the 2 alleles apart and chosen colonies were sequencing bidirectionally for exon 6 to 7 of ABO gene. The results showed that both A and B antigen were identified on red blood cells of the individuals and there was anti-A1 antibody in their serum. There was no 261G deletion and showed 297A/G, 467C/T, 526C/G, 657C/T, 703G/A, 742C/T, 796C/A, 803G/C, 930G/A heterozygotes by direct DNA sequencing. After cloning and sequencing, it was obtained the B101 and one novel A allele. The novel allele has one nucleotide change at 742 position C to T compared with A102, which results in an amino acid Arg to Cys at 248 position and was nominated as A213. It is concluded that C742T mutation of the α1, 3 N-acetyl-D-galactosaminyl-transferase gene can lead to A2 phenotype and with anti-A1 antibody in serum.
ABO Blood-Group System ; genetics ; immunology ; Alleles ; Antibodies, Anti-Idiotypic ; immunology ; Blood Donors ; Exons ; Genotype ; Heterozygote ; Humans ; Molecular Sequence Data ; Mutation ; N-Acetylgalactosaminyltransferases ; genetics ; Phenotype ; Sequence Analysis, DNA

BACKGROUND AND METHODS: To evaluate the role of specific antibodies to corn dust (CD) and their relationship to respiratory dysfunction, we detected serum specific IgE(slgE) and IgG4(slgG4) antibodies by ELISA in 42 employees working in the animal feed industry and 27 unexposed controls. RESULTS: Our survey revealed that 15 (34.9%) subjects had work-related respiratory dysfunction associated with or without nasal symptoms. Among these subjects, eight had airway hyper-responsiveness to methacholine. Significant differences were noted in slgE and slgG4 between exposed and unexposed groups (p = 0.04, p = 0.00 respectively), but no difference was noted in slgG (p = 0.1). Although there was no significant differences in the prevalence of specific IgE antibody between symptomatic (29%) and asymptomatic groups (19%, p = 0.55), the specific IgE levels were significantly higher in symptomatic workers than in asymptomatic workers (p = 0.03). Specific IgG antibody was detected in 1 (6%) symptomatic and 4 (15%) asymptomatic workers (p = 0.46). Specific IgG4 antibody was detected in 11 (73%) of symptomatic and 21 (78%) of asymptomatic workers (p = 0.90). The higher prevalence of slgG4 antibody was noted in workers with slgE antibody (p = 0.001). The correlation between slgG and exposure duration was significant (r = 0.36, p = 0.02). There was no association between the prevalence of slgE, slgG, and slgG4 to exposure intensity, smoking or atopic status. CONCLUSION: These results suggested that the existence of slgG and slgG4 might represent a response to CD exposure, and that some unexposed subjects had slgG to CD. Specific IgE might play a role in the development of respiratory symptoms.
Analysis of Variance ; Antibodies, Anti-Idiotypic/analysis* ; Asthma/immunology* ; Asthma/epidemiology ; Chi-Square Distribution ; Comparative Study ; Corn/adverse effects* ; Dust/adverse effects* ; Enzyme-Linked Immunosorbent Assay ; Human ; IgE/analysis* ; IgG/analysis* ; Male ; Occupational Diseases/immunology* ; Occupational Diseases/epidemiology ; Occupational Exposure/adverse effects ; Reference Values ; Respiratory Hypersensitivity/immunology* ; Skin Tests ; Smoking/immunology ; Substances: IgE ; Substances: IgG ; Substances: Antibodies, Anti-Idiotypic

OBJECTIVES AND METHODS: To confirm the local production of IgE antibody from the nasal polyp tissue, and to evaluate the difference between atopics and non-atopics, nasal polyp tissues were taken from both 10 atopic and 10 non-atopic subjects. The tissue total IgE (tlgE) level was measured by enzyme-linked immunosorbent assay (ELISA) and serum tlgE level by radio-immunoassay. The tissue albumin level was measured by nephelometry, and serum albumin level by Bromocresol green method. RESULTS: The polyp tissue tlgE/albumin as well as serum tlgE/albumin ratio were significantly higher in atopics than in non-atopics (p < 0.05), with no difference in the albumin level between the two groups (p > 0.05). Three non-atopic subjects had high polyp tissue tlgE/albumin (> 10). A significant correlation was noted between serum tlgE/albumin and polyp tlgE/albumin (r = 0.46, p = 0.04). The ratio of polyp tlgE/albumin to serum tlgE/albumin was greater than 1 in all of the non-atopic subjects and 7 of 10 atopic subjects. CONCLUSION: These findings support the hypothesis that IgE antibody could be locally produced from the nasal polyp tissue of non-atopic subjects as well as atopic subjects. The possibility of an isolated local production of IgE antibody was suggested.
Albumins/analysis ; Antibodies, Anti-Idiotypic/analysis* ; Biopsy, Needle ; Comparative Study ; Enzyme-Linked Immunosorbent Assay ; Female ; Human ; Hypersensitivity, Immediate/immunology* ; IgE/biosynthesis* ; IgE/analysis ; Male ; Nasal Polyps/pathology ; Nasal Polyps/immunology* ; Radioimmunoassay ; Reference Values ; Tissue Culture ; Substances: IgE ; Substances: Antibodies, Anti-Idiotypic ; Substances: Albumins

OBJECTIVETo investigate the protective immunity induced by the anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum in mice.

METHODSAn orthogonal table L(16) (4 x 2(12)) was selected as the experimental design. Eight-week-old Kunming outbred mice (male and female) were randomly divided into 16 experimental groups and 2 control groups. Control groups were injected with SP2/0 ascites intraperitoneally. Mice from each group were infected with 100 +/- 2 cercariae of Schistosoma japonicum in the abdominal skin and were sacrificed on the thirtieth day postchallenge. Adult worms were recovered and counted by perfusion of the left ventricle-portal vein. The SP2/0 ascites injected mice were used as controls and the percentage of protection was calculated.

RESULTSActive immunization of mice with NP30 could produce protection levels ranging from 22.36% to 50.46% depending on the different immunity protocols. The best immunization protocol was established from the results.

CONCLUSIONSActive immunization with NP30 can induce a degree of protection to infection with Schistosoma japonicum cercariae and NP30 is a potential vaccine candidate against Schistosoma japonicum.

Analysis of Variance ; Animals ; Animals, Outbred Strains ; Antibodies, Anti-Idiotypic ; immunology ; therapeutic use ; Antibodies, Monoclonal ; immunology ; therapeutic use ; Female ; Male ; Mice ; Schistosoma mansoni ; immunology ; Schistosomiasis mansoni ; immunology ; parasitology ; prevention & control ; Treatment Outcome ; Vaccination

OBJECTIVETo prepare anti-human IgG-dextran-adriamycin conjugate for immunotargeting of S180 sarcoma and assess its effects on the tumor weight and survival time of the tumor-bearing mice.

METHODSAnti-human IgG-dextran- adriamycin was synthesized by conjugating dextran and adriamycin with anti-human IgG. The immunoactivity of anti-human IgG-dextran-adriamycin was measured by enzyme-linked immunosorbent assay (ELISA), and the cytotoxicity of anti-human IgG, adriamycin, and the IgG-dextran-adriamycin conjugate against the tumor cells in vitro was evaluated using MTT assay. In mice bearing S180 sarcoma, the agents were tested for their effects against tumor cell growth and the survival time of mice.

RESULTSThe molar ratio of anti-mouse IgG, dextran, and adriamycin was 1:2.5:38 in the conjugate. The conjugates were shown to retain the immunoactivity of anti-human IgG, and possessed cytotoxicity to S180 cells in vitro. Administration of the conjugate and intratumor injection of human IgG resulted in a tumor suppression rate of 17.72%in mice bearing S180 sarcoma, but did not prolong the survival time of the mice.

CONCLUSIONThe anti-human IgG-dextran-adriamycin conjugate shows targeted antitumor effect against S180 sarcoma in mice.

Animals ; Antibodies, Anti-Idiotypic ; administration & dosage ; pharmacology ; Antibodies, Monoclonal ; administration & dosage ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cell Survival ; drug effects ; Dextrans ; administration & dosage ; pharmacology ; Doxorubicin ; administration & dosage ; pharmacology ; Female ; Immunoglobulin G ; administration & dosage ; pharmacology ; Immunotoxins ; administration & dosage ; pharmacology ; Mice ; Sarcoma 180 ; drug therapy ; pathology ; Survival Analysis ; Tumor Burden ; drug effects

OBJECTIVETo investigate the anti-human CEM lymphoma cell activities induced by TCR idiotypic DNA vaccine containing different antigen determinants in BALB/c mice.

METHODSThe specific rearranged gene fragment encoding TCRVbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into eukaryocytic expression vector pcDNA3, which was used as DNA vaccine and template for PCR amplifying different antigen determinant. Gene fragments encoding different antigen determinant were amplified and cloned into pcDNA3, separately. The experimental mice were immunized by intramuscular injection of the DNA vaccines. The specific anti-idiotype antibodies were detected by indirect immunofluorescence assay.

RESULTSTCRbetaV of CEM cell line contains five antigen determinants. Specific anti-idiotype antibody was detected in all of the six mice immunized with DNA vaccine containing all the five determinants (the highest titer was 1:480). Although the antibody could also be detected in four of the six mice immunized with DNA vaccine containing four of the five antigen determinants, the antibody titer was lower (the highest titer was 1:80). DNA vaccine containing two of the five determinants could not induce the specific antibody.

CONCLUSIONThe idiotypic DNA vaccine containing the whole TCRbetaV five antigen determinants could induce the specific anti-lymphoma idiotypic antibody in BALB/c mice.

Amino Acid Sequence ; Animals ; Antibodies, Anti-Idiotypic ; blood ; immunology ; Base Sequence ; Complementarity Determining Regions ; genetics ; immunology ; Epitopes ; genetics ; immunology ; HL-60 Cells ; Humans ; Lymphoma ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; immunology ; Sequence Analysis, DNA ; Tumor Cells, Cultured ; Vaccines, DNA ; genetics ; immunology

Swine hepatitis E virus (HEV) is widespread throughout pigs in both developing and industrialized countries. This virus is an important zoonotic agent and a public concern worldwide. Infected pigs are asymptomatic, so diagnosing swine HEV relies on detection of the virus or antibodies against the virus. However, several obstacles need to be overcome for effective and practical serological diagnosis. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) that used a purified recombinant capsid protein of swine HEV. The potential clinical use of this assay was evaluated by comparing it with a commercial kit (Genelabs Technologies, Diagnostics, Singapore). Results of the ELISA were highly correlated with those of the commercial kit with a sensitivity of 97% and specificity of 95%. ROC (receiving operator characteristic) analysis of the ELISA data produced a value of 0.987 (95% CI, 0.977~0.998, p < 0.01). The cut-off value for the ELISA was also determined using negative pig sera. In summary, the HEV-specific ELISA developed in the present study appears to be both practical and economical.
Animals ; Antibodies, Anti-Idiotypic/*analysis/blood/genetics ; Capsid Proteins/*genetics/metabolism ; Enzyme-Linked Immunosorbent Assay/*methods/veterinary ; Hepatitis E/diagnosis/immunology/*veterinary/virology ; Hepatitis E virus/genetics/*isolation & purification/metabolism ; Immunoglobulin G/blood/genetics ; ROC Curve ; Recombinant Proteins/genetics/metabolism ; Swine ; Swine Diseases/*diagnosis/immunology/virology