We evaluated tetanus specific IgG, IgM, IgG subclasses after DPT vaccination in infants and children. Tetanus toxoid specific IgG, IgM IgG subclasses were measured to characterize the isotope profile of antibody against tetanus toxoid. The values of the tetanus specific IgG in the positive group were significantly increased compared to those of the control group, and were significantly increased after two inoculation. Tetanus specific IgG was very low in adults and neonates. In our tetanus specific IgG subclasses study, forty-five of 56 cases (80%) showed predominantly IgG1 antibody responses to tetanus toxoid, while twenty-five of 56 cases (45%) showed IgG4 responses. Both IgG1 and IgG4 responses were demonstrated in 17 cases (30%). So we suggest that IgG was mainly involved in humoral immune response after DPT vaccination, and IgG1 may play an important role among IgG subclasses. IgG4, alone or together with IgG1, can also play a role in immune response to tetanus toxoid.
Antibodies, Bacterial/*biosynthesis ; Antibody Specificity ; Child ; Clostridium tetani/immunology ; Diphtheria-Tetanus-Pertussis Vaccine/*immunology ; Human ; Immunoglobulin G/biosynthesis/classification ; Immunoglobulin M/biosynthesis ; Infant

OBJECTIVETo determine the expression of immunoglobulins in HT-29 cells (an established colon cancer cell line, and explore their effect on the biological activities of the cancer cells.)

METHODSThe transcripts of variable regions of immunoglobulin heavy chains in HT-29 cells were detected by RT-PCR. Antisense CDR3 (specific to HT-29)-pIRES 1 neo vector was constructed, then transfected into HT-29 cells by electroporation. Programmed cell death and growth inhibition of HT-29 cells were detected by FCM and MTT, respectively.

RESULTSThe transcripts of Ig heavy chain (V(H) CDR3 region) were expressed in HT-29 cells. Moreover, they showed a monoclonal characteristic after being sequenced. After transfection of the antisense vector of CDR3 (specific to HT-29)-pIRES 1 neo, expression level of Ig in HT-29 cells was significantly decreased, and growth inhibition (P < 0.05) and apoptosis (P < 0.01) were induced.

CONCLUSIONThese results suggest that tumor derived Ig could promote the survival and growth of tumor cells.

Apoptosis ; Cell Proliferation ; Complementarity Determining Regions ; biosynthesis ; genetics ; DNA, Antisense ; genetics ; Electroporation ; Genetic Vectors ; HT29 Cells ; HeLa Cells ; Humans ; Immunoglobulin G ; metabolism ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin M ; metabolism ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; Immunoglobulins ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; genetics ; Transfection

OBJECTIVETo study the changes of immune function in mice offspring whose mothers were exposed to Rare Earth (RE)(NO(3))(3).

METHODSRE(NO(3))(3) was administered to mother mice after giving birth by gavage at dosages of 2, 20 and 200 mg/kg bw during breast-feeding period. The weights of spleen and thymus, the spleen plaque forming cells (PFC), the delayed type hypersensitivity (DTH) and the charcoal clearance of the offspring were determined.

RESULTSThe results obtained from the offspring after weaning showed that the body weight of offspring treated with 200 mg/kg RE(NO(3))(3) was 18.8% lower than that of the control group; at the dosage of 2 mg/kg, the number of IgM-PFC was increased by 82.7%; and at the dosage of 20 mg/kg the rate of clearance and clearance index were significantly higher than that of the control group. No difference in DTH was found in any treated group as compared to the control group. The results of offspring at three weeks after weaning showed that the number of IgM-PFC of the 20 and 200 mg/kg bw dose groups were 47.0% and 44.7% lower than that of control group respectively; the rate of clearance and clearance index of the 200 mg/kg group were significantly lower than that of the control group. No significant changes in DTH were observed in each exposed group.

CONCLUSIONRE(NO(3))(3) treatment affected the immune function of mice offspring which may caused by breast milk.

Animals ; Female ; Immunity ; drug effects ; Immunoglobulin M ; biosynthesis ; Metals, Rare Earth ; pharmacokinetics ; toxicity ; Mice ; Milk ; metabolism ; Organ Size ; drug effects ; Pregnancy

OBJECTIVESTo evaluate the sensitivity and specificity of IgM to recombinant antigen E2 of HEV envelope protein in the diagnosis of acute sporadic hepatitis E.

METHODSanti-E2 IgM was detected in sera samples from 176 cases of acute sporadic hepatitis E and 191 cases of acute non A-E hepatitis by ELISA and was compared with HEV IgM detected by some domestic and Genelabs (GL) kits. HEV RNA was also detected in sera positive for anti-E2 IgM. Logistic regression was used to analyze factors associated with the detection of anti-E2 IgM and HEV RNA.

RESULTSAnti-E2 IgM was found in 68.75% of the serum samples from the 176 patients of acute hepatitis E and the positive rate of HEV IgM detection by domestic kits was 56.25% (chi2 IgM = 6.49, P < 0.05). There were 37 (19.37%) anti-E2 IgM positive cases in those 191 sera of the acute non A-E hepatitis, out of which 11 cases were also detected as positive by the GL kit. Of the 158 anti-E2 IgM positive sera, HEV RNA was found in 81 (51.27%), among which 57.02% was positive in acute hepatitis E and 32.43% in acute non A-E hepatitis. No HEV RNA was found in the anti-E2 IgM negative sera from the cases of acute hepatitis E. By logistic regression analysis, no variance relative to the detection of anti-E2 IgM was found with the time interval from onset to hospitalization, age, levels of bilirubin, ALT and AST of the serum. Only the level of serum ALT was found being significantly associated with the detection of HEV RNA (P = 0.024).

CONCLUSIONS(1) anti-E2 IgM is a sensitive and specific serological maker for diagnosing an acute infection of HEV. (2) HEV is still the pathogen of some cases diagnosed clinically as non-A-E hepatitis. (3) Persistent HEV viremia is possibly an important factor influencing the severity of acute hepatitis E.

Adult ; Aged ; Aged, 80 and over ; Female ; Hepatitis E ; diagnosis ; Hepatitis E virus ; immunology ; Humans ; Immunoglobulin M ; biosynthesis ; Male ; Middle Aged ; Recombinant Proteins ; biosynthesis ; Sensitivity and Specificity ; Viral Envelope Proteins ; biosynthesis

OBJECTIVETo produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids.

METHODSrHuEPO was covalently coupled with bovine serum albumin (BSA) and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western blot.

RESULTSThe isotype of F3-mAb was found to be IgM with an affinity constant of 2.1 x 10(8) L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work.

CONCLUSIONSThe modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; Antibody Affinity ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Erythropoietin ; immunology ; Female ; Humans ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Weight ; Recombinant Proteins

OBJECTIVETo prepare monoclonal antibodies (McAbs) against human mesenchymal stem cells (hMSCs) and to study their biological characteristics.

METHODSBALB/C mice were immunized with pooled hMSCs. McAbs were prepared by hybridoma technique and their biological characteristics were analyzed by indirect immunofluorescence, immunohistochemistry and flow cytometry.

RESULTFive hybridoma cell lines were successfully established, which secret McAbs specifically against hMSCs. Investigations showed that all these McAb reacted only to hMSCs and had no cross-reaction to other human cells, the relative affinities of 5 McAbs were 1x10(6) (ZUB1), 1x10(5) (ZUB4), 1x10(6) (ZUC3), ZUE12 (1x10(5)) and 1x10(5) (ZUF10), respectively. Isotype analysis showed that ZUB1, ZUE12, ZUF10 against the same isotype, while ZUC3, ZUB4 against other two different isotypes alone. Flow cytometric analysis showed that the positive expression rate of cultured hMSCs was 87.39% (ZUB1), 88.07% (ZUB4), 88.12% (ZUC3), 69.89% (ZUE12) and 83.67% (ZUF10).

CONCLUSIONThe prepared five McAbs can specifically react against hMSCs, which can be used for selection and study of hMCSs.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibody Specificity ; Bone Marrow Cells ; cytology ; immunology ; Fluorescent Antibody Technique ; methods ; HL-60 Cells ; Humans ; Hybridomas ; secretion ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Mesenchymal Stromal Cells ; cytology ; immunology ; Mice ; Mice, Inbred BALB C ; Rats

We observed the time gap between oocyst shedding and antibody responses in mice (3-week-old C57BL/6J females) infected with Cryptosporidium parvum. Oocyst shedding was verified by modified acid-fast staining. The individually collected mouse sera were assessed for C. parvum IgM and IgG antibodies by enzyme-linked immunosorbent assay from 5 to 25 weeks after infection. The results showed that C. parvum oocysts were shed from day 5 to 51 post-infection (PI). The IgM antibody titers to C. parvum peaked at week 5 PI, whereas the IgG antibody titers achieved maximum levels at week 25 PI. The results revealed that IgM responses to C. parvum infection occurred during the early stage of infection and overlapped with the oocyst shedding period, whereas IgG responses occurred during the late stage and was not correlated with oocyst shedding. Hence, IgM antibody detection may prove helpful for the diagnosis of acute cryptosporidiosis, and IgG antibody detection may prove effective for the detection of past infection and endemicity.
Animals ; Antibodies, Protozoan/*biosynthesis/blood ; Cryptosporidiosis/*immunology ; Cryptosporidium parvum/*immunology/isolation & purification ; Enzyme-Linked Immunosorbent Assay ; Feces/parasitology ; Female ; Immunocompromised Host ; Immunoglobulin G/biosynthesis/blood ; Immunoglobulin M/biosynthesis/blood ; Mice ; Mice, Inbred C57BL ; Oocysts/immunology ; Specific Pathogen-Free Organisms ; Time Factors

Diagnosis of scrub typhus is difficult because its symptoms are very similar to other acute febrile illnesses, such as leptospirosis, murine typhus, and other viral hemorrhagic fevers. To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We have developed a chimeric recombinant antigen cr56 and two other recombinant antigens, r21 and kr56, from various serotypes of Orientia tsutsugamushi. They were tested for the detection of antibodies against O. tsutsugamushi in the patient's serum samples using enzyme-linked immunosorbent assay (ELISA) and dot-blot analyses. As of conventional immunofluorescence assay (IFA), when the mixture of these three recombinant antigens was used, both sensitivity and specificity of the recombinant antigens were increased up to 98% in IgM and IgG at ELISA and dot blotting. Additionally, both sensitivity and specificity by detection of IgM and IgG antibodies at rapid diagnostic test (RDT), using the mixture of three antigens and gold conjugated antibodies, were 99%. Our results suggest the use of mixture of these recombinant antigen proteins in ELISA or RDT is suitable as a diagnostic test for scrub typhus.
Antibodies, Bacterial/blood/chemistry/immunology ; Antigens, Bacterial/diagnostic use/genetics/metabolism ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect ; Gold/chemistry ; Humans ; Immunoassay ; Immunoglobulin G/blood ; Immunoglobulin M/blood ; Orientia tsutsugamushi/immunology/*metabolism ; Recombinant Proteins/biosynthesis/diagnostic use/genetics ; Scrub Typhus/*diagnosis ; Sensitivity and Specificity ; Serotyping

It is generally believed that liposomes modified with polyethylene glycol (PEG) have no or lower immunogenicity. However, based on many recent literatures, when the PEGylated liposomes were repeatedly applied to the same animal, the immune responses occurred. The first injection of PEGylated liposomes resulted in a reduction in the circulation time and an increase in hepatic and splenic accumulation of the second dose of PEGylated liposomes in a time-interval, which was called "accelerated blood clearance (ABC)" phenomenon. Such immunogenicity of PEGylated liposomes presents a barrier in the research of liposomal formulations and their use in the clinics. This review focused on the definition, the method of verification, the development of the reason for ABC phenomenon, influencing factors of ABC phenomenon, and discussed if other PEGylated nanocarriers also induce ABC phenomenon.
Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacokinetics ; Doxorubicin ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Immunoglobulin M ; biosynthesis ; blood ; Liposomes ; administration & dosage ; blood ; pharmacokinetics ; Liver ; metabolism ; Metabolic Clearance Rate ; Particle Size ; Polyethylene Glycols ; administration & dosage ; metabolism ; pharmacokinetics ; Serum Albumin, Bovine ; pharmacokinetics ; Spleen ; immunology ; metabolism