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1.
Chinese Journal of Biologicals ; (12): 859-865, 2024.
Article in Chinese | WPRIM | ID: wpr-1039279

ABSTRACT

@#Objective To develop and verify a reporter gene assay for the determination of antibody dependent cellular phagocytosis(ADCP)potency of Ig G2 monoclonal antibody(m Ab)against epidermal growth factor receptor(EGFR)by combining Design of Experiment(DOE)and one factor at a time(OFAT).Methods The Jurkat/NFAT-Re/FcγRⅡa stably transformed cell line was used as effector cells,while the A431 cell line as the target cells.The JMP software was used to optimize the seven key factors in the experiment by combining DOE and OFAT analysis,while the ratio of upper and lower asymptotes(D/A)was used as the statistic,and the reporter gene method was developed to evaluate the ADCP potency of Ig G2 anti-EGFR m Ab.The method was verified according to the general chapter<9401>of Chinese Pharmaco-poeia(Ⅲ/Ⅳvolume,2020 edition)and used to determine the biological potency of Ig G2 anti-EGFR m Ab injection.Results After three rounds of experiments,the reporter gene method to evaluate the ADCP potency of Ig G2 anti-EGFR m Ab was developed.The method showed a dose-response relationship and was consistent with the four-parameter regression equa-tion y=(A-D)/[1+(x/C)~B]+D.The range of seven key conditions was determined:the density of effector cells was(1.25-3.75)×10~4 cells/well,the density ratio of effector cells to target cells was 1.0-2.0,the incubation time of target cells was 20-40 min,the incubation time of administration was 15-30 min,the total time was 5.5-6.5 h,and the color time was 5-30 min with luciferase detection system(Bright-Glo)as the color agent.The method had good specificity.Six independent tests were run for the five potency levels,with the correlation coefficient r of 0.994 5 and the linear regression equation slope of 1.02.The relative potency of five potency levels respectively was(62.15±1.38)%,(78.53±2.82)%,(99.12±3.95)%,(123.27±4.59)%and(155.22±7.04)%,the range of relative biases was-2.9%-0.2%,and the range of generalized cross-validation(GCV)was 2.2%-4.6%.The method had good linearity,relative accuracy and precision in the range of 64%-156%.The mean value of the potency of IgG2 anti-EGFR m Ab in three tests was(101.5±2.8)%.Conclusion The reporter gene assay developed in this study can be used to evaluate the ADCP potency of IgG2 anti-EGFR mAb

2.
Colomb. med ; 53(2): e2014832, Jan.-June 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1404385

ABSTRACT

Abstract Background: Inborn errors of immunity, mainly Predominantly Antibody deficiencies with normal IgG levels are unrecognized in adults with lung diseases such as bronchiectasis or recurrent pneumonia. Objective: To determine IgM, IgA, IgG2 subclass deficiencies, and Specific antibody deficiency (anti-pneumococcal polysaccharide antibodies) in adults with non-cystic fibrosis bronchiectasis or recurrent pneumonia. Methods: Cross-sectional study. Consecutive patients with non-cystic fibrosis bronchiectasis or recurrent pneumonia were recruited in Cali, Colombia. IgG, IgA, IgM, and IgE, IgG2subclass and IgG anti-pneumococcal serum levels were measured. Results: Among the 110 participants enrolled, Antibody deficiencies with normal serum IgG levels were found in 11(10%) cases. IgA deficiency (3 cases), IgM deficiency (2 cases) and IgG2 deficiency (2 cases) were the most frequent primary immunodeficiencies. In addition, IgG2+IgA deficiency, Ataxia-telangiectasia, Hyper-IgE syndrome and Specific Antibody Deficiency(anti-polysaccharides) were found in one case each. Conclusions: Predominantly antibody deficiencies with normal IgG levels are an important etiology of non-cystic fibrosis bronchiectasis and recurrent pneumonia in adults.


Resumen Antecedentes: Los Errores Innatos de la Inmunidad principalmente las Deficiencias Predominantemente de anticuerpos con niveles normales de IgG no se conocen en adultos con enfermedades pulmonares como las bronquiectasias o la neumonía recurrente. Objetivo: Determinar las deficiencias de IgM, IgA y de subclase de IgG2 y la Deficiencia Específica de Anticuerpos (anticuerpos antineumocócicos de polisacáridos) en adultos con Bronquiectasias no Fibrosis Quística (BQnoFQ) o neumonía recurrente. Métodos: Estudio observacional prospectivo. Se reclutaron 110 pacientes consecutivos con BQnoFQ o neumonía recurrente en Cali, Colombia. Se midieron los niveles séricos de IgG, IgA, IgM e IgE, subclase IgG2 y anticuerpos anti-neumococo. Resultados: Se encontraron deficiencias de anticuerpos con niveles normales de IgG en el 10% de los sujetos; Cuatro casos con IgG2 baja, incluido 1 caso de deficiencia de IgG2 + IgA, 1 caso de ataxia-telangiectasia, 3 deficiencias de IgA (IgAD), 2 deficiencias selectiva de IgM (IgMD), 1 síndrome de Hiper-IgE (HIES-AR) y 1 deficiencia específica de anticuerpos. Ocho pacientes fueron diagnosticados con enfermedades relacionadas con la hipogammaglobulinemia IgG. Conclusiones: Las deficiencias predominantemente de anticuerpos con niveles normales de IgG son una etiología importante de BQnoFQ y neumonía recurrente en adultos. Los sujetos con bronquiectasias o neumonía recurrente requieren una evaluación exhaustiva de la respuesta inmune humoral y clínica.

3.
Article in Chinese | WPRIM | ID: wpr-950262

ABSTRACT

Objective: To study the role of antibodies in protection against Leishmania tropica (L. tropica) infection in the experimental model of BALB/c mice. Methods: BALB/c mice were vaccinated against L. tropica by soluble Leishmania antigen or recombinant L. tropica stress-inducible protein-1 (LtSTI1) of L. tropica, and against Leishmania major (L. major) by soluble Leishmania antigen. Monophosphoryl lipid A was used as an adjuvant. The L. tropica- or L. major-vaccinated mice were challenged by L. tropica or L. major, respectively. The levels of anti-Leishmania antibodies (IgG1 and IgG2a) were determined after vaccination and after challenge. Results: All vaccinated groups caused a higher antibody response in comparison with the control group. The L. major-vaccinated group showed lower IgG1 response than the control group after the challenge. Conversely, in L. tropica-vaccinated mice, the levels of antibodies were higher than the control group. Moreover, the group receiving rLtSTI1 and monophosphoryl lipid A showed higher levels of antibodies than those of the rLtSTI1 group. In vaccinated mice, antibody responses against L. tropica remained high until 16 weeks after the challenge. Conclusions: The higher levels of post-challenge antibodies are associated with protective vaccination against L. tropica infection of BALB/c mice. Our findings provide new insight into the association of antibody with vaccine-induced protective immunity against L. tropica infection. More studies are needed to clarify the role of antibody in protection against L. tropica.

4.
Article in Chinese | WPRIM | ID: wpr-865424

ABSTRACT

Objective: To study the role of antibodies in protection against Leishmania tropica (L. tropica) infection in the experimental model of BALB/c mice.Methods: BALB/c mice were vaccinated against L. tropica by soluble Leishmania antigen or recombinant L. tropica stress-inducible protein-1 (LtSTI1) of L. tropica, and against Leishmania major (L. major) by soluble Leishmania antigen. Monophosphoryl lipid A was used as an adjuvant. The L. tropica- or L. major-vaccinated mice were challenged by L. tropica or L. major, respectively. The levels of anti-Leishmania antibodies (IgG1 and IgG2a) were determined after vaccination and after challenge. Results: All vaccinated groups caused a higher antibody response in comparison with the control group. The L. major-vaccinated group showed lower IgG1 response than the control group after the challenge. Conversely, in L. tropica-vaccinated mice, the levels of antibodies were higher than the control group. Moreover, the group receiving rLtSTI1 and monophosphoryl lipid A showed higher levels of antibodies than those of the rLtSTI1 group. In vaccinated mice, antibody responses against L. tropica remained high until 16 weeks after the challenge. Conclusions: The higher levels of post-challenge antibodies are associated with protective vaccination against L. tropica infection of BALB/c mice. Our findings provide new insight into the association of antibody with vaccine-induced protective immunity against L. tropica infection. More studies are needed to clarify the role of antibody in protection against L. tropica.

5.
Rev. cuba. invest. bioméd ; 38(4): e302, oct.-dic. 2019. graf
Article in Spanish | LILACS, CUMED | ID: biblio-1093425

ABSTRACT

El diagnóstico de la meningoencefalitis por Angiostrongylus cantonensis se establece por la presencia de las larvas del helminto en el líquido cefalorraquídeo, pero esta evidencia es muy difícil de encontrar ya que las larvas son lábiles en este medio y no se hallan con frecuencia. Debido a que en Cuba, la presencia del parásito se remonta a 1981 y este parásito es el único que puede provocar esta enfermedad en el país, se realiza una revisión con el objetivo de revisar la literatura publicada sobre el tema para acopiar toda la evidencia que ayude al diagnóstico auxiliar de meningoencefalitis eosinofílica. Se propone que el estudio de la síntesis intratecal de IgE y C3c, unido al patrón de síntesis local de IgA+IgG principalmente y de IgG1+ IgG2 resultan las más indicadas. Existen otras proteínas que pudieran auxiliar como la síntesis intratecal de C4 y en menor proporción MBL(AU)


Diagnosis of meningoencephalitis due to Angiostrongylus cantonensis is based on the presence of helminth larvae in cerebrospinal fluid, but such evidence is very hard to find, since the larvae are labile in this medium and cannot be spotted easily. Based on the fact that presence of the parasite in Cuba dates back to 1981, and this is the only agent of the disease in the country, a review was conducted with the purpose of going over the published literature about the topic and gather evidence leading to the auxiliary diagnosis of eosinophilic meningoencephalitis. The study of the intrathecal synthesis of IgE and C3c, alongside the local synthesis pattern for IgA+IgG mainly and IgG1+IgG2, are proposed as the most appropriate. Other useful proteins are the intrathecal synthesis of C4 and to a lesser extent MBL(AU)


Subject(s)
Immunoglobulin A , Angiostrongylus cantonensis , Meningoencephalitis , Chronology as Topic
6.
Chinese Pharmaceutical Journal ; (24): 627-631, 2018.
Article in Chinese | WPRIM | ID: wpr-858366

ABSTRACT

OBJECTIVE: To establish a capillary zone electrophoresis(CZE) -based method for charge heterogeneity analysis of IgG2 monoclonal antibody. METHODS: The concentrations of TETA and EACA, pH of separation buffer and temperature of capillary were optimized, to obtain a method which can efficiently separate the charge variants of IgG2 mabs. The reproducibility and repeatability of the optimized CZE method were validated and a battery of mabs were evaluated. RESULTS: The separation effect was significantly influenced by TETA/EACA concentration, pH and separation temperature. Through a series of optimization, a CZE method which showed good resolution and precision(RSD of area percentage was below 3.0% was established and the method parameters were as below: 400 mmol•L-1 EACA/0.05% TETA, pH 6.2, separation under 30 kV voltage at 35℃. This method was also suitable for IgG1 and IgG4 mabscharge heterogeneity analysis. Based on the specific electrophorogram and migration time, it could also be used as an identification method for monoclonal antibodies. CONCLUSION: The optimized CZE method can efficiently separate the charge variants of IgG2 mabs and shows good precision and specificity, which can be used to analyze the charge heterogeneity and identification of monoclonal antibodies.

7.
Chinese Pharmaceutical Journal ; (24): 335-339, 2018.
Article in Chinese | WPRIM | ID: wpr-858403

ABSTRACT

OBJECTIVE: To establish the methods of characterizing and enriching the disulfide isoforms of panitumumab which is a human IgG2 mAb that has been used in the treatment of metastatic colorectal cancer (mCRC) and evaluate the biological activity difference among the isoforms, ie, IgG2-A, IgG2-B, and IgG2-A/B. METHODS: The disulfide isoforms of panitumumab were identified by reversed-phase chromatography. The isoform-A of panitumumab was enriched upon reduction-oxidation treatment when guanidine was added and isoform-B was enriched upon reduction-oxidation treatment without guanidine. Then the molecule structural difference of the isoform-A and isoform-B was analyzed by size-exclusion chromatography. At last, the biological activities of these isoforms were further investigated by cell-killing assay. RESULTS: The component proportions of isoform A, B and A/B in panitumumab were 38%, 32% and 30%, respectively. Under reduction-oxidation conditions, the disulfide isoforms converted to isoform-A when 0.9 mol•L-1 guanidine was used, whereas isoform-B was enriched in the absence of guanidine. And isoform-A was eluted earlier in SECHPLC, suggesting a larger apparent molecular size as compared with isoform-B. Furthermore, the in vitro biological activity measurement showed an increased activity of IgG2-A compared with IgG2-B in inhibiting the growth of DiFi cells. The IC50 s for IgG2-A and IgG2-B were 0.095 46 μg•mL-1 (95% CI 0.079 86 - 0.114 1 μg•mL-1) and 0.372 8 μg•mL-1 (95% CI 0.306 7-0.453 1 μg•mL- 1), respectively. CONCLUSION: The methods for characterizing and enriching the disulfide isoforms are established. Difference in the biological activity between isform-A and isoform-B is observed. The methods will provide technical assist to the process optimization and quality control of panitumumab.

8.
Immune Network ; : e10-2018.
Article in English | WPRIM | ID: wpr-740202

ABSTRACT

Interaction between pathogen-associated molecular patterns and pattern recognition receptors triggers innate and adaptive immune responses. Several studies have reported that toll-like receptors (TLRs) are involved in B cell proliferation, differentiation, and Ig class switch recombination (CSR). However, roles of TLRs in B cell activation and differentiation are not completely understood. In this study, we investigated the direct effect of stimulation of TLR1/2 agonist Pam3CSK4 on mouse B cell viability, proliferation, activation, Ig production, and Ig CSR in vitro. Treatment with 0.5 µg/ml of Pam3CSK4 only barely induced IgG1 production although it enhanced B cell viability. In addition, high-dosage Pam3CSK4 diminished IgG1 production in a dose-dependent manner, whereas the production of other Igs, cell viability, and proliferation increased. Pam3CSK4 additively increased TLR4 agonist lipopolysaccharide (LPS)-induced mouse B cell growth and activation. However, interestingly, Pam3CSK4 abrogated LPS-induced IgG1 production but enhanced LPS-induced IgG2a production. Further, Pam3CSK4 decreased LPS-induced germline γ1 transcripts (GLTγ1)/GLTε expression but increased GLTγ2a expression. On the other hand, Pam3CSK4 had no effect on LPS-induced plasma cell differentiation. Taken together, these results suggest that TLR1/2 agonist Pam3CSK4 acts as a potent mouse B cell mitogen in combination with TLR4 agonist LPS, but these 2 different TLR agonists play diverse roles in regulating the Ig CSR of each isotype, particularly IgG1/IgE and IgG2a.


Subject(s)
Animals , Mice , B-Lymphocytes , Cell Proliferation , Cell Survival , Hand , Immunoglobulin Class Switching , Immunoglobulin E , Immunoglobulin G , In Vitro Techniques , Pathogen-Associated Molecular Pattern Molecules , Plasma Cells , Receptors, Pattern Recognition , Recombination, Genetic , Toll-Like Receptors
9.
Article in Chinese | WPRIM | ID: wpr-694839

ABSTRACT

Objective To investigate serum IgG subclass concentrations in the adult population of Hunan region,and the effects of age,gender and lifestyle on them.Methods Serum IgG1,IgG2,IgG3,IgG4 and IgG concentrations from 170 adults making a health examination were detected by the immunonephelometric assay.Results The concentrations (mean or median [P25,P75]) of serum IgG1,IgG2,IgG3,IgG4 and IgG were (7.53 ± 0.14) g/L,3.99 (3.13,5.02) g/L,0.49 (0.30,0.70) g/L,0.53 (0.26,0.93) g/L and 12.2 (10.5,14.1) g/L,respectively.The serum IgG1/IgG,IgG2/IgG,IgG3/IgG and IgG4/IgG were (61.3 ±0.69)%,33.38% (27.8%,38.8%),3.97% (2.5%,5.3%) and 4.44% (2.1%,7.3%),respectively.The serum IgG3 concentrations and IgG3/IgG ratios in female adults were significantly higher than that in male adults (P =0.005 and 0.014).However,there were no significant difference in serum IgG1,IgG2 and IgG4 concentrations and IgG1/IgG,IgG2/IgG and IgG4/IgG ratios between male and female adults.The serum IgG3 concentrations in the 31-40 years old adults were significantly higher than that in the 41-50 years old (P =0.03),while there were no significant difference in serum IgG1,IgG2 and IgG4 concentrations between different age groups.The serum IgG1 concentrations in the adults with heavy smoking were significantly lower than that without smoking (P =0.023),while the serum IgG4/IgG ratios were the opposite (P =0.018).The serum IgG1 and IgG3 concentrations and IgG3/IgG ratios in the adults with midrange or heavy drinking were significantly lower than that without ethanol consumption (P =0.05,0.004 and 0.015,respectively).The serum IgG3 concentrations and IgG3/IgG ratios in the adults with low-risk metabolism syndrome were significantly higher than that with the high-risk (P =0.034 and 0.038).Conclusion Gender and age have the significant effect on serum IgG3 concentration.Heavy smoking may reduce serum IgG1 concentration and increase IgG4/IgG ratio.The decrease of serum IgG1,IgG3 and IgG3/IgG may be related to ethanol consumption.

10.
Anatomy & Cell Biology ; : 124-134, 2017.
Article in English | WPRIM | ID: wpr-21762

ABSTRACT

Asthma is characterized by chronic inflammation, goblet cell hyperplasia, the aberrant production of the Th2 cytokines, and eosinophil infiltration into the lungs. In this study, we examined the effects of baicalein, wogonin, and Scutellaria baicalensis ethanol extract on ovalbumin (OVA)-induced asthma by evaluating Th1/Th2 cytokine levels, histopathologic analysis, and compound 48/80-induced systemic anaphylaxis and mast cell activation, focusing on the histamine release from rat peritoneal mast cells. Baicalein, wogonin, and S. baicalensis ethanol extract also decreased the number of inflammatory cells especially eosinophils and downregulated peribronchial and perivascular inflammation in the lungs of mice challenged by OVA. Baicalein, wogonin, and S. baicalensis ethanol extract significantly reduced the levels of tumor necrosis factor α, interleukin (IL)-1β, IL-4, IL-5 and the production of OVA-specific IgE and IgG1, and upregulated the level of interferon-γ and OVA-specific IgG2a. In addition, oral administration of baicalein, wogonin, and S. baicalensis ethanol extract inhibited compound 48/80-induced systemic anaphylaxis and plasma histamine release in mice. Moreover, baicalein, wogonin, and S. baicalensis ethanol extract suppressed compound 48/80-induced mast cell degranulation and histamine release from rat peritoneal mast cells. Conclusively, baicalein and wogonin as major flavonoids of S. baicalensis may have therapeutic potential for allergic asthma through modulation of Th1/Th2 cytokine imbalance and histamine release from mast cells.


Subject(s)
Animals , Mice , Rats , Administration, Oral , Anaphylaxis , Asthma , Cytokines , Eosinophils , Ethanol , Flavonoids , Goblet Cells , Histamine Release , Histamine , Hyperplasia , Immunoglobulin E , Immunoglobulin G , Inflammation , Interleukin-4 , Interleukin-5 , Interleukins , Lung , Mast Cells , Ovalbumin , Ovum , Plasma , Scutellaria baicalensis , Scutellaria , Tumor Necrosis Factor-alpha
11.
Clinics ; Clinics;71(12): 687-694, Dec. 2016. tab, graf
Article in English | LILACS | ID: biblio-840027

ABSTRACT

OBJECTIVE: To investigate the transmission of anti-Staphylococcus aureus (Sa) IgG, IgG1 and IgG2 via placental transfer and the transfer of IgA via the colostrum according to maternal Sa carrier status at delivery. METHODS: We evaluated anti-Sa IgG, IgG1 and IgG2 in maternal and cord sera and IgA in colostrum from a case (n=49, Sa+) and a control group (n=98, Sa-). RESULTS: Of the 250 parturients analyzed for this study, 49 were nasally colonized with S. aureus (prevalence of 19.6%). Ninety-eight non-colonized subjects were selected for the control group. The anti-Sa IgG, IgG1 and IgG2 levels and the IgG avidity indexes in the maternal and cord sera did not differ between the groups, with a low transfer ratio of anti-Sa IgG to the newborns in both groups. The anti-Sa IgG2 titers were significantly higher than the IgG1 titers in the maternal and cord sera. Inversely, the transfer ratios were higher for anti-Sa IgG1 compared with IgG2; however, no differences between the groups were detected. The Sa-specific IgA levels and avidity indexes in the colostrum were equivalent between groups. CONCLUSIONS: Maternal Sa nasal colonization at delivery is not associated with higher antibody levels in the mother or newborns. The high titers of anti-Sa IgG2 found in the cord serum indicate a greater reactivity with non-protein antigens, which may further contribute to the susceptibility to staphylococcal infections at birth. The presence of IgA in the colostrum with avidity to S. aureus reinforces the importance of breastfeeding shortly after birth.


Subject(s)
Humans , Female , Pregnancy , Infant, Newborn , Adult , Placenta/immunology , Staphylococcus aureus/immunology , Breast Feeding , Immunoglobulin G/blood , Immunity, Maternally-Acquired/immunology , Antibodies, Bacterial/blood , Reference Values , Staphylococcus aureus/isolation & purification , Umbilical Cord/immunology , Immunoglobulin G/immunology , Enzyme-Linked Immunosorbent Assay , Cross-Sectional Studies , Colostrum/immunology , Statistics, Nonparametric , Antibodies, Bacterial/immunology
12.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;49(8): e5215, 2016. graf
Article in English | LILACS | ID: lil-787389

ABSTRACT

Angiotensin-converting enzyme (ACE) inhibitors have non-hemodynamic, pleiotropic effects on the immune response. The effects of ACE inhibitors on the production of cytokines and T-cell functions are well established. However, little is known on the effects of these medicines on humoral response to foreign antigens. In this study, we investigated the effect of enalapril treatment on ovalbumin (OVA)-specific IgG1 and IgG2c production in mice determined by ELISA. Two groups of 8-week-old C57BL/6 females mice (3–4/group) were subcutaneously immunized with OVA (10 μg/animal) in presence of Alhydrogel (1 mg/mouse) and boosted at day 21. The mice were treated with enalapril (5 mg/kg daily, po) or were left without treatment for one month. The animals were bled from the orbital plexus on days 0, 7, 14, 21, and 28 after the first immunization and the sera were stored at –20°C until usage. OVA-specific serum IgG1 and IgG2c were determined by ELISA using serum from each individual animal. The results showed that enalapril significantly increased anti-OVA serum IgG2c in the secondary response without affecting IgG1 synthesis. These data expand our understanding on the properties of enalapril on the immune response, including antibody production.


Subject(s)
Animals , Female , Angiotensin-Converting Enzyme Inhibitors/pharmacology , Enalapril/pharmacology , Immunity, Humoral/drug effects , Immunoglobulin G/blood , Ovalbumin/immunology , Antibody Formation/drug effects , Immunoglobulin G/immunology , Mice, Inbred C57BL , Models, Animal , Th1 Cells/immunology , Th2 Cells/immunology
13.
Chinese Journal of Immunology ; (12): 323-328, 2015.
Article in Chinese | WPRIM | ID: wpr-460374

ABSTRACT

Objective:To investigate the effects of chitin on atopic dermatitis in an OVA induced AD murine model.Methods:Twenty-eight BALB/c mice were randomly divided into three groups:the normal control group (N)(8),the chitin group(E) (10) and the AD model group(M)(10).The murine model of atopic dermatitis was established through intraperitoneal injection of OVA followed by repeated epicutaneous application of OVA on mice back skin( AD model group).During the set up of AD murine model,mice of the chitin group were given intragastric gavage of 3 mg/d for 4 weeks.At the end of the experiment, the mice were sacrificed and skin lesions were biopsied for histological study.HE and O-toluidine stained paraffin sections were observed under microscope.The spleen cells were cultured and challenged with OVA and chitin,respectively,the supernatant was obtained for cytokine determination.Serum levels of total and OVA-specific IgE and total IgG2a were determined with ELISA.Results:Chitin significantly inhibited skin inflammation induced by OVA.Compared with the AD model group,the thickness of the epidermis and dermis in the chitin group were obviously decreased.The numbers of dermal infiltrated inflammatory cells,eosinophils and mast cells were significantly decreased in the chitin group compared with the AD model group ( P<0.05-0.001 ).The serum level of total IgE and OVA-specific IgE were significantly lower in the chitin group than in the AD model group(P<0.05-0.001),while the serum level of IgG2a in the chitin group was significantly higher than that of the AD model group( P<0.001).The cultured spleen cells of the chitin group produced significantly higher levels of IL-12 and IFN-γ,but lower level of IL-4 compared with those of the AD model group after OVA challenge (P<0.05).Conclusion:Chitin can inhibit the inflammation and decease the seum level of IgE in the murine AD model.The antiallergic effect of chitin might be associated with the induced production of Th1 type cytokines by mice spleen cells.

14.
J Vector Borne Dis ; 2012 Sept; 49(3): 151-156
Article in English | IMSEAR | ID: sea-142840

ABSTRACT

Background & objectives: The challenge of malaria and efforts targeted at developing malaria vaccines triggered this study on the reactivity of IgG and its subclasses in the test serum specific to CSP. This work was directed at assessing the influence of age and gender on host humoral antibody against Plasmodium falciparum recombinant circumsporozoite antigen in Nigerian children. Methods: In all, 67 serum samples (>10,000 parasites/μl of blood) collected from malaria-infected children at the University College Hospital, Ibadan during the transmission season were analyzed by ELISA. Results: The mean absorbance values of IgG subclasses reactive against P. falciparum CSP appeared to be agedependent and ranged from 0.01 for IgG4 in younger children to 0.95 for IgG3 in older children. The sixty-seven subjects investigated in this study had significantly higher mean IgG1 and IgG3 than the uninfected controls (p <0.01). This follows the order IgG3 >IgG1>IgG2>IgG4 which confirmed the prevalence of the cytophilic antibodies (IgG1 and IgG3) in 65% of the malaria infected children over the non-cytophilic subclasses (IgG2 and IgG4). Similarly, there was low production of IgG4 and IgG2 levels in 35% of the subjects compared with control. IgG was detected in the serum of North American Subjects (NAS) which served as negative control for CSP-specific IgG subclasses. Although the NAS titre was lower than that of the malaria subjects in Nigeria, its IgG2 was, however, higher (0.16) than that of other subclasses. The mean absorbance values of total serum IgG subclass were higher than those of IgG subclasses specific to P. falciparum circumsporozoite antigen. The mean absorbance values of the total serum IgG subclass follows the order IgG2>IgG1>IgG4>IgG3. Interpretation & conclusion: Age and gender-dependent correlations of results suggest that acquired immunity could play a significant role in protection from malaria. Antibody levels are higher in male than female children of the same age group. Antibody levels also increase with age in both the male and female children.

15.
Chongqing Medicine ; (36): 2081-2083,2086, 2011.
Article in Chinese | WPRIM | ID: wpr-598290

ABSTRACT

Objective To investigate whether R-848 could inhibit IgE production of mice immunized with OVA plus ALUM in vivo.Methods BALB/c mice were immunized s.c on the back with PBS,OVA,OVA plus R-848,OVA plus CpG,OVA plus ALUM,and OVA plus ALUM in R-848 or CpG every two weeks for three times.The blood from the mice was harvested after the last immunization,and the concentration of OVA specific or total IgE,IgG1 and IgG2a in the sera was determined;the splenocytes from the immunized mice were harvested and cultured with or without OVA for 3 days,and the concentration of IL-4 and IFNγ in the supernatants was determined.Results Firstly,we investigated that R-848 as Th1 adjuvant could enhance the production of OVA specific IgG2a in mice immunized with OVA.Secondly,further study showed that R-848 could inhibit the production of OVA specific IgE and total IgE,and promote the production of OVA specific IgG2a in the sera of mice immunized with OVA plus ALUM.Moreover,the results of ELISA showed that R-848 inhibiting IgE production was related to its reducing IL-4 production and enhancing IFNγ production.Conclusion R-848 could inhibit IgE production of mice immunized with OVA plus ALUM in vivo.

16.
Biosalud ; 8(1): 37-46, ene.-dic. 2009. tab
Article in Spanish | LILACS | ID: lil-555158

ABSTRACT

Estudios etnofarmacológicos previos han demostrado el uso de las hojas de Phenax rugosus (Poir.) Wedd. y Tabebuia chrysantha G. Nicholson para el tratamiento de enfermedades infecciosas y de procesos inflamatorios. En esta investigación se obtuvieron los extractos acuoso y metanólico de cada una de estas plantas para valorar en ratas Wistar la capacidad de los mismos para inducir cambios significativos en los niveles séricos de anticuerpos del tipo IgM e IgG2b. Los resultados obtenidos muestran que dichos extractos no produjeron una respuesta que tuviera significancia estadística cuando se compararon las concentraciones de las inmunoglobulinas de los grupos tratados con las del grupo control.


Previous Ethnopharmacological studies have showed the use of Phenax rugosus (Poir..) Wedd. and Tabebuia chrysantha G. Nicholson leaves for the treatment of infectious diseases and in inflammatory processes. Aqueous and methanolic extracts of each plant was obtained in order to determine the capacity of these extracts to induce significant changes in the seric levels of IgM and IgG2b antibodies in Wistar rats. The results obtained show that these extracts do not produce a response with statistical significance when comparing the antibody levels between the control and treatments groups.


Subject(s)
Antibodies , Biological Products , Immunoglobulin G , Immunoglobulin M , Tabebuia
17.
Article in English | WPRIM | ID: wpr-169039

ABSTRACT

The mRNA expression of several cytokines was evaluated in splenocytes and mesenteric lymph node (MLN) cells of rats infected with Capillaria hepatica by reverse-transcription (RT)-PCR until week 12 after infection. IgG1 and IgG2a, which are associated with Th1 and Th2 response, respectively, were also assessed by ELISA. The results indicated that the majority of cytokines, including the Th1 (IL-2 and IFN-gamma) and Th2 cytokines (IL-4, IL-5 and IL-10) were expressed at maximal levels during the early stage of infection (after week 1-2), and the ELISA data also evidenced a similar pattern of changes in IgG1 and IgG2a. Th1 and Th2 cytokines responded in a similar fashion in this rat model. The expression of cytokines in splenocytes was significantly higher than that in MLN cells, thereby indicating that cytokine production is controlled more by spleen than by MLN. In addition, the observation that IFN-gamma expression increased unexpectedly at the time of maximal egg production (6 weeks after infection) indicated that IFN-gamma is a cytokine reacting against egg production. However, increased IL-5 expression occurring in tandem with worm activity indicated that the activity of C. hepatica might be controlled by IL-5 expression.


Subject(s)
Animals , Rats , Antibodies, Helminth/blood , Capillaria/immunology , Cytokines/biosynthesis , Disease Models, Animal , Enoplida Infections/immunology , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation , Immunoglobulin G/blood , Lymph Nodes/immunology , Lymphocytes/immunology , RNA, Messenger/analysis , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Spleen/cytology , Th1 Cells/immunology , Th2 Cells/immunology
18.
Article in English | WPRIM | ID: wpr-114843

ABSTRACT

The identification and characterization of antigens that elicit human T cell responses is an important step toward understanding of Leishmania major infection and ultimately in the development of a vaccine. Micropreparative SDS-PAGE followed by electrotransfer to a PVDF membrane and elution of proteins from the PVDF, was used to separate 2 novel proteins from L. major promastigotes, which can induce antibodies of the IgG2a isotype in mice and also are recognized by antisera of recovered human cutaneous leishmaniasis subjects. Fractionation of the crude extract of L. major revealed that all detectable proteins of interest were present within the soluble Leishmania antigens (SLA). Quantitation of these proteins showed that their expression in promastigotes is relatively very low. Considering the molecular weight, immunoreactivity, chromatographic and electrophoretic behavior in reducing and non-reducing conditions, these proteins are probably 2 isoforms of a single protein. A digest of these proteins was resolved on Tricine-SDS-PAGE and immunoreactive fragments were identified by human sera. Two immunoreactive fragments (36.4 and 34.8 kDa) were only generated by endoproteinase Glu-C treatment. These immunoreactive fragments or their parent molecules may be ideal candidates for incorporation in a cocktail vaccine against cutaneous leishmaniasis.


Subject(s)
Animals , Humans , Antigens, Protozoan/chemistry , Blotting, Western , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel/methods , Leishmania major/growth & development , Protein Isoforms/chemistry
19.
Article in English | WPRIM | ID: wpr-60517

ABSTRACT

Experimental murine models with high, intermediate and low levels of genetically based susceptibility to Leishmania major infection reproduce almost entire spectrum of clinical manifestations of the human disease. There are increasing non-comparative studies on immune responses against isolated antigens of L. major in different murine strains. The aim of the present study was to find out whether there is an antigen that can induce protective immune response in resistant and susceptible murine strains. To do that, crude antigenic extract of procyclic and metacyclic promastigotes of L. major was prepared and subjected to SDS-PAGE electrophoresis. Western-blotting was used to search for antigen(s) capable of raising high antibody level of IgG2a versus IgG1 in the sera of both infected resistant and susceptible strains. Two novel antigens from metacyclic promastigotes of L. major (140 and 152 kDa) were potentially able to induce specific dominant IgG2a responses in BALB/c and C57BL/6 mice. The 2 antigens also reacted with IgG antibody of cutaneous leishmaniasis patients. We confirm that 140 and 152 kDa proteins of L. major promastigotes are inducing IgG production in mice and humans.


Subject(s)
Mice , Humans , Female , Animals , Protozoan Proteins/immunology , Mice, Inbred C57BL , Mice, Inbred BALB C , Life Cycle Stages/immunology , Leishmaniasis, Cutaneous/immunology , Leishmania major/immunology , Immunoglobulin G/biosynthesis , Blotting, Western/methods , Antigens, Protozoan/immunology
20.
Article in Korean | WPRIM | ID: wpr-58174

ABSTRACT

In most cases, acute diarrhea in childhood heals spontaneously, but it may become the form of chronic diarrhea in immunodeficient children and then cause weight loss, dehydration, malabsorption and malnutrition. The immunodeficient diseases associated with chronic diarrhea include severe combined immunodeficiency syndrome, common variable immunodeficiency, acquired immunodeficiency syndrome, agammaglobulinemia or selective IgA deficiency. IgA deficiency is the most common primary immunodeficiency. Because many IgA deficient individuals seem to have compensated for their deficiency with increased IgM production and various nonimmunologic factors, the incidence of gastrointestinal involvement is not prominent. Some of those with IgA deficiency and recurrent infections have been found to also have IgG subclass deficiency. IgA deficiency with IgG2 and IgG4 subclass deficiency have high susceptability to infection and chronic diarrhea. IgG subclass deficiency, when present, is more likely to be found in association with a partial IgA deficiency rather than complete IgA deficiency. We report a 3-month-old male with intractable diarrhea accompanied by IgA, IgG2, and IgG4 deficiency.


Subject(s)
Child , Humans , Infant , Male , Acquired Immunodeficiency Syndrome , Agammaglobulinemia , Common Variable Immunodeficiency , Dehydration , Diarrhea , IgA Deficiency , Immunoglobulin A , Immunoglobulin G , Immunoglobulin M , Incidence , Malnutrition , Severe Combined Immunodeficiency , Weight Loss
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