Immunogenicity Studies of Various Experimental Vaccines in Chickens

In this paper, the main objective was to raise chickens’ antibodies against three crucial public health microorganisms: the human immunodeficiency virus-1, Salmonella spp, and Staphylococcus aureus. Immunogens were prepared from the said microorganisms. Chickens were vaccinated either orally or intramuscularly. After a booster immunization, mostly eggs were collected and assess for the presence of specific antibodies. The most important results were the production of a large amount of anti-HIV antibodies in chicken’s eggs, and also the synthesis of anti-protein a antibodies with the ability to inhibit the growth of S. aureus in vitro and to serve as anti-anti-idiotypic antibodies with the capacity of neutralizing the original antigen. Enzyme- linked immune absorbent assays detected the presence of these antibodies as anti-Salmonella antibodies that were critical in reducing the bacterial load in the stomach and caeca compared with a control group. The vaccines were effective and safe, but more laboratory work, and economics have to be carried out to start a human trial.

Role of lactadherin anti-idiotypic monoclonal antibodies Ab2β in the treatment of human rotavirus-induced diarrhea in a neonatal mouse model / 中国热带医学

@#Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Role of lactadherin anti-idiotypic monoclonal antibodies Ab2β in the treatment of human rotavirus-induced diarrhea in a neonatal mouse model / 中国热带医学

@#Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Tratamiento del asma severo con omalizumab: experiencia pediátrica en Chile y revisión de la literatura / Omalizumab treatment on severe asthma patients: pediatric experience in Chile and literature review

This paper presents clinical experience with Omalizumab treatment in 8 pediatric patients in Chile. All children presented difficult to control asthma despite receiving high intensity treatment, with low quality of life. All patients were studied in order to discard errors in asthma diagnosis and to evaluate asthma treatment adherence and inhalation technique. After evaluation, patients proven to have severe therapy resistant asthma were indicated treatment with Omalizumab. Significant clinical improvement was observed, with reduced asthma symptoms and number of exacerbations, as well as an improved quality of life. Omalizumab showed a good safety profile with mild and transient adverse reactions in 6 administrations of a total of 122.

Se presenta la experiencia clínica con el uso de Omalizumab en 8 pacientes pediátricos en nuestro país. Todos los pacientes presentaban asma sin control a pesar de recibir terapia de alta intensidad, asociado a una muy deficiente calidad de vida. La totalidad de los pacientes fueron sometidos en cada centro a un estudio exhaustivo para poder descartar error en el diagnóstico y se evaluó la adherencia y la técnica inhalada. Al comprobarse que estos pacientes tenían asma severo resistente a tratamiento se indicó Omalizumab, el cual produjo una mejoría clínica significativa. Se observó una reducción de las exacerbaciones y de los síntomas de asma acompañado de una mejoría de la calidad de vida, asociado a un buen perfil de seguridad. Se observaron reacciones adversas leves y transitorias en 6 administraciones de un total de 122.

Identification of T cell epitopes from ovarian cancer associated anti-idiotype antibody / 中华妇产科杂志

Objective To identify the T cell epitopes from ovarian cancer associated anti-idiotypic antibody 6B11in order to explore the molecular basis of 6B11 induced cellular immune responses against ovarian cancer.Methods Potential human leukocyte antigen(HLA)A0201 ligands were predicted by using SYFPEITHI algorithm and tested by the T2 binding assay for screening of HLA-A2 binding peptides from 6B11 complimentary determining region(CDR).Cytotoxic T lymphocytes(CTL)to 6B11 or peptides were generated by 3 rounds of in vitro stimulation with 6B11 or peptide-pulsod dendritic cells(DC),and then tested by 51Cr-release assay to ascertain the CTL epitope of 6B11.Cell proliferation assay was performed by using 6B11 specific CTL as responder cells and peptide-pulsed DC as stimulators to ascertain the helper T lymphocyte(Th)epitope of 6B11.Cytokine assay and interferon-γ ELISPOT assay were performed to verify the CTL and Th epitope of 6B11 further.Results Light chain CDR3 peptide(VL CDR3)of 6B11 induced specific CTL to kill HLA-A2 and target antigen positive ovarian cancer cells,which could be blocked by anti human major histocompatibility complex(MHC)Ⅰ antibody.Heavy chain CDR3 peptide(VH CDR3)of 6B11 stimulated the proliferation of 6B11-primed CTL,which could be blocked mainly by anti human MHC-Ⅱ antibody,and further experiments showed that part of the VH CDR3 peptide-primed CTL killed K562 cells.Peptides in VL CDR3 and VH CDR3 were the CTL and Th epitope mimicking the original antigen of 6B11 respectively.Collaboration of 6B11 CTL and Th epitope,or 6B11 CTL epitope and keyhole limpet hemocyanin(KLH),the former was more powerful in inducing specific cellular immune responses against ovarian cancer.6B11 or corresponding CTL and Th epitope specific CIL secreted high levels of interleukin-2 (1630,1503 ng/L)and interferon-γ(5620,5421 ng/L),while basal level of interleukin-4 was detected (253,274 ng/L).ELISPOT assay confirmed the existence of specific interferon-γ secreting cells in 6811 or epitopes specific CTL(196/1×106 T cell,184/1×106 T cell).Conclusions VL CDR3 and VH CDR3 peptides of 6B11 are the CTL and Th epitopes mimicking original antigen which could duplicate the activity of intact 6B11 to induce the cellular immune responses against ovarian cancer.The results have significant theoretical and practical value in application of anti-idiotypic antibody ag anti tumor vaccine.The acquired CTL and Th epitopes as constituents of future multiple peptides against ovarian cancer could be used in peptide vaccine based ovarian cancer therapy.

Screening and identification of recombinant anti-idiotypic antibody against monoclonal antibody MGb1 / 免疫学杂志

Objective T9 provide candidate molecules for developing recombinant anti-idiotypic antibody (anti-Id) vaccine of gastric carcinoma by selection of recombinant anti-Id to monoclonal antibody ( McAb) MGb1 directed against the cancer with phage display technique.Methods Balb/c mice were immunized with MGb1 and the mRNA was isolated from the spleens of the immunized mice. The VL and VH cDNAs of the antibody were amplified separately by RT-PCR and assembled into ScFv DNAs with a linker DNA. The ScFv DNAs were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage antibody ScFv library. After four rounds of biopanning to the library with MGb1, the MGb1-positive clones were selected from the enriched phages by ELISA. The types of the anti-Id ScFv displayed on the selected phage clones were preliminarily identified by competition ELISA. Results The VL and VH cDNAs was about 320 bp and 340 bp, respectively. The ScFv DNA were about 750 bp. After four rounds panning to the phage antibody ScFv library with MGb1, 18 MGb1-positive phage clones displayed anti-Id ScFv were selected from 50 pre-selected phage clones, among which 4 clones displayed β or γ type anti-Id ScFv. Conclusion The phagedisplayed anti-Id ScFvs to McAb MGb1 are successfully selected by recombinant phage antibody technique, which might lay a foundation for screening the anti-Id ScFv possessing the characteristics of inducing anti-gastric carcinoma immunity.

Potentiality of Anti-idiotypic Antibodies Mimicking GD2 to Induce Cellular Immunity

BACKGROUND: Disialoganglioside GD2 is a tumor-associated antigen that is overexpressed on tumor cells of neuroectodermal origin, such as melanoma, small cell lung carcinoma and neuroblastoma. Immunity against GD2 has anti-tumor activities, but GD2 is poorly immunogenic. Anti-idiotypic antibodies that mimic GD2 may induce more effective immune responses than GD2 antigen itself, because they are protein antigens and are known to be able to break immune tolerance. In our previous study, we produced anti-idiotypic antibodies mimicking GD2 (3A4 and 3H9), which induced humoral immunity. However, cellular immunity is essential to eradicate tumor cells in vivo as well as humoral immunity. In the present study, we investigated whether these anti-idiotypic antibodies 3A4 and 3H9 could induce cellular immunes responses. METHODS: BALB/C mice were immunized with anti-idiotypic antibody 3A4 or 3H9, or normal mouse IgG as a negative control. Lymphoproliferative responses, cytokine production responses, and delayed-type hypersensitivity reactions were measured in mice immunized with the anti-idiotypic antibodies. RESULTS: Both the anti-idiotypic antibody 3A4 and 3H9 induced GD2-specific lymphoproliferative responses and IFN-gamma production of lymph node lymphocytes in BALB/C mice. Only anti-idiotypic antibody 3H9 induced significant GD2-specific delayed-type hypersensitivity in the mice. CONCLUSION: These results show that anti-idiotypic antibodies 3A4 and 3H9 have the potentiality of inducing GD2-specific cellular immune responses that cannot be induced by the native antigen GD2 itself.

Generation of antiidiotypic antibody to monoclonal antibody MGd1 directed against gastric carcinoma by phage antibody library technique / 第三军医大学学报

Objective To pave the way for developing recombinant antiidiotypic antibody(antiId)vaccine of gastric carcinoma by generating phagedisplayed antiId to monoclonal antibody MGd1 directed against the cancer. Methods Balb/c mice were immunized i.p. with MGd1 conjugated with KLH, and mRNA was isolated from the spleens of the immunized mice. VH and VL DNAs of the antibody were amplified separately by RTPCR and assembled into ScFv DNAs with a linker DNA by PCR. The ScFv DNAs were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E.coli TG1. The transformed cells were infected with M13KO7 helper phage to yield phage antibody ScFv library. After four rounds of panning to the library with MGd1, the MGd1positive clones were selected by ELISA from the enriched phages. The types of the antiId ScFv displayed on the selected phage clones were preliminary identified by competition ELISA. Results The VH,VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. After four rounds of panning to the antibody library, 17 MGd1positive phage clones displayed antiId ScFv were selected from 40 enriched phage clones, among which 3 displayed ? or ? type antiId ScFv. Conclusion The successful generation of antiId ScFv to monoclonal antibody MGd1 by recombinant phage antibody library technology might lay a foundation for screening of novel candidate molecules for developing recombinant antiId vaccine of gastric carcinoma.

Characterization of Anti-anti-idiotypic Antibodies (Ab3) Induced by Immunization of Anti-idiotypic Antibodies (Ab2) Mimicking Disialoganglioside GD2

BACKGROUND: Disialoganglioside GD2 is a tumor-associated antigen that is overexpressed on tumor cells of neuroectodermal origin, such as melanoma and neuroblastoma. Anti-idiotypic antibodies that mimic GD2 may induce more effective immune responses than GD2 antigen itself, because they are protein antigens and are known to be able to break immune tolerance. In this study, to explore the potential of anti-idiotypic antibodies as tumor vaccines, the ability of anti-idiotypic antibodies (Ab2) to induce anti-anti-idiotypic antibodies (Ab3) that bind to the original antigen GD2 was investigated. METHODS: Six monoclonal anti-idiotypic antibodies (1A8, 1G5, 2B6, 3A4, 3D6, 3H9) to monoclonal antibody M2058, which is a monoclonal antibody to GD2, were produced in mice. Three (1A8, 3A4, 3H9) of them were selected based on their ability to inhibit the binding of Ab1 to D142.34 (murine melanoma cell expressing GD2). These 3 different Ab2 were injected into rabbits, and rabbit Ab3 induced by each of them were characterized. RESULTS: Ab3-containing sera from two rabbits immunized with 1A8, 3A4, or 3H9 bound significantly (P<0.05) to D142.34 but not to B78.96 (GD2-negative cell), and bound significantly (P<0.05) to isolated GD2 but not to GD1a. Ab3-containing sera from two rabbits immunized with 3A4 or 3H9 inhibited significantly (P<0.05) the binding of Ab1 M2058 to D142.34, and inhibited significantly (P<0.05) the binding of Ab1 M2058 to the Ab2. CONCLUSION: These results suggest that anti-idiotypic antibodies 3A4 and 3H9 have a potential to be used as vaccines against tumors expressing GD2 by inducing GD2-specific antibodies (Ab3).

Screening of anti-idiotypic antibody to monoclonal antibody MC3 directed against colorectal carcinoma by phage antibody library technology / 中国免疫学杂志

Objective:To generate phage displayed anti idiotypic antibody single chain variable fragments(anti Id ScFv)to monoclonal antibody MC3 directed against colorectal carcinoma.Methods:Balb/c mice were immunized i.p. with MC3(McAb against colorectal Carcinoma) conjugated with KLH,and mRNA was isolated from the spleens of the immunized mice.VH and VL DNAs of the antibody were amplified separately and assembled into ScFv DNAs with a linker DNA by PCR.The ScFv DNAs were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E.coli TG1.The transformed cells were infected with M13KO7 helper phage to yield phage antibody ScFv library.After four rounds of panning to the library with MC3,the MC3 positive clones were selected by ELISA from the preselected phages.Results:The VH,VL and ScFv DNAs were about 340,320 and 750 bp respectively.After four rounds of panning to the antibody library,15 MC3 positive phage clones displayed anti Id ScFv were selected from 50 enriched phage clones.Conclusion:The phage displayed anti Id ScFv to MC3 were successfully selected by phage antibody library technology,which might provide new putative candidate molecules for developing recombinant anti Id vaccine of colorectal carcinoma.

Inmunovaloración de anticuerpos IgM en muestras sospechosas de sarampión clínicamente

Se realiza el estudio de 60 sueros pareados sospechosos de Sarampión clínicamente llegados al Laboratorio Diagnóstico del Instituto de Medicina Tropical "Pedro Kourí" entre enero y mayo de 1996, procedentes de la vigilancia seroepidemiológica de la vacuna triple viral (sarampion, rubéola y parotiditis), a los cuales se les realizó la detección de anticuerpos hemaglutinantes a sarampión y rubéola, así como de IgM, con el estuche diagnóstico Clark Laboratories INC Measles IgM ELISA. Los casos positivos se confirmaron por las técnicas de inmunofluorescencia indirecta-IgM y neutralización. Se obtuvieron por inhibición de la hemaglutinación, 3 casos positivos a sarampión y rubéola, los cuales negativos a IgM de sarampión y a los que se les determinaron anticuerpos a Epstein Barr, Citomegalovirus y al virus herpes simple mediante inmunifluorescencia indirecta (IFI) la capacidad de estos virus de inducir respuesta policlonal. Por medio del ELISA IgM Clark se detectaron 6 casos positivos los cuales resultaron negativos por IFI.

60 matched sera clinically syspicious of measles that were received at the Diagnostic Laboratory of the "Pedro Kourí" Tropical Medicine Institute between January and May, 1996, coming from the seroepidemiological surveillance of the MPR vaccine were studied. The detection of measles and rubella hemagglutinant antibodies, as well as of IgM, was carried out with the Clark Laboratories INC. Measles IgM ELISA diagnostic kit. The positive cases were confirmed by the IgM-indirect immunofluorescence and neutralization. 3 positive cases to measles and rubella, which were negative to measles IgM, were obtained by hemagglutination inhibition. Antibodies against Epstein Barr, cytomegalovirus and herpes simplex virus were also determined by indirect immunofluorescence (IIF) due to the capacity of these viruses to induces polyclonal responses. 6 positive cases, which were negative by IIF, were detected by means of the above mentioned diagnostic kit.

Production of Monoclonal Anti-idiotypic Antibody to Monoclonal Anti-DNA Antibody / 대한면역학회지

It has been thought that autoimmune diseases like systemic lupus erythematosus and rhumatoid athritis are closely associated with anti-DNA antibodies (Abs). In studies of the control for anti-DNA Ab generation, an understanding of the regulatory mechanisms by anti-idiotypic Abs that influence the production of anti- DNA Abs would be facillitated by the availability of the hybridomas producing the pairs of DNA-specific and anti-DNA's idiotope-specific monoclonal antibodies (mAbs). We have produced a series of anti-DNA mAbs and then monoclonal anti-idiotypic Ab directed against idiotypic determinant of the 3D8 mAb that has the highest affinity to dsDNA and ssDNA among the anti-DNA mABs that we had obtained. The spleen cells of the MRL-Ipr/Ipr, autoimmune prone, mice were fused with P3X63Ag8.653 myeioma cells to obtain anti-DNA Ab secreting hybridomas. Out of the fourteen clones that showed strong binding to ssDNA, four clones had cross-reactivity with dsDNA whereas none of these clones reacted with left-handed z-DNA. The binding activities of the anti-DNA mAbs to various synthetic polynucleotide sequences were different respectively. Anti-idiotypic mAbs were generated by the fusion of myeloma cells and spleen cells from the Balb/c mice immunized with 3DB-Fab. We have produced two anti-idiotypic mAbs, B7 (IgG2a/k) and 02F3 (IgM/k), which were specific to 3DB-Fab and cloned the variable region of the heavy chain from the 02F3 hybridoma.

Cuantificación de IGG contra los componentes inmunogénicos de VA-ENGOC-BC en plasma de lactantes vacunados

Debido a las características particulares de la respuesta inmune en lactantes, así como a la eficacia mostrada por la vacuna cubana antimeningocóccica VA-MENGOC-BC, nos propusimos cuantificar la respuesta de la inmunoglobulina G contra los componentes inmunogénicos de los meningococos B y C presentes en la vacuna, en lactantes vacunados. Se tomó muestra por punción capilar a 109 lactantes entre 3 y 6 meses de edad antes de la vacunación a los 31,4 ± 2 días después de la primera y 32,3 ± días después de la segunda dosis vacunal. Se determinaron las concentraciones de inmunoglobulina G contra cada inmunógeno de la vacuna. Los niveles de inmunoglobulina G específica prevacunación, fueron elevadas contra el meningococo C. Se produjo un incremento estadísticamente significativo de anticuerpos para ambos inmunógenos después de la primera y segunda dosis, más marcado contra el meningococo C en la primera y para el meningococo B en la segunda, lo que apoya la presencia de memoria inmunológica.

Due to the particular characteristics of the immune response in infants and to the efficacy shown by the Cuban antimeningococcal vaccine denominated VA-MENGOC-BC, we propose ourselves to quantify the response of immunoglobulin G against the immunogenic components of the meningococci B and C present in the vaccine among the vaccinated infants. The sample was taken by capillary puncture from 109 infants between 3 and 6 months before vaccination, at 31.4±2 days after the first dose and at 32.3± days after the second one. The concentrations of immunoglobulin G against each immunogen of the vaccine were determined. The levels of prevaccination specific immunoglobulin G were elevated against meningococcus C. There was a statistically significant increase of atibodies for both immunogens after the first and second dose. It was more marked against meningococcus C in the first, and for meningococcus B in the secons one, which supports the presence of immunological memory.

CLONING AND SEQUENCE ANALYSIS OF THE LIGHT CHAIN VARIABLE REGION GENE OF MONOCLONAL ANTI-IDIOTYPIC ANTIBODY NP30 OF SCHISTOSOMA JAPONICUM / 中国寄生虫学与寄生虫病杂志

Objective] To amplify and sequence the light chain of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum. [Methods] By comparing the conserved regions at each end of the nucleotide sequences of murine germ line genes enco ding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of V L gene. The hybridoma cells secreting anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger′s method. The V L gene was compared with GenBank and published mouse V L genes. [Results] The full length of V L gene was 318 bp. The V L gene was a member of mouse Ig ? light chain subgroup IV and generated from rearrangement of germ line V and J? 4 genes. The V L gene sequence has been registered by GenBank(accession No. AF206720). [Conclusion] The obtained V L gene was a potentially functional gene of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum .

Active Immunotherapy with Anti-Idiotypic Antibody for Nasopharyngeal Cancer / 中国肿瘤生物治疗杂志

According to the immune network hypothesis proposed by Jeme, certain anti-idiotypic antibodies (Ab2) express three dimensional shapes which resemble the structure of natural antigens. Here we propose a study of active immunotherapy by Ab2 for patients with Nasopharyngeal Cancer (NPC) . Two Ab2,designated 2H4 and 5D3,against two Abl (FC_2 and HNL5)that recognize NPC associated antigen were generated. They could substitute NPC antigen to induce humoral and cellular immune response against NPC cells in syngeneic mice. Nineteen patients with NPC at stage IV were chosen for active immunotherapy. They were treated with Alum-precipitated 2H4 or 5D3 accompanying radiotherapy. Nine patients with radiotherapy alone were as control. Both anti-anti-idiotypic antibodies (Ab3) and anti-NPC antibodies (Abl') were increased and human anti-mouse antibodies (HAMA) occurred in nineteen patients of the experimental group; whereas the levels of Abl' did not rise in control group.Serum IL-2, IFN-? and TNF-? level were increased in most patients in experimental group. While in the control group, there was no difference of cytokine level between pretherapy and post- therapy.

Detection of Pemphigus Antibodies with Indirect ELISA and ABC ELISA / 中华皮肤科杂志

Objective To establish a new method for detecting pemphigus antibody (PAb). Methods PI 1 anti idiotype monoclonal antibodies against pemphigus and purified PAb from the serum of a patient with active pemphigus were used to establish an ELISA system for detecting the PAb. Results The result showed the purified PAb was IgG4 subclass, the sensitivity and specificity for the detection of PAb were high in the ELISA system, the sensitivity and specificity were not significantly different among the IIF, indirect ELISA and ABC ELISA. The standard curve for detecting the concentration of the purified PAb was primarily obtained in the study. Conclusion The ELISA system for the detection of PAb in the sera of patients is a good qualitative method, it might be of value in the clinical diagnosis of pemphigus. It is expected to quantitatively detect PAb of pemphigus patients in the future with the ELISA system established, which is directed to the IgG4 subclass of PAb, so it may be of value in the study of IgG4 subclass in the pathogenesis of pemphigus.

MOLECULAR MECHANISM OF APOPTOSIS OF CELLS IN EGG GRANULOMA INDUCED BY ANTI-IDIOTYPIC ANTIBODY NP30 / 中国血吸虫病防治杂志

Objective To study the molecular mechanism of apoptosis of cells in egg granuloma induced by anti-idiotypic antibody NP30 of Schistosoma japonicum.Methods BALB/c mice were randomly divided into two groups. The mice of the experimental group were immunized by injecting NP30 intraperitoneally for three times, while the mice of control group were injected normal saline intraperitoneally. The mice were sacrificed respectively on the 39th, 49th, 64th, 108th, 112nd day after challenge with schistosome cercariae. The expressions of apoptosis-related gene Bax, Bcl-2, death receptor Fas, FasL (Fas ligand) and c-Fos were examined by the S-P method of immunohistochemistry,and Bax, mRNA and Fas mRNA investigated by the in-situ hybridization. Results The expressions of Bax, Fas, FasL and c-Fos were positive in granuloma cells of both groups. The expressions of Bax and FasL in experimental group were higher than those in control group (P

Construction, expression and characterization of a mono-specific bivalent diabody derived from an anti-anti-idiotypic monoclonal antibody NP48 of Schistosoma japonicum / 中国血吸虫病防治杂志

Objective To construct a mono-specific bivalent diabody (scFv dimer) gene derived from an anti-anti-idiotypic monoclonal antibody NP48 of Schistosoma japonicum and to express and characterize the protein.MethodsThe mono-specific diabody gene (D) was constructed by SOE (splicing by overlap extension) and using Gly_4Ser as a linker to join the C-terminus of the V_H to the N-terminus of the V_L.D was linked with prokaryotic expression vector pBAD/g. The target protein expression in E.coli TOP10 was induced by arabinose. Then a purification procedure for the target protein was carried out. The antigen binding activity of expressed product was detected with Dot-ELISA. ResultsThe V_H-G_4S-V_L (D) gene was confirmed by sequencing. The pBAD/g-D recombinant were determined by digesting with endonucleases and expected bands were identified. There were less soluble target proteins in the supernantes and higher target proteins in the pellets as inclusion body when separating the D expression proteins. And the insoluble fraction was recovered as a soluble, correctly processed protein by solubilising with 8 mol/L Urea. The molecular weight of the target protein was about 27 kD. The binding activity of the target protein to anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum was verified by Dot-ELISA. ConclusionThe purified protein from the constructed recombinant pBAD/g-D could interact specifically with antigen NP30. So the constructed mono-specific diabody has the part characteristics of anti-anti-idiotypic monoclonal antibody NP48 of Schistosoma japonicum.

Effects of Anti Idiotypic Antibody NP30 on Modulation of Egg Granuloma Formation and Hepatic Fibrosis of Schistosomiasis * / 中国寄生虫学与寄生虫病杂志

Objective To study the effects of the monoclonal anti idiotypic antibody NP30 active immunization on egg granuloma formation and hepatic fibrosis in Schistosoma japonicum infection. Methods ICR mice were actively immunized with NP30 100 ?g ?3 ip. every 10 days while the mice in control group were injected with SP2/0 ascites ip. simultaneously. After cercariae challenging,the mice were killed at the 4th, 8th,12th, 16th, 20th and 24th week, respectively.Mouse livers were removed and stained histochemically with VG and subjected to immunohistochemical assay of collagen type Ⅰ,Ⅲ and fibronectin(FN).The volume of egg granulomas and the content of collagen type Ⅰ,Ⅲ and FN were determined quantitatively by NYD 1000 Image Analysis System. Results The volume of egg granulomas in NP30 immunized group was much smaller than that of control group from the 12th week after cercariae challenge. The cellular components of egg granulomas in NP30 immunized group were significantly different from those of the control group,exhibiting two types of atypical egg granulomas were found.VG stain revealed that the average optical density of collagen in hepatic granulomas of experimental group was lower than that of control group.Immunohistochemical assay revealed that the contents of collagen type Ⅰ,Ⅲ and fibronectin in egg granulomas of experimental group were lower than those of control group. Conclusion NP30 vaccination may induce both cellular and humoral protective immunity to modulate egg granulomas and suppress liver fibrosis of schistosomiasis japonica.

A STUDY ON THE MONOCLONAL ANTI-IDIOTYPIC ANTIBODY NP30 FOR THE DIAGNOSIS OF SCHISTOSOMIASIS JAPONICA / 中国寄生虫学与寄生虫病杂志

In this study the ability of the monoclonal anti-idiotypic antibody NP30 was tested as a substitute of diagnostic antigen in detecting antibody of Schistosoma japonicum from human sera by use of ELISA. The results showed that the seropositive rate was 98% with NP30 in the group of acute infection, which was comparable to 94% with gut associated antigens (GAA)and 98% with the soluble egg antigens (SEA); 87% with NP30 in the group of chronic infection which was comparable to 86% with GAA but lower than that of 98% with SEA. The false positive rate was about 3% for all three diagnostic antigens. The results also showed that the geometric mean titer (GMT) of antibody to NP30 was higher than that to GAA but lower than that to SEA in the acute infection group and the GMT of antibody to NP30 was lower than both those to GAA and to SEA in the chronic infection group,suggesting that the antibody to NP30 appeared earlier and decayed more quickly during the process of infection. The authors suggested that NP30 could be used for the diagnosis of schistosomiasis japonica.