Isolation of Theileria parasites from African buffalo (Syncerus caffer) and characterization with anti-schizont monoclonal antibodies

Parasitology ◽  
1987 ◽  
Vol 94 (3) ◽  
pp. 413-423 ◽  
Author(s):  
P. A. Conrad ◽  
D. A. Stagg ◽  
J. G. Grootenhuis ◽  
A. D. Irvin ◽  
J. Newson ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Lymphoblastoid Cell Lines ◽  
African Buffalo ◽  
Syncerus Caffer ◽  
Antigenic Diversity ◽  
Parent Clone ◽  
Parasite Populations ◽  
Limiting Dilution

Antigenic differences between intra-lymphocytic theilerial parasites isolated from the blood of 18 African buffalo and grown in vitro were assessed with anti-schizont monoclonal antibodies (mAbs). There was marked antigenic diversity both between isolates from different buffalo and between isolates taken at different times from the same buffalo. Many of the isolates from both wild and captive buffalo appeared to consist of mixed parasite populations. Some isolates were found by limiting dilution cloning and mAb testing to contain at least 3 or 4 distinct populations of Theileria. Once cloned, Theileria-infected lymphoblastoid cell lines retained their mAb profiles during prolonged in vitro cultivation and, when recloned, the subclones had the same mAb profile as their parent clone. The implications of these results for further studies on buffalo-derived theilerial parasites are discussed.

In Vitro Cultivation of Theileria lawrencei-Intected Lymphoblastoid Cell Lines Derived from a Buffalo (Syncerus caffer)

1974 ◽  
Vol 16 (1) ◽  
pp. 125-127 ◽  
Author(s):  
D.A. Stagg ◽  
C.G.D. Brown ◽  
J.G. Crawford ◽  
G.K. Kanhai ◽  
A.S. Young
Keyword(s):  
Cell Lines ◽  
In Vitro Cultivation ◽  
Lymphoblastoid Cell Lines ◽  
Syncerus Caffer

PRODUCTION OF HUMAN MONOCLONAL ANTIBODIESSPECIFIC FOR PLATELET MEMBRANE GLYCOPROTEIN IIIa

1987 ◽  
Author(s):  
Kenichi Furihata ◽  
Diane J Nugent ◽  
Amy L Bissonette ◽  
Elizabeth Vokac ◽  
Thomas J Kunicki
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Peripheral Blood Lymphocytes ◽  
Platelet Membrane ◽  
Lymphoblastoid Cell Lines ◽  
Membrane Glycoproteins ◽  
Human Monoclonal Antibodies ◽  
Microtiter Plates ◽  
Culture Supernatants ◽  
Limiting Dilution

Human monoclonal antibodies specific for platelet membrane glycoproteins (GPs) arepotentially important reagentsfor studies of the immunogenicity of membrane glycoproteins. A human monoclonalautoantibody, 5E5, reactive with plateletGPIIIa has been developed (Nugent, et al.,Blood, 1987, in press). In this report, we describe the production of additional human monoclonal antibodies specific for GPIIIa. Peripheral blood lymphocytes fromone patient with post-transfusion purpur(PTP) and one woman who had delivered an infant with neonatal alloimmune thrombocytopenic purpura (NATP) were used as a source of antigen-specific lymphocytes. A B-lympho-cyte-enriched population was transformed with Epstein Barr virus, strain B95-8, and cultured in microtiter plates. After two weeks, culture supernatants were screened by an antigen-capture ELISA wherein murine monoclonal antibody specificfor the GPIIb-IIIa complex was used to holdcorresponding antigen from a lysate ofnormal platelets. B-lymphoblastoid cell lines producing IgG and/or IgM antibodies were expanded and either cloned by limiting dilution technqiue or hybridized with a HAT-sensitive, ouabain-resistant heterohybrid fusion line, F6, using polyethylene glycol. Hybridomas were selected in medium containing HAT andouabain. After twoweeks, hybridomas producing anti-GPIIb and/or anti-GPIIIa antibody were cloned by limiting dilution. Culture supernatants from cloned B-lymphoblastoid cell lines and cloned hybridomas were rescreened by ELISA wherein affinity-purified GPIIIa or other platelet GP weredirectly conjugatedto microtiter plates. One IgM antibody produced by acloned B-lymphoblastoid cellline (CH16) andtwo IgG antibodies produced bycloned hybridomas(Del5.19 and Del5.23) were shownto react with GPIIIa but not other GP. Further characterization of these human monoclonal antibodies, produced continuouslyin culture now for four months, is in progress.

248 IN VITRO PRODUCTION OF CATTLE × BUFFALO HYBRID EMBRYOS USING BOVINE OOCYTES AND AFRICAN BUFFALO (SYNCERUS CAFFER CAFFER) EPIDIDYMAL SPERM

2007 ◽  
Vol 19 (1) ◽  
pp. 240
Author(s):  
O. D. Owiny ◽  
D. M. Barry ◽  
M. Agaba ◽  
R. A. Godke
Keyword(s):  
Bison Bison ◽  
Abnormal Development ◽  
African Buffalo ◽  
Syncerus Caffer ◽  
Epididymal Sperm ◽  
In Vitro Production ◽  
Cleavage Rate ◽  
Cell Stage ◽  
Bovine Oocytes

Interspecies hybridization of bovids occurs between domestic cattle and at least 3 other species: the American bison (Bison bison), yak (Bos grunniens), and banteng (Bos banteng). Birth of a cattle � buffalo hybrid was reported in Russia, but the report was never authenticated. Such hybrids could be important in improving livestock production and managing diseases that impede production in tropical Africa. We investigated hybridization between cattle and their closest African wild relative, the African buffalo (Syncerus caffer caffer). In an attempt to produce pre-implantation cattle � buffalo hybrid embryos in vitro, matured bovine oocytes were subjected to a standard IVF procedure with either homologous (n = 1166 oocytes) or heterologous (n = 1202 oocytes) buffalo epididymal sperm. After IVF, 67.2% of the oocytes inseminated with homologous sperm cleaved. In contrast, insemination with buffalo sperm resulted in a 4.6% cleavage rate. Cleavage was also slower in hybrids than in cattle embryos. Up to 52.2% of cleaved homologous embryos progressed to the morula stage compared with 12.7% for hybrids. No hybrid embryos developed beyond the 16-cell stage, whereas 40.1% of the cleaved bovine embryos developed to the blastocyst stage. Developmental anomalies such as polyspermy, uneven cleavage, vacuolization, and absence of nuclei in some blastomeres were common in the hybrid embryos. We conclude that interspecies fertilization of cattle oocytes with African buffalo sperm occurs in vitro and that the barrier to hybridization is in the early stages of embryonic development. Also, chromosomal disparity is the likely cause of fertilization abnormalities, abnormal development, and subsequent arrest, impairing the formation of pre-implantation hybrid embryos. Investigation into developmental abnormalities, including reciprocal hybridization and genetic studies of the hybrid embryos, are recommended.

The expression of nonagglutinating fimbriae and its role inProteus mirabilisadherence to epithelial cells

1997 ◽  
Vol 43 (8) ◽  
pp. 709-717 ◽  
Author(s):  
Douglas L. Tolson ◽  
Blair A. Harrison ◽  
Roger K. Latta ◽  
Kok K. Lee ◽  
Eleonora Altman
Keyword(s):  
Monoclonal Antibodies ◽  
Urinary Tract ◽  
Cell Lines ◽  
Proteus Mirabilis ◽  
Growth Conditions ◽  
Structural Abnormalities ◽  
Uroepithelial Cell ◽  
Uroepithelial Cells ◽  
Tract Infections

Proteus mirabilis is a common causative agent of human urinary tract infections, especially in catheterized patients and in those patients with structural abnormalities of the urinary tract. In addition to the production of hemolysin and urease, fimbriae-mediated adherence to uroepithelial cells and kidney epithelium may be essential for virulence of P. mirabilis. A single P. mirabilis strain is capable of expressing several morphologically distinct fimbrial species, which can each be favoured by specific in vitro growth conditions. The fimbrial species reported to date include mannose-resistant/Proteus-like fimbriae, ambient temperature fimbriae, P. mirabilis fimbriae, and nonagglutinating fimbriae (NAF). Here, using intact bacteria or purified NAF as immunogens, we have generated the first reported NAF-specific monoclonal antibodies (mAbs). Bacteria expressing NAF as their only fimbrial species adhered strongly to a number of cell lines in vitro, including uroepithelial cell lines. Binding of P. mirabilis was markedly reduced following preincubation with NAF-specific mAbs and Fab fragments. The presence of NAF with highly conserved N-terminal sequences on all P. mirabilis strains so far examined, combined with the ability of both anti-NAF mAbs and purified NAF molecules to inhibit P. mirabilis adherence in vitro, suggests that NAF may contribute to the pathogenesis of P. mirabilis.Key words: fimbriae, adherence, monoclonal antibodies, Proteus mirabilis, receptors.

scholarly journals The assessment of radiosensitivity in patients with ataxia-telangiectasia syndrome and in carriers of the mutated ATM gene using lymphoblastoid cell lines

2018 ◽  
Vol 72 ◽  
pp. 846-852
Author(s):  
Barbara Pietrucha ◽  
Hanna Gregorek ◽  
Edyta Heropolitańska-Pliszka ◽  
Bożena Cukrowska ◽  
Ewa Konopka ◽  
...  
Keyword(s):  
Cell Lines ◽  
Primary Immunodeficiency ◽  
Ataxia Telangiectasia ◽  
Ionising Radiation ◽  
Lymphoblastoid Cell Lines ◽  
X Rays ◽  
Primary Immunodeficiency Diseases ◽  
Immunodeficiency Diseases ◽  
Atm Gene

Introduction: Hypersensitivity to ionising radiation is most often observed in the course of primary immunodeficiency diseases, which are associated with dysfunctional DNA repair, especially with the repair of double-strand breaks. Due to phenotypic similarities between primary immunodeficiency diseases, radiosensitivity testing can prove useful in early differential diagnosis, when attempting to identify patients with increased toxic reactivity to radio- and chemotherapy, and can have an impact on the process of their preparation for stem cell transplantation. Aim: The aim of the study was to assess the radiosensitivity in vitro of patients with ataxia-telangiectasia (A-T) syndrome, and their parents, carriers of one copy of the mutated ATM gene. Material/Methods: Lymphoblastoid cell lines (LCLs) from 15 A-T patients (remaining under the care of the Immunology Clinic and Immunology Outpatient Clinic of the Children’s Memorial Health Institute) and 11 mothers and 11 fathers of A-T patients, were used for radiosensitivity assessment. A standard colony survival assay (CSA) was applied in the tests. Results: A markedly decreased survival fraction (SF) of LCLs after in vitro exposure to X-rays was observed in all A-T patients when compared to control cells. A clear diversification of radiosensitivity to ionising radiation was observed among obligate heterozygotes. SF for heterozygotes was between 1% and 53%, i.e. varied from the values in healthy individuals to the extreme values observed in A-T patients. Conclusion: The assessment of cell radiosensitivity in A-T patients using CSA may be a useful additional test for confirming a clinically suspected disease. In heterozygous carriers, it can be an indicator of increased risk of carcinogenesis, and in both A-T patients and their parents can be helpful in making decisions with regard to radio- and/or chemotherapy.

scholarly journals Hybridoma technology; advancements, clinical significance, and future aspects

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Sanchita Mitra ◽  
Pushpa Chaudhary Tomar
Keyword(s):  
Monoclonal Antibodies ◽  
Clinical Significance ◽  
Cell Lines ◽  
Tumor Antigens ◽  
Specific Antigen ◽  
Hybridoma Cells ◽  
Main Body ◽  
Highly Sensitive ◽  
Hybridoma Technology

Abstract Background Hybridoma technology is one of the most common methods used to produce monoclonal antibodies. In this process, antibody-producing B lymphocytes are isolated from mice after immunizing the mice with specific antigen and are fused with immortal myeloma cell lines to form hybrid cells, called hybridoma cell lines. These hybridoma cells are cultured in a lab to produce monoclonal antibodies, against a specific antigen. This can be achieved by an in vivo or an in vitro method. It is preferred above all the available methods to produce monoclonal antibodies because antibodies thus produced are of high purity and are highly sensitive and specific. Main body of the abstract Monoclonal antibodies are useful in diagnostic, imaging, and therapeutic purposes and have a very high clinical significance. Once hybridoma cells become stable, these cell lines offer limitless production of homogenized antibodies. This method is also cost-effective. The antibodies produced by this method are highly sensitive and specific to the targeted antigen. It is an important tool used in various fields of research such as in toxicology, animal biotechnology, medicine, pharmacology, cell, and molecular biology. Monoclonal antibodies are used extensively in the diagnosis and therapeutic applications. Radiolabeled monoclonal antibodies are used as probes to detect tumor antigens in the living system; also radioisotope coupled antibodies are used for therapeutic target specific action on oncogenic cells. Short conclusion Presently, the monoclonal antibodies used are either raised in mice or rats; this poses a risk of disease transfer from mice to humans. There is no guarantee that antibodies thus created are entirely virus-free, despite the purification process. Also, there are some immunogenic responses observed against the antibodies of mice origin. Technologically advanced techniques such as genetic engineering helped in reducing some of these limitations. Advanced methods are under development to make lab-produced monoclonal antibodies as human as possible. This review discusses the advantages and challenges associated with monoclonal antibody production, also enlightens the advancement, clinical significance, and future aspects of this technique.

scholarly journals B lymphoblastoid cell lines with normal and defective O-glycosylation established from an individual with blood group Tn

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 973-979 ◽  
Author(s):  
CG Gahmberg ◽  
L Peltokorpi ◽  
LC Andersson
Keyword(s):  
Blood Group ◽  
Cell Lines ◽  
Helix Pomatia ◽  
Lymphoblastoid Cell Lines ◽  
Membrane Glycoproteins ◽  
Leukocyte Antigen ◽  
Galactosyl Transferase ◽  
The Common ◽  
Common Leukocyte Antigen

Abstract Individuals with the Tn blood group contain terminal serine/threonine- linked N-acetylgalactosamine residues in their blood cells. This is due to lack of UDP-D-galactose: D-N-acetyl galactosamine beta-D-galactosyl transferase from part of their red cells and probably from their leukocytes. We have established B lymphoblastoid cell lines from such an individual by in vitro infection of his lymphocytes with Epstein- Barr virus. The original line contained a mixture of cells reactive and nonreactive with Helix pomatia lectin (Hp). These cells were subcloned after staining with fluorescent Hp by a fluorescence-activated cell sorter (FACS) into homogeneous, phenotypically stable lines of Hp- positive (Hp+) and Hp-negative (Hp-) cells. The molecular differences between the membrane glycoproteins were characterized by carbohydrate- specific surface labeling techniques, Hp affinity chromatography, polyacrylamide slab gel electrophoresis and glycopeptide/oligosaccharide analysis. The major O-glycosidic membrane glycoprotein (GP105) was retained on Hp-Sepharose columns only from Hp+ cells, whereas the common leukocyte antigen (GP160–200) was partially retained on Hp columns from both lines. These proteins were isolated by immune precipitation with monoclonal antibodies and characterized. The results show that the GP105 glycoprotein from Hp+ cells contains terminal N-acetylgalactosamine residues but also more complex oligosaccharides. The common leukocyte antigen showed different electrophoretic mobilities in Hp+ and Hp- cells. UDP-galactose D-N- acetyl galactosamine beta-galactosyl transferase was almost absent in the Hp+ cells. These cell lines are useful for studies on the functional role and regulation of the biosynthesis of O-glycosidic carbohydrates.

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