Abstract:
:We have developed an antigen presenting system based on the capsid protein of the Flock House virus (FHV) and used it to display, in different positions on its external surface, two neutralizing epitopes found at residues 735-752 of HIV-1 gp160. We have compared the immunoreactivity of these FHV chimeric proteins and of the corresponding synthetic peptide using a panel of neutralizing mouse monoclonal antibodies (mAbs) directed against two distinct sequences (IEEE and ERDRD) contained in this epitope of the gp41 region. We have observed that both the FHV chimeric protein and the synthetic peptide are clearly detected in ELISA procedures by the mAbs recognizing the sequence IEEE. The denaturation of these recombinant proteins had little effect on the recognition pattern of this group of monoclonals, suggesting minor conformational requirements for the display of this epitope. The FHV chimeric proteins were also recognized by the mAbs directed against the ERDRD epitope, whereas the corresponding synthetic peptide was not recognized. In this case, denaturation of these recombinant proteins completely abolished the reactivity of the second group of mAbs, arguing for the existence of strong conformational constraints. Additionally, we have investigated whether an isolated loop structure from the FHV protein was sufficient to provide the conformational requirements for the presentation of these epitopes. These experiments have shown that the stabilized loop structure, although improving the presentation of both epitopes, is not as efficient as the native loop in the intact FHV protein. The data obtained with these mAbs support the recently observed limitations in the use of synthetic peptides for the screening of the immune response against conformational epitopes. The establishment of appropriate tools able to present epitope sequences in a structure resembling the native conformation will be useful for accurate epidemiological studies and for the design of new epitope-specific vaccines.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Buratti E,Tisminetzky SG,Scodeller ES,Baralle FEdoi
10.1016/0022-1759(96)00097-xsubject
Has Abstractpub_date
1996-10-16 00:00:00pages
7-18issue
1-2eissn
0022-1759issn
1872-7905pii
0022-1759(96)00097-Xjournal_volume
197pub_type
杂志文章abstract::A method for cloning hybridomas is described which involves transfer of single cells with a capillary tubing connected to a suction apparatus. This method enables the efficient recovery of antibody-producing clones of differing abundances from the parent hybridoma cultures. The 4 cell lines submitted to this cloning p...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90084-5
更新日期:1985-04-22 00:00:00
abstract::A method for separating IL-1 from plasma inhibitors by silica extraction has been developed and coupled to a highly sensitive bioassay using the LBRM TG6 cell line and the I1-2 dependent HT2A cell line. Using this assay we have detected IL-1 activity in plasma from patients undergoing elective surgery. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90399-7
更新日期:1988-04-06 00:00:00
abstract::The detection of site-directed anti-idiotypic antibodies is usually based on their ability to inhibit the binding of antigen to idiotype, in either solid- or fluid-phase radioimmunoassays. Passage of serum over antigen-coupled immunosorbents for the purpose of removing the idiotypes from complexes with putative anti-i...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90172-4
更新日期:1987-06-26 00:00:00
abstract::We developed a dual wavelength spectrophotometric assay that permitted beta-lactamase and beta-galactosidase activities to be measured concurrently in a single sample. We also constructed a target cell, E. coli ML-35p, that was substantially cryptic for its periplasmic beta-lactamase and cytoplasmic beta-galactosidase...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90414-0
更新日期:1988-04-06 00:00:00
abstract::A rapid quantitative flow cytometric (FCM) assay for the combined kinetic measurement of phagocytosis and intracellular killing of fluorescein-isothiocyanate (FITC) labelled Candida albicans is described. The fraction of phagocytosing leukocytes and the numbers of attached or internalized Candida albicans per phagocyt...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90451-4
更新日期:1984-08-03 00:00:00
abstract::A procedure to enhance incorporation of radioactive iodine into cell-surface proteins is described. The effects of chemical modification of cells with the iodinatible haptens N-succinimidyl-3-(4-hydroxyphenyl) propionate (Bolton-Hunter reagent) and methyl-p-hydroxybenzimidate HC1 (Wood reagent) are compared. The Wood ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90177-2
更新日期:1985-02-28 00:00:00
abstract::The attachment of cells to plastic tissue culture plates is a widely used criterion of cell viability in microcytotoxicity assays. When cell suspensions of primary human colon carcinoma and melanoma cells which were of low initial viability (assessed by trypan blue exclusion) were allowed to adhere to tissue culture p...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90039-9
更新日期:1981-01-01 00:00:00
abstract::There is an emerging trend in the pharmaceutical industry to evaluate a variety of surrogate biomarkers in Phase I/II clinical studies with the intention of determining potential activity of drugs early in clinical development. A number of cytokines expressed in pathological conditions are currently being considered a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00543-9
更新日期:2002-02-01 00:00:00
abstract::Studies describing the induction of apoptosis for CD4 mAbs do not delineate between epitope-dependent and Fc-driven epitope cross-linking induced cell death. Keliximab and clenoliximab are two CD4 mAbs that differ only in their heavy chain isotypes, being an IgG1 and a modified IgG4, respectively. These antibodies sup...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00447-1
更新日期:2001-11-01 00:00:00
abstract::We have previously shown a polymorphism of human Fc receptors for mouse IgG1 using an EA rosette technique in which human erythrocytes sensitized with a murine IgG1 monoclonal antibody against glycophorin A acted as indicator cells. We now describe a method to quantitate this EA rosetting using the pseudoperoxidase ac...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90223-7
更新日期:1987-07-16 00:00:00
abstract::Glycosylation of the Fc domain is an important driver of antibody effector function. While assessment of antibody glycoform compositions observed across total plasma IgG has identified differences associated with a variety of clinical conditions, in many cases it is the glycosylation state of only antibodies against a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2015.06.005
更新日期:2015-10-01 00:00:00
abstract::Human interleukin-7 (IL-7) is a cytokine that appears to be critical for early T- and B-cell development and although IL-7 is currently under investigation as a therapeutic agent in a variety of hematolymphopoietic disorders, there have been few instances of the detection or investigation of this cytokine using a biol...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00237-9
更新日期:2000-03-06 00:00:00
abstract::There is a need for simple and sensitive assays to assess innate and adaptive immune responses to microbial agents and vaccines. Herein, we describe a whole blood method allowing to measure the induction of cytokine synthesis at the mRNA level. The originality of this method consists in the combination of PAXgene tube...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(03)00074-7
更新日期:2003-05-01 00:00:00
abstract::A method is described for isolation of antibody-forming cells with the help of cluster formation. This was achieved by incubating spleen cells from immunized mice with sheep erythrocytes at 37 degrees C. During incubation rabbit-anti-mouse immunoglobulin serum was present in order to facilitate the formation of cluste...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(76)90103-4
更新日期:1976-01-01 00:00:00
abstract::Hybridoma cells producing antibody reactive with poly(A) were isolated following fusion of spleen cells from a BALB/c mouse immunized with poly(A).poly(dT) and myeloma cells. The antibody was purified from ascitic fluid by the formation of immune complexes with poly(A). The antibody reacted with poly(A) and some homop...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90269-d
更新日期:1993-08-26 00:00:00
abstract::Ia molecule (Ek,b beta:Ek alpha or Ak beta:Ak alpha)-containing planar membranes were constructed with cholesterol and a 9:1 molar ratio of the phospholipids dipalmitoyl phosphatidylcholine and dilinoleoyl phosphatidylcholine. This lipid composition was found to be optimal for the stimulation of T cell hybridomas of d...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90432-7
更新日期:1987-04-02 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90083-j
更新日期:1993-01-04 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90363-2
更新日期:1987-01-26 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90135-6
更新日期:1981-01-01 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90279-5
更新日期:1990-12-31 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90186-3
更新日期:1985-02-28 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(76)90076-4
更新日期:1976-01-01 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90463-1
更新日期:1986-02-12 00:00:00
abstract::Genital infections with herpes simplex virus type 2 (HSV-2) are a source of considerable morbidity and are a health concern for newborns exposed to virus during vaginal delivery. Additionally, HSV-2 infection diminishes the integrity of the vaginal epithelium resulting in increased susceptibility of individuals to inf...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2016.09.004
更新日期:2016-12-01 00:00:00
abstract::An antibody binding-inhibition test is described, which allows the detection of P. falciparum in red blood cells (RBC) infected in vitro, using a crossreacting, monoclonal anti-P. berghei antibody and P. berghei coated microtiter plates. Experiments carried out to determine the coating efficiency of various P. berghei...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90231-5
更新日期:1985-09-03 00:00:00
abstract::The pronase-enhanced release of 14C-labeled DNA was used to assay cytotoxicity of murine natural killer cells. YAC-1 lymphoma cells were labeled with [14C]thymidine and incubated with increasing numbers of spleen cells from nude mice. At harvest, nuclease-free pronase was used to digest damaged target cells. Increases...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90152-0
更新日期:1980-01-01 00:00:00
abstract::Murine lymphocytes readily undergo spontaneous and glucocortocoid-induced apoptosis in vitro. It has been previously demonstrated that during apoptosis, many cell types including lymphocytes, enzymatically cleave their DNA, thus demonstrating a sub-G0 DNA peak when stained with propidium iodide and analyzed by flow cy...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00216-2
更新日期:1995-12-27 00:00:00
abstract::The antigen receptor expressed on most T lymphocytes, the T cell receptor (TCR), is a multisubunit complex containing TCR alpha,beta and CD3-gamma,delta, epsilon and zeta chains, in the form alphabetadeltaepsilongammaepsilonzetazeta. The intracellular transport of TCR proteins is routinely measured by the extent of pr...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00103-9
更新日期:1999-10-29 00:00:00
abstract::We found that some plasma proteins such as haptoglobin (Mr 200,000) and alpha 1-acid glycoprotein (Mr 40,000) do not bind at all to nitrocellulose membranes in classical transfer conditions. In contrast, these proteins can be clearly identified when a charged nylon membrane is disposed on the anodal side of the nitroc...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90226-8
更新日期:1988-03-16 00:00:00
abstract::Immune complexes have been implicated in the pathogenesis of a number of infectious diseases. The predominant immunoglobulin class associated with circulating immune complexes is IgG, although immune complexes containing IgM have been described. The role of IgM immune complexes in disease pathogenesis has been difficu...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(87)80005-4
更新日期:1987-08-24 00:00:00