Analysis of cellular interactions in limiting dilution cultures.


:Limiting dilution (LD) cultures are often used to study cellular heterogeneity in responses of murine splenocytes to specific or polyclonal activation. LD titration curves often reveal a nonlinear dependence of response on input cell dose. Although 'zigzag' shaped curves of this kind are often interpreted and analyzed as resulting from interactions among three distinct cell types, we observe that a more parsimonious two cell model, including a cell type that can generate both positive and negative effects, provides better fit to a wide range of experimental data. We have developed mathematical models for the accurate estimation of the frequencies of both interacting cell types and of the parameters for their multi-hit interaction. We show examples of LD cultures in which specific experimental manipulations alter the frequency of only one of the two cell types, or alter the interaction parameters without a change in responder frequency. We also provide a simplified method for approximation of the model parameters using graphical approaches and simple algebra. Lastly, we present an improved method for calculation of the effect generated per responder cell in microclonal cultures.


J Immunol Methods


Dozmorov IM,Lutsenko GV,Sidorov LA,Miller RA




Has Abstract


1996-02-05 00:00:00














  • Selection of hapten-specific single-domain antibodies from a non-immunized llama ribosome display library.

    abstract::Picloram-specific variable fragments (V(HH)s) of heavy chain antibodies (HCAbs) were selected from a nai;ve-llama library using ribosome display technology. A cDNA library of V(HH)s was constructed from lymphocytes of a non-immunized llama and engineered to allow in vitro transcription and translation. With no stop co...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Yau KY,Groves MA,Li S,Sheedy C,Lee H,Tanha J,MacKenzie CR,Jermutus L,Hall JC

    更新日期:2003-10-01 00:00:00

  • Human antibodies responsible for binding inhibition and polymerization inhibition of human immunodeficiency virus type 1 reverse transcriptase.

    abstract::Using a solid-phase non-radioisotopic (non-RI) reverse transcriptase (RT) assay, antibodies inhibiting human immunodeficiency virus type 1 (HIV-1) RT activity (RTI antibody) were investigated for their ability to inhibit binding of RT to a template-primer and DNA polymerization. The RTI antibody inhibited the binding ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Odawara F,Sano K,Otake T,Okubo S,Nakano T,Takasaki T,Ueba N,Misaki H,Nakai M

    更新日期:1996-12-15 00:00:00

  • Suspended mouse peritoneal macrophages. Preparation and properties.

    abstract::Since macrophages (MPH) are able to adhere firmly to solid surfaces, the recovery of viable and functional MPH has proven to be extremely difficult. We have developed a simple method using agarose coating for preparing MPH and culturing the cells in suspension. Their properties were tested over 72 h. The oxidative bur...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Château MT,Rabesandratana H,Caravano R

    更新日期:1991-09-20 00:00:00

  • Evaluation of T7 and lambda phage display systems for survey of autoantibody profiles in cancer patients.

    abstract::In the current study we attempted to evaluate the suitability of T7 Select 10-3b and lambdaKM8 phage display systems for the identification of antigens eliciting B cell responses in cancer patients and the production of phage-displayed antigen microarrays that could be exploited for the monitoring of autoantibody prof...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Kalniņa Z,Siliņa K,Meistere I,Zayakin P,Rivosh A,Abols A,Leja M,Minenkova O,Schadendorf D,Linē A

    更新日期:2008-05-20 00:00:00

  • Rapid in vitro micro-cytotoxicity tests for the detection and quantitation of neutralizing antibodies to both viruses and toxins.

    abstract::A generally applicable method was developed for the rapid and quantitative detection of both toxin- and virus neutralizing antibodies. The method was optimized for three different biological agents, i.e., Shigella toxin, influenza viruses (A/Beying, A/Taiwan and B/Yamagata) and Chikungunya virus. The in vitro micro-cy...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: van de Water C,van Dura EA,van der Stap JG,Brands R,Boersma WJ

    更新日期:1993-12-03 00:00:00

  • A reliable method for reconstituting thymectomized, lethally irradiated guinea pigs with bone marrow cells.

    abstract::We developed a reliable method for reconstituting thymectomized, lethally irradiated guinea pigs. Injection of 2.5-10 X 10(7) syngeneic bone marrow cells into adult thymectomized, lethally irradiated guinea pigs produced survival of 46-100% of treated animals. Gentamycin sulfate (5 mg/kg of body weight) for 10 days wa...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Terata N,Tanio Y,Zbar B

    更新日期:1984-05-25 00:00:00

  • ELISA kits based on monoclonal antibodies do not measure total IL-1 beta synthesis.

    abstract::The paper by Herzyk et al. which appears in this issue of the Journal of Immunological Methods is an important and long overdue contribution. The study clearly and convincingly demonstrates that two widely used commercial ELISA kits fail to detect over 90% of the 31 kDa human IL-1 beta precursor (pro-IL-1 beta) in var...

    journal_title:Journal of immunological methods

    pub_type: 评论,杂志文章


    authors: Dinarello CA

    更新日期:1992-04-08 00:00:00

  • Ultrastructural localisation of lipopolysaccharide-binding sites with peroxidase-conjugated lipopolysaccharides.

    abstract::The localisation of lipopolysaccharide-binding sites on erythrocytes with peroxidase-coupled LPS is described. LPS was isolated from Fusobacterium nucleatum (Fus MC-8) by phenol-water extraction. The LPS was coupled to horseradish peroxidase by the two-step method of Avrameas and Ternynck (1971). The biological and se...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Nygren H,Dahlen G

    更新日期:1979-01-01 00:00:00

  • Selective assay for CyPA and CyPB in human blood using highly specific anti-peptide antibodies.

    abstract::Cyclophilins A and B (CyPA and CyPB) are known to be the main binding proteins for cyclosporin A (CsA), a potent immunosuppressive drug. Due to the high homology between the two proteins, antibodies to CyPB were found to cross-react with CyPA. In order to avoid this phenomenon, we raised specific antibodies against pe...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Allain F,Boutillon C,Mariller C,Spik G

    更新日期:1995-01-13 00:00:00

  • Cobra venom factor: improved method for purification and biochemical characterization.

    abstract::A method to purify cobra venom factor (CVF) from cobra venom by sequential column chromatography is described which yields a product virtually free of phospholipase A2, a common contaminant of CVF preparations. The separation of phospholipase A2 from CVF was achieved by chromatography on Cibacron blue-agarose, a resin...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Vogel CW,Müller-Eberhard HJ

    更新日期:1984-10-12 00:00:00

  • A new adhesion assay using buoyancy to remove non-adherent cells.

    abstract::A new adhesion assay was developed that utilizes buoyancy, rather than washing or centrifugation, to remove non-adherent cells. Biotinylated cells were added to wells containing cell monolayers or purified protein substrates. Non-adherent cells were then removed by floatation on a dense Percoll solution. The adherent ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Goodwin AE,Pauli BU

    更新日期:1995-12-01 00:00:00

  • Bovine gut-associated lymphoid tissue--morphologic and functional studies. I. Isolation and characterization of leukocytes from the epithelium and lamina propria of bovine small intestine.

    abstract::A successful technique for the isolation of highly pure suspensions of viable leukocytes from the small intestine of cattle is described. Procedures ranging from mechanical mincing to enzymatic digestion of tissues were compared. The most reliable and reproducible procedure was the sequential treatment of tissues with...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Nagi AM,Babiuk LA

    更新日期:1987-12-04 00:00:00

  • The effect of antigen suicide on numbers of cells binding defined antigenic determinants.

    abstract::Lymph node cells from guinea pigs immunized to oxidized ferredoxin (O-Fd) were treated in an antigen suicide procedure designed to inactivate lymphocytes binding the haptenic peptide determinants of the ferredoxin molecule. O-Fd-induced DNA synthesis and cells binding either peptide determinant were examined both befo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Pearson T,Levy J,Kilburn D,Fairhurst K

    更新日期:1976-03-01 00:00:00

  • Solid-phase preparation of vimentin-type intermediate filaments for immunoassays.

    abstract::A simple method is described for the preparation of vimentin-type intermediate filaments for solid-phase immunoassays. Bovine aortic endothelial cells were grown to confluency in multiwells, and extracted with Triton X-100 and a high-salt buffer. Residual protein in the wells consisted of 53% vimentin as judged by SDS...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Hansson GK

    更新日期:1985-12-27 00:00:00

  • Development of an RNA-based kit for easy generation of TCR-engineered lymphocytes to control T-cell assay performance.

    abstract::Cell-based assays to monitor antigen-specific T-cell responses are characterized by their high complexity and should be conducted under controlled conditions to lower multiple possible sources of assay variation. However, the lack of standard reagents makes it difficult to directly compare results generated in one lab...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Bidmon N,Kind S,Welters MJP,Joseph-Pietras D,Laske K,Maurer D,Hadrup SR,Schreibelt G,Rae R,Sahin U,Gouttefangeas C,Britten CM,van der Burg SH

    更新日期:2018-07-01 00:00:00

  • Allele-specific PCR analysis for detection of the gld Fas-ligand point mutation.

    abstract::The discovery of a naturally occurring missense point mutation in the gene encoding Fas-ligand (FasL/CD95L) in generalized lymphoproliferative disease (gld) mice has lead to the characterization of FasL as an important mediator of apoptosis. Further analysis of FasL function can be facilitated by crossing the gld muta...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Hoek RM,Kortekaas MC,Sedgwick JD

    更新日期:1997-12-15 00:00:00

  • Ligand binding analysis of interleukin-2 receptor complexes using surface plasmon resonance.

    abstract::In this study we have employed surface plasmon resonance to examine the kinetic binding constants of a new class of soluble cytokine receptor complexes. The solution assembly of both homomeric and heteromeric interleukin-2 receptor ectodomain complexes has been achieved using coiled-coil molecular recognition. These c...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Wu Z,Johnson KW,Ciardelli TL

    更新日期:1995-06-14 00:00:00

  • Increased exosome production from tumour cell cultures using the Integra CELLine Culture System.

    abstract::Exosomes are nanometer-sized vesicles, secreted from most cell types, with documented immune-modulatory functions. Exosomes can be purified from cultured cells but to do so effectively, requires maintenance of cells at high density in order to obtain sufficient accumulation of exosomes in the culture medium, prior to ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Mitchell JP,Court J,Mason MD,Tabi Z,Clayton A

    更新日期:2008-06-01 00:00:00

  • The use of luminous bacteria for determination of phagocytosis.

    abstract::The existing methods for phagocytosis evaluation are inadequate for assessing all the real events occurring during phagocytosis, or for continuously following the kinetics of the process. Our purpose is to establish the use of luminous bacteria as an object for phagocytosis. The bioluminescence test offers an easy and...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Barak M,Ulitzur S,Merzbach D

    更新日期:1983-11-25 00:00:00

  • Addressing soluble target interference in the development of a functional assay for the detection of neutralizing antibodies against a BCMA-CD3 bispecific antibody.

    abstract::Proper evaluation of immunogenicity during clinical development of biotherapeutics is a major challenge to bioanalytical scientists, in part due to matrix interference in anti-drug antibody (ADA) and neutralizing antibody (NAB) assays. If not addressed, matrix interference could confound the immunogenicity assessment ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Wang Y,Luong M,Guadiz C,Zhang M,Gorovits B

    更新日期:2019-11-01 00:00:00

  • A computational study of the interaction of the foot and mouth disease virus VP1 with monoclonal antibodies.

    abstract::Foot and mouth disease is caused by a non-enveloped virus (FMDV), which disposes several antigenic sites at the surface of their capsid proteins. The most relevant and immunodominant antigenic site of FMDV (site A or AnSA) includes a key virus-cell interaction element (RGD motif) located in the Viral Protein 1 (VP1), ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Marrero R,Limardo RR,Carrillo E,König GA,Turjanski AG

    更新日期:2015-10-01 00:00:00

  • A method for the quantification of a colored or fluorescent signal in enzyme immunoassays by photodensitometry.

    abstract::A colored or fluorescent signal is generally evaluated with the naked eye, or by means of different more or less sophisticated and costly instruments. Photodensitometry is an additional technique which is both inexpensive and simple to perform. This technique can satisfactorily quantify a signal without the use of eit...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Labrousse H,Avrameas S

    更新日期:1987-10-23 00:00:00

  • Generation of novel killer hybridomas derived from proliferation-suppressed somatic cell hybrids between YACUT T cell lymphoma and normal lymphocytes activated in secondary mixed lymphocyte cultures.

    abstract::Somatic cell hybridization between the YACUT T cell lymphoma cell line with normal lymphocytes activated in secondary mixed lymphocyte cultures (MLCs) consistently yielded IL-2-dependent CD4- CD8 alpha+ beta- Fc gamma RIII+ hybrids with cytotoxic function. The hybrids expressed T cell receptors other than that of YACU...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Kubota K,Nakazato K,Tamauchi H,Sasahara T,Katoh H

    更新日期:1996-06-10 00:00:00

  • From big flow cytometry datasets to smart diagnostic strategies: The EuroFlow approach.

    abstract::The rise in the analytical speed of mutiparameter flow cytometers made possible by the introduction of digital instruments, has brought up the possibility to manage progressively higher number of parameters simultaneously on significantly greater numbers of individual cells. This has led to an exponential increase in ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章,评审


    authors: Pedreira CE,Costa ESD,Lecrevise Q,Grigore G,Fluxa R,Verde J,Hernandez J,van Dongen JJM,Orfao A,EuroFlow.

    更新日期:2019-12-01 00:00:00

  • Increased immunogenicity of a non-amphipathic protein (BSA) after inclusion into iscoms.

    abstract::Bovine serum albumin (BSA) was used as a non-amphipathic model protein to be included into iscoms. Pretreatment at an acidic pH (2.5) was used to reveal hydrophobic regions, after which BSA could be integrated. In immunization experiments in mice the BSA iscoms induced long lasting and considerably higher serum antibo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Morein B,Ekström J,Lövgren K

    更新日期:1990-04-17 00:00:00

  • Measurement of immunoperoxidase activity. A rapid and reproducible immunoassay for quantitation of cellular antigens.

    abstract::A procedure is described making it possible to obtain quantitative data with the immunohistochemical peroxidase anti--peroxidase (PAP) technique. After applying PAP on fixed fibroblast preparations, peroxidase enzymic activity is measured on solubilized immune-complexes. The results show this technique to be sensitive...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Van Leuven F,Cassiman JJ,Van den Berghe H

    更新日期:1978-01-01 00:00:00

  • Time-resolved fluorescence using a europium chelate of 4,7-bis-(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA). Labeling procedures and applications in immunoassays.

    abstract::We describe optimal conditions for protein labeling with a new fluorescent probe, 4,7-bis-(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA). The labeled proteins are suitable for time-resolved fluorometric applications because BCPDA forms fluorescent complexes with Eu3+ which exhibit very long fluo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Diamandis EP,Morton RC

    更新日期:1988-08-09 00:00:00

  • Production and characterization of monoclonal antibodies raised against recombinant human granzymes A and B and showing cross reactions with the natural proteins.

    abstract::The human serine proteases granzymes A and B are expressed in cytoplasmic granules of activated cytotoxic T lymphocytes and natural killer cells. Recombinant granzyme A and granzyme B proteins were produced in bacteria, purified and then used to raise specific mouse monoclonal antibodies. Seven monoclonal antibodies (...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Kummer JA,Kamp AM,van Katwijk M,Brakenhoff JP,Radosević K,van Leeuwen AM,Borst J,Verweij CL,Hack CE

    更新日期:1993-07-06 00:00:00

  • Production of Alexa Fluor 488-labeled reovirus and characterization of target cell binding, competence, and immunogenicity of labeled virions.

    abstract::Respiratory enteric orphan virus (reovirus) has been used to study many aspects of the biology and genetics of viruses, viral infection, pathogenesis, and the immune response to virus infection. This report describes the functional activity of virus labeled with Alexa Fluor 488, a stable fluorescent dye. Matrix assist...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Fecek RJ,Busch R,Lin H,Pal K,Cunningham CA,Cuff CF

    更新日期:2006-07-31 00:00:00

  • In situ tetramer staining.

    abstract::The development of MHC tetramer staining has opened the doors to multiple avenues of new research [Science 274 (1996) 94]. In this review, we will discuss the development and application of in situ MHC tetramer (IST) staining. We describe two independently developed IST staining methodologies and discuss current uses,...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章,评审


    authors: Skinner PJ,Haase AT

    更新日期:2002-10-01 00:00:00