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














  • Influence of specimen age and anticoagulant on flow cytometric evaluation of granulocyte oxidative burst generation.

    abstract::The oxidative burst generation capacity of granulocytes can be reliably detected by a flow cytometric procedure using lysed whole blood, dihydrorhodamine 123 (DHR), and phorbol myristate acetate (PMA). This assay is used to detect chronic granulomatous disease (CGD) and the CGD carrier state. To assess the feasibility...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Prince HE,Lapé-Nixon M

    更新日期:1995-12-15 00:00:00

  • Production of a monoclonal antibody reactive with poly(A) and its application in the isolation of poly(A)+ RNA.

    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


    authors: Kitagawa Y,Ishibashi K

    更新日期:1993-08-26 00:00:00

  • Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper discs. I. A description of the technique and of methods for the analysis of colonies.

    abstract::A novel technique for establishing short term clones of antigen- or mitogen-activated splenic B lymphocytes is described. Spleen cells are plated onto the surface of filter paper discs and subsequently stimulated by antigen or mitogen in situ; activated B cells proliferate and differentiate into pure colonies of cells...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Kelsoe G

    更新日期:1985-02-11 00:00:00

  • Antibodies and radioimmunoassays for phosphoserine, phosphothreonine and phosphotyrosine. Serologic specificities and levels of the phosphoamino acids in cytoplasmic fractions of rat tissues.

    abstract::For antibody production, the O-phosphorylated derivative of tyrosine, threonine, or serine was covalently linked to succinylated bovine albumin via the carbodiimide reaction. Each conjugate was then complexed with methylated bovine albumin for immunization of rabbits. To determine binding, the corresponding O-phosphor...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Levine L,Gjika HB,Van Vunakis H

    更新日期:1989-11-30 00:00:00

  • An improved clonogenic culture method for thymic epithelial cells.

    abstract::A clonogenic assay system for thymic epithelial cells (TECs) is of crucial importance for identifying thymic epithelial stem and/or progenitor cells, evaluating their activities, and understanding the mechanisms of thymic involution. However, current systems are not sufficiently sensitive at detecting and quantifying ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Sekai M,Wang J,Minato N,Hamazaki Y

    更新日期:2019-04-01 00:00:00

  • An improvement on the Bollman method of restraining and collecting thoracic duct lymph from the rat.

    abstract::A method of obtaining rat thoracic duct lymphocytes is described, including a review of the surgical technique, with several modifications, and including an outline of a newly designed rat restrainer that helps to triple thoracic duct output compared to the Bollman restrainer. The new restrainer allows the rat increas...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Reich DJ,Keller SE

    更新日期:1988-06-13 00:00:00

  • CD4 mAb induced apoptosis of peripheral T cells: multiparameter subpopulation analysis by flow cytometry using Attractors.

    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


    authors: Fishman-Lobell J,Tsui P,Reddy M,DiPrinzio R,Eichman C,Sweet RW,Truneh A

    更新日期:2001-11-01 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

  • Sensitivity and specificity of immunohistological methods on freeze-dried paraffin sections.

    abstract::The application of freeze-dried paraffin sections for immunohistology eliminates many of the problems associated with the cryostat technique: the sections are thin sufficiently, the cell morphology is improved, there are no diffusion artifacts and the intensity of immunohistological reactions is superior to that obtai...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Nemes Z,Thomázy V,Szeifert G

    更新日期:1982-01-01 00:00:00

  • Sensitive time-resolved fluoroimmunoassay of nerve growth factor and the disappearance of nerve growth factor from rat pheochromocytoma PC12 cell culture medium.

    abstract::A sensitive time-resolved fluoroimmunoassay of nerve growth factor (NGF) has been developed. The method is based on the unique property of the lanthanides for delayed fluorescence, which reduces substantially the endogenous fluorescence of biological substances, because the excitation of the sample and detection of th...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Arumäe U,Neuman T,Sinijärv R,Saarma M

    更新日期:1989-08-15 00:00:00

  • Differential binding properties of protein A and protein G for dog immunoglobulins.

    abstract::We studied the binding of dog immunoglobulins G, A, M and E to protein A and protein G. Passive cutaneous anaphylaxis (PCA) testing was used for the measurement of dog IgE and enzyme-linked immunosorbent assays (ELISA) were used for the measurements of dog IgG, IgA and IgM. Protein A from lyophilized cells of Staphylo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Peng ZK,Simons FE,Becker AB

    更新日期:1991-12-15 00:00:00

  • Development of monoclonal antibody-based immunoassays for detection of Helicobacter pylori neutrophil-activating protein.

    abstract::Neutrophil-activating protein (NAP) is a major virulence factor expressed by Helicobacter pylori isolates associated with severe chronic gastroduodenal inflammation and peptic ulcers. NAP is one of the main protective antigens and a target for vaccine development against Helicobacter infection. In addition, NAP is a p...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Iankov ID,Penheiter AR,Carlson SK,Galanis E

    更新日期:2012-10-31 00:00:00

  • In vitro derivation of macrophage from guinea pig bone marrow with human M-CSF.

    abstract::The guinea pig has a storied history as a model in the study of infectious disease and immunology. Because of reproducibility of data and availability of various reagents, inbred mice have since supplanted the guinea pig as the animal model-of-choice in these fields. However, several clinically-significant microorgani...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Yu KO,Porcelli SA,Shuman HA

    更新日期:2013-03-29 00:00:00

  • Production and characterization of monoclonal antibodies against non-A1c glycated hemoglobin.

    abstract::Hybridomas secreting monoclonal antibodies specific for hemoglobin nonenzymatically glycated in the non-A1c position were produced by fusion of SP 2/0 myeloma cells with spleen cells from BALB/c mice immunized with nonenzymatically glycated hemoglobin prepared from human erythrocytes. Wells containing hydridomas secre...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Steward LA,Wu VY,Shea E,Cohen MP

    更新日期:1991-07-05 00:00:00

  • Accurate and statistically verified quantification of relative mRNA abundances using SYBR Green I and real-time RT-PCR.

    abstract::Among the many methods currently available for quantifying mRNA transcript abundance, reverse transcription-polymerase chain reaction (RT-PCR) has proved to be the most sensitive. Recently, several protocols for real-time relative RT-PCR using the reporter dye SYBR Green I have appeared in the literature. In these met...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Marino JH,Cook P,Miller KS

    更新日期:2003-12-01 00:00:00

  • Humanised mice have functional human neutrophils.

    abstract::The differences between murine and human neutrophils mean that findings in mice may not translate to humans, and therefore an in vivo model with human neutrophils would be an important methodological advance. We generated humanised mice by injecting human cord blood derived CD34+ stem cells into irradiated NOD-scid-γc...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Coughlan AM,Freeley SJ,Robson MG

    更新日期:2012-11-30 00:00:00

  • Enumeration of t rosette-forming cell population in rabbit peripheral blood using human, sheep, guinea-pig, rat, mouse and chicken red blood cells under different experimental conditions.

    abstract::Attempts were made to estimate peripheral T-lymphocytes in rabbits by their ability to form rosettes with sheep, human, guinea-pig, rat, mouse and chicken RBC. The reaction was studied under different conditions of pH and temperature. The test was also carried out in the presence or absence of glutaraldehyde. With she...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Vakharia DD,Vakharia MR,Rao SS

    更新日期:1977-01-01 00:00:00

  • Long term cultures of human monocytes in vitro. Impact of GM-CSF on survival and differentiation.

    abstract::In vitro differentiation of human monocytes (Mo) provides large amounts of mature and functionally competent macrophages (M phi) which may be used as potentially powerful anticancer agents for adoptive immunotherapy. Granulocyte macrophage-colony stimulating factor (GM-CSF) was evaluated for its ability to influence l...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Eischen A,Vincent F,Bergerat JP,Louis B,Faradji A,Bohbot A,Oberling F

    更新日期:1991-10-25 00:00:00

  • Determination of immunoglobulin classes of specific antibodies secreted by single cells. A cluster formation assay.

    abstract::A method is described to identify specific antibody-forming cells and their immunoglobulin classes by a multilayer cluster formation. Cells producing specific antibodies are mixed with erythrocytes coated with the corresponding antigen in presence of rabbit anti-human immunoglobulins. Clusters are formed only if the m...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Steinitz M

    更新日期:1980-01-01 00:00:00

  • Cell mediated immunity to herpes simplex in man. IV. A correlation of lymphocyte stimulation and inhibition of leukocyte migration.

    abstract::The cell mediated immune response to herpes simplex virus was assessed by a modification of the leukocyte migration inhibition test, developed to increase its sensitivity and correlated with the in vitro stimulation of lymphocytes. The introduction of a preincubation step allowed us to demonstrate significant inhibiti...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Russell AS,Kaiser J,Lao VS

    更新日期:1976-01-01 00:00:00

  • Methods for assessing complement activation in the clinical immunology laboratory.

    abstract::Complement activation is a key component of the pathogenesis of immune-mediated tissue damage in many diseases. Assessment of complement activation in current practice is largely based on the measurement of intact C3 and C4 or the determination of complement haemolytic function. These parameters reflect activation onl...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章,评审


    authors: Porcel JM,Peakman M,Senaldi G,Vergani D

    更新日期:1993-01-04 00:00:00

  • Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation.

    abstract::LRP6 is a cell surface molecule that plays a critical role in the Wnt signaling pathway, and is implicated in numerous human diseases. Studies of cellular signaling mediated by LRP6 have relied on overexpression experiments, due to the lack of good monoclonal antibodies (mAbs) reactive with native LRP6 ectodomain. By ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Yasui N,Mihara E,Nampo M,Tamura-Kawakami K,Unno H,Matsumoto K,Takagi J

    更新日期:2010-01-31 00:00:00

  • Comparative analysis of flow cytometric methods for apoptosis quantitation in murine thymocytes and human peripheral lymphocytes from controls and HIV-infected persons. Evidence for interference by granulocytes and erythrocytes.

    abstract::The present article reports a multiparametric cytofluorimetric analysis of apoptosis in murine thymocytes and human PBMC from healthy donors or HIV-infected patients. We have evaluated four previously described cytofluorimetric methods of apoptosis quantification, each of them detecting distinct cellular alterations o...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Lecoeur H,Gougeon ML

    更新日期:1996-10-30 00:00:00

  • The concentrations of glutamine and ammonia in commercially available cell culture media.

    abstract::The amino acid glutamine is an essential nutrient for cells in culture. In aqueous solutions such as liquid culture media, glutamine spontaneously decomposes into ammonia. In this study, we examined the toxicity of ammonia for two different cell lines. In mouse hybridoma cell cultures, viable cell counts were reduced ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Heeneman S,Deutz NE,Buurman WA

    更新日期:1993-11-05 00:00:00

  • Establishment of the first WHO International Standard for etanercept, a TNF receptor II Fc fusion protein: Report of an international collaborative study.

    abstract::Etanercept, a recombinant human tumor necrosis factor (TNF) receptor Fc fusion protein is an effective treatment option in adults with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis or plaque psoriasis and paediatrics with juvenile idiotypic arthritis and plaque psoriasis. Patent expiration in Europ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Wadhwa M,Bird C,Dilger P,Rigsby P,Jia H,Gross MEB,participants of the study.

    更新日期:2017-08-01 00:00:00

  • Analysis of amplified T cell receptor V beta transcripts by a non-isotopic immunoassay.

    abstract::We have recently described a new colorimetric DNA enzyme immunoassay (DEIA) for detecting specific hybrids of complementary nucleic acids. This technology is based on an antibody that selectively recognizes double, but not single stranded DNA and therefore reveals the hybridization event independently from the DNA seq...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Bettinardi A,Imberti L,Sottini A,Primi D

    更新日期:1992-01-21 00:00:00

  • A reliable, rapid and inexpensive two-color fluorescence assay to monitor serum cytotoxicity in xenotransplantation.

    abstract::Removal and/or neutralization of preformed anti-pig antibodies in non-human primate blood have been shown to prevent the hyperacute rejection of transplanted pig organs. The purpose of this study was to establish a suitable in vitro method that would allow for screening and comparison of various agents and methods pot...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Neethling FA,Koscec M,Oriol R,Cooper DK,Koren E

    更新日期:1999-01-01 00:00:00

  • Characterisation of monoclonal antibodies against human interleukin-10 and their use in an ELISA for the measurement of this cytokine.

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    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: De Groote D,Marchant A,Fauchet F,Jadoul M,Dehart I,Gérard C,Gevaert Y,Lopez M,Gathy R,Franssen JD

    更新日期:1994-12-28 00:00:00

  • A paper radioimmunosorbent test (PRIST) for the detection of larva-specific antibodies to Toxocara canis in human sera.

    abstract::A paper radioimmunosorbent test (PRIST) was shown to be sensitive and reproducible when used with excretory/secretory antigen of Toxocara canis second stage larvae. Whatman No. 50 filter paper (5 mm discs) gave the most consistent and clear results with antigen at a concentration of 100 micrograms/ml, and could be sto...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Smith HV,Quinn R,Bruce RG,Girdwood RW

    更新日期:1980-01-01 00:00:00

  • Double fluorescence technique for measurement of complement-fixing antibody to lymphocyte subsets.

    abstract::A double fluorescent antibody method for quantitating human complement-fixing antibody to lymphocyte subclasses has been developed. The indicators in this system are a C6-deficient serum as a non-lytic source of complement, rhodamine-labeled anti-C3 and fluorescein-labeled murine monoclonal antibodies to human lymphoc...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章


    authors: Ozturk GE,Hayward AR,Weil R 3rd,Kohler PF

    更新日期:1985-12-17 00:00:00