Automated reading of HLA-A,B,C typing and screening

Bruning, J. W.; Claas, F. H. J.; Kardol, M. J.; Lansbergen, Q; Naipal, A.; Tanke, Hans J.

doi:10.1016/0198-8859(82)90135-5

Cited by 52 publications

(23 citation statements)

References 7 publications

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“…Two microliters of B cell supernatants or hybridoma supernatants was tested for HLA-A2 Abs by CDC (13), with mononuclear cells (5000/well from three donors typed as HLA-A2,Ϫ; HLA-A1,A2; and HLA-A2 negative) as targets. Results were expressed as percentage of target cell lysis, where 100% lysis was caused by mAb FK5, detecting a marker common to all leukocytes.…”

Section: Complement-dependent Cytotoxicity (Cdc)mentioning

confidence: 99%

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

Mulder

Eijsink

Kardol

et al. 2003

The Journal of Immunology

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Characterizing the individual B cells that participate in the production of anti-HLA Abs requires isolation and culture of these cells and a suitable assay for detection of Abs produced in these B cell cultures. We previously showed that B cell precursors, programmed for anti-HLA Ab secretion, are present at measurable frequencies in peripheral blood of women immunized by pregnancy. In this study, we show that tetrameric HLA-A2, although designed for characterization of CTLs, provides a suitable affinity ligand for isolation of allospecific B cells, which subsequently can be induced to produce HLA-A2 Ab in a CD40-driven culture system. The validity of this concept was established by assaying human hybridomas, producing anti-HLA Abs, for specific tetrameric HLA-A2 binding. The availability of anti-HLA Ab-producing B cell cultures that are established without immortalization will be of value when T-B cell interaction is studied at an alloantigen-specific level.

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Section: Complement-dependent Cytotoxicity (Cdc)mentioning

confidence: 99%

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

Mulder

Eijsink

Kardol

et al. 2003

The Journal of Immunology

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“…Autologous EB-LCL was generated by cultivation with RPMI-1640 medium supplemented with 10% FCS for 5-6 weeks. The cells revealed B-cell type having surface phenotype CD3-CD19t HLA-A, B and DR typing of the autologous EB-LCL and PBL from the IM patient was performed by Propidium iodide method (Bruning et al 1982 …”

Section: Target Cellsmentioning

confidence: 99%

CD8+ T cell subsets of cytotoxic T lymphocytes induced by Epstein-Barr virus infection in infectious mononucleosis.

Ebihara

Sakai

Koyama

1990

Tohoku J. Exp. Med.

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Mononuclear peripheral blood lymphocytes (PBL) from patient with infectious mononucleosis (IM) were tested in a 51Cr-release assay for cytotoxicity against autologous and allogeneic lymphoblastoid cell line (LCL), or Epstein-Barr virus (EBV)-genome positive and negative cell line. In acute phase, PBL lyse an autologous LCL as well as allogeneic LCL (Wa cells). High levels of cytotoxicity were observed in the combinations between effector and target cells sharing HLA-Class 1 product. EBV-genome positive Daudi and Raji cells which lack HLA-Class 1 antigen and have mismactched HLA-Class 1 antigen, respectively showed resistance to killing. EBV-genome negative tumor cells except NK sensitive K562 cells were not killed by IM lymphocytes. However, the IM lymphocytes without atypical form in convalescent phase failed to show killing activity against autologous and allogeneic LCL. These findings suggest that cell surface membrane antigen structure on EBV-infected LCL may be able to explain the recognition and triggering of lysis of target cells by HLA-Class 1 restricted cytotoxic T cells (CTL) from acute IM. Phenotypic analysis of PBL with atypical form from IM was made by two-color flow cytometry. The data demonstrate that CD8+ T cells quantitatively represent the major population of lymphocytes expanded during acute IM. Furthermore, approximately 70% of these CD8+ T cells express HLA-DR on these surface, suggesting that they have undergone activation. However, IL 2R (CD25 antigen) expression was not significantly elevated on activated T cells. The salient profile on cytofluorographs of an acute IM was the increased number of CD3+CD19-, CD8+CDllb-, CD8+CD28+ and CD8+S6F1+ cells. However, CD3-CD19+, CD8+CDllb+, CD8+S6F1-, CD4+Leu8-and CD25+HLA-DW antigens were little expressed. Increased number of CD8~CDllb-, CD8+CD28+ and CD8+ S6F1+ cells, which are regarded as CTL were reduced according to the improvement of the clinical symptoms and laboratory findings. These results together with HLA typing analysis suggested a possibility HLA-Class 1 restriction of the CTL with surface phenotype of CD8+CDllb-, CD8+CD28+, and CD8+S6F1t cytotoxic T cells (CTL) ; CD8 T cells ; HLA ; Epstein-Barr virus

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“…The microcytotoxicity test with propidium iodide (Sigma) was used according to the method of Bruning et al [16]. Two microliters of ap ropriately diluted mAb, and 2 p1 of cell susTerasaki plate, under an oil drop.…”

Section: Microcytotoxicity Testmentioning

confidence: 99%

An attempt to produce “pre‐T” cell hybridomas and to identify their antigens

Aihara

Bühring²,

Aihara

et al. 1986

Eur J Immunol

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With the aim of identifying some of the stages in the development of pre-T cells (cells of the T cell lineage before they enter the thymus), we have produced a large number of hybridomas by the fusion of BALB/c bone marrow cells, bone marrow cells from BALB/c-nu/nu mice, BALB/c fetal liver cells and BALB/c fetal thymocytes with the AKR thymoma BW5147. The hybridomas were selected for the expression of the Thy-1.2 antigen of the normal cell donor and for their ability to produce interleukin 2 (IL 2) upon co-culture with irradiated normal spleen cells. A set of these hybridomas is described in this communication. The hybridomas were then used to immunize rats and to generate monoclonal antibody-producing B cell hybridomas. Most, if not all, of the immunizing hybridomas were derived from pre-T cells as evidenced by the fact that they produce IL 2, and express some of the T cell markers (the Thy-1.2, Ly-1, Ly-2 or L3T4 antigens). The monoclonal antibodies were tested on a panel of pre-T cell hybridomas and on normal cells obtained from spleen, lymph nodes, thymus and bone marrow. The testing was carried out by the microcytotoxicity assay and flow cytometric analysis. Three groups of antibodies could be distinguished. Some antibodies were broadly reactive, being positive with virtually all the clones in the pre-T cell panel and with a substantial fraction of normal lymphoid cells. The identity of the antigens detected by these antibodies remains unknown but they do not seem to correspond to any of the known cell surface markers. Other antibodies reacted only with some of the pre-T cell clones and did not react at all with normal lymphoid cells obtained from adult animals. Finally, other antibodies still reacted only with a minor subpopulation of thymocytes or of thymocytes and bone marrow cells, as well as some of the pre-T cell clones; they did not react with spleen and lymph node cells. These antibodies might be specific for cells in the prethymic phase of the T cell differentiation pathway. They should prove useful for the identification of pre-T cell markers and hence for the isolation of pre-T cells and their functional analysis.

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Automated reading of HLA-A,B,C typing and screening

Cited by 52 publications

References 7 publications

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

Identification, Isolation, and Culture of HLA-A2-Specific B Lymphocytes Using MHC Class I Tetramers

CD8+ T cell subsets of cytotoxic T lymphocytes induced by Epstein-Barr virus infection in infectious mononucleosis.

An attempt to produce “pre‐T” cell hybridomas and to identify their antigens

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