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Biomedical subjects

D Kabelitz

Publications and source records attributed to D Kabelitz.

At least 181 records · Page 10Linked to original sources
PubMed1985

Allorestricted cytotoxic T cells. Large numbers of allo-H-2Kb-restricted antihapten and antiviral cytotoxic T cell populations clonally develop in vitro from murine splenic precursor T cells.

Cytotoxic T lymphocyte (CTL) responses of splenic T cells from C57BL/6 B6) mice and mutant H-2Kbm1 (bm1) mice to haptenic (trinitrophenyl [TNP] ) and herpes simplex virus (HSV) determinants in the context of an allogenic (wild-type or mutant) H-2Kb molecule were analyzed in a modified limiting dilution system. In the B6-anti-bm1TNP mixed leukocyte reaction (MLR), estimated frequencies for precursors of CTL clones that lysed bm1TNP targets ranged from 1/120 to 1/400; in the bm1-anti-B6TNP MLR, estimated frequencies of precursors of CTL clones that lysed B6TNP targets ranged from 1/500 to 1/1,300. Estimated frequencies for precursors of CTL clones that lysed the respective unmodified and TNP-modified allogeneic targets were two- to three-fold lower. Lytic specificity patterns determined by split-well analysis showed that at least 20-30% of the generated CTL populations (selected for a high probability of clonality) in both MLR displayed allorestricted lysis of TNP-modified concanavalin A blast targets. In the B6-anti-bm1HSV MLR, estimated frequencies for precursors of CTL clones that lysed bm1HSV targets ranged from 1/70 to 1/300; in the bm1-anti-B6HSV MLR, estimated frequencies for precursors of CTL clones that lysed B6HSV targets ranged from 1/300 to 1/1,200. Again, estimated frequencies for precursors of CTL clones that lysed the respective noninfected and virus-infected allogeneic targets were two- to fourfold lower. Of the CTL populations selected for a high probability of clonality at least 30-60% displayed allorestricted lysis of virus-infected lipopolysaccharide blast targets in both MLR. It is concluded that a large fraction of clonally developing CTL populations stimulated with TNP-modified or HSV-infected allo-H-2Kb-bearing cells displayed an allorestricted pattern of recognition. It was further evident that the estimated frequencies of splenic precursors that generated allorestricted CTL clones was two- to threefold higher than the estimated frequencies of precursors that gave rise to the respective alloreactive CTL populations.

Animals↗
PubMed1985

A rapid colorimetric assay for the determination of IL-2-producing helper T cell frequencies.

Interleukin 2 (IL-2) activity is tested in conditioned media by assessing its ability to support proliferation of selected IL-2 dependent T cell lines, conventionally measured by [3H]thymidine incorporation. Here, we compare this [3H]thymidine uptake test for measuring IL-2 activity with a rapid and sensitive colorimetric method which is based on the ability of viable cells to cleave 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). The sensitivity of the colorimetric method was dependent on the indicator cell line used, being greatest with the cytotoxic T cell line 16 (CTLL-16). The colorimetric method is at least as sensitive as [3H]thymidine uptake tests, does not rely on radioactivity, and is ideally suited to screen large numbers of individual samples for IL-2 activity. The latter point was demonstrated by calculating IL-2-producing helper T cell frequencies in heterogeneous murine lymphocyte populations: in this assay, splenic T cells were clonally expanded under limiting dilution conditions and supernatants conditioned by these in vitro growing T cell clones were tested for IL-2 activity with the colorimetric method. This allowed us to obtain reliable estimates of the frequency of progenitor cells of IL-2-producing T cell clones in various populations.

Animals↗
PubMed1985

Recombinant interleukin 2 rapidly augments human natural killer cell activity.

Recombinant human interleukin 2 (r-IL-2) rapidly stimulated human natural killer cell activity in vitro. Augmentation of NK activity occurred within 1 hr of preincubation with r-IL-2. Responsive killer cells were typical NK cells as shown by cell fractionation procedures. These included Percoll density gradient separation and depletion of OKT3+ T cells by an indirect rosetting method. Analysis with a panel of polyclonal and monoclonal antibodies against alpha and gamma interferon revealed that this early enhancement of NK activity by r-IL-2 was independent of the production of both types of interferon.

Antibodies↗
PubMed1985

Modulation of natural killing by tumor promoters: the regulatory influence of adherent cells varies with the type of target cell.

We have analyzed the regulatory effects of two classes of tumor promoters, phorbol diesters and indole alkaloids, on human natural killer (NK) cell activity in vitro. In accordance with previous reports, we found that 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibited natural killing against K 562 targets by unseparated mononuclear cells. Here, suppression of NK required the presence of adherent cells (macrophages). Contrary to the results obtained with K 562, tumor promoter-induced suppression of NK activity tested against U 937, another cell line of known NK susceptibility, was independent of the presence of adherent cells. Thus, NK cytotoxicity of effector cells rigorously depleted of adherent and Ia-positive cells was still inhibited when assayed against U 937, while it was generally enhanced when tested against K 562. Identical results were obtained with teleocidin and dihydroteleocidin B, two members of the recently discovered indole alkaloid class of tumor promoters. Therefore, we demonstrate that the regulatory effect of tumor promoters on human NK activity (suppression or stimulation) is determined not only by macrophages at the effector cell level but also by the type of target cell under study.

Carcinogens↗
PubMed1985

Human cytotoxic T lymphocytes. I. Limiting-dilution analysis of alloreactive cytotoxic T-lymphocyte precursor frequencies.

A limiting-dilution system was established to measure the frequency of alloreactive cytotoxic T-lymphocyte precursors (CTL-p) in human peripheral blood T cells. Culture medium supplemented with recombinant interleukin-2 enabled clonal expansion of all CTL-p stimulated by allogeneic peripheral blood or spleen cells. The range of CTL-p frequencies in fully HLA-mismatched responder-stimulator combinations was 1:240 to 1:1230. Split-well analysis of individual microwells showed that the cytotoxic T-cell clones generated under limiting-dilution conditions showed exquisite specificity for the stimulating alloantigens. Alloreactive CTL-p were enriched in the OKT4- T-cell subset. This limiting-dilution system was highly reproducible and can thus be applied to investigate human cytotoxic T-cell precursor frequencies in various clinically relevant situations.

Cells, Cultured↗
PubMed1985

In vitro maturation of B cells in chronic lymphocytic leukemia. I. Synergistic action of phorbol ester and interleukin 2 in the induction of Tac antigen expression and interleukin 2 responsiveness in leukemic B cells.

Recent evidence indicates that interleukin 2 (IL 2), formerly thought to serve as growth factor exclusively for activated T cells, is directly involved in human B cell differentiation. We have investigated the role of IL 2 and IL 2 receptors (as defined by monoclonal anti-Tac antibody) in the phorbol ester-induced in vitro maturation of leukemic B cells from patients with chronic lymphocytic leukemia (CLL). Peripheral blood lymphocytes from B cells from CLL patients with high (greater than 10(5)/microliters) white blood cell counts were depleted of residual T lymphocytes and low-density cells (primarily macrophages) by consecutive steps of E rosetting, complement-mediated lysis of OKT3+ and OKT4+ cells, and Percoll density gradient centrifugation. No OKT3+ T cells were detectable in these cell populations before or after culture. When incubated for 3 days with phorbol ester plus recombinant human IL 2 (rIL 2), 12 to 57% of highly purified B cells from four of five tested patients expressed Tac antigen. Both phorbol ester and rIL 2 were required for maximal Tac antigen expression. Functional studies revealed that phorbol ester-activated (but not resting) CLL B cells responded to rIL 2 with [3H]thymidine incorporation and with enhanced secretion of IgM. Tac+ B cells were isolated in two cases on a fluorescence-activated cell sorter. In one patient, stimulation of Tac+ B cells with rIL 2 resulted in enhanced [3H]thymidine incorporation but no change in IgM secretion, as compared with Tac- B cells; in the second patient, stimulation of Tac+ B cells with rIL 2 did not result in [3H]thymidine uptake, but did result in significant IgM secretion. These findings indicate that certain leukemic B lymphocytes can be induced to express IL 2 receptors and respond to IL 2. The use of resting clonal B cell populations arrested at distinct stages of differentiation may help to better define the stage(s) at which IL 2 acts directly on B cells to induce proliferation and/or terminal differentiation.

Antibodies, Monoclonal↗
PubMed1985

Phorbol ester-induced differentiation of chronic B lymphocytic leukaemia cells--regulatory impact of autologous and allogeneic accessory cells.

Phorbol ester (TPA) induction of chronic lymphocytic leukaemia (CLL) cells can be used as a model system for the study of human B cell differentiation. We have analysed the role of accessory cells and the correlation to target cell surface Ig phenotype in TPA-induced morphological and functional differentiation of 12 CLL populations. A more than three-fold increase in secreted IgM was seen in 10 of 12 cases, with the strongest responses in patients having monoclonal serum IgM. CLL populations negative for or having only weak surface mu chain expression were less inducible. The impact of autologous and allogeneic accessory cells on TPA induction was studied in cell enrichment/depletion experiments using both physical and cytotoxic antibody techniques. CLL cells physically depleted of autologous E+ and monocytic (light density fraction) cells still responded to TPA. This response could be enhanced by allogeneic E- light fraction cells. Further depletion of autologous accessory cells by treatment of the E- high density fraction CLL cells with a panel of monoclonal antibodies plus complement demonstrated the permissive role of one or two populations of autologous cells expressing low avidity E receptors and the T3, T4 and T8 antigens. Augmenting T cells of similar phenotype were found among allogeneic cells from normal individuals. Thus, TPA-induced IgM secretion in biopsy B-CLL cells is regulated by minute numbers of autologous helper T cells. Furthermore, the Ig secretory response of CLL populations seems to be correlated with the surface Ig the surface immunoglobulin phenotype of the leukaemic cells.

Aged↗
PubMed1984

Modulation of natural killer susceptibility by indole alkaloid tumor promoter dihydroteleocidin B.

The indole alkaloid tumor promoter dihydroteleocidin B (DHTB) was shown to reduce the natural killer (NK) cell susceptibility of two established cell lines, U937 and K562. The decrease in NK susceptibility correlated with the induction of differentiation as documented by positive benzidine staining in the erythroleukemia K562 and by antibody-dependent cellular cytotoxicity effector function in the histiocytic lymphoma line U937, respectively. In contrast, DHTB treatment did not alter the NK sensitivity of the NK-resistant B-lymphoblastoid-cell line RPMI 8866. Cold target inhibition experiments suggested that both effector-target recognition and post-recognition steps were affected by DHTB. These results lend further support to the notion that NK susceptibility of a given tumor cell may vary with the stage of differentiation.

Alkaloids↗
PubMed1984

Phorbol ester treated chronic B lymphocytic leukaemia cells induce autologous T cell proliferation without generation of cytotoxic T cells.

The ability of the tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) to modify the stimulatory capacity of leukaemic B cells in autologous mixed leucocyte reactions (AMLR) was studied. T lymphocytes from patients with chronic lymphocytic leukaemia (CLL) did not proliferate when stimulated with untreated autologous non-T cells. In contrast, vigorous 3H-thymidine incorporation was seen in six of eight cases when CLL stimulator cells were pre-incubated for 3 days with TPA. No cytotoxic T cells against TPA treated or untreated autologous target cells were generated in such AMLR. These results indicate that enhancement of AMLR stimulatory capacity may be among the characteristic features of TPA driven in vitro differentiation of CLL cells.

B-Lymphocytes↗
PubMed1983

Density separation of chronic lymphocytic leukemia cells: low-density non-T cells are efficient stimulator cells in allogeneic and autologous mixed leukocyte reaction.

Unseparated mononuclear cells and E rosette-depleted non-T cells from a majority of patients with chronic lymphocytic leukemia (CLL) were found to be weak stimulators in the allogeneic mixed-leukocyte reaction (MLR). However, a minor population (1-5%) of highly active stimulator cells could be isolated from all patients studied by buoyant density centrifugation using discontinuous gradients of Percoll. The same low-density non-T-cell fraction also stimulated autologous T-cell proliferation in the autologous mixed-leukocyte reaction (AMLR). In contrast, Percoll-separated high-density non-T cells, including the leukemic B-cell pool, were completely inactive as stimulators in AMLR. These results suggest that the presence of efficient stimulator cells among CLL mononuclear cells may be masked by the large number of nonstimulating leukemic B cells.

Cell Separation↗
PubMed1983

Phorbolester-treated human lymphocytes are susceptible to natural killer cell-mediated cytolysis.

The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) was found in this study to render normal human lymphocytes susceptible to allogeneic and autologous natural killer (NK) cell lysis. Both T and non-T cells became susceptible after culture for 3 days in the presence of 1 ng/ml TPA. Effector cells were nonadherent mononuclear cells of low density, enriched for large granular lymphocytes and HNK-1+ cells. Activation of effector cells with interferon increased lysis of TPA-treated lymphocytes. The present system may provide a new model to study NK cell target structures.

Caenorhabditis elegans Proteins↗
PubMed1982

Immune functions of human synovial cells. Phenotypic and T cell regulatory properties of macrophage-like cells that express HLA-DR.

Normal and rheumatoid synovial cells have been analyzed in frozen sections and in suspension. HLA-DR-expressing, macrophage-like cells are demonstrated in normal synovial intima and in rheumatoid tissue. Suspended normal synoviocytes equaled peripheral blood non-T lymphocytes as stimulators of mixed lymphocyte reactions, whereas adherent rheumatoid synovial cells were extremely efficient as such stimulators and in presenting soluble antigens to autologous T lymphocytes. This HLA-DR-dependent T lymphocyte regulation might provide a cellular basis for the HLA-D haplotype-arthritis associations.

Antibodies, Monoclonal↗
PubMed1982

Inhibition of autologous mixed lymphocyte reaction by aggregated IgG molecules.

Human autologous and allogeneic mixed lymphocyte reactions (MLR) were studied using various T cell subsets as responding cells. T cells expressing Fc gamma receptor for IgG (TG) were weak responder while T cells with Fc mu receptor and T cells lacking Fc receptor responded vigorously in both types of MLR. Percoll-separated T cells of low density (TL) enriched for TG cells were, however, excellent responders in autologous MLR, whereas T cells of high density (TH) were the major responders in allogeneic MLR. Continuous presence of or short-term pulse with aggregated IgG significantly reduced the autologous MLR of unfractionated T and TL cells while largely sparing proliferation in allogenic MLR. These findings are discussed with respect to diminished autologous MLR reactivity in certain autoimmune diseases.

Autoimmune Diseases↗
PubMed1982

Alloantigen-induced suppressor- and memory cells in chronic lymphocytic leukemia.

T cell function was evaluated in patients with B cell type of chronic lymphocytic leukemia (CLL). Unseparated peripheral blood lymphocytes (PBL) and T cells from CLL patients stimulated in a primary allogeneic MLR were able to inhibit significantly a second MLR between the original responder (CLL) and stimulator (normal PBL) cell donors. Furthermore, it is shown the T lymphocytes from patients with CLL develop immunologic memory during the course of a primary MLR as evidenced by an enhanced response in secondary MLR. These results are discussed with respect to recently described imbalances of T cell subpopulations in CLL.

B-Lymphocytes↗
PubMed1982

Evidence in support of a self-perpetuating HLA-DR-dependent delayed-type cell reaction in rheumatoid arthritis.

Originating from observations on similarities between the rheumatoid synovial tissue and skin lesions in delayed-type hypersensitivity reactions--similarities as to massive infiltrates of "helper" T lymphocytes close to HLA-DR-expressing macrophage/dendritic cells--a notion is formed on the importance of local macrophage-dependent helper T-cell activation in the rheumatoid joint similar to that in a delayed-type skin reaction. In vitro studies on suspended synovial cells have been used to test and qualify these ideas. It is shown that (i) HLA-DR-expressing cells in normal synovial intima can, like epidermal Langerhans cells, mediate T-cell activation; (ii) the large numbers of rheumatoid synovial HLA-DR-expressing macrophage-like/dendritic cells are heterogeneous and mediate either efficient activation or suppression of T-lymphocyte proliferation, and (iii) specificity of rheumatoid T cells can be analyzed with the help of autologous synovial antigen-presenting cells; a specific anti-collagen type II response is reported in three patients.

Arthritis, Rheumatoid↗
PubMed1982

Density distribution profiles of T cells: TM, TG and TA cells and response patterns in autologous versus allogeneic MLRs.

Discontinuous Percoll gradients have been used to obtain selected human peripheral blood T lymphocytes without having to resort to interactions with immune complexes in the fractionation of Tm, Tg and Ta cells. Here, we could show that Ta cells represent a heterogeneous population with no distinct density profile, in contrast to light (Tg) and heavy (Tm) cells. Enriched, heavy Tm cells could be shown to be excellent responders in allogeneic MLR while failing to react in autologous MLR. In contrast, T cells of light density preferentially respond in autologous compared with allogeneic MLR.

Animals↗
PubMed1981

Enrichment of MLR-activated human suppressor cells by discontinuous density-gradient centrifugation.

6-day allogeneic mixed lymphocyte cultures (MLR) were fractionated on a four-step discontinuous Percoll gradient. The various fractions were added as regulatory cells to freshly established micro-MLRs between the original or third-party cell donors. Unspecific suppressor activity was highly enriched in gradient fraction III (density 1.069--1.062 g/ml), consisting mainly of blast-like cells. Suppressor cells were non-adherent T cells as determined by their spontaneous rosette formation with sheep erythrocytes and their susceptibility to lysis by anti-T-cell globulin plus complement. Mitomycin C treatment diminished their inhibitory capacity. In addition, gradient fraction III was also enriched with alloantigen-activated memory cells. By contrast, unprimed alloreactive cells were recovered from gradient fraction IV (density greater than or equal to 1.070 g/ml), consisting mainly of small, resting lymphocytes.

Adult↗