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T Diamantstein

Publications and source records attributed to T Diamantstein.

At least 181 records · Page 10Linked to original sources

Inhibition of IL 2-dependent proliferation of rat T lymphoblasts by the monoclonal antibody ART62 which reacts with MHC class 1 antigens.

During the course of studies designed to obtain monoclonal antibodies (mAb) that recognize the rat interleukin 2 receptor, a mouse IgG1 mAb (ART62) was identified which inhibits the interleukin 2 (IL 2)-dependent proliferation of rat T lymphoblasts without affecting the binding of IL 2 to such cells. In order to characterize the cell surface components that react with the mAb ART62, T lymphoblasts were surface-labeled with 125I, and the radioactive molecules were immunoprecipitated by the antibody analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The mAb ART62 precipitated two major components of 48,000 m.w. and 12,000 m.w., respectively, which were different from those which react with the anti-IL 2-receptor antibody ART18, a molecule of 50,000 to 55,000 m.w. Sequential immunoprecipitation studies revealed that the mAb ART62 reacts with the MHC class 1 antigen that reacts with the classical anti-rat MHC class 1 mAb OX18, and vice versa. In contrast to the mAb ART62, OX18 that does not affect and several other mAbs known to inhibit the rat MLR failed to inhibit IL 2-dependent proliferation of rat T lymphoblasts. In contrast to the anti-IL 2 receptor antibody ART18, ART62 effectively inhibited IL 2-driven proliferation even when added to cells already committed to proliferate by IL 2-IL 2 receptor interaction. These data raise the possibility that MHC class 1 antigens could be involved in the chain of reactions mediating the signals required for cell proliferation.

Animals↗

TCGF(IL 2)-receptor inducing factor(s). II. Possible role of ATL-derived factor (ADF) on constitutive IL 2 receptor expression of HTLV-I(+) T cell lines.

Interleukin 2 (IL 2) receptor (IL 2-R) is constitutively expressed on T cell lines established from the patients with adult T cell leukemia (ATL), which is a human T cell leukemia lymphoma virus (HTLV-1)(+) T4(+)-leukemia endemic in Japan, the United States, and other countries. Many of these cell lines continuously produce an acidic lymphokine, ATL-derived factor (ADF), which preferentially induces the synthesis and expression of IL 2-R on a sensitive HTLV-1(-) non-T cell line (YT). The induced IL 2-R was characterized by the binding of 125I-IL 2 and flow cytometry by using fluoresceinated anti-human IL 2-R monoclonal antibodies (anti-Tac). Scatchard analysis with 125I-IL 2 showed ADF induced high-affinity receptor sites on YT cells. To test the possibility that ADF produced by HTLV-1(+) T cells is involved in the abnormal expression of IL 2-R, we studied the effect of ADF on an HTLV-1(+) IL 2-dependent T cell line (ED) in which the beta-chain gene of the T cell antigen receptor (T beta) was rearranged. Unlike IL 2-independent HTLV-1(+) cell lines that constitutively expressed Il 2-R, the IL 2-R expression on ED cells declined in the absence of crude IL 2 or recombinant IL 2. When either ADF or recombinant IL 2 was added to the culture of ED cells, there was a dose-dependent enhancement of IL 2-R expression in 24 hr. ADF and IL 2 showed a synergism in the IL 2-R induction, and both factors were needed to induce the maximal receptor expression in these T cells. The lack of IL 2 production by ADF-treated YT, as well as ED cell line suggested IL 2 may not be involved in the IL 2-R induction by ADF. Northern blot hybridization with human IL 2-R cDNA probe showed the increase of IL 2-R mRNA in YT cells after ADF-treatment. ADF also enhanced IL 2-R expression of a rat T cell line transformed by HTLV-1(TARS-1), as demonstrated with anti-rat IL 2 receptor monoclonal antibodies (ART-18). An ADF-like IL 2-R-inducing factor was also detected in the conditioned medium of two HTLV-1(+) rat T cell lines (TARL-2 and TART-1), which constitutively expressed a higher number of Il 2-R than TARS-1 cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗

[The effect of cefaclor, doxycycline and other oral antibiotics on phagocytosis of polymorphonuclear granulocytes and PHA-induced stimulation of blood lymphocytes in man].

The influence of the orally applicable antibacterial agents, cefaclor (Panoral), doxycycline, erythromycin, and cotrimoxazole on phagocytosis by human neutrophil granulocytes was investigated. Further, the influence of cefaclor, erythromycin, and doxycycline on PHA stimulation of human blood lymphocytes was studied. Doxycycline (1 mg/ml) had strongly suppressive effects on phagocytosis, whereas cefaclor (1 mg/ml cotrimoxazole (0.1 mg/ml), and erythromycin (0.1 mg/ml) did not suppress phagocytosis. PHA-induced lymphocyte stimulation was strongly inhibited by doxycycline (10 micrograms/ml), whereas cefaclor (10 micrograms/ml) did not suppress at all. Erythromycin (10 micrograms/ml) exerted some inhibition (average 12.6%). These results are discussed with respect to immunoneutrality of antibacterial agents.

Anti-Bacterial Agents↗

Functional analysis of mononuclear cells infiltrating into tumors. II. Differential ability of mononuclear cells obtained from various tissues to produce helper factors that are involved in the generation of cytotoxic cells.

The requirement of CGF in the generation of cytotoxic cells against syngeneic tumor cells (T-9) and in the rejection of transplanted T-9 cells has been investigated. Spleen cells obtained from sensitized rats showed strong cytotoxicity against 51Cr-labeled T-9 cells upon incubation with CGF for 48 hr. Human recombinant IL 2 and rat IFN failed to generate cytotoxic cells from spleen cells of sensitized rats. CGF are produced by spleen cells upon inoculation of T-9 cells into sensitized rats as a host in vivo immune response. Production of CGF preceded the appearance of cytotoxic cells in regional lymph node and tumor tissues. In those rats, inoculated tumor cells were eventually rejected. In contrast, spleen cells failed to produce CGF upon inoculation of T-9 cells in unsensitized rats. Cytotoxic cells were not detected in unsensitized rats, and inoculated tumor grew in those rats. Thus, CGF is likely to be involved in the generation of cytotoxic cells and in the rejection of inoculated syngeneic tumor cells. A Mono Q anion-exchange column with an FPLC system allowed the chromatographic separation of CGF from IL 1, IL 2, IL 3, and CSF.

Animals↗

Immunohistochemical localization of cells reacting with monoclonal antibodies directed against the interleukin-2 receptor of murine, rat and human origin.

Recently, species specific monoclonal antibodies, directed against the interleukin-2 (IL-2) receptor of murine, rat and human origin, have been produced. In this study we demonstrate with immunohistological methods that cells reacting with these antibodies are present in normal primary and secondary lymphatic organs. The cells are exclusively localized in T cell-dependent areas, and their number increased as a result of immunization.

Animals↗

Induction of interleukin 2 responsiveness in thymocytes by synergistic action of interleukin 1 and interleukin 2.

Thymocyte cultures from C3H/HeJ mice were stimulated for proliferative responses with purified preparations of interleukin 1 (IL 1) and interleukin 2 (IL 2). Synergistic responses were obtained in the absence of mitogen. In the presence of excess IL 2, the thymocyte proliferation response was strictly dependent on the amount of IL 1 in the cultures. Antibodies to IL 1 inhibited the response in a dose-dependent manner. The combination of IL 1 plus IL 2 induced the appearance of IL 2 receptors on murine thymocytes as detected with a monoclonal antibody directed against the IL 2 receptor. Neither IL 1 nor IL 2 alone had this effect. The thymic subpopulation found to become IL 2 responsive upon IL 1 stimulus was the peanut agglutinin-negative (PNA-) medullary fraction.

Animals↗

Developmentally controlled expression of IL 2 receptors and of sensitivity to IL 2 in a subset of embryonic thymocytes.

At various times of gestation murine fetal thymocytes were tested for IL 2 receptor (IL 2-R) and T cell differentiation antigen expression. The majority of 14 to 15 day fetal thymocytes were IL 2-R and Thy-1 antigen positive, yet negative for the Lyt and L3T4 marker. A subset of IL 2-R-positive fetal thymocytes could be induced by recombinant IL 2 to proliferate over at least 10 days. Growth of these proliferating cells could not be enhanced by syngeneic feeder cells nor suppressed by monoclonal anti-I-A or anti-I-E antibodies. No antigen-specific effector functions could be induced in the proliferating Thy-1, IL 2-R-positive cells. As a whole, the results suggest a developmentally controlled rather than antigen-induced expression of IL 2-R during embryogenesis of thymocytes.

Animals↗

Induction of IL 2 receptor expression and cytotoxicity of thymocytes by stimulation with TCF1.

We investigated the role of T cell cytotoxicity inducing factor 1 (TCF1) in the induction of a cytotoxic T cell response. We found that help-deficient thymocyte cultures supplied with saturating amounts of purified IL 2 did not develop CTL in a 5-day culture. The expression of cytotoxicity was dependent on the addition of TCF1 derived from the T cell hybridoma K15. TCF1 also induced proliferation of thymocytes in the presence of IL 2. Only the PNA- thymocyte subpopulation responded to TCF1 with proliferation and cytotoxicity in the presence of IL 2. The monokine IL 1 also induced proliferation in this subpopulation but failed to induce cytotoxicity. IL 1 was further distinguished from TCF1 by inhibition of IL 1-induced but not TCF1-induced proliferation by anti-IL 1 antibodies. In addition, using anti-IL 2 receptor antibodies (AMT 13), we showed that TCF1 in the presence of IL 2 substantially increased IL 2 receptor expression in thymocytes. IL 1 had the same effect on induction of IL 2 receptor expression as TCF1. Because some effects of IL 1 and TCF1 are distinct and some overlap, we discuss whether IL 1 and TCF1 induce different subsets of PNA- thymocytes.

Animals↗

Partial characterization of the putative rat interleukin 2 receptor.

T lymphoblasts of rat origin were (a) surface labeled with 125I and (b) internally labeled with 3H-marked sugars. Cell lysates were purified by immunoabsorption using the putative anti-rat interleukin 2 (IL2) receptor monoclonal antibody ART18 . The purified material was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoretic analysis. Two specific membrane components were detected: a 50-kDa major and a 36-kDa minor component in reducing and a 45-kDA major and a 72-kDa minor component in nonreducing conditions, respectively. Both components were found to be susceptible to trypsinization and to neuraminidase treatment. 3H-labeled sugars were incorporated into the major component. The results indicate that the rat IL2 receptor is either a 50-kDa glycoprotein or a 36-kDa molecule, or that both components are part of the receptor molecule.

Animals↗

Detection and localization by the monoclonal anti-interleukin 2 receptor antibody AMT-13 of IL 2 receptor-bearing cells in the developing thymus of the mouse embryo and in the thymus of cortisone-treated mice.

During embryonic development of the mouse, before expressing classical T cell markers, the blast cells colonizing the thymus react with the monoclonal antibody AMT-13 shown previously to detect interleukin 2 receptors. The proportion of AMT-13+ cells decreases as gestation time increases. On the other hand, the proportion of Thy-1+, Lyt-1+ and Lyt-2+ cells increases during ontogenesis. On the 19th day of gestation when the thymus architecture is comparable to the adult thymus, the AMT-13+ cells become localized in the subcapsular area of the cortex. In the adult thymus after cortison treatment the regenerating cells express the AMT-13 antigen. The AMT-13 antigen presumably the interleukin 2 receptor is the first marker of the early embryonic thymocytes reported until now that may be related to cellular function.

Animals↗

Studies on the interleukin-2 receptor, its generation and dynamics using monoclonal anti-interleukin-2 receptor antibodies.

There is increasing evidence suggesting that the monoclonal antibodies ART-18, AMT-13 and anti-Tac recognize species-specific antigenic determinants of the interleukin-2 (IL-2) receptors of rat, mouse and human origin, respectively. In order to compare directly the molecules (glycoproteins) recognized by these antibodies, concanavalin A (ConA) activated T-lymphocytes of the respective species were surface labeled with 125I, after which the materials immunoprecipitated by the appropriate anti-IL-2 receptor antibodies were subjected to SDS-PAGE analysis. The noncross-reacting antibodies ART-18 and AMT-13 both precipitated a 50-55-kD molecule. The anti-Tac-reactive material (the putative human IL-2 receptor) is considerably different (60-65 kD) from those precipitated by antibodies ART-18 and AMT-13 (the putative rat and mouse IL-2 receptors). An indirect binding assay using the anti-mouse IL-2 receptor antibody AMT-13 showed that, after addition of ConA to spleen cell cultures, T-lymphocytes expressed IL-2 receptors before the onset of the ConA-induced DNA synthesis. The ConA-induced expression of the IL-2 receptor is apparently a transient event. IL-2 receptor bearing cells progressively lost their receptors (within 6 days) when recultured in the absence of ConA. Cells re-exposed to ConA regained IL-2 receptors. Short exposure of T-cells (thymocytes) to ConA or the nonmitogenic compound phorbol myristate acetate (PMA) is not sufficient to trigger IL-2 receptor expression. Murine thymocytes incubated with PMA for 30 min or with ConA for 4 hr (mitogen-pulsed T-cells) failed to bind the anti-IL-2 receptor antibody AMT-13 and to absorb IL-2 activity present in semipurified IL-2 preparations, but they proliferated vigorously in response to the same IL-2 preparations. The IL-2 preparations, when absorbed with thymocytes, lost: (1) the capacity to generate IL-2 receptors, and (2) the capacity to induce proliferation of mitogen-pulsed cells; but they retained the capacity to induce proliferation of T-lymphoblasts. These results suggest the existence of a factor, IL-2 receptor inducing factor (RIF), present in the IL-2 preparations. It is postulated that RIF is a prerequisite for the acquisition of IL-2 receptors and consequently for IL-2 responsiveness by lectin-activated cells.

Animals↗

Resistance to 4-hydroperoxycyclophosphamide of T cells involved in cell mediated antibacterial immunity.

4-Hydroperoxycyclophosphamide is a derivative of cyclophosphamide which in vitro is converted into an active metabolite of cyclophosphamide. This compound was used to define the relative susceptibilities of T cells involved in the immune response of mice to the intracellular pathogen, Listeria monocytogenes. L. monocytogenes-specific T cell proliferation and interleukin production in vitro, as well as adoptive protection and delayed-type hypersensitivity in vivo, all proved to be markedly resistant to the action of 4-hydroperoxy-cyclophosphamide, indicating a great homogeneity within the cellular immune response to intracellular pathogens.

Animals↗

Interleukin 2 receptors on an insulin-specific T cell line: dynamics of receptor expression.

Long-term cultured T blasts with specificity for bovine insulin (BK-BI-1.2), which cease growing in IL 2 supplemented medium and require periodic antigenic challenge to resume proliferation, were selected as a model system to analyze the regulation of the growth of activated T cells. The use of AMT-13, a monoclonal antibody (mAb) directed at the murine IL 2 receptor, in indirect binding experiments and in FACS analysis allowed us to examine the time-dependent expression of IL 2 receptors on BK-BI-1.2 blasts after antigenic stimulation. The data reveal a transitory expression of IL 2 receptors, attaining maximal levels on day 2 after antigenic induction and having declined to low levels by day 6. mAb 10-2.16, reactive with I-Ak, did not inhibit T cell-proliferative capacity when the cells were subcultured in IL 2. This result suggests that, once induced to maximal levels by antigen, the transitory expression of IL 2 receptors on the descendent cells is not dependent on the continual presence of antigen-presenting cells. Thus, the progressive loss of IL 2 receptors apparently is not due to a mechanism operating by clearance of receptors from the cell surface on completion of each cell cycle, leading to dependency of the descendent cells on repeated contact with antigen for renewed receptor expression. The disappearance of IL 2 receptors from the surface of antigen-stimulated T cells might provide a basis for the control of immunologic specificity in vivo.

Animals↗

A rat monoclonal antibody that binds specifically to mouse T lymphoblasts and inhibits IL 2 receptor functions: a putative anti-IL 2 receptor antibody.

To obtain an anti-interleukin 2 (IL 2) receptor antibody, we immunized rats with phorbol myristate acetate-pulsed mouse T lymphoblasts. The spleen cells of the rats were fused with rat myeloma cells. Several stable clones of hybridoma cells were obtained that produced monoclonal antibodies (mAb) reacting specifically with mouse T lymphoblasts. One of these mAb, AMT-13, showed characteristics of a putative anti-IL 2 receptor antibody. As tested by FACS analysis, the mAb AMT-13 binds to murine T lymphoblasts or to IL 2-dependent T cell lines, but not to B lymphoblasts, thymocytes, splenocytes, lymph node cells, or bone marrow cells. The mAb AMT-13 inhibited in a species-specific and dose-dependent manner: i) the capacity of mouse T lymphoblasts to absorb IL 2 activity, and ii) the capacity of mouse T lymphoblasts to proliferate in response to IL 2. The function of T lymphoblasts of rat origin was not affected by the mAb AMT-13. The time course of the acquisition by mitogen-stimulated spleen cells of the capacity to absorb IL 2 activity was paralleled by that of their capacity to bind the mAb AMT-13. Preliminary biochemical analysis of the antigen recognized by AMT-13 revealed a major component with an apparent m.w. of 50,000 to 60,000. According to these data, the mAb AMT-13 seems to be directed against an antigenic determinant of the murine IL 2 receptor molecule.

Animals↗

The characteristics of a monoclonal antibody that binds specifically to rat T lymphoblasts and inhibits IL 2 receptor functions.

In order to obtain an anti-interleukin 2 (IL 2) receptor antibody, we immunized mice with phorbol myristate acetate-pulsed rat T lymphoblasts. The spleen cells of the mice were fused with myeloma cells. Several stable clones of hybridoma cells were obtained that produced monoclonal antibodies (mAb) reacting specifically with rat T lymphoblasts. Only one of these mAb, the mAb ART18, showed characteristics of a putative anti-IL 2 receptor antibody. This mAb was produced on a large scale, purified, and characterized. As tested by a binding assay, 125I-labeled mAb ART18 bound to rat T lymphoblasts (7.5 X 10(4) binding sites per cell), but not to thymocytes or spleen cells of rat origin, or to lymphoblasts, thymocytes, or spleen cells of murine origin. Only marginal binding to lipopolysaccharide-stimulated rat lymphoblasts was detected. The mAb ART18 inhibited in a species-specific and dose-dependent manner i) the capacity of rat T lymphoblasts to absorb IL 2 activity and ii) the capacity of rat T lymphoblasts to proliferate in response in IL 2. The function of T lymphoblasts of murine origin was not affected by the mAb ART18. The time course of the acquisition by mitogen-stimulated spleen cells of the capacity to absorb IL 2 activity was paralleled by that of their capacity to bind 125I-labeled mAb ART18. According to these data, the mAb ART18 seems to be directed against an antigenic determinant of the IL 2 receptor molecule.

Animals↗

Studies on T lymphocyte activation. II. Monoclonal antibody inhibiting the capacity of rat T lymphoblasts to absorb and to respond to IL-2: an anti-IL-2 receptor antibody?

We describe the production and some of the properties of a monoclonal antibody which inhibits binding of Interleukin 2 (IL-2) to rat T lymphoblasts as well as the capacity of these lymphoblasts to proliferate in response to IL-2 of rat or murine origin. According to these criteria, the antibody seems to be directed against an antigenic determinant on the IL-2 receptor. The antibody, obtained from hybridomas derived from mice immunized with phorbol-myristate-acetate-pulsed rat T lymphoblasts, exhibited species specificity, indicating that IL-2 receptors of various species are not necessarily identical, even if they can respond to IL-2 from different species.

Animals↗