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Biomedical subjects

K S Zier

Publications and source records attributed to K S Zier.

At least 37 records · Page 2Linked to original sources

Expression of class II antigens by subsets of activated T cells.

Gene products coded for within the HLA complex play an important role in the control of immune responses. Class I antigens, coded for by the HLA-A, B, and C loci, are expressed by virtually all mononuclear blood cells. Class II antigens, coded for by the DR, DQ, and DP loci, have a more limited tissue distribution. They are expressed by B cells, monocytes, and by activated, but not by resting, T cells. The class II molecules of B cells and antigen-presenting cells have long been of interest to immunologists, since they are involved in the presentation of antigen, in communication between T cells and B cells and between T cells and adherent cells, and in susceptibility to certain diseases. The class II antigens expressed by activated T cells, however, remain largely uncharacterized in terms of their specificity, functional significance, and molecular nature. We have studied the expression of DR and DQ antigens by activated T cells and then examined the expression of DR versus DQ antigens by Leu 2a and Leu 3a subsets of mitogen-activated populations. Our results demonstrated that, as for class II-positive macrophages, the intensity of staining with monoclonal antibodies directed against DR antigens was much greater than that obtained with those directed against DQ antigens. Interestingly, the percentages of Leu 2a- and Leu 3a-positive cells which expressed DR antigens were quite similar, as were the percentages of Leu 2a and Leu 3a cells which expressed DQ. Thus, there does not seem to be preferential expression of DR versus DQ antigens by mitogen-activated T-cell subsets. Finally, though both DR-positive-DQ-positive and DR-positive-DQ-negative populations were detected, few or no DR-negative-DQ-positive cells were observed in these populations.

Antigens, Differentiation, T-Lymphocyte↗

Generation of antigen specific cytotoxic T lymphocytes (CTL) directed against class II molecules expressed by activated T cells.

Mitogen-activated T cells were used to provide a source of Class II antigens to CTL originally stimulated against mononuclear cells expressing both foreign Class I and Class II determinants. Our results indicated that 7-10-day-old activated T antigen-presenting cells, which shared only Class II antigens with the original priming cell, were able to stimulate the differentiation of CTL-recognizing Class II determinants. The use of 14-day-old activated T cells as target cells in the CML assay, compared with 72 h PHA blasts or 7-day-old activated T cells, enabled a more sensitive detection of the anti-Class II CTL.

Antigen-Presenting Cells↗

Presentation of Class I antigens on T cells can induce the generation of antigen-specific CTL.

Cytotoxic T lymphocyte (CTL) lines generally require a source of specific antigen to continue to proliferate in vitro. We previously showed that populations of mononuclear cells grown in medium containing interleukin 2 (IL 2) and composed largely of activated T cells were able to present Class I alloantigen to CTL lines. On the basis of these findings we were interested to know whether T cells themselves were able to present antigen or whether this was a function of the small number of contaminating non-T cells. To answer this question, populations of activated or resting mononuclear cells were rigorously depleted of non-T cells before use as antigen-presenting cells. We observed that populations composed of greater than 99% T cells were able to support the differentiation of antigen-specific CTL. These results were confirmed by using cells from an established T cell line. The proliferation of the same lines, however, was less than that of lines grown in the presence of antigen-presenting cells containing some non-T cells. These results suggest that although T cells can present Class I alloantigens to CTL, they may be less effective in triggering cell division than populations containing a source of non-T cells.

Antigen-Presenting Cells↗

Proliferative T cell clones directed at a human minor antigen.

Clones directed against a human minor antigen were isolated from an interleukin-2-dependent cytotoxic T cell line. As expected, some clones exhibiting cytotoxicity for HLA-identical minor antigen-positive cells were detected. In addition, some clones were detected which, though not cytotoxic, were able to proliferate in the primed lymphocyte typing (PLT) test in response to stimulation by cells from some HLA-identical sibs and certain DR 2+/ve unrelated cells. These clones expressed the Leu 3a cell surface antigen. DR 2 was one of the class II determinants expressed by the responding cell used to generate the cytotoxic T lymphocyte line. These results are consistent with the interpretation that helper T cells are able to recognize minor antigens in the context of self-class II HLA determinants and respond in PLT.

Anemia, Aplastic↗

Generation of CTL lines using cultured T cells (CTC) as feeder cells. I. Specificity for class I determinants.

Lines of cytotoxic T lymphocytes (CTL) may be grown in long-term culture in medium supplemented with Interleukin-2 (IL-2). In most cases a source of feeder cells is also necessary. Because they are available in large numbers, we tested whether cultured T cells (CTC) would serve as feeder cells. We confirmed the observation of others that CTL grown in medium supplemented with IL-2, but in the absence of feeder cells, grew poorly and became less cytotoxic with time. CTL lines directed class I HLA antigens and grown with either x-irradiated peripheral blood lymphocytes (PBL) or x-irradiated CTC proliferated well, were similarly cytotoxic, and demonstrated the same antigenic specificity. By using cells from members of a family in which one offspring had inherited a recombinant chromosome, it was possible to show that sharing of the Class I determinants between the original stimulating cells in the primary culture and the CTC feeder cells was sufficient to enable the maintenance of strongly cytotoxic CTL lines.

Cell Division↗

Decreased synthesis of interleukin-2 (IL-2) in insulin-dependent diabetes mellitus.

Synthesis of interleukin-2 (IL-2) by lymphocytes from 26 insulin-dependent diabetic subjects (IDDM) was compared with that by lymphocytes from 24 nondiabetic control subjects. The control group produced 1.001 +/- 0.071 U/ml (mean +/- SEM). The IDDM group, containing patients diagnosed between 5 days and 10 yr before testing, produced only 0.59 +/- 0.050 U/ml (P less than 0.002). IL-2 synthesis by 6 non-insulin-dependent diabetic subjects (NIDDM) was not decreased (mean +/- SEM, 1.20 +/- 0.04 U/ml). Moreover, decreased levels of IL-2 production was found with lymphocytes of patients in good control, as well as those in poor control. These data suggest that decreased IL-2 synthesis is specific for IDDM, not explainable solely as a consequence of poor metabolic control, and thus, might be involved in the pathogenesis of the disease.

Adolescent↗

The use of cytotoxic T cell lines to detect the segregation of a human minor alloantigen within families.

A cytotoxic T cell (CTL) line, which detected a minor alloantigen provisionally called W was generated in vitro with lymphocytes from a multiply transfused individual, S1. Lymphocytes from S1 were first stimulated with cells from an unrelated known from previous studies to express the minor antigen. The primary CTL were then restimulated with cells from a W +/ve HLA identical sib, S2, in the presence of IL-2. As in previous work, recognition of the W antigen by these CTL was restricted by HLA-B7. Antigen assignments of W + W -, based upon cold target inhibition studies, confirmed previous assignments which had depended upon the ability of lymphocytes either to stimulate the generation of or to be killed by anti-W CTL effectors. Testing of lymphocyte targets from members of several unrelated families in which HLA-B7 segregated showed that the CTL lines could detect the expression of W on cells of individuals in the general population. In 3 of 5 cases, members of an HLA identical sib pair differed for W. These results open up the possibility of designing studies using CTL lines to determine whether differences for minor alloantigens play a role in clinical transplantation.

Binding, Competitive↗

Recognition of human minor alloantigen(s) by cytotoxic lymphocytes in vitro.

Lymphocytes from an extensively transfused patient with aplastic anemia were induced to cytotoxicity against target cells from several HLA-matched siblings by in vitro stimulation with allogeneic cells. Effective stimulating cells shared HLA-B7 with the patient, but not all B7 individuals were effective. An additional factor, which was found to segregate in both the patient's and an unrelated sibship, was also necessary. Segregation of this minor alloantigen, W, was also revealed among the patient's HLA-matched sibs by differential susceptibility to lysis by effectors from the patient. The ratio of six positive to four negative siblings suggests that the antigen difference might be coded by a single locus. Lymphocytes from a normal sib, who like the patient is lacking the minor antigen, could not be induced to cytotoxicity against positive targets. Thus in vivo sensitization of the donor of the responding cells appears to be necessary for the demonstration of the cytotoxic response to the minor antigen in vitro. No correlation was observed between the segregation pattern of W and of known blood group antigens, and no cytotoxic antibody to W was detected in the patient's serum in several trials.

Adult↗

Cytotoxic T lymphocyte lines (CTLL) against a human minor alloantigen.

Cytotoxic T-cell lines were generated following in vitro culture of lymphocytes from a patient suffering from aplastic anemia together with those of his HLA-identical brother, a repeated transfusion donor. The segregation pattern within the family of the determinant(s) detected by these cytotoxic cells strongly suggested that a minor alloantigen(s) was being detected. Testing of the effectors on a panel of unrelated individuals indicated that it was best seen in association with HLA-B7, which was common to both the patient and his sibling donor.

Anemia, Aplastic↗

Functional and antigenic properties of cultured T cells in the cell mediated lympholysis (CML) assay.

Cultured T cells (CTC) were expanded in Interleukin-2 (IL-2) and used as reagents in cell mediated lympholysis (CML). CTC were able to induce the generation of primary cytotoxic effector cells and to function as 51Cr labeled target cells and cold target inhibitors. Since large numbers of CTC can be produced from as few as 1-2 X 10(6) lymphocytes within a short period of time, these reagents will enable CML studies to be performed in a broader range of situations, including those with were heretofore impossible.

Antigens, Surface↗

Generation of discriminatory PL against HLA-D antigens not detected by serology.

A family was studied in which the father and the mother were both homozygous for the HLA-DR antigens. All children responded to the parents in MLC, even though each had a copy of the parental DR antigen, implying that they recognized a difference. When the father or the mother was sensitized in PLT against any child, all siblings could restimulate equally well. However, by sensitizing a family member with one copy of the DR phenotype against another family member with the alternate form of the phenotype, discriminatory PL could be produced. This may help define the subtypic specificities of PL antigens.

Epitopes↗

Cell-mediated immune responses between HLA-identical siblings: recognition of antigenic changes associated with acute myelogenous leukaemia.

Cell-mediated immune reactions between a patient suffering from acute myelogenous leukaemia (AML) and an HLA-identical sibling were studied in order to characterize the in vitro reactions in MLC and CML prior to bone marrow transplantation. Our results indicated that antigenic differences were detectable between the blasts and the remission lymphocytes. While the normal sibling did not respond in MLC to her HLA-identical sister's remission lymphocytes, there was an anti-blast response. This proliferative response, however, did not lead to the development of detectable cytotoxic cells capable of destroying blast cells. Unrelated individuals, on the other hand, responded strongly both in MLC and CML to the allogeneic tumour blasts and remission lymphocytes of the patient and the lymphocytes of the healthy sibling. The kinetics and magnitude of the MLC response to blast cells was different from that to remission lymphocytes. This response indicated that the blast cells expressed antigenic differences which were recognized in MLC by both the HLA-identical sibling and unrelated individuals. Furthermore, these tumour cells were capable of sensitizing allogeneic, but not syngeneic lymphocytes to become cytotoxic, though they seemed to be more resistant to destruction in CML than normal cells.

Adult↗

Restimulation in secondary MLC by autologous non-T cells.

Lymphocytes sensitized in mixed leucocyte culture (MLC) were restimulated with non-T or complete cells. Cells primed against autologous non-T cells and restimulated with autologous cells gave responses with the kinetics of a secondary response. In contrast, cells primed against either type of autologous cell responded to restimulation by allogeneic cells with the kinetics of a primary MLC response. Following priming against alloantigens good secondary responses against both complete and non-T allogeneic restimulators were seen. Restimulation with autologous non-T cells, but not autologous complete cells, also led to secondary responses. The observation of different responses to antigens expressed on non-T autologous lymphocytes, in comparison to complete cells, implies that the secondary MLC involves more than response against alloantigens. Allogeneic and autologous MLC may be qualitatively different phenomena, the latter representing the recognition of identifying structures between interacting populations of cooperating cells.

Autoantigens↗

Linear density gradient separation of human lymphocyte subsets. I. Analysis by mixed leukocyte culture and cell-mediated lympholysis responses.

Linear density gradients were used to separate either unsensitized human peripheral blood lymphocytes or cells sensitized in mixed leukocyte culture (MLC), at different time points in the immune response. Testing of cells from the various density fractions for their ability to respond in MLC and cell-mediated lympholysis (CML) revealed that (a) activity between individual fractions differed, as well as activity between individual fractions and a suspension of unfractionated cells, (b) although precursor cells of MLC and CML were maximally enriched in closely associated light density regions, their respective distribution profiles were strikingly different, and (c) the density of cells responding in MLC and CML differed depending upon the point of the immune response at which the cells were separated. Whereas effector cells were of light density relative to the precursor cells, putative memory cells for CML exhibited a high density.

Cell Differentiation↗