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S L Wee

Publications and source records attributed to S L Wee.

At least 37 records · Page 2Linked to original sources
PubMed1989

Cell-type-specific cytotoxicity of anti-CD4 and anti-CD8 ricin immunotoxins against human alloreactive T-cell clones.

Potent T-cell subset-directed immunotoxins (ITs) were generated by conjugating the anti-CD4 monoclonal antibody (MoAb) G17-2 and the anti-CD8 MoAb G10.1 to the ribosome-inhibitory protein, ricin. The cell-type-specific cytotoxicities of the generated ITs were evaluated at the clonal level using human alloreactive T-cell clones. The kinetics of anti-CD4 ricin-induced inactivation of protein synthesis in target CD4+ cloned T-cells was first order with no detectable lag period and a maximum rate of 0.07 logs per hour (t10 = 13.6 hours; first-order rate constant/K = 0.17 hr-1). The alloantigen specific lytic function of the CD4+ cytolytic T-cell clone JMAC28 was acutely sensitive to anti-CD4 ricin, and no residual lytic activity against allogeneic targets was detectable 24 hours after treatment with as little as 0.5 mmol/L anti-CD4 ricin. Notably, both anti-CD4 ricin and anti-CD8 ricin elicited a selective and dose-dependent inhibition of clonal proliferation of target T-cell clones with a maximum kill of greater than 3 logs at 5 nmol/L. No significant "bystander effects" were observed for non-target cells. Bone marrow progenitor cells CFU-GM, BFU-E, and CFU-GEMM were only minimally affected by either IT. We conclude that these ITs show considerable potential for effective depletion of T-cell subpopulations from allogeneic donor marrow grafts for clinical graft-versus-host disease (GVHD) prophylaxis.

Antigen-Antibody Complex↗
PubMed1989

Fc-receptor for mouse IgG1 (Fc gamma RII) and antibody-mediated cell clearance in patients treated with Leu2a antibody.

A pilot study was performed to explore the clinical potential of Leu2a antibody in reversing acute renal allograft rejection. Anti-Leu2a, a murine IgG1 monoclonal antibody (mIgG1 mAb), is specific for the CD8 molecule that is expressed in high density on class I reactive T cells. Of the 6 recipients treated with anti-Leu2a, two responded with a complete reversal of rejection with long-term allograft function maintained for over a year. In two other recipients, acute rejection was initially reversed, but later rejection episodes resulted in allograft failure at 2-3 months posttreatment. Rejection in the two other recipients showed no response to Leu2a mAb treatment. Specific depletion of peripheral blood CD8+ cells occurred in four of the six recipients. Even in this small series, it was evident that cell clearance was neither necessary nor sufficient for reversal of rejection. However, a complete correspondence between cell clearance and the ability of the recipients' mononuclear cells to undergo mitogenic response to the mIgG1 anti-CD3 (Leu4) mAb in vitro was noted. Binding of IgG1 mAb to the Fc-receptor (Fc gamma RII/CD32) expressed on blood monocytes is known to be essential for the T cell mitogenic response to soluble mIgG1 CD3 mAb in vitro. Our data suggest that binding to Fc gamma RII may be an essential step in the process of mIgG1 Leu2a mAb-mediated cell clearance in vivo.

Animals↗
PubMed1987

Role of CD2 and IL 1 beta in T cell responsiveness to IL 2.

We have evaluated the effects of an anti-CD2 monoclonal antibody (moAb), Leu-5b, on inhibition of lymphocyte responsiveness. Given previous reports that anti-CD2 moAb prevent production of interleukin 2 (IL 2), as well as up-regulation of IL 2 receptor, we chose a test system in which lymphocytes were activated by exogenously added recombinant IL 2 (rIL 2). Our studies thus allowed us to evaluate other possible mechanisms by which anti-CD2 moAb could act. We found that Leu-5b not only blocked the proliferative response of human peripheral blood leukocytes (PBL) to IL 2, but also the development of cytotoxicity as assayed in lectin-dependent cytotoxicity (LDCC). Addition of recombinant interleukin 1 beta (rIL 1 beta) reconstituted the proliferative response in the presence of Leu-5b but did not allow development of cytotoxicity. Our findings are consistent with the concept that Leu-5b delivers a signal to IL 2-responsive cells that prevents their subsequent proliferation and development of cytotoxicity, but that only the former event, i.e., proliferation, is reconstituted by the addition of IL 1 beta.

Antibodies, Monoclonal↗
PubMed1985

Loss and re-acquisition of lytic function by cloned cytotoxic T lymphocytes: role of specific antigen and interleukin 2.

We have previously reported the isolation of 2 human allospecific cytotoxic T-lymphocyte (CTL) clones that can undergo a reversible functional change from a highly lytic phase to a nonlytic quiescent phase and again to a lytic phase. The entire process was found to be regulated by recombinant interleukin 2 (rIL-2). We have now extended these studies in 3 ways. First, we show that specific alloantigens can also function as a signal to reactivate lytic function in the reverted CTL. However, in contrast to CTL reactivated with rIL-2, the antigen-reactivated CTL apparently fail to undergo subsequent cell division. Second, we have also found that this reversion phenomenon is not the same for all CTL tested, as 2 other CTL clones were found that did not revert to the non-lytic phase when cultured for up to 60 h in rIL-2-free medium. Third, the expression of Tac (IL-2 receptor) was also studied throughout the process of reversion and reactivation. rIL-2 added to the cell that had reverted to a non-lytic phase induced, an increase in the expression of Tac receptors during the reactivation phase, to levels greater than those expressed on cells continuously cultured in rIL-2-supplemented medium.

Antigens, Surface↗
PubMed1985

Human alloreactive CTL clones: loss and reacquisition of specific cytolytic activity can be regulated by "recombinant" interleukin 2.

In this communication, we demonstrate that human alloreactive CTL at the clonal level, can undergo reversible processes vis-a-vis lytic function: from a highly lytic phase to a nonlytic quiescent phase and back to a lytic phase. "Reversion" and reacquisition of function can be controlled by removing or adding back a chromatographically purified product of a human interleukin 2 gene (rIL 2) that appears to serve, either directly or indirectly, as a lysis-inducing signal for these CTL.

Cell Line↗
PubMed1985

B lymphocyte regulation of human hematopoiesis.

Epstein Barr virus (EBV)-transformed B lymphoblastoid cell lines (BLCL) were derived from seven different individuals. The ability of BLCL supernatants to stimulate hematopoietic colony formation in vitro was tested in a conventional stem cell assay system. Supernatants promoted the growth of single (GM, E, MK) as well as multi-lineage (GEMM) colonies in bone marrow cultures. Our results indicate that EBV-transformed B lymphocytes produce cytokines that affect in vitro stem cell proliferation and differentiation. These studies demonstrate the regulatory potential of activated B lymphocytes in human hematopoiesis.

Adult↗
PubMed1984

Functionally distinct human T cell clones that produce lymphokines with IL-2-like activity.

Various functionally distinct human T cell clones derived from in vitro mixed leukocyte cultures are found to secrete lymphokines with detectable Interleukin-2 (IL-2)-like activity upon antigenic stimulation. These lymphokine producing clones are not only dependent on exogenous growth factors provided in the form of crude phytohemagglutinin (PHA)-induced spleen conditioned medium for growth but can themselves be driven to proliferate by their own lymphokines. The induction of lymphokine production appears to be antigen-specific and the lymphokines secreted are believed to contain nonantigen and nonspecies specific IL-2-like activity. We show here that IL-2-like lymphokines are produced by a subset of T lymphocyte clones, i.e., the help-independent cytotoxic T cells clones that have the capacity to proliferate to, as well as to lyse, the original sensitizing cells. In addition, other T cell clones capable of producing active lymphocytes include those clones that have the T helper cell characteristics, i.e., can undergo antigen-induced proliferation but are not cytotoxic; in the presence of lectin (PHA), however, some helper T cells (Th) but not others, can express lectin-dependent cell-mediated lysis. Finally, yet another subset of T lymphocytes, the help-dependent cytotoxic T cell clones that cannot proliferate to antigenic stimulation, was found to produce no detectable IL-2-like activity.

Clone Cells↗
PubMed1984

HLA loss variants of a B27+ lymphoblastoid cell line: genetic and cellular characterization.

Variants of a lymphoblastoid cell line, LCL 526 (SB3 MB1 DR1 B44 C5 A2/SB4 MT4 DR4 B27 C2 A24), which lost various HLA specificities were selected with monoclonal antibodies and complement using a method developed by Kavathas et al. (PNAS 77:4251, 1980). Using alpha B27 monoclonals, 8 B27 only loss mutants and 4 B27 haplotype multiple loss mutants were generated. The parental LCL 526 and two of the B27- mutants were used to select alpha B27 CTLs. The selection of six A2 loss, one A2-C5 loss, and 14 A2 haplotype multiple loss variants as well as secondary selection on haplotype loss variants to obtain A null, B null, DR null, and total A, B, C, null variants is also described. The usefulness of these mutants for the study of the relationship between B27 and disease and as two new haplotypes for immunologic, genetic, and molecular research is discussed. These mutants are available to other researchers.

Antibodies, Monoclonal↗
PubMed1984

A cytolytic human T lymphocyte clone differentially recognizing HLA-B27 subtypes.

A cytolytic human T cell (CTL) clone, designated F/M-F159, has been produced, the lytic specificity of which distinguishes subtypes of HLA-B27. This was demonstrated in cell-mediated lympholysis (CML) assays of: 1) a panel of target cells from unrelated donors, 75 B27 + and 36 B27-; 2) six families, including 20 B27 + and 14 B27- individuals; and 3) B27 + and B27- variants of a B27+ lymphoblastoid cell line (LCL). Specificity of F/M-F159 for HLA-B27 was confirmed by blocking studies with monoclonal antibodies. Lysis of B27 + targets reactive with the anti-B27 monoclonal antibody B27M2 was 30-104%, while lysis of B27 +, B27M2- targets was 4-22%. Lysis of B27- targets expressing HLA-Bw47, known to be cross-reactive with the B27M2 antibody, was 10 to 19%, while lysis of all other B27- targets was less than or equal to 10%. Clone F/M-F159 lysed B27 + targets, and failed to lyse B27- targets, irrespective of the clinical status of the cell donors. It is concluded that F/M-F159 recognizes an epitope present on the majority of serologically identified HLA-B27 molecules and that this epitope is closely related to, but not identical with, the epitope recognized by the antibody B27M2. These findings are interpreted as supporting a direct role for HLA-B27 in disease pathogenesis.

Antibodies, Monoclonal↗
PubMed1982

Helper cell-independent cytotoxic clones in man.

We report here a class of helper cell-independent cytotoxic T cell (HITc) clones in man that can proliferate in response to antigenic stimulation as well as mediate cytotoxicity. HITc appear to be rare among clones derived from primary in vitro allosensitized culture, but constitute the majority of clones derived from cells sensitized to autologous Epstein-Barr virus-transformed lymphoblastoid cell lines. The implications of the derivation and function of HITc clones are discussed.

Cell Line↗
PubMed1981

Early detection and specificity analysis of human cytolytic T lymphocyte (CTL) colonies generated in soft agarose culture: a potential assay for definition of CTL defined (CD) determinants.

In this communication we describe and early, large-scale screening assay for the detection of colonies with varied cytolytic specificity. The colonies are generated in soft agarose culture from day 3 mixed lymphocyte culture (MLC) alloactivated cells. A cell mediated lympholysis (CML) assay utilizing as few as 500 target cells has made it possible to prescreen for cytolytic T lymphocyte (CTL) colonies and to test for antigen specificities as early as 11 and 14 days, respectively, after MLC priming. Large numbers of colonies, from 80 to over 150, have been prescreened against a specific sensitizing target cell, and as many as 30 CTL colonies have been simultaneously tested against a panel of multiple targets carrying defined HLA-A, -B, -C, -DR antigens to evaluate antigen specificity. All CTL colonies are lytic against the specific sensitizing target cell and do not lyse the target autologous to the responder. Some are found to be operationally specific in that they lyse only those target cells which share HLA serologically defined (SD) antigens with the sensitizing cells, and other show cytolytic patterns which are not correlated with known HLA-SD antigens. These observations support, at a much finer level of analysis, the possible distinction between SD and CTL defined (CD) determinants.

Cells, Cultured↗
PubMed1980

Family of large surface proteins on human lymphocytes.

Cell surface molecules of T-rosette-positive human peripheral blood lymphocytes were labelled by lactoperoxidase and examined on 5% polyacrylamide gels. Several bands, ranging in molecular weight from 220,000 to 170,000 daltons, were seen. Analysis of the tryptic and chymotryptic digests of the 187K, 200K, 210K and 220K proteins indicated that they are of similar peptide composition. Study of clones derived from this population indicated that individual cells within a population differed in expression of these proteins.

Animals↗
PubMed1980

In vitro studies on the control of thyroid autoantibody synthesis.

A current hypothesis suggests that autoimmune responses are prevented in normal individuals by a population of suppressor cells. We have investigated this using a system in which thyroid autoantibodies are synthesised in culture by peripheral blood mononuclear cells (PBM) from patients with Hashimoto's thyroiditis. A group of 8 Hashimoto patients and 17 normal subjects were used in the study. PBM from the group of patients did not differ significantly from those of the control group in terms of their ability to synthesise IgG in culture, to respond to allogeneic cells or to PHA. (Nor were any genetic differences observed between the two groups in terms of HLA-A, B or C antigens.) In 6 co-culture experiments involving equal numbers of PBM from patients and normal donors, inhibition of thyroid antibody production was not observed. Additionally, T-cell enriched populations from 2 normal donors were unable to specifically inhibit thyroid antibody synthesis by Hashimoto B-cell enriched fractions from 2 Hashimoto patients. Consequently, we were unable to show suppressor cell activity in normal PBM and it is suggested that these cells may need to be activated by antigen before their presence can be demonstrated.

Adult↗
PubMed1978

Diminished mixed lymphocyte response in ankylosing spondylitis.

A diminished mixed lymphocyte response was reported by Nikbin et al. (1976) among patients with ankylosing spondylitis, their asymptomatic relatives and also normal controls carrying the B27 antigen. In the present communication, the responses in 48 ankylosing spondylitis patients and 45 controls were examined in mixed lymphocyte cultures tested against a 'standard stimulator' made up of pooled lymphocytes. A significantly diminished response is confirmed among the ankylosing spondylitis patients, but not in the control group carrying the B27 antigen. The diminished mixed lymphocyte response therefore appears to be more directly associated with the disease than with the B27 antigen, and possibly represents a specific T-cell defect associated with the pathogenesis of the disease.

Female↗