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Biomedical subjects

J A Hank

Publications and source records attributed to J A Hank.

At least 73 records · Page 4Linked to original sources
PubMed1989

Specific recognition of human leukemic cells by allogeneic T cell lines.

Clinical and experimental data suggest a role for the immune response in preventing leukemic relapses following allogeneic bone marrow transplantation the graft-versus-leukemia (GVL) effect. In the context of an allogeneic BMT, a number of different immune mechanisms mediated by donor cells may be responsible for the GVL effect. We have approached this question by using limiting dilution cultures of alloactivated human lymphocytes to analyze the in vitro allogeneic cytolytic response against fresh allogeneic leukemia. Initial results in the limiting dilution assays with split culture analyses demonstrated frequent alloreactive cytolytic T lymphocyte precursors that destroyed remission peripheral blood lymphocytes and leukemic cells from the allogeneic leukemic patient. These assays also demonstrated frequent lymphokine-activated killer (LAK) cell precursors that lysed both the LAK sensitive Daudi line and the allogeneic leukemia. In these experiments, isolated cultures also showed cytolytic activity directed against the allogeneic leukemic blasts without activity against remission PBL, or the LAK-sensitive Daudi cell line. Two T cell lines (ABL1 and ABL2) isolated from an LDA, demonstrated this form of specificity, mediating destruction specifically against the allogeneic acute lymphoblastic leukemic cells. Both cell lines ABL1 and ABL2 were CD3+, TCR alpha beta +, and CD4+. These 2 cell lines mediated little or no cytotoxicity against a large panel of other targets tested (natural killer sensitive and resistant cell lines, allogeneic PBL, and allogeneic fresh leukemic blasts). Antibody-blocking experiments revealed a role for the CD3-TCR receptor of both cell lines in lysis of leukemic cells; the CD4 and MHC class II molecules were clearly involved in the lysis by the ABL1 cell line. Specificity of recognition for the allogeneic leukemic blasts was further confirmed by unlabeled target competitive inhibition studies. The mechanism of the preferential lysis of leukemia by the alloactivated T cell lines described in this paper remains uncertain. Nevertheless, these leukemic-specific populations provide a means by which the human GVL effect may be further studied in vitro.

Antibodies, Monoclonal↗
PubMed1989

Phase 1 clinical trial of recombinant interleukin-2: a comparison of bolus and continuous intravenous infusion.

The toxicologic, biologic, and clinical effects of recombinant interleukin-2 (IL-2) were tested in 25 patients with cancer. Escalating doses from 10(3) to 10(7) U/m2 per day were given by either daily bolus injection (BI) or continuous infusion (CI) for 7 consecutive days. Dose-limiting toxicities included a decline in performance status, systolic hypotension, and fever, which reversed promptly with discontinuation of therapy. The maximum tolerated dose of IL-2 by BI for 7 days was 3 X 10(6) U/m2 per day and for CI was 10(6) U/m2 per day. Significant changes in the number, phenotype, and function of circulating peripheral blood lymphocytes occurred at doses greater than or equal to 10(6) U/m2 per day by both administration schedules. With the initiation of therapy, a decline in the number of circulating peripheral blood lymphocytes (PBL) was seen in patients treated by either BI or CI. Additionally, the in vitro cytotoxic activity of these PBL against K562 was markedly decreased. Within 24 h of completing BI or CI, a rebound increase in the number of circulating PBL was seen. The phenotype of the circulating PBL after completion of treatment showed a significant (p greater than or equal to 0.05) increase in the numbers of OKT-3+, OKT-8+, OKT-10+, OK-Ia+, OKM1+, and OKT-11+ for patients treated by CI. Those patients treated by BI had a significant increase in Ia+ and OKT10+ cells. At IL-2 doses greater than or equal to 10(5) U/m2 per day, the PBL obtained following treatment with rIL-2 demonstrated in vitro cytotoxic capacity against K562 target cells that was significantly enhanced over pretreatment levels. This study demonstrates that IL-2 can be given by CI or BI in a non-ICU setting with acceptable dose-dependent toxicity. Upon completion of treatment an increase in the number of activated cells could be detected. Although no clinical responses occurred, the generation of endogenous activated PBL capable of enhanced cytotoxicity is encouraging. Future studies will explore the use of multiple courses of treatment with IL-2 to determine if therapeutic efficacy can be accomplished.

Antibodies, Monoclonal↗
PubMed1989

Prospects for clinical cancer treatment using interleukin-2.

Even if many of these immunologic approaches to anticancer therapy show the desired effect, with a greater percentage of patients having antitumor responses (hopefully without need for ICU level toxicity), it may be unrealistic to hope that these biological treatments will significantly prolong the survival of most patients with large progressively growing tumors. It is possible that the large, yet finite, number of cells with LAK activity that can be activated endogenously, or infused into a patient, may only be able to destroy a finite number of neoplastic cells. If so, the smaller the number of neoplastic cells at the time of in vivo LAK induction therapy, the more likely the chance for effective eradication of all tumor cells. Experimental animal studies support this postulate (46). Clinical regimens utilizing IL-2 therapy, alone or combined with other agents, with documented immunologic and antitumor activity, will need to be tested in a large number of patients in randomized adjuvant trials. This testing will probably require the involvement of large cooperative oncology trials groups, in order to determine the potential prolongation of survival by any adjuvant IL-2 approach. A vast number of regimens could presently be suggested for combining these separate approaches in groupwide trials (especially when issues of dose, route, and scheduling are considered). Trials in murine models must continue rapidly and be interpreted with caution in generating regimens to be tested clinically. Further treatment improvements, to be identified in Phase I and Phase II clinical trials, are still required to allow large groupwide randomized Phase III trials to test tolerable and effective IL-2-containing regimens likely to significantly prolong survival.

Combined Modality Therapy↗
PubMed1989

Antigen-specific recognition of autologous leukemia cells and allogeneic class-I MHC antigens by IL-2-activated cytotoxic T cells from a patient with acute T-cell leukemia.

Culturing of leukemic blood lymphocytes from a patient with acute T-cell lymphoblastic leukemia (T-ALL) with interleukin-2 (IL-2) yielded T-cell line AK-1 with a remarkable cytotoxic specificity. This line mediated strong lysis of tumor target lines expressing major histocompatibility complex (MHC) class I antigens, such as Raji, CEM, and Molt-4 cells, but no killing of K562 and Daudi cells, which are deficient in MHC class I. In contrast, lymphokine-activated killer (LAK) cells from normal donors destroyed all these tumor targets, without MHC restriction. Line AK-1, originating from residual normal T cells present in the leukemic blood, lysed autologous leukemic blasts and peripheral blood lymphocytes (PBL) from many but not all allogeneic individuals but failed to kill autologous remission lymphocytes. Destruction of the autologous leukemic targets by AK-1 could be inhibited by unlabeled competitor target cells that were lysed by AK-1, but not by target cells that were not lysed. This suggests that AK-1 specifically recognized an alien determinant on the autologous ALL cells, crossreactive with allogeneic MHC class I antigens. This reactivity with some degree of tumor specificity may be a leukemic equivalent to responses reported for populations of tumor infiltrating lymphocytes (TIL) seen in some solid tumors.

Adult↗
PubMed1988

Repetitive weekly cycles of interleukin-2. II. Clinical and immunologic effects of dose, schedule, and addition of indomethacin.

Clinical trials with high doses of interleukin 2 (IL-2) have shown antitumor responses, but many of the patients have experienced severe and occasionally life-threatening toxic effects. Preclinical studies indicate that modifications in IL-2 dose, route, and schedule can influence both immune activation and antitumor effects. This study evaluated the clinical tolerance to and immunologic modifications induced by four repetitive weekly cycles of IL-2, with two dose levels (1 X 10(6) and 3 X 10(6) U/m2 per day) of IL-2 and three different daily administration schedules [bolus, continuous, or combined (bolus and continuous)], with and without indomethacin treatment. Patients in all treatment groups experienced acceptable, non-life-threatening toxic effects and immune system stimulation characterized by rebound lymphocytosis with increased numbers of natural killer and lymphokine-activated killer cells and enhanced direct cytolytic function. These immune changes were significantly enhanced by the repetition of IL-2 cycles beyond the first week of therapy. At an IL-2 dose of 3 X 10(6) U/m2 per day, bolus IL-2 was less immunostimulatory than continuous-infusion IL-2. The combined regimen (with half of each daily dose given as a bolus and half as a 24-hr infusion) was as stimulatory as continuous-infusion IL-2 and also induced antitumor effects. Finally, the addition of indomethacin to this regimen did not significantly modify in vitro or in vivo immune response parameters but appeared to worsen the systemic toxic effects of renal dysfunction and capillary leakage. These results suggest that continuous or combined infusion of IL-2 at 3 X 10(6) U/m2 per day on this schedule should be considered for further testing in phase II trials or in combination with other therapeutic modalities.

Adult↗
PubMed1988

Clinical and immunological effects of recombinant interleukin 2 given by repetitive weekly cycles to patients with cancer.

Eleven patients received four consecutive weekly cycles of human recombinant interleukin 2 (IL-2) by continuous infusion for 4 days/week. Two dose levels were tested, 1 and 3 X 10(6) units/m2/day. Toxicities experienced by most patients included fever, rigors, fatigue, anemia, eosinophilia, and liver function abnormalities. All side effects from treatment reversed and no severe or life-threatening problems occurred. A marked lymphocytosis was seen following the 4 weeks of therapy. Fresh lymphocytes obtained during this lymphocytosis mediated enhanced destruction in vitro of a natural killer cell-resistant tumor cell line (Daudi). The increase in the absolute number of circulating lymphocytes and their enhanced ability to mediate direct lysis of Daudi targets resulted in a greater than 100-fold mean increase in cytotoxic potential by the end of IL-2 treatment. One patient, with renal carcinoma, who was treated at 3 X 10(6) units/m2/day experienced a sustained measurable response with greater than 50% regression of pulmonary and hepatic metastases. Five patients were retreated with a second course of IL-2, lasting 4 weeks. This therapy was well tolerated in four of these five patients, with similar immunological changes occurring. No further antitumor responses were seen in these patients. Thus, a relatively well tolerated immunotherapy regimen using IL-2 can induce dramatic increases in lymphocyte number and augment their in vitro antitumor reactivity.

Cytotoxicity, Immunologic↗
PubMed1988

In vivo induction of the lymphokine-activated killer phenomenon: interleukin 2-dependent human non-major histocompatibility complex-restricted cytotoxicity generated in vivo during administration of human recombinant interleukin 2.

The availability of purified human recombinant interleukin 2 (IL-2) has enabled clinical trials to test its in vivo effects. We report here the immunological effects of 7 consecutive days of IL-2 treatment given to 25 patients with cancer in a clinical Phase I study. Peripheral blood lymphocytes obtained from patients following therapy with IL-2 had enhanced proliferative responses to IL-2 and enhanced direct cytotoxic activity on K562 target cells. This lytic activity was further augmented by the addition of IL-2 during the 51Cr release assay. Fresh peripheral blood lymphocytes from some patients who had just completed treatment at the higher IL-2 dose levels were able to kill both the natural killer-resistant Daudi cell line and fresh tumor cells while pretreatment samples and peripheral blood lymphocytes from healthy controls were not. This lytic activity was best detected when IL-2 was present in the in vitro effector assay. These results demonstrate that the administration of IL-2 to patients with cancer induces a population of effector cells able to directly destroy natural killer-resistant target cells, when assayed in the presence of IL-2.

Cells, Cultured↗
PubMed1988

Clinical adoptive chemoimmunotherapy with allogeneic alloactivated HLA-haploidentical lymphocytes: controlled induction of graft-versus-host-reactions.

A total of 13 cancer patients were treated with Adoptive Chemoimmunotherapy (ACIT) using alloactivated HLA haploidentical lymphocytes. Donor lymphocytes were activated in vitro using a pool of irradiated allogeneic lymphocytes (MLC-cells) and some further expanded by culturing in T-cell growth factor (TCGF-cells). The first 6 patients received i.v. cyclophosphamide (CPM) followed 24 h later by escalating doses of MLC-cells, then 7 days later they received an infusion of TCGF-cells. Minimal toxicity was seen. The next 7 patients received CPM (800 mg/m2) and a combined MLC and TCGF-cell infusion (total cell dose ranged from 0.79 x 10(10) to 2.26 x 10(10)). Of these 7 patients, 3 developed mild graft-versus-host reaction (GVHR) which resolved without treatment, and 2 patients had progressive GVHR which was arrested by methylprednisolone (2 mg/kg). Peripheral blood lymphocytes from these 2 patients, during the GVHR, had increased activated T-cells (OKT-10+ and OK-Ia+). In vitro expansion, in TCGF, of these activated T-cells enabled HLA typing to prove they were of donor origin. Only 1 clinical antitumor response was observed in the first 6 patients. The results of this study indicate that this form of ACIT can be given to patients with acceptable toxicity. Self-limited or easily controlled GVHR may be induced and primed donor cells persisting in the circulation are probably responsible. Further testing is required to determine whether the immune response induced by this form of ACIT may be therapeutically effective.

Child↗
PubMed1988

CD4+ proliferative T-cell clones reactive to autologous Epstein-Barr virus-transformed B cells (EBV-LCL) can be restricted by HLA class I molecules.

In this study, we show that an HLA-loss variant, EBV-LCL .180, which lacks HLA-DR, DP, DQ and B determinants but expresses HLA-A and C molecules, can activate autologous proliferative T cells which recognize EBV-LCL antigens (or LYDMA). T-cell clones isolated from bulk culture (A.180) of T cell primed with autologous variant .180 when tested for their proliferative reactivity to .180 and three autologous class I-loss variants, each lacking specific class I determinants, showed requirement for class I HLA molecules. Clones A.180.Cl and A.180.F3D respond to the A2+, B-null, C+ mutant .53, but not to the A-null, B5+, C+ mutant .144, or the A-null, B-null, C+ mutant .184, indicating that these clones are restricted by an HLA-A2 determinant. These proliferative T-cell clones express the CD4 marker and are noncytotoxic, even in the presence of concanavalin A (Con A) lectin. In coculture experiments, A.180.Cl was further shown to provide lymphokine "help" for EBV-LCL-specific, autologous cytotoxic T-cell clones. These results suggest the repertoire of HLA determinants employed by EBV-LCL-specific proliferative T cells, in addition to the previously shown HLA-DR, DP, and DQ determinants, also includes class I molecules.

Antigens, Differentiation↗
PubMed1988

Transient decrease in IL-2-responsive lymphocytes 24 hours after initiation of continuous IL-2 infusion in cancer patients.

Cancer patients were treated with recombinant interleukin-2 (IL-2) in a Phase I clinical trial. Patients were given four repetitive weekly cycles of four days of continuous i.v. IL-2 infusions followed by 3 days of observation. A transient 80% decrease in the number of circulating peripheral blood lymphocytes (PBLs) was noted 24 h after initiation of the IL-2 infusion. The in vitro IL-2 induced proliferative response, and natural killer (NK) activity of the PBLs recovered at this time was only 10-20% of that by the same number of PBLs obtained prior to IL-2 therapy. This effect was transient and rebound increases in circulating lymphocytes expressing high NK and lymphokine-activated killer functions were demonstrated at the end of a 4 day IL-2 infusion. To study further whether the drop in lymphocyte activity observed at initiation of IL-2 therapy was due to activation of a suppressor mechanism, patient PBLs isolated 24 h into the IL-2 infusion were mixed with their pre-IL-2 therapy PBLs at different ratios and assayed in NK and proliferation assays. These mixing experiments did not prove suppression to be the mechanism for the decreased response; rather, the PBLs obtained 24 h into IL-2 therapy merely diluted out the in vitro response of PBLs obtained prior to therapy. Although there was a considerable drop in circulating PBLs 24 h after initiation of each of three subsequent weekly IL-2 treatment cycles, these remaining lymphocytes in the peripheral blood were functional in both NK and IL-2 proliferative assays. These PBLs obtained 24 h into each of the subsequent three cycles showed a progressive increase in their in vitro NK and IL-2-induced proliferative activity, reaching levels two to three times higher than that of pretherapy PBLs by the fourth cycle. Thus, IL-2 caused a transient disappearance of lymphocytes from the circulation at the initiation of each cycle, but lymphocyte function was impaired only in the first IL-2 cycle. These data suggest that resting IL-2 responsive cells initially leave the circulation upon exposure to IL-2, but that such cells become activated and some remain detectable in the circulation when subsequent weekly cycles of IL-2 are given.

Blood Cell Count↗
PubMed1988

The in vitro function of lymphocytes from 25 cancer patients receiving four to seven consecutive days of recombinant IL-2.

Twenty-five cancer patients received human recombinant interleukin-2 (IL-2) for 4 to 7 consecutive days in a Phase I trial. IL-2 was administered either as a daily intravenous bolus infusion (lasting 15 min), or as a continuous infusion lasting 24 h each day. Prior studies have demonstrated that in vivo administration of IL-2 at high doses is associated with changes in the phenotype of circulating peripheral blood lymphocytes (PBL) (determined with monoclonal antibodies), and the induction of augmented in vitro natural killer activity (NK) by PBL obtained following in vivo IL-2. We have noted that fresh lymphocytes obtained after 4-7 consecutive days of IL-2 (greater than or equal to 10(6) U/m2/day) show an augmented ability to kill NK-sensitive and -insensitive target cells (K562 and Daudi targets, respectively), especially when tested with IL-2 present during the 4-h 51Cr release assay. We have further analyzed lymphocytes in a battery of in vitro proliferative and cytotoxic assays. We present here the summary of this quantitative analysis of their responses in both antigen-specific and -nonspecific immune responses. Striking in vitro changes were observed in the proliferative response to IL-2 and in cytotoxicity stimulated by (or requiring) in vitro IL-2. Proliferative responses to antigens and to mitogens were not as dramatically altered following in vivo IL-2, nor were allospecific or allo-activated cytotoxic interactions. These studies indicate that the most striking changes in lymphocyte function measured in vitro following in vivo IL-2 are seen in those functions requiring IL-2.

Cytotoxicity, Immunologic↗
PubMed1987

Nonrestricted cytotoxicity mediated by interleukin 2-expanded leukocytes is inhibited by anti-LFA-1 monoclonal antibodies (MoAb) but potentiated by anti-CD3 MoAb.

Previous results have shown that in addition to their ability to kill tumor cell lines, peripheral blood leukocytes (PBL) expanded in interleukin 2 (IL-2) can also destroy normal PBL targets. Cold target competition results show that PBL and tumor cells can be destroyed by the same population of IL-2-expanded leukocytes (IEL), with better killing observed for tumor cell targets. Since cytolytic activity of IEL is nonspecific, differential binding of target cells by IEL could determine how well each target cell type can be killed. The binding affinity of IEL, in turn, could be influenced by the accessory molecules expressed on effector and target cells. We tested the effect of MoAb to LFA-1, CD2, CD3, CD4, CD8, and HLA molecules on killing mediated by IEL. Anti-LFA-1 inhibited strongly the killing of normal PBL and to a lesser extent the killing of tumor cells. Anti-CD2, CD3, CD4, CD8, and HLA class I molecules did not inhibit the nonspecific killing; rather, anti-CD3 potentiated the killing of PBL, K562, and Daudi cells. These results support the notion that qualitative and quantitative variations in LFA-1-mediated binding of target cells by IEL could result in differential killing of targets. The possibility of using anti-CD3 to selectively potentiate the killing of tumor cells is discussed.

Antibodies, Monoclonal↗
PubMed1987

Differential antigen presentation by heat-treated peripheral blood mononuclear cells and Epstein-Barr virus-transformed lymphoblastoid cell lines (EBV-LCL): heated EBV-LCL present alloantigen and soluble antigen but are deficient in the stimulation of autologous EBV-LCL primed T cells.

Heat-treated PBM (1 hr at 45 degrees C) cannot present soluble Candida albicans antigens (CAN) or stimulate in the mixed lymphocyte culture (MLC) reaction. This is despite their continued expression of serologically defined class II MHC antigens. In contrast, heat-treated EBV-LCL present soluble CAN and stimulate allogeneic T cells in the MLC. Heated EBV-LCL stimulate strong secondary responses from allogeneic alloprimed T-cell lines in the primed lymphocyte test (PLT), while heated PBM stimulate only weak secondary allogeneic responses. To test whether this difference was due to a subtle difference in the thermal stability of the functional expression of class II MHC antigens on PBM and EBV-LCL cells, the EBV-LCL cells were heated for 1 hr at temperatures from 45 degrees C to 60 degrees C. Even after treatment at 60 degrees C, the heated EBV-LCL strongly stimulated alloreactive T cells in MLC and PLT reactions. Heated EBV-LCL are not nonspecifically mitogenic, as they do not stimulate autologous T-cell lines primed to alloantigens. However, the weak response of alloprimed T-cell lines to heated allogeneic PBM can be augmented by coculturing with autologous heated EBV-LCL, suggesting heated EBV-LCL maintain a metabolic activity necessary for allogeneic stimulation that is deficient in heated PBM. While heated EBV-LCL stimulate allogeneic alloprimed T-cell lines, they no longer stimulate autologous EBV-LCL primed T-cell lines; irradiated EBV-LCL stimulate both strongly. This suggests the involvement of a heat labile antigenic or metabolic factor in the T-cell recognition of autologous but not allogenic EBV-LCL.

Antigen-Presenting Cells↗
PubMed1987

Status and potential of interleukin-2 for the treatment of neoplastic disease.

Interest in interleukin-2 results from the fact that it works in vivo on the cell-mediated arm of the immune response and in vitro it is effective on naive lymphocytes. Although it has no direct cytotoxic effect on neoplastic cells, it does help mediate the destruction of tumor cells. Current clinical trials are attempting to determine optimum dose and duration of treatment, since the toxicity of IL-2 appears to increase in proportion to dosage. The authors summarize our knowledge to date.

Animals↗
PubMed1987

Phase 1 clinical evaluation of recombinant interleukin-2.

The availability of large quantities of pure recombinant IL-2 has permitted clinical studies of its effects in patients with cancer. Based on the rapid half-life (3-5 minutes) and the need for prolonged exposure to IL-2 to maximally stimulate responsive cells we conducted a Phase 1 clinical trial designed to compare the clinical tolerance, immunologic effects and antitumor activity of IL-2 given by CI and daily BI for 7 consecutive days. The MTD of IL-2 based on completion of 7 days of treatment was found to be 3 X 10(6) U/m2/day for BI and 10(6) U/m2/day for CI. These results are approximately equivalent to those reported by others (Lotze, 1985 and Atkins, 1986), provided corrections are made for activity (2.3 Cetus IL-2 units = 1 BRMP IL-2 Unit). Although dose limiting toxicity was observed at the higher doses of IL-2 tested, these doses were tolerated for a shorter period of time (3-4 days) with striking immunologic effects (Sondel et al. 1987). As such, shorter treatment schedules utilizing higher doses of IL-2 warrant further investigation. The toxicity associated with the administration of IL-2 is considerable and clearly dose related. Of particular clinical importance is the fever, hypotension, fluid accumulation and decrease in PS. The etiology of these toxicities appears to be related to the release of endogenous cytokines following activation of the patient's immune system. It is also possible that the in vivo activation by IL-2 of cells mediating NRC may play some role in the toxicity associated with IL-2 (Sondel, 1986). It remains unclear whether immunosuppressive methods such as the use of cyclophosphamide or steroids will enable blockade of the immune mediated toxicity without also diminishing its therapeutic value. Further studies examining methods to control the severe toxicity associated with high doses of IL-2 are needed. The future role that IL-2 may play in treating cancer is unclear. Whether IL-2 alone can have significant antitumor activity may depend on factors such as tumor bulk, the patient's inherent ability to generate cytotoxic cells or the susceptibility of a particular malignancy. As such further studies will need to examine the importance of dose, timing, schedule, method of IL-2 administration as well as tumor burden in a wide variety of cancers.(ABSTRACT TRUNCATED AT 400 WORDS)

Aged↗