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Biomedical subjects

W E Samlowski

Publications and source records attributed to W E Samlowski.

At least 37 records · Page 2Linked to original sources
PubMed1991

Effects of supravital fluorochromes used to analyze the in vivo homing of murine lymphocytes on cellular function.

A number of supravital fluorochromes are available to study lymphocyte homing in vivo. These include fluorescein isothiocyanate (FITC), which binds to cell surface proteins; Hoechst 33342, which binds to AT rich regions of cellular DNA; and the lipid bilayer incorporated dyes PKH-2 and PKH-26. The relative advantages and disadvantages of each of these probes for analyzing murine lymphocyte homing are as yet poorly understood. We evaluated the effects of each dye on labeling efficiency, as well as cell viability, homing, mitogen responsiveness and cytotoxicity. PKH-26 provided long-term labeling (up to 15 days) with the least detrimental effects on cellular function. Detection of FITC in vivo was impaired by tissue autofluorescence, and the dye acted as a co-mitogen for lectin or IL-2-induced proliferation. Hoechst 33342 eluted from cells over a few hours and inhibited lymphocyte proliferation. PKH-2 had detrimental effects on cell viability and resulted in the down-modulation of peripheral lymph node homing receptor expression. None of the probes interfered with the induction of cytotoxic lymphocytes by IL-2. While these fluorochromes are easy to use and provide powerful tools to analyze the lymphocyte localization in vivo, our experiments demonstrate that important limitations are imposed by each probe that need to be considered by investigators during the design and interpretation of experiments.

Animals↗
PubMed1991

Reduction of testosterone synthesis after high dose interleukin-2 therapy of metastatic cancer.

We have investigated the effect of high dose iv bolus interleukin-2 (IL-2) therapy on sex hormone and adrenal steroid concentrations in six men treated for metastatic renal cell carcinoma or malignant melanoma. Blood concentrations of testosterone, 17 beta-estradiol, LH, FSH, cortisol, dehydroepiandrosterone (DHEA), and DHEA sulfate (DHEA-S) were measured before and after a 5-day course of IL-2 therapy. Cortisol levels rose and DHEA-S decreased insignificantly. DHEA declined, reaching a nadir (P less than 0.001) on day 6, and testosterone decreased significantly on day 2 and reached a nadir on day 6 (P less than 0.0001). Concentrations of both steroids then gradually rose. Estradiol rose on day 4 (P less than 0.001) and then declined. Neither LH nor FSH was affected significantly, although there was a rise in the mean level of LH after IL-2 therapy. Our results suggest that high dose IL-2 therapy in men affects both adrenal and testicular androgen production without inhibiting pituitary trophic hormone secretion. These effects of IL-2 on plasma sex steroids may be the result of cytokines stimulated by IL-2 therapy, rather than direct responses to IL-2.

Adult↗
PubMed1990

Human tumor cell line resistance to chemotherapeutic agents does not predict resistance to natural killer or lymphokine-activated killer cell-mediated cytolysis.

Cancer cells selected for resistance to natural product chemotherapeutic agents typically display cross-resistance to a variety of structurally and mechanistically diverse agents, a phenomenon known as multidrug resistance. Preliminary studies involving cells selected for multidrug resistance in vitro have suggested that the development of resistance to these agents might simultaneously confer resistance to some forms of immunotherapy. Using human tumor cell line models, we have investigated the relationship between either intrinsic or selected multidrug resistance and sensitivity to natural killer (NK) or lymphokine-activated killer (LAK) cell-mediated cytolysis. We compared the NK and LAK cell susceptibility of three human tumor cell lines displaying distinct mechanisms of selected drug resistance with that of the parental drug-sensitive lines. We also evaluated the NK and LAK susceptibility of five established renal cell carcinoma lines, all of which were found to be intrinsically resistant to doxorubicin and vinblastine. The drug-resistant cell lines were variably sensitive to NK-mediated lysis. In contrast, all drug-resistant cell lines tested were LAK cell sensitive. The NK and LAK cell-mediated cytolytic sensitivities of the drug-resistant cell lines correlated well with those of the drug-sensitive parental lines, suggesting that susceptibility to lysis was related intrinsically to each tumor type, and not to the resistance phenotype. We attempted to correlated the NK sensitivity of these cells with the cell surface expression of Class I or II histocompatibility antigens, or the presence or absence of the membrane inhibitor of complement-mediated reactive lysis. None of these phenotypic markers were found to predict NK resistance. We therefore conclude that these cells, which are either spontaneously resistant to commonly utilized antitumor agents or are multidrug resistant as a result of drug exposure in vitro, remain sensitive to LAK cell-mediated cytolysis. Our studies suggest that interleukin 2-induced LAK cells may be useful in the therapy of some chemotherapy-resistant cancers.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1990

Phase II evaluation of mitoxantrone in advanced pancreatic carcinoma: a Southwest Oncology Group study.

Patient with advanced adenocarcinoma of the pancreas and no prior chemotherapy were treated on a Phase II trial of mitoxantrone. Doses were adjusted for hepatic dysfunction as defined by bilirubin. Twenty-four patients with a bilirubin less than or equal to 1.5 mg% received mitoxantrone 12 mg/m2 i.v. repeated every three weeks. Myelosuppression in the form of leukopenia was the major toxicity. There were no responses in twenty-four evaluable patients.

Adenocarcinoma↗
PubMed1990

Mechanisms of pertussis toxin inhibition of lymphocyte-HEV interactions. I. Analysis of lymphocyte homing receptor-mediated binding mechanisms.

The molecular mechanisms by which pertussis toxin (PTX) inhibits lymphocyte homing to peripheral lymph nodes (PLN) remain poorly understood. PTX-treated lymphocytes express homing receptors, yet cannot extravasate into PLN in vivo. Methylation of PTX, a procedure known to inactivate the B-oligomer of the toxin, restored high endothelial venule (HEV) binding capacity. In vitro studies established that toxin exposure inhibited the accessory role of LFA-1 in HEV binding. In contrast, PTX-exposed lymphocytes exhibited normal MEL-14-mediated HEV binding. Analysis of membrane fluidity revealed a 20% decrease in fluorescence polarization in PTX-exposed lymphocytes. On the basis of the current experiments, we propose a "zipper" model of lymphocyte-HEV interaction, in which lateral mobility of adhesion receptors in the cell membrane toward a site of endothelial contact is necessary to maintain adhesion against the shear force due to blood flow. PTX inhibits these processes by decreasing membrane fluidity, and by altering accessory adhesion molecule function.

Animals↗
PubMed1990

Vascular leak syndrome associated with interleukin-2: chest radiographic manifestations.

Adoptive immunotherapy with interleukin-2 (IL-2) is associated with a generalized vascular leak syndrome. Pulmonary edema is a common occurrence and is rarely responsible for acute respiratory failure requiring assisted ventilation. The authors have performed a retrospective review of chest radiographs in 19 patients undergoing the priming course of high-dose IL-2 therapy for metastatic melanoma and renal cell carcinoma. This study was primarily designed to evaluate the prevalence and patterns of pulmonary edema and pleural effusions. During the first 5 days of therapy, alveolar edema was identified in 21% (n = 4) and signs of interstitial edema in 53% (n = 10) of patients. Pleural effusions were seen in 42% (n = 8). No patient in this series required assisted ventilation during this period. However, two patients subsequently developed fatal, drug-related myocardial injury. IL-2 toxicity is a well established cause of self-limited, increased-permeability pulmonary edema.

Capillary Permeability↗
PubMed1989

Changes in homing receptor expression on murine lymphokine-activated killer cells during IL-2 exposure.

The effects of IL-2 on the expression of homing receptors by lymphocytes of NK or lymphokine activated killer (LAK) cell derivation has not yet been evaluated. We developed a murine model to evaluate the potential of LAK cells to localize into peripheral lymph nodes since LAK cells are used to treat human cancers which have metastasized to these tissues. Using a frozen section binding assay, LAK cell adhesion to the lymph node microvasculature was easily demonstrable. Inhibition studies demonstrated that LAK cell binding to lymph nodes was mediated by mechanisms previously described in T cells. LAK cell surface expression of the 85- to 95-kDa homing receptor recognized by the antibody MEL-14 on LAK cells was assessed by indirect immunofluorescence. The percentage of cells which bound MEL-14 decreased slightly over 3 days of IL-2 exposure (from 73 to 60%), particularly in the large granular lymphocyte (cytotoxic effector) subpopulation (45% MEL-14+). The expression of another homing-related molecule, leukocyte function-associated Ag-1, markedly increased during activation of LAK cells. Despite the expression of these homing receptors, we observed almost no LAK cell localization into lymph nodes in vivo. Furthermore, IL-2 pretreatment of recipient animals did not increase the adhesion of LAK cells to lymph node microvasculature or enhance their extravasation. IL-2 activation of non-T, non-B lymphocytes results in significant changes in both the expression and function of cell surface homing receptors. Our results indicate that in vitro analysis does not always predict in vivo localization potential.

Animals↗
PubMed1989

Molecular mechanisms of lymphocyte extravasation. III. The loss of lymphocyte extravasation potential induced by pertussis toxin is not mediated via the activation of protein kinase C.

The present study evaluated whether protein kinase C (PKC) activation was involved in the lymphocytosis promoting properties of pertussis toxin (Ptx). The exposure of mouse lymphocytes to phorbol esters (as a means to selectively activate PKC) caused a depression in their subsequent capacity to localize into lymph nodes and Peyer's patches in vivo. This pattern of inhibition was quite similar to that observed with lymphocytes treated with Ptx. The mechanisms responsible for the observed decreases in localization to lymphoid organs caused by these two agents, however, appeared to be distinct. Exposure of lymphocytes to PMA was followed by a time and dosage-dependent decrease in the surface density of MEL-14 defined homing receptors. Ptx-treated lymphocytes retained normal density of this homing receptor. Consequently, PMA-treated lymphocytes lost their capacity to bind to high-endothelial venules in in vitro lymph node binding assays while Ptx-treated cells retained normal high-endothelial venule binding potential. We conclude from this study that: 1) the activation of PKC in lymphocytes by PMA can alter their recirculation properties via mechanisms that diminish their expression of surface receptors which support extravasation into lymph node and mucosal lymphoid tissues, and 2) even though Ptx has been reported to elevate the rate of inositol phosphate turnover in lymphocytes, the loss of extravasation potential of Ptx-treated lymphocytes is not mediated via the modification of surface homing receptors as observed in cells exposed to the known PKC activator, PMA.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1989

Hypocholesterolemia in patients treated with recombinant interleukin-2: appearance of remnant-like lipoproteins.

Reversible acute hypocholesterolemia was observed during treatment of metastatic cancer with high-dose intravenous recombinant interleukin-2 (IL-2). Further analysis revealed virtual disappearance of high-density lipoproteins (HDLs) and marked reduction in the concentration of low-density lipoproteins (LDL); the remaining LDL and intermediate-density lipoproteins (IDL) were enriched in triglyceride relative to cholesterol and had broad-beta electrophoretic mobility, properties reminiscent of remnant lipoproteins. These changes differ qualitatively and quantitatively from those previously reported for other cytokines such as tumor necrosis factor (TNF) or the interferons (IFNs).

Cholesterol↗
PubMed1989

Peripheral lymph node helper T-cell recovery after syngeneic bone marrow transplantation in mice prepared with either gamma-irradiation or busulfan.

The optimum marrow ablative regimen for preparing recipients of bone marrow transplantation (BMT) has not been established. gamma-Irradiation, but not busulfan, produces a characteristic microvascular injury pattern which results in depressed capacity of normal lymphocytes to localize into the lymph nodes of syngeneic murine BMT recipients. Since peripheral lymph nodes are important sites for initiation and amplification of immune responses, the preparative regimen might delay recovery of regionally compartmentalized immune functions after BMT. We evaluated the effects of busulfan and gamma-irradiation on the phenotypic and functional reconstitution of helper T-cell function within the peripheral lymph nodes of BMT recipients. Both marrow ablative regimens caused a protracted delay in regeneration of peripheral lymph node CD4+ T cells. Specific helper T-cell functions, such as contact hypersensitivity and alloantigen responses, remained significantly depressed in the lymph nodes of irradiated mice for prolonged periods (up to 60 weeks). These responses recovered more rapidly in busulfan-treated BMT recipients. In contrast, the capacity of peripheral lymph node T cells to provide "help" for antigen-specific immunoglobulin production was only transiently depressed by either preparative regimen. Our experiments confirm the hypothesis that the marrow ablative regimen, particularly gamma-irradiation, may contribute to the period of immunodeficiency which follows BMT. The pattern of immune recovery observed suggests that preparative total body irradiation (TBI) may selectively depress the regional recovery of the TH1 [interleukin-2 (IL-2) and gamma-interferon (gamma-IFN) secreting] lymphocyte subset.

Animals↗
PubMed1989

Severe myocarditis following high-dose interleukin-2 administration.

We observed a patient who developed unusually severe myocardial dysfunction during therapy for renal cell carcinoma with high-dose interleukin-2 (IL-2). Although cardiac isoenzymes became markedly elevated shortly after the completion of 14 doses of IL-2 (100,000 U/kg of IL-2 every eight hours), serial electrocardiograms revealed only nonspecific changes. Echocardiography documented diffuse myocardial dysfunction, with akinesis of the anteroseptal region, suggestive of myocardial infarction. This anteroseptal hypokinesis persisted over a five-day period. The patient died unexpectedly and postmortem evaluation of the heart revealed a severe, diffuse lymphocytic, and eosinophilic myocarditis with myocyte necrosis, especially prominent in the anteroseptal region. This is the first histologically confirmed case of myocarditis reported, to our knowledge, in association with IL-2 therapy for cancer.

Carcinoma, Renal Cell↗
PubMed1988

Effect of ionizing radiation on thymic epithelial cell function. I. Radiation-spared thymic epithelial grafts expedite the recovery of T cell function in lethally irradiated and fetal liver reconstituted mice.

A murine model system was developed to determine whether ionizing radiation has a detrimental influence on thymic epithelium, cell function. Normal mice were lethally irradiated, grafted intracamerally with normal fetal thymic epithelium, and then reconstituted with fetal liver cells. These animals were compared with a group of animals who received their thymic grafts before the irradiation protocol. Analysis of the reconstitution of T cell function in peripheral lymph nodes and spleens at various times post transplantation demonstrated that animals with radiation-spared thymic grafts had superior proliferative responses to T cell mitogens and alloantigens. It was also determined that the capacity of these animals to elicit contact hypersensitivity responses was significantly greater when compared with animals whose thymic grafts had been radiated. The observed difference in T cell function could not be ascribed to a difference in the rate of export of mature T cells from the thymic grafts since the absolute number of Thy-1+, L3T4+, or Lyt-2+ lymphocytes present in the peripheral lymphoid compartment of our two groups of animals was equivalent. Immunohistologic analysis of the thymic grafts demonstrated a marked reduction in the medullary compartment of the repopulated grafts that had been exposed to ionizing radiation. The results of this study suggest: 1) that irradiation of the thymic microenvironment during marrow ablative preparative regimens may be in part responsible for some of the immune alterations observed in marrow transplant recipients, and 2) that our model system may provide a valuable tool for delineating the roles played by medullary and cortical epithelial cells of the thymus on the T cell maturation and education processes.

Animals↗
PubMed1987

Flecanide-induced immune neutropenia. Documentation of a hapten-mediated mechanism of cell destruction.

Immune-mediated granulocytopenia due to cardiac antiarrhythmic medications is a rare, but potentially dangerous, event. This article characterizes the first case, to our knowledge, of severe granulocytopenia associated with the administration of flecanide acetate, a new class I antiarrhythmic drug. Immunologic studies determined that flecanide was capable of binding to the surface of normal neutrophils. The patient's serum contained an IgG antibody that could specifically bind to the haptenized neutrophils, presumably mediating enhanced destruction of mature granulocytes both in the serum and within the bone marrow. Cessation of flecanide therapy resulted in resolution of the granulocytopenia. The titer of antineutrophil antibody in the patient's serum decreased to background levels within the next five months. Similar antibodies were not found in serum from nonsensitized individuals. The capacity of flecanide to bind to normal neutrophils may prove to be a significant risk factor for the subsequent development of antineutrophil antibodies and agranulocytosis in patients receiving this drug. Careful hematologic monitoring of all patients who are receiving this medication is, therefore, strongly urged.

Aged↗
PubMed1987

Recovery of contact hypersensitivity responses following murine bone marrow transplantation: comparison of gamma-irradiation and busulfan as preparative marrow-ablative agents.

Bone marrow transplantation (BMT) is often followed by significant morbidity and mortality due to protracted immunodeficiency. We have hypothesized that the bone marrow-ablative regimen may delay the recovery of normal immune function following transplantation by impairing the interaction of host endothelial cells with circulating graft-derived lymphocytes. This report compares the relative effects of busulfan (an alkylating drug) and gamma-irradiation on the tissue-specific localization potential of lymphocytes and the eventual recovery of immune function within syngeneic murine transplant recipients. Localization of normal lymphocytes into peripheral lymph nodes of irradiated BMT recipients was markedly less (less than 50%) than in busulfan-treated or normal mice over the first 2 months post-BMT. This finding correlated with irradiation-induced endothelial cell edema and microvascular occlusions within lymphocyte-receptive areas of the nodal microvasculature. The effect of both preparative regimens on the recovery of contact hypersensitivity (CHS) was also analyzed. This response recovered more quickly (between 1 and 2 months) in busulfan-pretreated animals. Further experiments demonstrated that the decrease in CHS responsiveness appeared, in part, related to a depression in the capacity of lymphocytes to localize into skin sites of antigen deposition within irradiated mice. The impairment of tissue-specific lymphocyte localization may represent a novel mechanism by which whole body irradiation can contribute to delayed immunologic reconstitution following bone marrow transplantation.

Animals↗