Unplanned intraoperative and postoperative hemodilution: oxygen transport and consumption during severe anemia.
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Adult↗
Biomedical subjects
A Lichtenstein
Publications and source records attributed to A Lichtenstein.
Stimulation of the respiratory burst of murine peritoneal inflammatory neutrophils by conjugation with tumor cells.
Murine peritoneal neutrophils (PMNs), elicited by i.p. injection of formalin-killed Corynebacteria parvum, spontaneously lyse teratocarcinoma targets through the secretion of reactive oxygen intermediates. Examination of effector-target interactions at the single cell level revealed that PMNs conjugated to tumor cells were 3-fold more frequently stained by nitroblue tetrazolium compared to nonconjugating PMNs suggesting that tumor targets stimulated a potent tumor-lytic respiratory burst. This notion was confirmed by the detection of superoxide and hydrogen peroxide generation from PMNs as well as a luminol-dependent chemiluminescent response following conjugation with viable tumor targets. Generation of superoxide was dependent upon the presence of dihydrocytochalasin B. In addition to teratocarcinoma cells, comparable stimulation was achieved by conjugation with YAC and P815 targets but not thymocytes. Reactive oxygen intermediate release was also achieved by mixing peritoneal PMNs with heat-killed tumor cells. In contrast to bacteria-induced effectors, PMNs elicited by i.p. injection of thioglycollate were incapable of responding following conjugation with tumor targets although they were competent for reactive oxygen intermediate release when stimulated by phorbol myristate acetate. Teratocarcinoma targets were sensitive to concentrations of H2O2 that could be achieved by PMNs following contact. These data indicate that Corynebacteria-elicited inflammatory PMNs lyse their bound tumor targets by a mechanism similar to a stimulus-secretion model.
Animals↗
Antitumor effect of the streptococcal preparation OK-432 in a murine model of ovarian cancer.
The antitumor effects of the streptococcal preparation OK-432 were analyzed in a murine ovarian teratocarcinoma (MOT) model. Administration of OK-432 i.p. prevented tumor outgrowth in 75% of mice challenged with 10(3) MOT cells i.p. 24 h previously. Treatment was less successful in mice challenged with 10(4) or 10(5) cells, preventing tumor growth in 25% of the former and only 5% of the latter group. Tumor-challenged mice cured by injections of OK-432 were not rendered resistant to a subsequent challenge with 10(3) MOT cells 75 days after initial treatment. Only the i.p. route of administration was effective as i.v. OK-432 did not prolong survival of tumor-challenged mice. An antitumor response was detected as early as 24 h after i.p. treatment. This correlated temporally with an influx of neutrophils into the peritoneal cavity. Peritoneal cells obtained between 6 and 24 h after treatment were capable of lysing MOT targets in vitro. A single cell cytotoxicity assay demonstrated that peritoneal neutrophils, elicited by i.p. injection of OK-432, could bind to and lyse MOT targets. These data indicate that OK-432 is effective against small tumor cell inocula in this murine model of ovarian cancer and, furthermore, that the neutrophilic response into the peritoneal cavity plays a role in tumor rejection.
Animals↗
Evidence for peripheral blood B lymphocyte but not T lymphocyte involvement in multiple myeloma.
Because there is controversy regarding whether subsets of peripheral blood lymphocytes (PBLs) are part of the malignant clone in patients with multiple myeloma, we studied this question by immunoglobulin and T cell receptor gene analysis. Southern blot analysis with antibody probes demonstrated clonal immunoglobulin gene rearrangements in PBLs of seven of nine patients that were identical to those seen in their marrow plasma cells. Circulating plasma cells were not detected in any of these patients. In contrast, no patient demonstrated clonally rearranged T cell receptor genes. In one sequentially studied patient, PBLs obtained at diagnosis when he had stage I (Durie-Salmon) contained only germline DNA, while analysis of PBLs at relapse (stage III) revealed a clonally rearranged band. These data confirm the notion that circulating lymphocytes in patients with myeloma are part of the malignant clone and, furthermore, these malignant cells are of B cell rather than T cell lineage.
Aged↗
Antitumor and immunologic effects of a pyridine-extracted fraction of Propionibacterium acnes.
The antitumor and immunological effects of a pyridine extractable fraction of Propionibacterium acnes were tested in a murine ovarian teratocarcinoma (MOT) model. Previous studies have demonstrated that tumor rejection in this model depends upon sequential activation of tumoricidal neutrophils (PMNs) followed by cytostatic macrophages. The pyridine extract significantly prolonged the survival of mice challenged with 10(3) or 10(4) MOT cells but had little impact on a 10(5) tumor inoculum. In vivo cytoreduction occurred within the first 24 h following IP treatment which correlated temporally with the influx of tumoricidal PMNs into the peritoneal cavity. Immunotherapy failure in mice challenged with 10(5) MOT cells occurred between 48 and 72 h after treatment when macrophage chemotaxis into the peritoneal cavity was initiated. Although injection of unfractionated bacteria activated MOT-cytostatic macrophages, the pyridine extract was deficient in this regard. Intraperitoneal injection of the pyridine extract resulted in an early (day +1) depression and late (day +5) enhancement of peritoneal NK cytotoxicity. These data suggest that the retention of neutrophil-activating moieties in the pyridine extract are sufficient for antitumor effects against low tumor inocula while the depletion of macrophage-activating determinants results in diminished effects against larger tumor cell challenges.
Adjuvants, Immunologic↗
Spontaneous tumor cytolysis mediated by inflammatory neutrophils: dependence upon divalent cations and reduced oxygen intermediates.
The role of divalent cations and reactive products of the respiratory burst were investigated in spontaneous tumor lysis mediated by inflammatory neutrophils (PMNs). Murine peritoneal PMNs, obtained five hours after intraperitoneal injection of bacteria, conjugated and lysed teratocarcinoma cells in chromium release and single-cell cytotoxicity assays. The presence of extracellular magnesium was required and was sufficient for tumor cell binding to PMNs. Postbinding lytic events depended upon the simultaneous presence of extracellular calcium and magnesium. Catalase and superoxide dismutase inhibited postbinding lytic events, indicating that production of reduced oxygen moieties was important. Scavengers of hydroxyl radicals could inhibit tumor cell binding, but none could affect postbinding lytic events. Neither could inhibitors of myeloperoxidase decrease tumor lysis. The ability of conjugating PMNs to lyse their bound targets correlated with their reduction of nitro blue tetrazolium (NBT). Optimal concentrations of phorbol myristate acetate (PMA) markedly increased the NBT positivity of PMNs and the killing of bound tumor cells. Even with optimal stimulation of the respiratory burst, however, there was still a significant number (19%) of bound targets that escaped lysis, suggesting active resistance to oxygen-mediated tumor cell injury.
Animals↗
In vitro tumor cell cytolysis mediated by peptide defensins of human and rabbit granulocytes.
We examined the activity of defensins, cysteine-rich cationic peptides that are abundant in the cytoplasmic granules of human and rabbit granulocytes, against various tumor targets. The three human defensins, HNP-1, HNP-2, and HNP-3, lysed human and murine targets in chromium release and dye exclusion assays. Defensin-mediated tumor cell lysis was concentration-dependent, inhibited by serum, and dependent on temperature-sensitive events. Lysis was first detected by three hours of incubation and it reached a plateau between eight and 14 hours. In vitro exposure of murine teratocarcinoma cells to HNP 1-3 abrogated their oncogenicity in vivo. Nonmalignant target cells were also susceptible to defensin-mediated lysis. Four rabbit granulocyte defensins exerted marked (NP-1, NP-2) or moderate (NP-3a, NP-3b) cytotoxic activity, whereas defensin NP-5 was not cytotoxic. When tumor cells were incubated with human defensins in combination with hydrogen peroxide, synergistic cytotoxicity was detected. As defensins are released from granulocytes by various stimuli, their release could contribute to extracellular cytotoxicity which is independent of reactive oxygen intermediates.
Animals↗
Intraperitoneal immunotherapy of epithelial ovarian carcinoma with Corynebacterium parvum.
Corynebacterium parvum was administered intraperitoneally to 21 patients with epithelial ovarian cancer. Nineteen patients had surgically measurable disease and two received adjuvant therapy. Surgically confirmed responses were documented in six of 19 patients (31.6%), with two complete responses (10.5%) and four partial responses (21.1%). Three patients (15.8%) had stable disease, and 10 patients (52.6%) had disease progression. The mean survival of the patients who had a complete response was 35.5 months; the four patients who had a partial response the mean survival was 26.6 months, and of the nonresponders the mean survival was 12.6 months (p less than 0.02). The mean survival of the entire group was 18.2 months. Initial response and patient survival correlated with the amount of disease pretreatment. Thus six responding patients had less than or equal to 5 mm maximum diameter tumors, that is, minimal residual disease. Toxicity in the 86 courses of therapy included abdominal pain in 78% of cases, fever in 56%, nausea in 40%, and vomiting in 22%. Stimulation of cytotoxic lymphocytes resulted from the administration of C. parvum, which induced a significant increase of both intraperitoneal natural killer lymphocyte cytotoxicity and antibody-dependent cell-mediated cytotoxicity in six of nine patients tested; these two types of cytotoxicity correlated with response to therapy and may be partially responsible for the surgically documented tumor regression. While the clinical usefulness of intraperitoneal C. parvum is limited because of its toxicity, intraperitoneal immunotherapy may prove useful in patients with minimal residual ovarian cancer when more refined agents become available.
Adenocarcinoma, Mucinous↗
Use of a self-generating percoll gradient and single cell cytotoxicity assay to identify tumor-lytic properties of inflammatory neutrophils.
Within a murine model of regional immunotherapy, the cytolytic potential of peritoneal neutrophils could not be confirmed or quantified using routine techniques of cell separation and chromium release assays. We, therefore, developed procedures for the enrichment of neutrophils and estimation of the frequency of killer cells. Peritoneal exudate cells from mice injected with Corynebacterium parvum were fractionated on a self-generating Percoll gradient to enrich for neutrophils and deplete macrophages. A significant enrichment of neutrophils (greater than 90%) was obtained in a band corresponding to a density of 1.088 with a recovery of 35-50% of input. Neutrophil-enriched cell populations were then mixed with tumor cells to examine neutrophil-target interactions at the single cell level. Conjugates of neutrophils and tumor targets were obtained and the majority were lytic. With the aid of trypan blue staining and safranin counterstaining, it was possible to distinguish effector cells from targets and neutrophils from other host cells. The frequency of conjugates was dependent upon the effector to target cell ratio and was not affected by changes in temperature (range 4-30 degrees C). The post-binding lytic events were initiated rapidly after conjugation and tumor lysis was completed within 30 min. The lytic events occurred optimally between 25 degrees and 37 degrees C. The present studies support the role of neutrophils in tumor lysis following administration of an immunoadjuvant. The techniques described are important to further study the role of neutrophils in disease states as well as the underlying mechanisms of neutrophil-mediated tumor cytotoxicity.
Animals↗
Anti-tumor effect of inflammatory neutrophils: characteristics of in vivo generation and in vitro tumor cell lysis.
Inflammatory neutrophils elicited by intraperitoneal injection of Corynebacterium parvum, thioglycollate or proteose peptone were capable of lysing different murine and human tumor targets in a short-term chromium-release assay. A single-cell cytotoxicity assay, which evaluated effector-target cell interactions at the single-cell level, confirmed a PMN-mediated tumor-lytic effect. Optimal lysis was achieved by PMNs obtained 6 hr after injection of C. parvum and 16 hr after injection of thioglycollate. In vitro, loss of tumor cell membrane integrity occurred extremely rapidly following conjugation with inflammatory PMNs (beginning within 15 min of the binding step). By 45 min, the lytic event was completed. Addition of catalase or superoxide dismutase to the cytotoxicity assays prevented tumor lysis in a concentration-dependent fashion, indicating that hydrogen peroxide and superoxide, products of the PMN respiratory burst, are mediators of the lytic reaction.
Animals↗
Rejection of murine ovarian cancer following treatment with regional immunotherapy: correlations with a neutrophil-mediated activation of cytostatic macrophages.
Rejection of the murine ovarian teratocarcinoma (MOT) in C3HeB/FeJ mice, following intraperitoneal (ip) treatment with Corynebacterium parvum (C. parvum), is abrogated by injections of silica. We, therefore, investigated whether C. parvum-elicited macrophages affect MOT targets in vitro. Tumor-cytostatic, but not cytolytic, macrophages were detected in normal and tumor-challenged mice treated with C. parvum. The dose responsiveness and kinetics of macrophage activation strongly correlated with tumor rejection. A pyridine extract of C. parvum, possessing greatly diminished tumor rejection properties, was significantly less effective in activating macrophages. Cytostatic macrophage activation and prevention of tumor outgrowth also followed treatment in C3H/HEJ mice, a strain with a known deficiency in cytolytic macrophage function. Peritoneal neutrophils, obtained 6 hr after treatment with C. parvum, were capable of activating cytostatic macrophages when reinjected ip into normal mice. These results indicate a critical role for tumor cytostatic macrophages in this immunotherapy model and suggest their activation is mediated by inflammatory neutrophils.
Animals↗
Intraperitoneal recombinant alpha 2-interferon for 'salvage' immunotherapy in persistent epithelial ovarian cancer.
Fourteen patients with epithelial ovarian cancer were treated with intraperitoneal (i.p.) administration of alpha-recombinant interferon (rIFN-alpha 2) after documentation of persistent disease at second-look laparotomy and combination chemotherapy. After therapy, 11 patients had a surgical re-evaluation which confirmed 4 complete responses (36%), 1 partial response (9%), and disease progression in 6 (55%). Five of 7 patients (71%) with minimal residual disease (MRD, i.e. less than 5 mm) had a surgically-documented response, whereas there was none in the 4 patients whose tumors were greater than or equal to 5 mm. Fever greater than or equal to 38 degrees C was seen in 58%, greater than or equal to 39.0 degrees C in 18%; nausea and vomiting in 37%, and abdominal pain in 22%. There was no consistent alteration in peripheral WBC's during treatment, while i.p. monocytes and lymphocytes showed a significant boost on day 1 after each dose of rIFN-alpha 2. Natural killer (NK) lymphocyte cytotoxicity was elevated in the i.p. cavity fluid obtained from most patients on day 1 after treatment, while blood NK values showed considerable variability. Pharmacokinetic studies showed i.p. levels of rIFN-alpha 2 were 30-1000 times blood levels. I.p. rIFN-alpha 2 may act by increasing concentrations of drug and augmenting regional host cells in patients with MRD ovarian cancer.
Adult↗
Intraperitoneal recombinant alpha-interferon for "salvage" immunotherapy in stage III epithelial ovarian cancer: a Gynecologic Oncology Group Study.
Fourteen patients with persistent epithelial ovarian cancer documented at second look laparotomy after combination chemotherapy were treated with 146 cycles of alpha-recombinant interferon (rIFN-alpha 2) administered i.p. The initial dose was 5 X 10(6) units which was escalated weekly to 50 X 10(6) units over 4 weeks and then continued weekly for a total of 16 weeks. Eleven patients underwent surgical reevaluation after therapy which confirmed four pathological complete responses (36%), one partial response (9%), and disease progression in six patients (55%). Five of seven patients (71%) with residual tumor less than 5 mm had a surgically documented response, whereas there was no response in the four patients whose tumors were greater than or equal to 5 mm. Three patients were evaluable for clinical response only: one patient who refused surgery had a complete clinical response with total resolution of ascites; one had stable disease; and one had disease progression. Fever greater than or equal to 38 degrees C was seen in 58%, fever greater than or equal to 39.0 degrees C was seen in 18%, vomiting in 37%, abdominal pain was reported in 22%, and one patient had infectious peritonitis. Peripheral white blood cell counts and i.p. washings were obtained pretreatment and on days 1, 3, and 7 after treatment. While there was no consistent alteration in peripheral white blood cell counts, the numbers of i.p. monocytes and lymphocytes showed a significant boost on day 1 after each dose of rIFN-alpha 2. Natural killer lymphocyte cytotoxicity was elevated in the i.p. cavity fluid obtained from most patients on day 1 after treatment, while blood natural killer lymphocyte cytotoxicity values showed considerable variability. Pharmacokinetic studies show that i.p. levels of rIFN-alpha 2 were 30-1000 times blood levels. rIFN-alpha 2 i.p. may act by increasing concentrations of drug and augmenting regional host cells in patients with minimal residual ovarian cancer.
Adult↗
Peripheral blood lymphocytes with receptors for a determinant common to B-lymphoblastoid cell lines and acute myelogenous leukemia blasts.
We investigated whether the rosetting of B-lymphoblastoid cell lines (B-LCL) by peripheral blood lymphocytes (PBLs) reflected possible interactions between lymphoid cells and immature cells of the hematopoietic system. Rosette formation could be blocked by the addition of soluble antigen extracted from B-LCL or blasts obtained from patients with acute myelogenous leukemia (AML). This inhibition was specific for AML blasts (similarly extracted material from melanoma lines had no inhibitory effect) and for the B-LCL receptor (leukemic extracts had no effect on surface receptors for sheep red blood cells (E) or antibody-sensitized red blood cells (EA)). The B-LCL receptor is present on leukemic Sezary T-cells as well as normal T-cells and its sensitivity to various enzymatic treatments is markedly different from that of E and EA receptors. In addition, B-LCLs derived from in vitro EB-viral infection of a normal donor's B lymphocytes were significantly rosetted by that donor's autologous PBLs. These data suggests the B-LCL receptor, present on mature T-cells, can recognize self determinants on myeloblasts and B-LCL. Further investigation will determine whether this interaction can affect the function of rosetted target cells.
Antibodies, Monoclonal↗
Lymphocyte cytotoxicity in the peritoneal cavity and blood of patients with ovarian cancer.
After an intensive course of combination chemotherapy, 16 patients with minimal residual ovarian cancer that was documented at second-look laparotomy, had an indwelling Tenckhoff catheter placed and underwent multiple peritoneal saline lavages. Lymphocyte-enriched populations from the peritoneal cavity and peripheral blood were obtained by density gradient centrifugation and examined for cell-surface phenotype and a variety of immune functions, including natural killer cytotoxicity and antibody-dependent cell-mediated cytotoxicity. Phenotypic characterization revealed that peritoneal lymphocytes consisted primarily of T cells and cells bearing receptors for the crystallizable fragment of immunoglobulin G (IgG) (crystallizable fragment-receptor), and contained a very low number of B cells. Peritoneal natural killer lymphocyte cytotoxicity and antibody-dependent cell-mediated cytotoxicity were very low in all but two patients. Incubation of peritoneal lymphocytes with Corynebacterium parvum and interferon in vitro did not result in augmented cytotoxicity against susceptible targets. Supernatants from cultured peritoneal cells of all patients markedly inhibited natural cytotoxic activity of normal donor blood lymphocytes. These results suggest that lymphocytes collected from the peritoneal cavity of patients with minimal residual ovarian cancer are deficient in natural and antibody-dependent cytotoxic effector function. This deficiency may influence the host's ability to control the spread and proliferation of tumor cells in the peritoneal cavity.
Adenocarcinoma↗
Activation of peritoneal lymphocyte cytotoxicity in patients with ovarian cancer by intraperitoneal treatment with Corynebacterium parvum.
Following an intensive course of combination chemotherapy, patients with minimal residual ovarian cancer were treated with increasing concentrations of intraperitoneal Corynebacterium parvum to assess whether or not natural killer (NK) cells could be activated. Immunotherapy was administered every 2 weeks, initially with a dose of 250 micrograms/m2, which was progressively escalated as tolerated. Each treatment induced a peritoneal cellular exudate which consisted primarily of neutrophils 48h after injection and of lymphocytes and macrophages at 7 and 14 days after injection. Peritoneal NK cytotoxicity increased during treatment in six of nine patients tested. NK precursor cells susceptible to in vitro activation with either C. parvum or interferon became detectable during treatment in all five patients tested. In four of these five, precursors were detected prior to the development of enhanced spontaneous NK activity. In four patients serially studied, peripheral blood NK activity increased during therapy. These results indicate that regional intraperitoneal therapy with C. parvum can enhance nonspecific antitumor cytotoxic mechanisms within the peritoneal cavity.
Cytotoxicity, Immunologic↗
Inhibition of natural killer cell lysis by natural killer-inhibitory substance: mechanism of suppression.
The mechanism of suppression of NK-mediated lysis by a soluble product of peritoneal cells (NK-IS, natural killer-inhibitory substance) was investigated. Pretreatment of effector cells resulted in depressed NK lysis while pretreatment of targets had no effect, indicating suppression is due to alterations in effector cell function rather than changes in target cells. NK-IS had no effect on the formation of conjugates between effectors and NK-susceptible targets. When NK-IS was added to effector-target cell mixtures after the binding step had been successfully completed, ensuing lysis was significantly depressed, confirming that NK-IS inhibited a postbinding lytic event. The degree of suppression caused by NK-IS was directly related to the duration of exposure to the inhibitory molecule. In addition, a preliminary temperature-dependent step of binding to and/or intracellular entry of NK-IS into effectors is required before suppression can occur. NK-IS prevents the activation of NK cell lysis by interferon and Corynebacterium parvum and effectively inhibits lysis mediated by already activated effectors. The potent suppression of NK lysis and prevention of interferon and C. parvum-mediated activation of NK lysis by a soluble product of peritoneal cells may explain the extremely low level of NK effector cell function within the peritoneal cavity.
Animals↗
Augmentation of NK activity by Corynebacterium parvum fractions in vivo and in vitro.
Biochemically modified whole cell preparations and derived fractions of Corynebacterium parvum (C. parvum) were evaluated for the ability to augment natural killer cell cytoxicity in vivo and in vitro in rats. Unfractionated C. parvum enhanced peritoneal cell (Pc) NK activity in a dose dependent fashion. This activity appeared to be enriched in insoluble light residue material obtained from hot phenol water extraction. Enhancement of Pc cytotoxicity was significantly greater at all time points tested in rats injected with light residue when compared to rats injected with comparable amounts (by dry weight) of unfractionated organisms. In addition, pyridine extractable material and HCl modified preparations were capable of boosting Pc NK activity following intraperitoneal (I.P.) injection. Periodate treatment abrogated C. parvum's ability to boost Pc cytotoxicity and insoluble residue material obtained from pyridine extraction was likewise devoid of NK-enhancing properties. Culture of rat spleen cells overnight with unfractionated C. parvum, light residue and pyridine residue materials enhanced NK cytotoxicity whereas HCl and periodate modified whole cell preparations as well as phenol and pyridine extractable material were incapable of boosting cytotoxicity in vitro. In vitro augmentation by culturing with light residue was dependent on the presence of adherent cells in rat spleen cell populations. Pyridine extracts boost cytotoxicity in vivo and have no effect in vitro while the opposite is true of pyridine residue material suggesting different mechanisms of NK augmentation by C. parvum between in vitro and in vivo systems.
Animals↗