Factors determining the actual received dose intensity in a program of multicyclic dose-intensive alternating chemotherapy with sequential stem cell support.

Dose intensity has been related to clinical outcome in several solid tumors. We studied the influence of clinical and cellular parameters on dose intensity received in a series of 53 patients with metastatic breast cancer or advanced ovarian cancer. They received courses of cisplatin 120 mg/m(2) plus etoposide 600 mg/m(2) alternating every 14 days with ifosfamide 8 g/m(2) plus paclitaxel 200--350 mg/m(2). Blood stem cell support was administered after every course except for the first one. Patients with excellent mobilization underwent immunomagnetic selection of CD34+ cells. We found a significant inverse correlation between the CD34+ cell dose infused and the delay for the administration of the next cycle. A CD34+ cell dose between 1.5 and 5 x 10(6)/kg per cycle was found to be feasible and was followed by a median delay of 1 day (not different from doses above 5 x 10(6)/kg). Three factors independently predicted the actually received dose intensity in a multiple regression model (R(2) = 0.4): previous autologous transplantation, eligibility for immunomagnetic selection (excellent response to mobilization) and median CD34+ cell dose received along the treatment.

[Intensive chemotherapy in the treatment of patients with metastatic breast cancer]. / Quimioterapia intensiva en el tratamiento de pacientes con cáncer de mama metastásico.

PURPOSE: Phase II study with intensive chemotherapy and autologous stem cells support in patients with metastatic breast cancer. METHODS: Forty-nine patients were treated with high-doses of two cytotoxic drugs and support with stem cells obtained from several leukapheresis without movilitation. The cells were reinfused forty-eight hours after finishing the administration of chemotherapy. RESULTS: Twenty-one patients (47%, CI-95%: 32.4-63.3%) achieved a complete remission. The objective responses rate was 73% (CI-95%: 57.2-85%). Overall and progression-free survival up to 4 years were 31% and 20%, respectively. Ten patients remain progression-free among 17 and 46 months. The most frequent extramedullary toxicity was hepatic and renal. Three patients (6%) died during the procedure. CONCLUSIONS: Intensive chemotherapy with hematopoietic support yields, with a moderate toxicity, a high objective response and complete remission rate. A small group of patients achieves a long progression-free survival.

Photo-induced toxic epidermal necrolysis caused by clobazam.

Toxic epidermal necrolysis (TEN) is a life-threatening disease, the pathogenesis of which remains largely unknown. We describe a 23-year-old woman under treatment with clobazam who developed lesions of TEN in light-exposed areas. Patch and photopatch tests with clobazam were negative. The cellular phenotype and cytokines were studied in blister fluid. The cellular infiltrate was composed mainly of T lymphocytes with a predominant cytotoxic phenotype. There was an increase in the level of tumour necrosis factor (TNF)-alpha in blister fluid compared with the control (a patient with bullous pemphigoid).

Characterization of peripheral blood steady-state progenitor cells preserved in liquid culture conditions with or without GM-CSF and IL2.

The cellular characteristics of steady-state peripheral blood progenitor cell (PBPC) apheresis, including total number of lymphomononuclear cells, CD34 and CFUs, was evaluated in a group of 26 chemo-radiotherapy patients as well as in a group of 23 surgically resected cancer patients. Three-to seven-day incubation in standard liquid culture conditions with growth factors (IL2, GM-CSF or both) correlated with a statistically significant increase in CD34+ and CD56+ cell populations compared with incubation without growth factors, especially when both GM-CSF and IL2 were used. In addition, an increase in CD33+, CD13+ and HLA-DR+ cell populations was observed after 3-7 days incubation with GM-CSF. The basal culture control exhibited a decrease in CD33+ and CD13+ cell populations while CD34+ and CD56+ cell populations were maintained. These results were similar in the treated and untreated groups of patients. The infusion of GM-CSF and IL2 preincubated PBPC after intensive chemotherapy was associated with a rapid hematological recovery with a median time duration for WBC < 500/uL, WBC < 1.000/uL and platelets < 20.000/uL of 7.9 days, 14.9 days and 10.7 days respectively. We conclude that a short GM-CSF and IL2 preincubation of steady-state PBPC is associated with an increase in cell populations exhibiting the immune and progenitor cell phenotypes and correlates with an early hematological recovery after intensive chemotherapy.

High-dose BCNU and autologous progenitor cell transplantation given with intra-arterial cisplatinum and simultaneous radiotherapy in the treatment of high-grade gliomas: benefit for selected patients.

A phase II study of postoperative high-dose carmustine (HDBCNU), intracarotid cisplatin (CDDP), and radical radiotherapy in patients with high-grade glioma was performed. Patients underwent 4-6 consecutive days of blood hematopoietic progenitor cell (HPC) apheresis without prior mobilization. Chemotherapy included intracarotid CDDP, 60 mg/m2, and BCNU, 900 mg/m2. HPC were infused 48 h after HDBCNU. Whole brain irradiation, up to 50 Gy, was started on the 8th day after HPC infusion. With a median follow-up time of 44 months, median overall survival was 15.5 months. Eight patients (23.5%) are alive free of disease 2-6 years after treatment (seven out of 25 patients with glioblastoma multiforme and one out of nine patients with anaplastic astrocytoma). Survival was influenced by young age, good performance and complete surgical resection. Two patients (5.8%) died of therapy-related complications. Acute hematological toxicity of HDBCNU was moderate, with a full recovery on day 26. No acute pulmonary or hepatic toxicity was found. Late severe neurological toxicity was observed in one third of patients surviving beyond 2 years. We conclude that HDBCNU, 900 mg/m2, intracarotid CDDP and radical radiotherapy appear to benefit some patients with high-grade gliomas, and phase III studies should preferentially select young patients with resectable tumors.

Activation of human lymphomononuclear cells by peptides derived from extracellular matrix proteins.

A series of peptides of 15 amino acids with sequences contained in human extracellular matrix (ECM) proteins (fibronectin, laminin A, laminin B1, tenascin, undulin, alpha 1-chain of type IV and VIII collagen and alpha 2-chain of type VIII collagen) have been synthesized. The selected structures conformed to the following pattern: (i) Pro at position 6, (ii) Leu, Lys, Ile, Val, Ala or Gly at position 2, (iii) Glu or Asp at position 11. Fibronectin and the indicated peptides, when present in cultures of lymphomononuclear cells from healthy donors, promoted stimulation of monocytes manifested by a release of IL-1 alpha, IL-beta, IL-6 and TNF alpha; an increase in the percentage of cells expressing CD14, CD16, CD11b and CD14/CD16; an increase in cytotoxicity against HT-29. Cytotoxicity against K562 and Daudi cells (targets of NK and LAK cells) was also observed together with an increase in the percentage of cells expressing CD56, CD56/CD16 (corresponding to NK cells), and CD56/CD8 (corresponding to NK-like lymphocytes), indicating a stimulation of lymphocytes. Activated monocytes and lymphocytes contained a large number of granules with DNAse activity. These results suggest that at least some of the immunological properties of ECM proteins could be accounted for by motifs fulfilling a characteristic sequence pattern shared by all of them.

Immunomodulation induced by synthetic peptides derived from Staphylococcus aureus protein A.

Peptides from 10 to 22 amino acids containing sequences encompassed by Staphylococcus aureus protein A were synthesized. Some of these peptides, when present in cultures of lymphomononuclear cells from healthy donors or from cancer patients (melanoma, breast carcinoma, non-Hodgkin lymphoma and renal cell carcinoma) promoted: (i) changes in the phenotype of the lymphomononuclear population, (ii) stimulation of monocytes (release of IL-1 and TNF-alpha), and (iii) an increase in cytotoxicity against K562, Daudi and HT-29 cells. Isolated monocytes responded also to those peptides with a release of IL-1 and TNF alpha and an increase of cytotoxicity against HT-29 cells. It was found that the active peptides had the following structural pattern: a length of at least 15 amino-acid residues with a proline at position 6, valine, leucine, isoleucine, glycine, alanine or lysine at position 2, and glutamic or aspartic acid at position 11. Replacement of Pro at position 6 with any other residue turned the peptide inactive. Replacement of residues at positions 2 and 11 with amino-acid residues other than those required for activity resulted in compounds with a marked decrease in the immunomodulating properties described, or lacking these properties altogether.

[Depression of the mononuclear phagocyte system caused by high doses of narcotics]. / Depresión del sistema mononuclear-fagocítico provocada por altas dosis de morfínicos.

We evaluated in human monocytes the effect of high doses of alfentanyl on the expression of vimentin filaments, the phagocytic activity and the membrane display of HLA-DR molecules in the subjects undergoing surgery. The study was performed on 30 patients, ASAI-II. The patients received 100 mcg/kg i.v. of Alfentanil and the maintenance of anaesthesia was made with Alfentanil (2-3 mcg/kg/min.). The patients were randomized in two groups. The patients were ventilated with N2O:O2 (1:1) (Group I) or air: O2 (1:1) (Group II). After surgery, all patients of the Group II received Naloxone (0.2-0.4 mg). Central venous blood samples were obtained before induction, one and two hours after induction of anaesthesia and at the end of surgery. Separation of monocytes was performed according to Boyum technique. CD35 and HLA-DR molecules and vimentin filaments were studied by indirect immunofluorescence method using monoclonal antibodies. Percentage of positive cells were read with a cytofluorometer. The phagocytic function of monocytes was determined by ingestion of latex particles. Cortisol and ACTH plasma levels were determined by RIA. High doses of Alfentanyl depress phagocytic function and membrane display of CD35 and HLA-DR molecules in monocyte and induce marked changes in the organization of vimentin filaments in these cells in patients undergoing surgery. This monocytic depression was more marked in the patients ventilated with N2O. In our results there was uninhibition of ACTH and cortisol plasma levels responses to surgical stress by Alfentanil administration. Since the effects of Alfentanil were reversed by Naloxone, an opioid receptor mechanism seems to mediate these events.

[Spontaneous tumor regression. Report of 2 cases]. / Regresión tumoral espontánea. A propósito de dos casos.

Two cases of spontaneous tumor regression (STR) occurring in a patient with non Hodgkin lymphoma and in another patient with squamous carcinoma of the lung are presented. Both cases fulfill the criteria of STR defined by Everson and Cole. Recent results obtained in basic and clinical studies have indicated that immunological mechanisms could play an important role in STR. The mediator effects more frequently referred are: 1) generation of antineoplastic cytotoxic cells; 2) production of immunoregulatory cytokines by lymphocytes and monocytes, and 3) possible cross reaction between tumor and bacterial antigens. These mechanisms of action are discussed in relation to the presented cases.

Opioid peptides modulate the organization of vimentin filaments, phagocytic activity, and expression of surface molecules in monocytes.

It is theorized that intermediate filaments are important in the modulation of membrane activity and cell motility; however, their functions are unknown. The assembly and organization of these filaments are under hormonal regulation. We investigated in human monocytes the in vitro effects of Met-enkephalin, Leu-enkephalin, and beta-endorphin on the expression of immunoreactive cytoskeletal vimentin filaments. We simultaneously examined their effect on the phagocytosis of Candida albicans and on the membrane display of surface molecules. The three opioid peptides markedly reduced the expression of vimentin filaments, the phagocytic activity, and the display of HLA-DR molecules at concentrations of 10(-6), 10(-8), and 10(-10) M. On the other hand, the intravenous administration of fentanyl, a synthetic opiate agonist, to patients undergoing surgery induced similar changes in monocytes. In other experiments, 10(-8) M beta-endorphin also decreased the expression of CR3 but did not influence the display of CD13, a surface protein of unknown function. Expression of vimentin filaments correlated directly with the display of HLA-DR antigens and CR3 and with the phagocytic activity. The results of this paper indicate that opiates and opioids, neuropeptides known to be released during stress, can directly depress several monocyte functions. Furthermore, from these data it may be speculated that intermediate filaments may regulate the membrane expression of some surface molecules and the phagocytic process.

[Changes caused by staphylococcal protein A and gamma interferon on the natural cytotoxic activity in patients with cancer]. / Modificaciones por proteína A estafilocócica e interferon-gamma en la actividad citotóxica natural de pacientes con cáncer.

The effects on natural killer (NK) activity of in vitro incubation of effector cells with staphylococcal Protein A (SPA) and gamma interferon (gamma IFN) in patients with different types of tumors have been compared. The modifications induced in NK activity by these two immunomodulators were assessed using the K-562 cell line and autologous tumoral cells as targets of natural cytotoxicity. The results obtained show that both SPA and gamma-IFN are good inducers of NK activity against the K-562 cell line. SPA always shows itself more efficient than gamma-IFN in increasing the basal level of NK activity. The cytotoxicity against autologous tumoral cells was also greatly increased by the two immunomodulators assayed, SPA being also in this type of assay more efficient than gamma-IFN. Taken the data as a whole the increases induced by SPA and gamma-IFN are parallel both against K-562 and autologous tumoral cells. However when the different types of tumours are considered separately some discrepancies between NK activity against K-562 and autologous tumoral cells appear. The effects of SPA as a modifier of NK activity against autologous tumoral cells looks clear. In our experiments incubation of effector cells with SPA behaves more effectively than the well established gamma-IFN model in the induction of this type of activity. The intimate mechanisms by which SPA exerts its action are quite interesting and merit further investigation.

[Role of the contact plasma system in human physiopathology]. / Papel del sistema de contacto plasmático en la fisiopatología humana.

The possible implications of the plasma contact system in several human pathological conditions have been reviewed. The mechanism of activation and the biochemistry of the different components of this physiological system are described. The activation of the plasma contact system and its important physiological consequences are considered. The implication of the plasma contact system in the pathogenesis of different human and experimental pathological conditions (shock, disseminated intravascular coagulation, extracorporeal circulation...) is reviewed. A particular consideration is given to the pathogenesis of glomerulonephritis where the plasma contact system has been more directly implicated.

[Protein A: an immunomodulator in cancer patients]. / Proteína A: inmunomodulador en pacientes con cáncer.

Protein A defined as one of the components of the wall of Staphylococcus Aureus Cowans 1 has shown its effects as modifier of the immune response: it induce changes in cellular receptors, polyclonal activation of T and B lymphocytes, liberation of lymphokines, increase in the production of gamma-interferon (probably as the result of activation of NK cells). In the present work we have studied NK activity and its modifications by Protein A and gamma-interferon in blood of 50 patients with solid tumours. The method used was the Cr 51 liberation assay. The effectors cells were non-adherent lymphocytes treated with different doses and times of incubation of Protein A and gamma-interferon. As target cells we utilized the cellular line K-562 for NK activity and autologous tumoral cells isolated from surgical specimen for tumour-specific cytotoxicity. The results obtained allow us to state the following conclusions: 1. NK activity against K-562 cells is not always a specific parameter of such activity. 2. Tumour-specific cytotoxicity can vary according to the histological type of tumour. 3. Protein A is a potent inducer of tumour-specific cytotoxicity in a higher degree than gamma-interferon.

Lymphocyte populations and subpopulations in some lymphoproliferative dermatoses.

We present a study of lymphocyte populations and subpopulations as seen in histological cuts of skin with malignant and benign lymphoproliferative alterations using the immunological and histochemical techniques later described. Disclosed are the results of samples obtained of lymphocyte populations which were used to help differentiate between said alterations. It is proven that in T cell lymphomas. The infiltrate is dominated by markers for EA IgM which correlates with the results of other authors in peripheral blood.