Phase I/II trial of autologous activated macrophages in advanced colorectal cancer.

Autologous activated macrophage (AAM) therapy is an adoptive cellular therapy based on ex vivo differentiation and activation of autologous peripheral blood monocytes. This study was undertaken to evaluate the tolerance, efficiency and biological effects of AAMs in chemoresistant progressive colorectal cancers. From January 1993 to May 1995, 15 patients were treated. Mononuclear cells were collected six times by weekly apheresis, cultured for 7 days, and activated with interferon-gamma. AAMs were then separated by elutriation and re-infused intravenously, with a mean total of 7.95 x 10(9) macrophages per patient. Clinical tolerance was good: toxicity consisted only of a World Health Organisation grade 2 fever after 28% of the infusions. Responses were not seen in the 14 evaluable patients, as expected with very bulky tumours: in 11, the tumours continued to progress, but disease was stabilised in 3 patients who experienced progression-free survival for 14, 12 and 12 weeks, respectively.

[Peripheral stem cell collection, search for predictive factors: a multicenter study. Working Group on Transfusion and Therapeutic Techniques of the French Blood Transfusion Society ]. / Collection de cellules souches périphériques, recherche de facteurs prédictifs: une étude multicentrique. Groupe de travail Technique transfusionnelle et thérapeutique de la SFTS.

AIMS: A multicentric study involving 12 centers was made to investigate the results of peripheral stem cell collection carried out between 1996 and 1997 from 655 patients with hemopathic syndromes or malignant tumors, The aim of this investigation was to determine the predictive factors for transplant quality, and to thereby optimize collection procedures. PATIENTS AND METHODS: Information sheets were completed for 1,346 cytapheretic sessions, i.e., 655 grafts. The samples were taken after induction chemotherapy and exposure to hematopoeitic colony-stimulating growth factors (except the LMCs). Each graft was defined as being of good or bad quality depending on the number of CD34+ cells that it contained. Based on the data available in the literature, a workgroup consensus was reached that a level of CD34+ cells +/- 2.10(6)/kg recipient body weight constituted a good transplant criterion. The 2 subgroups (good graft versus lower quality graft) were compared by univariate analysis followed by discriminant multivariate analysis. RESULTS: It was established that a number of parameters were significantly linked to the criterion of collection quality; however, 3 predictive factors emerged from the multivariate analysis--the level of circulating CD34+ cells; the number of cytaphereses; the number of blood volumes treated. CONCLUSION: It was concluded that the level of circulating CD34+ cells seems to be an essential aspect in predicting the quality of the transplant and the number of cytaphereses required to obtain a sufficiently rich collection. Moreover, it also appears that at least 2 blood volumes should be treated to optimize the results.

[Effect of contrast media products on the erythrocyte aggregation and deformability in vitro]. / Influence des produits de contraste sur l'agrégation et la déformabilité erythrocytaire in vitro.

It has been suggested that contrast media could interfere with red blood cell aggregation, hyperosmolar media leading to an inhibition of red blood cell aggregation whereas non ionic products might induce a sludge phenomenon. We present an in vitro study accompanying two contrast media: 1) Ioxitalamate of sodium and meglumin (ionic, hyperosmolar). 2) Iopaminol (non ionic). In their effect on hemorheological parameters of red blood cell aggregation. Blood samples have been obtained from 7 healthy donors. Contrast media have been tested at increased contrast media (O.1, 1, 2, 10, 100 mg/ml of Iodine in final concentration). The following parameters have been studied: hematocrit, fibrinogen level, erythrocyte aggregation using the Erythro-aggregometer*, whole blood viscosity at 3 different shear rates (0.87, 18.74, 118 sec.-1) using Low Shear 30*. Deformability of red blood cell was assessed by ektacytometry. Osmolarity was controlled in each sample. Results show an inhibition effect of both contrasts media on red blood cell aggregation. There is a concomitant decrease of blood viscosity at low shear rates. On the contrary, apparent viscosity increases at high shear rates in parallel with the contrast media concentration. This effect is more pronounced with ioxitalamate above a concentration of 10 mg/ml. Ektacytometric parameters are not modified by contrast media and this could indicate a complete reversibility of the media-induced alteration on red blood cell. In order to precise the prothrombotic effect on contrast media, hemorheological studies have to be completed by the assessment of their effect on hemostatic parameters.

[Effects of contrast media products on erythrocyte aggregation. An ex vivo study]. / Influence des produits de contraste sur l'agrégation érythrocytaire. Une étude ex vivo.

It has been suggested (Fisher and al., Rainiko and al.) that contrast media could interfere with a red blood cell aggregation, low-osmolarity and non ionic contrast media leading to red blood cell aggregates in vitro. Clinical significance of this phenomenon is not known. We have studied 21 consecutive patients admitted in X ray department for pyelography. Sodium and meglumine ioxitalamate (Telebrix 38 R; Guerbet) osmolarity 2 100 mOsm kg--viscosity 8.5 cp at 37 degrees C is used for contrast. Red blood cell aggregation is studied by erythraggregometer Sefam. Samples are collected before (T0), 10 and 30 minutes (T10, T30) after contrast infusion. Hemogram and coagulation parameters are measured in parallel. Results show a significant increase of aggregation times at T10, with a trend towards baseline at T30. Dissociation shear rates follow the same fluctuations. We note a concomitant diminution of both fibrinogen and hematocrit levels, well-correlated with red blood cell aggregation. These results suggest a process of hemodilution which could explain the diminution of red blood cell aggregation with this type of contrast medium.

[Interference of anaesthesia and transfusion on rheological parameters (author's transl)]. / Interférence de l'anesthésie et de la transfusion sur les paramètres rhéologiques.

This study was designed to investigate the rheological and plasmatic parameters from multiple transfused patients and patients undergoing surgery non transfused. Blood filtrability compared with the erythrocyte electrophoretic mobility shows the perturbance resulting from the delay of conservation and the importance of the transfused blood. The 2-3 DPG, ATP and oxygen-hemoglobin affinity suggest the excellent recuperation of the erythrocyte metabolic function in vitro. The rheological parameters seems to be a good mean in the supervision of red massive transfusion blood cell.

[Contact (prekallikrein) and fibrinolytic (plasminogen/antiplasmin value) system in the physiopathology of vascular complications of diabetes]. / Apport de l'étude du système contact (prékallicréine) et fibrinolytique (rapport plasminogène/antiplasmine) à l'approche physiopathologique des complications vasculaires du diabète.

Both micro and macro-angiopathy involve interaction of blood vessel and plasma factors which may be altered in diabetes. Little is known concerning changes in contact and fibrinolysis factors as prekallikrein, plasminogen and fast antiplasmin in diabetes. We tested also the plasminogen/antiplasmin ratio in 151 diabetics and 64 normal subjects (18 to 74 years old). Our conclusion is:--the absence of correlation between the clinical characteristic of diabetes and prekallikrein level,--the decrease of plasminogen/antiplasmin ratio in complicated diabetes,--in complicated diabetes, correlation between antithrombin III, VIII factor and fibrinogen levels, perhaps, in relation with inflammation signs.

[Biological diagnosis of postoperative thrombopenia]. / Diagnostic biologique d'une thrombopénie post-opératoire.

The recognition of the postoperative thrombopenia is important because thrombopenia give an hemorrhage risk and modify the tolerance to preventive anticoagulant treatments. Some thrombopenias can be iatrogenic: thrombopenias by transfusion of a large volume of preserved blood, or by hemodilution, but require a substitution, therapy are easily diagnosed. Post-transfusional thrombopenias require an antibody analysis (especially for antiPLA1). The search for drugs interactions is often complex; heparin induced thrombopenia constitutes a severe but fortunately a rare complication of the heparin therapy. Other thrombopenias are related to operative complications this is the case of consumption-coagulopathies due to infections, the release of thromboplastin from tissues, hepatic cirrhosis, pancreatitis, etc. The evaluation of hemostasis verifies clinical hypotheses and guides the treatment. Thrombopenias can be due to various disorders revealed or occurring during an operation. Although the concurrences are rare, they do not always preclude the possibility of finding a collagen disease (connectivitis), a thrombocytogenetic thrombotic purpura, and especially an idiopathic thrombopenic purpura. In any case, diagnosis is easier if the preoperative platelet levels are known. Thus, platelet counts should be included in pre and postoperative evaluations.

[Post-transfusion thrombopenia treated with gammaglobulins]. / Thrombopénie post-transfusionnelle traitée par les gammaglobulines.

Thrombopenia and a haemorrhagic syndrome occurring on the eighth postoperative day were probably related to a post-transfusion purpura, confirmed by the discovery of antiplatelet antibodies not belonging to the PLA1 antigen system. The stabilization of this haemorrhage and a full recovery were obtained by giving high doses of intravenous human gammaglobulins. Blood replacement only rarely induces postoperative immuno-allergologic thrombopenia; heparin is the best known causative agent. It is concluded that platelets should never be given so long as the responsible agent is not formally identified.

Comparative interest of two coaxial viscometers: ecktacytometer and low shear 30.

We have studied some rheological parameters in different clinical status by using two coaxial viscometers. The Contraves Low Shear sinus 30 allows available shear rates range from gamma (128s-1 to 0,0175 s-1), and the parameters studied were viscoelastiticy, thixotropy and blood viscosity. The Ecktacytometer+ is another method for measuring cell deformation in couette flow. The aim of our study was to compare these two technics, to consider their clinical interest and finally to visualize an eventual correlation.

Separation of dendritic cells from highly purified human monocytes by counterflow centrifugation induces tissue factor expression.

BACKGROUND: In vitro generation of dendritic cells (DCs) from human monocytes represents a promising tool in immunotherapy. However, it is not known whether the separation of DCs from monocytes induces tissue factor expression and therefore may trigger coagulation in patients receiving these DC preparations. The aim of this study is thus to analyze tissue factor expression on monocyte-derived DCs and to compare their ability to trigger thrombin generation to that of macrophages obtained from the same monocytes. STUDY DESIGN AND METHODS: Human monocytes are separated by leukapheresis and washed by using counterflow centrifugation in sterile, endotoxin-free conditions. Macrophages are grown from human monocytes in the presence of GM-CSF alone and immature DCs are grown in the presence of GM-CSF plus IL-4 for 5 days with fetal calf serum (IDC-FCS). Immature DCs are also grown from human monocytes for 7 days in the presence of GM-CSF plus IL-4 with human group AB serum (IDC-HS). The addition of prostaglandin E(2) and TNFalpha in this culture medium at Day 5 leads to mature DCs (MDC-HS). Tissue factor mRNA expression is studied by RT-PCR analysis. Tissue factor antigen is measured by ELISA in cell lysates and by direct flow cytometry. The procoagulant activity of intact cells is assessed by using an amidolytic assay or a chronometric assay. RESULTS: IDC-FCS express tissue factor mRNA and antigen and trigger thrombin generation. Procoagulant activity of IDC-FCS is dependent on both tissue factor expression and exposure to anionic phospholipid. Monocyte-derived macrophages cultured for 5 days with GM-CSF alone express lower levels of tissue factor mRNA, tissue factor antigen, and procoagulant activity than IDC-FCS. IDC-HS and MDC-HS also express high levels of tissue factor mRNA and antigen and support procoagulant activity. CONCLUSION: Monocyte-derived DCs express a high level of functional tissue factor and support procoagulant activity. This finding should be taken into account in clinical trials.

[Serum antibodies against hepatitis C: study of 4,100 blood donors from the northeast of France]. / Anticorps sériques contre le virus de l'hépatite C: étude de 4,100 donneurs de sang du nord-est de la France.

Between June 1989 and February 1990 the prevalence of antibodies to hepatitis C virus (anti-HCV) was studied in blood donations from 4,100 donors tested in north east France. 37 samples were found reactive for anti-HCV (prevalence of anti-HCV: 0.90%) without any significant difference in sex (males: 0.94%; females: 0.87%) and age distributions. 178 (4.34%) of the 4,100 donors were found anti-HBc positive, 5 of these donors being anti-HCV positive (13.51% of all anti-HCV positive donors). 52 donors (1.27%) had raised alanine aminotransferase (ALT) levels (greater than or equal to 2 N: 2 times the M +/- 2 SD value): 3 were found anti-HCV positive (8.11% of all anti-HCV positive donors). Association of the 2 surrogate markers is poorly sensitive since it detects only 8 (21.62%; males: 4, females: 4) of all anti-HCV positive donors. Furthermore, it appears weekly specific since it discards 230 blood samples of which 222 (96.52%) were anti-HCV negative. The 2 surrogate markers are complementary to one another and none of the anti-HCV positive donors had both anti-HBc antibodies and raised ALT. The mean ALT level is significantly higher in anti-HCV positive donors as compared to seronegative (M +/- 1 SD: 51 +/- 82 U/l versus 24 +/- 17 U/l). In anti-HCV positive donors, a marginal (r = 0.34) though statistically significant (p less than 0.05) positive correlation was found between ALT level and anti-HCV ratio.(ABSTRACT TRUNCATED AT 250 WORDS)

Adherent-free generation of functional dendritic cells from purified blood monocytes in view of potential clinical use.

There is increasing interest in dendritic cells (DC) that are capable of initiating antitumor immune responses. An in vitro cell differentiation method has recently been developed that uses GM-CSF and IL-4 to generate human DC from adherent blood mononuclear cells cultured on tissue culture plastic. These cells are competent for antigen uptake but express relatively low levels of co-stimulatory molecules and thus correspond to immature resident tissue DC. We have adapted this method to consider some variables that are pertinent to clinical use, including a large scale differentiation of functional DC in a culture system suitable for clinical use. We report here that sizable numbers of monocytes purified by elutriation from blood leukocytes and cultured in Teflon bags develop with high efficiency into typical DC, as defined by morphology and membrane phenotype. When compared with usual adherent DC, cells generated under our adherent-free conditions exhibited lower CD1a expression and antigen capture capacity, but maintained the ability to present soluble antigens to T cells. They neoexpressed a high level of the co-stimulator molecule B7-2 (CD86) and was potent accessory cells for T cell proliferation, but they lacked the CD83 marker of DC full maturation. This study may constitute a prerequisite step for clinical investigations in tumor immunotherapy.

[Autologous haematopoietic stem cell transplantation. Preliminary results of a regional multicentric study (author's transl)]. / Greffe de cellules-souches hématopoïétiques autologues. Premiers résultats d'un protocole régional (author's transl)]

Autologous bone marrow transplantation represents a new approach to the treatment of malignant diseases when conventional therapy has failed. For this reason, the authors have collected bone marrow from 46 patients, including 24 with acute leukaemia, 7 with chronic myeloid leukaemia, 10 with lymphosarcoma and 5 with solid tumours. The mean of total cryopreserved CFU-c was 8.5 X 10(6) (range: 0.2-25). Ten cases of autologous bone marrow transplantation are reported. Seven patients had been prepared with high dosage chemotherapy alone (TACC) and three with chemotherapy combined with total body irradiation. Haematopoiesis restarted within 9 to 15 days in 5 patients and within 22 to 34 days in the other 5. Complete remission was obtained in all 5 patients with acute myeloid leukaemia grafted during their first relapse, the longest remission up to now being 390 days. One patient with chronic granulocytic leukaemia is still in second chronic phase after 360 days. Stem cells were transplanted early in the course of a T-lymphosarcoma, during complete remission; maintenance chemotherapy was withdrawn, and the chances of success of this treatment alone are being evaluated. The kinetics of blood and bone marrow CFU-c populations after transplantation were studied in 4 cases and were found to correlate closely with haematopoietic recovery following ablative bone marrow therapy. Stem cell transplantation can only be justified in acute leukaemia if it is carried out immediately after complete remission to consolidate the results and, hopefully, to prolong the remission.

Autologous bone marrow transplantation in acute myeloid leukemia in relapse or in complete remission.

We report ten cases of acute myeloid leukemia treated by intensive therapy followed by autologous bone marrow transplantation. Seven patients were in first relapse, and three were in complete remission. The conditioning regimen consisted of either chemotherapy alone (6-thioguanine, cytarabine, lomustine, and cyclophosphamide [TACC; eight patients]) or cyclophosphamide and total-body irradiation (two patients). All the patients in first relapse achieved complete remission (CR). The median remission duration was 9.9 months (range, 5-14), and the median survival was 14.4 months (range, 9-23.8). Of the three patients autografted during CR, one relapsed at Month 5 and two others remain in CR and are well at 18+ and 18.3+ months. High-dose chemotherapy followed by autologous bone marrow transplantation seems to be a valuable treatment for acute myeloid leukemia, if it is used immediately after the CR to consolidate the remission.