[Culturing of normal and tumor cells of the human prostate]. / Culture de cellules normales et tumorales de prostate humaine.

Cell lines of neoplastic human prostate gland tissue are uncommon. We studied the condition of long term prostatic cell culture; 18 primary cultures of epithelial prostatic cells and 15 subcultures were maintained for several months. The optimum medium for primary prostatic cell cultures contains: little calcium, epithelial growth factor (EGF), phosphoethanolamine, spermine and cholera toxin, the cell characterisation for cultivation of normal, hyperplastic and neoplastic prostate gland tissue in subcultures uses monoclonal antibody against cytokeratin, prostatic acid phosphatase and prostatic specific antigen.

[Initial results in man of immunolymphoscintigraphy of cancer of the prostate]. / Premiers résultats chez l'homme de l'immunolymphoscintigraphie dans le cancer de prostate.

The loco-regional investigation of carcinoma of the prostate usually comprises ilio-obturator lymphadenectomy. This procedure carries a significant morbidity. Immunolymphoscintigraphy may provide a non-invasive alternative to this operation. Monoclonal (MAB) 227 A anti-prostatic acid phosphatase antibodies have been produced and selected for their affinity and specificity. This MAB was fragmented to its F (ab')2 form and marked with Iodine 123. Fifteen patients with prostate cancer were given 100 to 400 micrograms of the MAB by periprostatic perineal injection. The region was scanned 1 hr, 3 hrs, 6 hrs and 24 hrs after the injection. The results of the immunolymphoscintigraphy were compared with the histology of the ganglia removed at surgery or needle biopsy guided by CT scanning. In 13 cases the results were concordant for the two techniques (10 negatives and 3 positives). Two patients showed extra-prostatic fixation whilst the histology remained negative. Immunolymphoscintigraphy may therefore provide a simple method of detecting local metastases of carcinoma of the prostate if these initial results are confirmed.

[Monoclonal anti-prostatic acid phosphatase antibodies (production and characterization)]. / Anticorps monoclonaux anti-phosphatase acide prostatique (production et caractérisation).

5 monoclonal antibodies have been produced by Köhler and Milstein. The production and the characteristics of the monoclonal antibodies are presented. They are all IgG1 and have high specificity against prostatic cells producing prostatic acid phosphatase. These monoclonal antibodies can be used to perform immunoscintigraphies and immunolymphoscintigraphies to titrate prostatic acid phosphatase serum levels and to identify the prostatic origin of a tumor.

[Analysis of various isoferritins with monoclonal antibodies]. / Analyse de différentes isoferritines par des anticorps monoclonaux.

Balb/c Mice were immunized with splenic isoferritin. Spleen cells from immunized Mice were fuzed with SP2O myeloma cells. Four monoclonal antibodies designated respectively M29, M211, M386 and B8 were selected. Various isoferritins were analysed by immunodiffusion (Ouchterlony technic). With these monoclonal antibodies and with Rabbit polyclonal sera: human basic and acidic isoferritins and Horse spleen ferritin. Ferritin could be precipitated by these monoclonal antibodies. These results confirm that the ferritin molecule consists of repeating antigenic sites. No immunoreactivity difference could be detected between acidic and basic human ferritine. Species specificity was recognized. The high affinity constant of the M29 monoclonal antibody allowed development of a standardized radioimmunossay of ferritin.

Granulocyte progenitor compartments after allogeneic bone marrow grafts.

Bone marrow granulocyte colony forming cells (CFU-C) were measured in HLA compatible sibling donor-recipient pairs. The regeneration of granulocytes and CFU-C compartments were also studied in order to evaluate haemopoietic recovery. The number of nucleated bone marrow cells in the donation was 23 +/- 4 X 10 cells, which recipients received (3.5 +/- 0.4) X 10(8) nucleated cells/kg and (1.19 +/- 0.32) X 10(5) CFU-C/kg. This produced a bone marrow reconstitution of (67 +/- 26) X 10(5) CFU-C/kg by day 30. There was a significant correlation between CFU-C/kg and (1) granulocyte count on day 30 (P equal less than 0.05) and (2) the first day of reappearance of neutrophils in the blood (P equal less than 0.05). These results indicate that the speed and completeness of granulocyte regeneration can be predicted by measurement of the size of the CFU-C inoculant in the bone marrow graft.

Survival of CFU-C in recipient blood after bone marrow transplantation.

In six patients undergoing allogeneic bone marrow transplantation granulocyte colony and cluster forming units were detectable in the blood up to 24 hours after intravenous marrow infusion. The mean surviving fraction 30 minutes after the end of the infusion was 0.3 for colonies and 0.42 for total aggregates. CFU-C declined logarithmically with time, but a small secondary rise in blood CFU-C occurred 2--6 hours after the end of the marrow infusion. Three patients were plasma-exchanged immediately before the marrow graft for major donor-recipient ABO incompatibility. The proportion of total groups remaining in the blood at 30 minutes was significantly higher in the plasma-pheresed patients (P = less than 0.05), and the secondary rise in CFU-C was less marked, but no correlation of the blood CFU-C decay curve with the subsequent fate of the graft was observed.

Bone marrow culture in aplastic anemia.

Blood and bone marrow granulocyte colony forming units (CFUc) were assayed in 46 patients with aplastic anemia, and the serum was examined for its inhibitory action on normal CFUc growth. All patients showed a gross reduction in colonies and clusters in incidence and absolute number in the bone marrow and blood. Two proliferative abnormalities of CFUc in aplastic anaemia were identified: a significantly higher than normal cluster to colony ratio (P less than 0.05) and a higher than normal ratio of granulocytes to total aggregates in the bone marrow. Eleven out of 34 patients tested had serum inhibitory to normal CFUc. These patients were indistinguishable from the rest on haematological and CFUc culture characteristics, and no correlation between the results of CFUc assay and haematological severity was found. The results suggest that the CFUc is abnormal in aplastic anaemia, the reduction in pool size being related to a failure of self-renewal, but an immunological role in the pathogenesis of aplastic anaemia remains unproven. The close relationship of CFUc incidence to the percentage of granulocyte precursors in the marrow, together with the failure of the CFUc assay to predict clinical severity, limits the practical use of the assay to the confirmation of diagnosis in aplastic anaemia.

Effect of antilymphocyte globulin on granulocyte precursors in aplastic anaemia.

Granylocyte colony forming units (CFU-C) were studied in 22 patients with severe aplastic anaemia before and after treatment with antilymphocyte globulin (ALG). Nine patients showed a clinical response to ALG characterized by a rise in the granulocyte count to over I X 10(9)/1 within 30 d. These patients were distinguished in vitro from non-responders by an increase in CFU-C numbers after incubation of bone marrow cells with ALG, and by the presence of inhibitors of normal CFU-C in the serum in six out of seven patients tested. In responding patients bone marrow CFU-C rose while most non-responding patients showed no change or a fall in CFU-C after treatment. In addition in three out of four responding patients examined serum inhibitors disappeared after treatment. The horse ALG used in this study also stimulated normal CFU-C in vitro. This evidence is contrary to the hypothesis that ALG stimulates CFU-C in aplasia by inactivating an abnormal suppressor lymphocyte population. The nature of the stimulation by ALG remains unclear. But in practice the effect of ALG on bone marrow cells and study of CFU-C inhibitors in serum could be used to select patients likely to respond to ALG treatment.

Variations in granulocyte colony forming cell numbers in adult blood.

Blood granulocyte colony forming units (CFU-C) were studied in normal adults to establish: (i) a normal range; (ii) variability due to the culture technique; (iii) variability of blood CFU-C within individuals. Thirty men studied had 98 (range 8--300) CFU-C X 10(3)/1, and 28 women studied had 44 (range 0--260) CFU-C X 10(3)/1. This difference was significant (P less than 0.001). There was also a significant sex difference in the total number of cells forming colonies and clusters per litre; and in the incidence of colony formers and of cluster formers in buffy coat and mononuclear cell blood fractions. CFU-C were assayed in four subjects over a 10 week period. When buffy coat cells were used as a source of colony stimulation the week to week variation in the combined growth of the four subjects was wide (+/- 36%) but with conditioned medium the variation was smaller (+/- 14%). In all subjects colony and cluster growth varied in the same way (r = 0.77, P = .001) but there was no correlation with the total leucocyte count. A 3--4 week cyclical change in CFU-C/1 was found which was independent of the variation inherent in the technique. The physiological significance of the sex difference and the apparent cyclical changes in blood CFU-C are not explained, but the results emphasize the wide fluctuations in CFU-C that may occur in normal individuals.