Metabolic activity of phagocytosing granulocytes in chronic granulocytic leukemia: ultrastructural observation of a degranulation defect.

The functional capacities of granulocytes in patients with chronic granulocytic leukemia are still a subject of controversy, probably due to the heterogeneity of the abnormalities observed from patient to patient. For a better definition of these abnormalities, 14 patients with untreated chronic granulocytic leukemia were studied. The patients were divided into three groups on the basis of the functional activities of their phagocytosing granulocytes. In four patients (group I), the granulocytes were normal in respect to particle ingestion, nitroblue tetrazolium (NBT)-stimulated reduction, cyanide-insensitive oxygen (O2) consumption, superoxide anion (O2-)-stimulated production, hydrogen peroxide (H2O2) production, and iodination. They also had a normal myeloperoxidase (MPO) content. In four patients (group III), the granulocytes were significantly defective in all of these activities. In the six remaining patients (group II), all the initial metabolic steps of the phagocytosing granulocytes (ingestion, NBT reduction, O2 consumption, O2-production, H2O2 production) were normal, as were the MPO content of the granulocytes, while iodination was strikingly decreased. These metabolic features suggested a degranulation defect which was observed ultrastructurally in the only patient studied among these six. The phagocytosing granulocytes of this patient did not degranulate and no deposits of MPO activity were seen in the phagosomes.

Bilirubin and paranitrophenol glucuronyl transferase activities and ultrastructural aspect of the liver in patients with chronic hemolytic anemias.

The hepatic ultrastructural aspect and the hepatic bilirubin (Bil-GT) and paranitrophenol (PNP-GT) glucuronyl transferase activities were studied in twenty subjects with a chronic hemolytic anemia (HA) and five subjects with an aplastic anemia. In chronic HA: the hepatic ultrastructural aspect showed mitochondrial abnormalities affecting structure (paracrystalline inclusions), size (giant mitochondria) and shape (irregular mitochondria); there was a decrease of Bil-GT activity in 80 percent of the patients. No connection could be shown between the HA etiology and the degree of the enzymatic activity decrease. The physiopathological mechanism of this enzymatic activity decrease is unknown; no deficiency of PNP-GT was observed; in chronic aplastic anemias there does not seem to be any modification, either of Bil-GT activity, or of hepatic PNP-GT activity. It is concluded that Bil-GT is often decreased in HA and that neither Bil-GT measurement nor the ultrastructural aspect of the liver distinguish chronic HA from Gilbert's syndrome.

Bilirubin and paranitrophenol glucuronyl transferase activities of the liver in patients with Gilbert's syndrome An attempt at a biochemical breakdown of the Gilbert's syndrome.

Hepatic bilirubin (Bil-GT) and paranitrophenol glucuronyl transferase (PNP-GT) activities were measured in 26 subjects with Gilbert's syndrome (GS) and in one subject with a Crigler-Najjar type 2. Firstly, the results allowed us to distinguish three groups of patients in GS. In the first group, Bil-GT activity decreased by 25% of that of the controls and PNP-GT activity was normal. In the second group, Bil-GT decreased by 25% of that of the controls and PNP-GT decreased by 50% of that of the controls. In the third group, Bil-GT decreased by 50% of that of the controls and PNP-GT activity was normal. Secondly, the results showed in the Crigler-Najjar type 2 that Bil-GT activity was 25% of that of the controls and PNP-GT was 10% of that of the controls. From these results the following hypothesis has been raised: (1) the subjects of the third GS group were probably heterozygous to the homozygous disease which affected the subjects of the first GS group, and (2) the subjects of the second GS group were most likely heterozygous to the homozygous disease which affected our Crigler-Najjar type 2. However, in the present state of our knowledge, the scheme of GS classification which we propose requires confirmation.

Quantitative iodination of human blood polymorphonuclear leukocytes.

It was shown by Pincus and Klebanoff that a correlation existed between leukocytic iodination measured in vivo and microbicidal leukocytic activity. We have analyzed the results of this test in relation to time and in the presence of variable quantities of polymorphonuclear leukocytes (PMN). The values observed per time and PMN unit proved to be equivalent in the presence of 2.5 X 105 PMN or 5.0 x 105 PMN per 0.5 ml of incubation medium, measured after 10, 20 and 30 minutes or in the presence of 1.0 x 106 PMN, measured after 10 minutes. That is to say iodination is proportional to leukocyte concentration and incubation time. Increase of either the quantity of cells or the incubation time, beyond the area we defined, reduced iodination per cell and per unit of time. Concerning the patients with an insufficient iodination, we have studied 2 parameters in the presence of 5.0 x 105 PMN: 1) initial iodination measured after 10 and 20 minutes and 2) stability of iodination measured after 60 minutes. These two parameters were equally affected in two cases with myelofi-rosis, 3 patients with acquired refractory anaemia, one with chronic lymphoid leukaemia, one with erythroleukaemia, one with hairy cell leukaemia, one with systemic mastocytosis and almost complete myeloperoxidase dificiency, one with sickle cell disease, two with liver diseases and two with chronic myeloid leukaemia. The iodination at the 60th minute was more affected than at the 10th minute with a patient with myelofibrosis and 4 other patients with acquired refractory anaemias. The significance of these differences is not well understood; however the meaning of the decrease in the iodination of whatever type is that a PMN anomaly exists directly related to the myeloperoxidase H2O2 halogenation system, or to one of the stages of engulfment and/or metabolic events preceeding it and leading to the production of H2O2. This test, with the alterations we introduced, is suggested as a test for detection of functional PMN abnormalities.