Impact of CD34 subsets on engraftment kinetics in allogeneic peripheral blood stem cell transplantation.

Our objective was to evaluate, probably for the first time, the impact of CD34 subsets on engraftment kinetics in allogeneic PBSC transplantation (PBSCT). PBSC graft components were analyzed in 62 cases for the absolute count/kg of total CD34+ and the following subsets: DR- and +, CD71+/-, CD38+/-, CD33+/- and CD61+/-. Time to ANC >0.5 and >1 x 10(9)/l and platelets >20 and >50 x 10(9)/l was reported. The median value for each parameter was used to discriminate rapid from slow engraftment. Four parameters showed significant predictive power of early neutrophil engraftment, namely CD34+ /DR- (P = 0.002), CD34+/38- (P = 0.02), CD34+/CD61- (P = 0.04) and total CD34+ cell dose (P = 0.04). Four parameters showed significant predictive power of early platelet engraftment, namely CD34+/CD61+ (P = 0.02), CD34+ /CD38- and total CD34+ cell dose (P = 0.04) and CD34+ /CD71- (P = 0.05). Comparing patients who received > to those who received < the threshold dose(s), only CD34+ /CD38- lost its significance for neutrophil engraftment; and only CD34+ /CD61+ retained its significance for platelet engraftment (P = 0.03); furthermore, the former group required significantly fewer platelet transfusions (P = 0.018). We concluded that in allogeneic PBSCT, the best predictor of early neutrophil engraftment is the absolute CD34+ /DR- and for early platelet engraftment is the absolute CD34+ /CD61+ cell dose.

Cytochemical demonstration of 'platelet' peroxidase at the light microscope level.

A reliable and reproducible modified method was developed for the cytochemical demonstration of the so-called 'platelet' peroxidase at the light microscopy (LM) level. Interestingly, not only normal platelets and megakaryocytes showed the peroxidase activity, but also normal lymphocytes. As with the ultrastructural platelet peroxidase (PPO) method, myeloperoxidase (MPO) was also demonstrated in granulocytes. Peroxidase activity was exhibited by blast cells of acute myeloid leukaemia, including megakaryoblasts of megakaryoblastic leukaemia, by hairy cells of hairy cell leukaemia, by centroblasts of centroblastic lymphoma and by plasma cells of plasma cell leukaemia. The enzyme activity was also demonstrated in the mast cells of systemic mastocytosis. It seems that different forms of peroxidase are present in most haemic cells. Fixation and conditions of incubation are probably the determining factors for their demonstration. The method may allow further cytochemical discrimination at the LM level between blast cell populations which are negative for MPO by standard methods.

Cytochemistry of gamma-glutamyltransferase in haemic cells and malignancies.

An improved cytochemical method for the demonstration of gamma-glutamyltransferase is described. The enzyme was demonstrated in almost all normal leucocytes and in platelets. There was markedly reduced activity in most lymphoproliferative disorders. In a single case of Hodgkin's disease, T-lymphocytes showed slight to moderate activity contrasting with the marked activity displayed by Reed-Sternberg cells. The plasma cells of multiple myeloma and plasma cell leukaemia showed activity equal to or stronger than that of their normal counterparts. In acute myeloid leukaemia the positivity varied widely in blast cells, but was consistently increased in monoblasts of acute monoblastic leukaemia. gamma-Glutamyltransferase may serve as a differentiation marker in the study of granulocytic and lymphocytic cell lineages.

Histochemistry of dipeptidyl aminopeptidase (DAP) II and IV in reactive lymphoid tissues and malignant lymphoma.

A modified histochemical method was used to show the presence of dipeptidyl aminopeptidase (DAP) II and IV in fixed, freeze dried, cryostat sections of tonsils, lymph nodes, and skin. In 14 reactive tonsils and lymph nodes both enzyme reactions were largely confined to T dependent areas where scattered positive lymphocytes were shown in the paracortical zones, while lymphocytes of germinal centres (B dependent areas) were negative. In either site some macrophages showed strong positivity for both enzymes. In 23 lymph node and two skin biopsy specimens of non-Hodgkin's lymphoma the neoplastic lymphocytes of 12 B cell lymphomas were completely unstained, whereas in the 13 cases of T cell lymphoma the neoplastic lymphocytes showed variable reactions with positivity for DAP II in eight and for DAP IV in seven, both reactions being positive in four and negative in two. Touch imprints of a lymph node from a case of Hodgkin's disease showed that the Reed-Sternberg cells were unreactive for both enzymes. The histochemistry of DAP II and IV may supplement other histochemical and immunological markers in the cytological classification of lymphomas.

Cytochemical method for demonstrating magnesium-activated adenosine triphosphatase in haemic cells.

A modified cytochemical method was developed for demonstrating magnesium-activated adenosine triphosphatase in haemic cells. The enzyme was shown in a variety of normal and leukaemic, unfixed cells. The high sensitivity of this enzyme to all fixatives tried in this study hampered further exploration and characterization of this enzyme.

Comparative cytochemical study of dipeptidyl aminopeptidase (DAP) II and IV in normal and malignant haemic cells.

Modified cytochemical methods were used to show dipeptidyl aminopeptidases (DAP) II and IV in peripheral blood buffy coat preparations and bone marrow smears. In 23 normal buffy coats both enzymes were confined to lymphocytes. DAP II was found in T and B lymphocytes (about 80%) while DAP IV was restricted to T lymphocytes only (around 46%). In 11 normal bone marrows DAP II was found in 53% of the lymphocytes, as well as in plasma cells, macrophages, and occasional myeloblasts. DAP IV was found only in lymphocytes (around 32%). DAP II activity, but not DAP IV activity, was present in all of the mast cells in a case of systemic mastocytosis. Whereas DAP II was found, to a variable extent, in leukaemic myeloblasts, monoblasts, proerythroblasts, and in megakaryoblasts in 52 cases of acute myeloid leukaemia, DAP IV was not shown. Variable positivity to DAP II and DAP IV was found in the lymphoblasts in seven cases of acute lymphoblastic leukaemia, in 14 cases of B chronic lymphocytic leukaemia, and in three cases of non-Hodgkin's lymphoma. DAP II activity was variable compared with DAP IV activity, which was constantly reduced. Virtually all of the myeloma cells (96%) all of the myeloma cells (96%) in five cases of multiple myeloma and two cases of plasma cell leukaemia were DAP II positive and DAP IV negative. In 10 cases of hairy cell leukaemia most hairy cells were positive to DAP II (74%) with no demonstrable DAP IV activity. In a single case of Sézary's syndrome around 90% of the helper T cells were positive to DAP II with no DAP IV activity.