Nicoletta Polli

Cushing's Disease and Pregnancy

The placenta behaves as an important endocrine organ and is responsible for several changes in the hypothalamo-pituitary-adrenal (HPA) axis. The placenta produces both CRH and ACTH in progressively increasing amounts throughout gestation [1,2] but HPA axis homeostasis is protected by a parallel increase in CRH-binding protein [3]. ACTH, in turn, stimulates adrenal cortisol secretion and plasma and urinary free cortisol levels rise accordingly but are buffered by an increased hepatic production of cortisol-binding protein [2,4]. Cortisol secretion during pregnancy follows the usual pulsatile and circadian rhythmicity [5] even during the third trimester of gestation when levels may reach twice the normal limit [4]. The HPA axis, however, becomes somewhat resistant to physiologic modulators. Indeed, 1 μg/kg CRH is insufficient to evoke significant ACTH and cortisol responses in the third trimester of gestation [6] and higher doses (2 μg/kg) are needed [7]. Moreover, pregnant women usually fail to adequately suppress cortisol secretion after low dose dexamethasone administration suggesting a resetting of maternal feedback setpoint [4].

Selective purging by human interleukin‐2 activated lymphocytes of bone marrows contaminated with a lymphoma line or autologous leukaemic cells

Summary The ability of recombinant interleukin 2 (rIL2) activated lymphocytes (LAK) to purge BM samples contaminated by tumour cells was evaluated. Human BM mononuclear cells were contaminated with 10% of the lymphoma line CA46 and then cultured in liquid medium containing 1000 U/ml of rIL2 and/or LAK autologous to the used BM. At the end of coculture the growth of residual tumour cells and of CFU‐GM were evaluated by clonogenic assay. No tumour cell growth was observed in 5/5 independent experiments after 18 h of coculture with LAK. No significant inhibition of CFU‐GM growth was also noted. Subsequently, the effect of LAK on BM obtained from four leukaemic patients and contaminated with 20–50% of their own AML and ALL cells was studied using MAb as a tool for identifying leukaemic cells. LAK eliminated 24–78% of contaminating cryopre‐served uncultured autologous leukaemic cells. In five cases the BM was contaminated by a low (2%) amount of ALL cells. In these patients the monoclonal heavy chain rearrangement typical of ALL was no longer visible after coculture with LAK. Evidence for selective tumour cytotoxicity by LAK was confirmed by using autologous BM cells as hot and cold targets in a 51Cr release assay. Finally, successful haematologic reconstitution of lethally irradiated BALB/c mice was obtained using syngeneic BM cocultured with LAK. These results support the investigational use of rIL2 and LAK in the treatment of human leukaemia.

Distinct morphophenotypic features of chronic B-cell leukaemias identified with CDlc and CD23 antibodies

Abstract: Morphological criteria usually applied to diagnose various subtypes of B‐cell chronic lymphoid leukaemia are largely subjective. Immu‐nophenotyping of 61 relevant cases using a selected panel of monoclonal antibodies (mAb), showed that CDlc and CD23 mAb were able to separate B‐cell chronic lymphocytic leukaemia (B‐CLL) from other chronic B‐cell lymphoproliferative diseases. Lymphocytes of B‐CLL were CDlc‐, CD23+, whereas those of other types of chronic B‐cell leukaemia were CDlc+/‐, CD23‐, and CD38 +/‐. Non‐B‐CLL cases had a significantly higher amount of large peroxidase‐negative (unstained) cells analyzed with an automated blood cell counter (Technicon H6000). This type of volumetric assessment allowed a separation between typical and “atypical” B‐CLL, which otherwise were both CDlc‐, and CD23+. These combinations of phenotypic markers corresponded to well‐defined haematopathologic entities, conventionally diagnosed on peripheral blood (PB) and bone marrow smears, and on histologic sections of lymph nodes and spleen.

Biological and clinical features of acute lymphoblastic leukaemia with cytoplasmic granules or inclusions: description of eight cases

We describe eight patients (four children and four adults) with an acute lymphoblastic leukaemia (ALL) with cytoplasmic granules or inclusions. The incidence of this variant of acute leukaemia in our whole series of patients with ALL is 1.8%. The granules or inclusions were usually positive for aspecific esterases (ANAE) and/or acid phosphatase, and the immunophenotype was in all cases typical of a CALLA positive B‐lineage ALL (CD10+, CD19+ and/or CD24+, DR+, TdT+, anti‐T‐, anti‐My‐, SIg‐). In one paediatric case, CD33 was unusually coexpressed. Ultrastructural investigations were performed in one case and demonstrated large granules containing vesicles, usually membrane bound, in the majority of blast cells. In the two cases analysed, Ig heavy chain gene rearrangement was detected. In this series of patients prognosis was poor since three never achieved a complete remission, four relapsed and only one is still in first continuous remission.

Bone marrow and tissue expression of gpIIb/IIIa, LFA-1, Mac-1 and gp150,95 glycoproteins.

Monoclonal antibodies (MAbs) against platelet glycoprotein gpIIb/IIIa and the leucocyte adhesion molecules LFA‐1, Mac‐1, and gp 150,95 α chain (CD11a,b,c) and β chain (CD18) have been tested in normal and leukaemic bone marrows, in different human tissues, and in a patient with leucocyte adhesion deficiency (LAD). The effect of these MAbs on platelet aggregation was also tested. GpIIb/IIIa showed widespread distribution, while reactivity of CD11/18 antibodies was limited to haematopoietic cells. Platelets and megakaryocytes were reactive with one CD11a (25.5.2), and with no CD11b/c or CD18 MAbs. GpIIb/IIIa was present on the platelets of the patient with LAD, whereas 25.5.2, (CD11a) bound to his platelets but not to his leucocytes. These data indicate that LFA‐1, Mac‐1, and gp150,95 are not present on human platelets, but they suggest the existence of crossreacting epitopes on gpIIb/IIIa, which is consistent with the hypothesis that these molecules belong to a supergene family of adhesion molecules.

Scanning Immunoelectron Microscopy of Hairy Cell Leukemia

Scanning electron microscopy has shown a typical cell surface morphology in hairy cell leukemia. Scanning immunoelectron microscope techniques, utilizing monoclonal antibodies and colloidal gold particles, have recently become available. Eight patients with hairy cell leukemia have been studied with a panel of monoclonal antibodies of which B1, BA1, OKM1, anti-TAC and LeuM5 were shown to be suitable for scanning immunoelectron microscopy and reactive with hairy cells. The combination of typical cell surface features with reactivity for the B1 and LeuM5 monoclonal antibodies permits accurate diagnosis of hairy cell leukemia, particularly when low percentages of hairy cells are present as in hypocellular or in interferon-treated patients.

Leukaemoid reaction with megakaryocytic features in newborns with Down's syndrome

A leukaemoid reaction was observed in 3 newborns with Downs syndrome. Thrombocytopenia was present in 2, requiring platelets transfusions in 1, and red cell transfusions were necessary in 2 patients. Blast cells characterization by specific monoclonal antibodies showed a prevalence of megakaryoblasts in all 3 cases. This feature was confirmed in 2 of them by the demonstration of platelet peroxidase (PPO) activity under transmission electron microscopy (TEM). A spontaneous remission of the leukaemoid picture was observed after 2–3 months. However, in 1 case a relapse of the myeloproliferative disorder with the same features of the blast cell population was diagnosed after 16 months. Chemotherapy with low‐dose Ara‐C, started because of a relevant clinical involvement, induced a complete remission.

Morphologic, immunologic, and cytogenetic characteristics of secondary acute unclassifiable leukemia in Hodgkin's disease

Blast cells from five cases of secondary unclassifiable leukemia following therapy for Hodgkins disease were studied by cytochemical, immunological and cytogenetic analyses. Cytochemical and immunological reactivity were in accordance with poorly differentiated, myeloid blasts. The four cases in which karyotype analysis was performed showed specific chromosomal abnormalities. No evidence of multiple lineage involvement was found. Problems in classifying these cases of secondary ANLL were due to the high grade of undifferentiation of the blast cells. Their low cytochemical reactivity with markers of myeloid differentiation was similar to what may be observed in patients with acute undifferentiated leukemia or with chronic myeloid leukemia in blast crisis.

Low Insulin-Like Growth Factor I and Leukopenia in Anorexia Nervosa

OBJECTIVE Considering that leukopenia and anemia are commonly observed in anorexia nervosa (AN) and that growth hormone (GH) and insulin-like growth factor-I (IGF-I) markedly influence the activation, growth and survival of hemopoietic cells, we sought for possible relationships between hematologic parameters and the GH-IGF-I axis in a group of patients with AN. METHOD Twenty patients were studied. Leukocyte and erythrocyte counts, as well as baseline serum GH levels and IGF-I standard deviation score (SDS) values, were determined in each participant and correlations between parameters were searched. RESULTS Leukocyte and erythrocyte counts, as well as IGF-I SDS values, were significantly lower, conversely GH was significantly higher in AN patients than in normal weight participants. In patients, IGF-I SDS values were positively correlated with leukocyte count and BMI, whereas no correlation was found between IGF-I SDS and hemoglobin or erythrocytes. CONCLUSION The demonstration of a positive correlation between leukocyte number and circulating IGF-I in AN suggests a likely pathogenetic role of IGF-I deficiency in this hematologic abnormality.

Acute lymphoblastic leukaemia with cytoplasmic granules or inclusions

The description by Cantu-Rajnoldi et al in the Journal (1989, 73, 309-314) of eight cases of acute lymphoblastic leukaemia with cytoplasmic granules or inclusions assumes a homogeneity of the group which may not be justified. A variety of structures, both membrane- and non membrane- bound in leukaemic lymphoblasts may have the light microscopic appearances of azurophil granules, including lysosomal type granules, parallel tubular arrays, collections of virus-like vesicles, detached masses of nuclear chromatin detached nuclear clubs and other unidentified structures