Junko Fukumori

Granulocyte‐colony stimulating factor‐induced proliferation of primary adult T‐cell leukaemia cells

Granulocyte‐colony stimulating factor (G‐CSF) is known to induce proliferation and differentiation of granulocyte progenitors, and is widely used to treat neutropenia induced by intensive chemotherapy for malignant lymphoma or adult T‐cell leukaemia/lymphoma (ATL). G‐CSF is thought not to stimulate malignant lymphoid cells. In the present study we examined the ability of G‐CSF to induce in vitro growth of primary ATL cells from 14 patients (nine acute‐type, two chronic‐type and three lymphoma‐type), and we analysed the in vivo counts of ATL cells in patients who received G‐CSF for neutropenia. FACS analysis using phycoerythrin‐labelled recombinant G‐CSF demonstrated that ATL cells from 11/14 patients express some G‐CSF receptor (G‐CSFR), with a range between 5.4% and 87.3%. Cells expressing G‐CSFR also expressed CD4. Reverse polymerase chain reaction (PCR) analysis demonstrated expression of G‐CSFR messenger RNA in G‐CSFR expressing cells. Leukaemic cells derived from seven (four acute‐type, one chronic‐type and two lymphoma‐type) of the 14 patients proliferated in vitro in response to G‐CSF, as measured by [3H]thymidine incorporation; maximum responses were at G‐CSF concentrations of 10–100 ng/ml. Nine of 14 patients receiving rG‐CSF for neutropenia were analysed retrospectively for ATL cell numbers. Four patients whose primary tumour cells proliferated in response to rG‐CSF in vitro showed a significant increase in ATL cell count after administration of rG‐CSF (P =0.038), whereas five patients whose leukaemic cells did not proliferate in vitro showed no significant increase in ATL cell count. G‐CSF can stimulate proliferation of ATL cells which may complicate therapy for this disease.

IL-2-induced growth of CD8+ T cell prolymphocytic leukemia cells mediated by NF-κB induction and IL-2 receptor α expression

Abstract The binding of interleukin-2 (IL-2) to its receptor on normal T cells induces nuclear expression of nuclear factor κB (NF-κB), activation of the IL-2 receptor (IL-2R) α chain gene, and cell proliferation. In the present study, the role of IL-2R signaling in the growth of CD8 + T cell prolymphocytic leukemia (T-PLL) cells has been investigated. Flow cytometry revealed that primary leukemia cells from a patient with CD8 + T-PLL expressed IL-2Rα and β chains, and the cells showed a proliferative response and an increase in IL-2Rα expression on culture with exogeneous IL-2. Northern blot analysis failed to detect IL-2 mRNA, suggesting that IL-2 may act in a paracrine manner in vivo . Electrophoretic mobility-shift assays revealed that recombinant IL-2 increased NF-κB binding activity in nuclear extracts of the leukemia cells, and Northern blot analysis showed that IL-2 increased the abundance of mRNAs encoding the NF-κB components c-Rel and KBF1 in these cells. IL-2 binding analysis demonstrated that IL-2 markedly increased the number of low affinity IL-2Rs on the leukemia cells, without an effect on the number of high-affinity IL-2Rs. These results show that IL-2 is capable of inducing the nuclear expression of NF-κB in primary CD8 + T-PLL cells, and that this effect is mediated, at least in part, at a pretranslational level.

Granulocyte-colony stimulating factor induces differentiation and apoptosis of CD2, CD7 positive hybrid leukemia cells in vivo and ex vivo

We report a case of a 70-year-old man with a hybrid leukemia treated successfully with granulocyte-colony stimulating factor (G-CSF) combined with a cytocine arabinoside regimen through the induction of differentiation of leukemic cells into monocytoid cells resulting in apoptosis. The leukemic cells demonstrated a TCR-gamma rearrangement, and expressed CD2, CD7, CD33 and G-CSF receptors but not CD11b on the cell surface nor non-specific esterase in cytoplasm. Several days following the administration of G-CSF, the cells with monocytoid characteristics such as CD11b and cytoplasmic non-specific esterase appeared in the peripheral blood replacing the blastic cells. The cells were shown to be derived from the same clone of the leukemic cell because of the identical TCR-gamma gene rearrangement. The short-term culture of leukemic cells with G-CSF induced the differentiation into a monocyte lineage, resulting in apoptosis. Although there is no denying the possibility that cytosine arabinoside is partly responsible, our results strongly suggest that G-CSF plays the main role in differentiation of leukemic cells into a monocyte lineage inducing apoptosis in vivo in this patient.

Interleukin-2-mediated growth of leukemic cells in lymph nodes of patients with adult T-cell leukemia/ lymphoma

We investigated the effect of interleukin-2 (IL-2) on tumor growth of primary adult T-cell leukemia/lymphoma (ATL) cells in biopsied lymph node cells obtained from 14 patients (seven [corrected] with acute-type disease, one with chronic-type disease and six [corrected] with lymphoma-type disease). Biological activity of IL-2 in culture supernatants of the cells was detected in six out of 12 cases. The IL-2 mRNA in the lymph node cells was detected in four out of nine patients by northern blotting. However, it was detected in all nine patients examined by reverse polymerase chain reaction (PCR) method. Lymph node cells from 12 out of 14 patients showed a high or moderate proliferative response to IL-2; the remaining two patients showed a slight response. These results suggest that malignant growth of primary tumor cells in lymph nodes may be associated with the IL-2-IL-2 receptor system in patients with ATL more frequently than had been previously thought.

Trial of Inpatient Medicine Diary for Patient Education.

We prepared a new type of medicine diary in which inpatients should fill their conditions by themselves.We delivered these diaries to inpatients, who were hospitalized in the Second Department of Surgery and the First Department of Internal Medicine, and examined the matters filled in these diaries.As the results of discussion on its usefulness, the followings were obtained.1) The inpatient medicine diary clearly reflected the number of drugs administered and the frequency of changing prescription, which suggested the usefulness.2) The inpatient medicine diary reflected increase and decrease in the amount of drugs administered and withdrawal of administration rightfully and with real time.This suggests that it should be a quite useful method for us to give the instructions efficiently.3) The inpatient medicine diary is effective and useful in making up good communication with patients.

Challenge Tests with Non-Steroidal Antiinflammatory Drugs in Aspirin-Induced Asthma Patient.

Non-steroidal antiinflammatory drug (NSAID) challenge tests were given to patient with aspirin-induced asthma (AIA) to observe the changes of the forced expiratory volume in one second (FEV1.0) between before and after challenges.PL granules for children (PL®) as acid NSAID, that consists of 45 mg salicylamide, 25mg acetaminophen and 10 mg anhydrous caffein per gram, mepirizole and tiaramide hydrochloride as basic NSAID, and syoseiryuto as chinese traditional medicine were used.FENT1.0 was measured before and several times after oral challenges of NSAIDs and syoseiryuto, and decrease of more than 20% of the initial value (basal value) was defined as a positive response.PL, Acid NSAID, showed clear positive response 30 min after challenge, but basic NSAID mepirizole and chinese traditional medicine syoseiryuto showed negative response.On the other hand, tiaramide hydrochloride, that is also basic NSAID, showed positive response when administered 40 mg although negative in a dose of 20 mg.These results suggest that the administration of NSAID to patient with AIA, even in a case of basic NSAID, must be done carefully, and it should be carried out as challenge tests in those cases.