In vitro expansion of CD34+/CD41+ cells from human peripheral blood CD34+/CD41- cells: role of cytokines for in vitro proliferation and differentiation of megakaryocytic progenitors.

The aim of this study is to clarify the transitional change of the proliferation and differentiation of human peripheral blood CD34+ cells to megakaryocytic lineage, focusing on its clinical application. We developed a rapid system to purify human peripheral blood CD34+ cells from healthy volunteers, which produced CD34+ cells with a 90% purity. The purified CD34+ cells predominantly consisted of CD41- cells, and the rate of coexpression of CD41 was 0.6% +/- 0.5%. When the purified cells were cultured in liquid phase for 10 days in the presence of recombinant human stem cell factor (rSCF: a ligand for c-kit), interleukin-3 (rIL-3), and thrombopoietin (rTPO: a ligand for Mpl), the number of CD34+/CD41+ cells increased to 19% +/- 7% of total expanded cells on day 4 (4 days of liquid culture) and then gradually decreased to 2.2% +/- 0.6% on day 10. The absolute number of CD34+/CD41+ cells increased and reached a plateau on day 6, and 1.7 +/- 0.6 x 10(5) CD34+/CD41+ cells were produced by 1 x 10(5) CD34+/CD41- day 0 cells. The CD34-/CD41+ cells appeared on day 6, continuously increased in number until day 10, and constituted the main population of expanded cells on day 10, with a value of 38% +/- 18%. On day 10, 19.5 +/- 10.6 x 10(5) of CD34-/CD41+ cells were produced by 1 x 10(5) CD34+/CD41- day 0 cells. The deletion of rTPO from this cytokine combination decreased the number of CD34+/CD41+ and CD34-/CD41+ cells, after days 6 and 8, respectively. Day 0 cells required rIL-3 for promoting colonies containing megakaryocytes, whereas rTPO alone promoted almost no megakaryocytic colonies from day 0 cells. Thus, a combination of IL-3 and SCF expands CD34+/CD41+ cells from CD34+/CD41- cells, and TPO mainly acts to increase CD34-/CD41+ cells. This study suggests that if the expansion of CD34+/CD41+ is performed in vitro, the 6 days' culture of peripheral blood CD34+/CD41- cells with a combination of IL-3 and SCF with TPO provides the most rapid and stable products of CD34+/CD41+ cells for the rapid recovery of platelets in patients with peripheral blood stem cell transplantation.

[Autoimmune thrombocytopenia following syngeneic peripheral blood stem cell transplantation].

A 35-year-old man with non-Hodgkin's lymphoma (NHL) (follicular small cleaved, B cell, stage IVB) received double myeloablative chemotherapy with syngeneic peripheral blood stem cell transplantation (PBSCT). Although platelet recovery was delayed until day 29 after the second transplantation, thereafter trilineage hematopoietic reconstitution was achieved. The evaluation after PBSCT did not detect any residual tumor. The patient was in good health until day 138, when his platelet count suddenly began falling; on day 150, it had fallen to 1.5 x 10(4)/microliter, and the patient was re-admitted for treatment. The bone marrow was normocellular with a normal count and megakaryocyte structure. Other examinations, including serological tests and computed tomography of the neck, chest, abdomen, and retroperitoneum, did not indicate a recurrence of NHL or reveal the cause of thrombocytopenia. The patient's platelet-associated IgG (PAIgG) level was at 70.9 ng/10(7) platelets (normal range: 9-25 ng/10(7) platelets); a diagnosis of thrombocytopenia due to an autoimmune mechanism such as idiopathic thrombocytopenic purpura (ITP) was made. Prednisolone therapy increased the platelet count and reduced the PAIgG level. Thrombocytopenia with an ITP-like mechanism rarely occurs more than 100 days after autologous or syngeneic stem cell transplantation, and should be taken into consideration as a late complication of PBSCT.

Effective high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation in a patient with the aggressive form of cytophagic histiocytic panniculitis.

A 20-year-old Japanese man developed generalized, subcutaneous, painless nodules, fever, abnormal liver function, serosal effusions, hepatosplenomegaly, lymphadenopathy and anemia. Skin biopsies revealed lobular panniculitis with a morphologically benign histiocytic infiltration and prominent phagocytosis. Atypical T lymphocytes were also present in the skin and liver. The diagnosis given was aggressive cytophagic histiocytic panniculitis (CHP) or aggressive subcutaneous panniculitic T cell lymphoma (SPTCL). He received cyclophosphamide, doxorubicin, and vincristine on day 1, prednisolone on days 1-5, and etoposide on days 1, 3 and 5 (CHOP-E), with the support of granulocyte colony-stimulating factor. This regimen was repeated every 2 weeks and complete clinical remission (CCR) was attained after three cycles of CHOP-E. As the clinical course of aggressive CHP is recurrent and often fatal, he was given high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation (APBSCT), after five cycles of CHOP-E. He has remained in CCR for 12 months after APBSCT. High-dose chemotherapy followed by APBSCT is considered to be one of the most beneficial therapies for patients with aggressive CHP and aggressive phase SPTCL.

[Outpatient chemotherapy with continuous infusion of 5-fluorouracil (CI 5-FU) and intravenous bolus leucovorin (IVB LV) in advanced gastrointestinal cancer].

This study was designed to evaluate the efficacy, toxicity, and quality of life (QOL) of outpatient chemotherapy with 5-FU and LV in advanced gastrointestinal cancer. Treatment consisted of CI 5-FU 450-500 mg/body/day, days 1-28, IVB LV 30 mg/body q wk. 5-FU was administered with the Baxter infusor (0.5 ml/hr). LV was not administered if Grade 2 mucositis occurred. Seven patients (pts) with advanced gastrointestinal cancer (esophageal -2, gastric -1, colorectal -4) have been treated to maintain the efficacy of prior inpatient chemotherapy, and two pts with colorectal cancer instead of oral UFT. And to evaluate toxicity and QOL, six pts (bile duct -1, gastric -2, colorectal -3) treated as adjuvant chemotherapy were added. The median duration of response of the nine pts was 3.03 months. Because of mucositis, the administration of LV was restricted. The mean administration of it was 2.1 times (62 mg). Grade 3 or 4 toxicities of mucositis were seen in 40% of the cases. Grade 1 or 2 skin toxicities were seen in all pts. Mean score of QOL in these outpatients chemotherapy was 78.0 +/- 11.5, ranked between inpatient chemotherapy (5-FU+CDDP 59.0 +/- 13.8) and oral UFT (91.8 +/- 6.1). In conclusion, this schedule of 5-FU and LV combination offers a high patient QOL and is suitable for advanced gastrointestinal cancer treatment.

[Pharmacokinetics of lidocaine in perfused rat liver].

Lidocaine pharmacokinetics were studied using perfused rat livers under several conditions. 1) In the low perfusion group (perfusion rate = 1/2 of control group), the elimination rate constant and clearance of lidocaine in the first phase were reduced, but in the second phase, metabolism of lidocaine was more active than in other groups. 2) High albumin concentration in the perfusate did not significantly affect lidocaine metabolism. 3) Low pH of the perfusate (pH = 7.10) did not affect the metabolism of lidocaine. 4) Acute and chronic liver damage inhibited the lidocaine metabolism, which may be due to decreases in viable hepatocytes, effective hepatic blood flow and mitochondrial P-450. 5) Halothane in the concentration of 2.5% reduced the lidocaine metabolism. This was probably due to inhibition of hepatocyte activity by halothane because increases in GOT, LDH and lactate in the perfusate and a decrease in the O2 consumption by the rat liver were observed.