Keiji Nakano

Mobilization of Bone Marrow Cells by G-CSF Rescues Mice from Cisplatin-Induced Renal Failure, and M-CSF Enhances the Effects of G-CSF

Cisplatin, which is a broadly used anticancer drug, is widely known to induce acute renal failure as a result of renal tubular injury. This article examines whether G-CSF and/or M-CSF rescues mice from renal failure induced by cisplatin. BALB/c mice received intraperitoneal injections with or without G-CSF and/or M-CSF for 5 d (from day -5 to day -1). The day after the last injection of G-CSF and/or M-CSF (day 0), the mice received an intraperitoneal injection of cisplatin. When pretreated with G-CSF or G-CSF + M-CSF, the mice showed longer survival and lower serum creatinine and blood urea nitrogen levels than mice that had been received injections of M-CSF or saline. Histologically, pretreatment with G-CSF or G-CSF + M-CSF attenuated the damage to renal tubules induced by cisplatin. BALB/c mice that had received a transplant of bone marrow cells of enhanced green fluorescent protein (EGFP)-transgenic mice ([EGFP-->BALB/c] mice) were treated with or without G-CSF and/or M-CSF, followed by injection of cisplatin as well as above. [EGFP-->BALB/c] mice that were treated with G-CSF or G-CSF + M-CSF showed a significantly higher number of EGFP(+) tubular epithelial cells in the kidney than mice that were treated with only M-CSF or saline. These results suggest that bone marrow cells mobilized by G-CSF accelerate the improvement in renal functions and prevent the renal tubular injury induced by cisplatin and that M-CSF enhances the effects of G-CSF.

Macrophage colony-stimulating factor (M-CSF), as well as granulocyte colony-stimulating factor (G-CSF), accelerates neovascularization

It has been reported that bone marrow cells (BMCs) differentiate into endothelial cells of blood vessels, and that granulocyte colony‐stimulating factor (G‐CSF) mobilizes progenitors in the BMCs to the peripheral blood, while macrophage colony‐stimulating factor (M‐CSF) augments the production of monocytes. We examined whether M‐CSF augments the differentiation of BMCs into endothelial cells of blood vessels using a hindlimb‐ischemic model. Either G‐CSF or M‐CSF, or both, was administered to the hindlimb‐ischemic mice for 3 days. Both M‐CSF and G‐CSF augmented the differentiation of BMCs into endothelial cells of blood vessels through vascular endothelial cell growth factor (VEGF), resulting in early recovery of blood flow in the ischemic limbs.

Treatment and transfer of emphysema by a new bone marrow transplantation method from normal mice to Tsk mice and vice versa.

We have recently established a new bone marrow transplantation (BMT) method in which bone marrow cells are injected into the intrabone marrow (IBM). In the present study, we used an animal model for emphysema (tight‐skin [Tsk] mouse) to examine whether IBM‐BMT could be used to treat emphysema in Tsk mice. IBM‐BMT was carried out from C3H mice into Tsk mice (8–10 weeks old) that had already shown emphysema. Six months after transplantation, the lungs of all the Tsk mice treated with IBM‐BMT [C3H→Tsk] showed similar structures to those of normal mice, whereas the [Tsk→Tsk] mice showed emphysema, as seen in age‐matched Tsk mice. Next, we attempted to transfer emphysema from Tsk mice to C3H mice by IBM‐BMT. Six months after IBM‐BMT, the [Tsk→C3H] mice showed emphysema. These results strongly suggest that emphysema in Tsk mice originates from defects of stem cells in the bone marrow.

Hepatocyte Growth Factor Delivered by Ultrasound-Mediated Destruction of Microbubbles Induces Proliferation of Cardiomyocytes and Amelioration of Left Ventricular Contractile Function in Doxorubicin-Induced Cardiomyopathy

At present, there is no curative strategy for advanced cardiomyopathy except for cardiac transplantation, which is not easily performed, mainly due to a shortage of donors. It has been reported that myocardial progenitor cells exist even in the postnatal heart, suggesting that myocardial progenitor cells could proliferate under some situations and might improve cardiac function in cardiomyopathy‐induced hearts. In this study, recombinant human hepatocyte growth factor (rhHGF) was delivered using ultrasound‐mediated destruction of microbubbles (UMDM) into the cardiomyopathy‐induced heart by doxorubicin (20 mg/kg). Intravenous injection of rhHGF (IV‐rhHGF) alone or UMDM alone failed to improve the morphology or the function of the cardiomyopathy‐induced heart, but (IV‐rhHGF + UMDM) treatment significantly improved the heart morphologically and functionally, and repetitive treatments of (IV‐rhHGF + UMDM) enhanced the effects. The number of bromodeoxy‐uridine‐positive cardiomyocytes significantly increased in the (IV‐rhHGF + UMDM)–treated hearts compared with the untreated hearts. Moreover, Sca‐1+ myocardial progenitor cells express c‐Met, a receptor for HGF. These results suggest that (IV‐rhHGF + UMDM) treatment could morphologically and functionally improve the heart in the case of doxorubicin‐induced cardiomyopathy through the proliferation of the myocardial progenitor cells.

Simultaneous Injection of Bone Marrow Cells and Stromal Cells into Bone Marrow Accelerates Hematopoiesis In Vivo

We have previously demonstrated that stromal cells can support the proliferation and differentiation of hematopoietic cells in vitro and in vivo and that a major histocompatibility complex restriction exists between hematopoietic stem cells and stromal cells. We have also found that intra–bone marrow (IBM) injection of allogeneic bone marrow cells (BMCs) leads to more rapid reconstitution of hematopoietic cells than intravenous injection. In the present study, we examine the effect of simultaneous injection of stromal cells and BMCs into the same bone marrow on the recovery of donor hematopoietic cells and demonstrate that simultaneous IBM injection of BMCs plus stromal cells is more effective in reconstituting recipients with donor hematopoietic cells than intravenous injection of BMCs plus stromal cells or IBM injection of BMCs alone.

A New Strategy for Treatment of Malignant Tumor: Intra‐Bone Marrow–Bone Marrow Transplantation Plus CD4− Donor Lymphocyte Infusion

Donor lymphocyte infusion (DLI) is clinically used for the treatment of malignant tumors. We have found recently that intra‐bone marrow–bone marrow transplantation (IBM‐BMT) can be used to treat various autoimmune diseases, even when radiation doses are reduced. In addition, recently we have found that IBM‐BMT can prevent not only graft failure but also graft‐versus‐host disease (GvHD). Based on these findings, we attempted to prevent and treat the progression of a tumor (Meth‐A cell line: BALB/c‐derived fibrosarcoma) by DLI plus IBM‐BMT. When the tumors had grown to approximately 10 × 10 mm, the tumor‐bearing BALB/c (H‐2d) mice were irradiated with 5 Gy, and whole spleen cells from C57BL/6J (B6) (H‐2b) mice (as DLI) were then intravenously injected into the BALB/c mice. Simultaneously, bone marrow cells (BMCs) from B6 mice were injected directly into the bone marrow cavity of the BALB/c mice (IBM‐BMT). The tumors decreased in size, but the mice died of GvHD. However, when CD4+ T‐cell–depleted spleen cells were used for DLI, the recipients showed only mild GvHD and survived longer, due to the slow growth of the tumor. In contrast, when CD8+ T‐cell–depleted spleen cells were used for DLI, the recipients showed more severe GvHD than those injected with whole spleen cells. These results suggest that IBM‐BMT plus DLI (the depletion or reduction of a certain cell population like CD4+ T cells) could be helpful to suppress both GvHD and tumor growth.

Autopsy case of primary choriocarcinoma of the urinary bladder.

Choriocarcinomas usually develop in the uterus and ovaries in the female, being extremely rare in the extragenital organs in the male. Extragenital choriocarcinomas in the male usually develop in the mediastinum or retroperitoneum. The frequency of choriocarcinoma in the urinary bladder is extremely low. The purpose of the present paper was to report an autopsy case of choriocarcinoma in the urinary bladder in the male. An 81‐year‐old male patient with macrohematuria was first diagnosed with transitional cell carcinoma (TCC). At autopsy a hemorrhagic necrotic tumor, which was found in the urinary bladder with metastatic lesions in the lungs, was diagnosed as choriocarcinoma microscopically. There was no evidence for choriocarcinoma derived from any other organs than the urinary bladder, although there were metastatic lesions in both lungs and the direct invasion into the prostate. From these findings it is concluded that the tumor was a primary choriocarcinoma in the urinary bladder in a male patient. Choriocarcinoma of the urinary bladder is very rare, but the prognosis is extremely poor in comparison with TCC even in the urinary bladder. Therefore, it is essential to clearly discriminate between choriocarcinomas and TCC.

Predictive value of IgE/IgG4 antibody ratio in children with egg allergy

BackgroundThe aim of this study was to investigate the role of specific IgG4 antibodies to hen’s egg white and determine their utility as a marker for the outcome of oral challenge test in children sensitized to hen’s eggMethodsThe hen’s egg oral food challenge test was performed in 105 sensitized children without atopic dermatitis, and the titers of egg white-specific immunoglobulin G4 (IgG4) and immunoglobulin E (IgE) antibodies were measured. To set the cut-off values of IgG4, IgE, and the IgE/IgG4 ratio for predicting positive results in oral challenges, receiver operating characteristic curves were plotted and the area under the curves (AUC) were calculated.ResultsSixty-four of 105 oral challenges with whole eggs were assessed as positive. The AUC for IgE, IgG4, and IgE/IgG4 for the prediction of positive results were 0.609, 0.724, and 0.847, respectively. Thus, the IgE/IgG4 ratio generated significantly higher specificity, sensitivity, positive predictive value (%), and negative predictive value (%) than the individual IgE and IgG4. The negative predictive value of the IgE/IgG4 ratio was 90% at a value of 1.ConclusionsWe have demonstrated that the egg white-specific serum IgE/IgG4 ratio is important for predicting reactivity to egg during food challenges.

Allogeneic Intrabone Marrow-Bone Marrow Transplantation plus Donor Lymphocyte Infusion Suppresses Growth of Colon Cancer Cells Implanted in Skin and Liver of Rats

We have recently found that allogeneic intrabone marrow‐bone marrow transplantation (IBM‐BMT) + donor lymphocyte infusion (DLI) using CD4+ cell‐depleted spleen cells (CD4− cells) can prevent graft‐versus‐host disease (GvHD) but suppress tumor growth (Meth A: fibrosarcoma) in mice. In the present study, we show that allogeneic IBM‐BMT + DLI using CD4− cells also has suppressive effects on the growth of colon cancer cells implanted not only in the skin but also in the liver of rats. First, we examined the effects of allogeneic IBM‐BMT + DLI on the subcutaneously inoculated ACL‐15 (rat colon cancer cell line). Lethally irradiated Fischer rats (F344 rats) were transplanted with T‐cell‐depleted bone marrow cells (BMCs) from Brown Norway (BN) rats. Simultaneously, DLI was performed using whole spleen cells (whole cells), CD4+ cell‐depleted spleen cells (CD4− cells) or CD8+ cell‐depleted spleen cells (CD8− cells) of BN rats. Although allogeneic IBM‐BMT + DLI suppressed tumor growth, a considerable number of rats treated with allogeneic IBM‐BMT + DLI using whole cells or CD8− cells died due to GvHD. In contrast, allogeneic IBM‐BMT + DLI using CD4− cells also suppressed tumor growth, but there was no GvHD. Based on these findings, we next examined the effects of allogeneic IBM‐BMT + DLI using CD4− cells on the cancer cells implanted in the liver. Allogeneic IBM‐BMT + DLI using CD4− cells via the portal vein significantly prolonged the survival. These results suggest that allogeneic IBM‐BMT + DLI using CD4− cells could become a new strategy for the treatment of solid tumors.

An Innovative Approach to Bone Marrow Collection and Transplantation in a Patient with β-Thalassemia Major: Marrow Collection Using a Perfusion Method Followed by Intra-Bone Marrow Injection of Collected Bone Marrow Cells

Using small animals (mice and rats) and monkeys, we have found that the combination of bone marrow collection using the perfusion method (PM) and intra-bone marrow-bone marrow transplantation (IBM-BMT) of the collected cells is safe and effective in treating various intractable diseases. Based on these findings, we attempted to apply this method to humans. We report here the first case of a patient (6 years old) with β-thalassemia major who underwent allogeneic BMT using this new PM + IBM-BMT method. The white blood cell counts of the patient gradually increased to more than 1500/µL by day 47 and continued to increase, reaching the highest level (8600/µL) on day +55. Fluorescence in situ hybridization data on day +33 showed that 98% of the peripheral blood cells were from the donor. Notably, there were no symptoms of graft-versus-host disease (GvHD). However, on day +56, the patient regrettably died of asphyxia resulting from sticky sputum. There was no evidence of infection (in the lung or liver) or GvHD (in the skin) by necropsy. We hope that this case report will help make our new strategies more readily available for the treatment of patients with various intractable diseases.