Leukopenia In Mice Research Articles

Integrated transcriptomic and metabolomic profiles reveal the protective mechanism of modified Danggui Buxue decoction on radiation-induced leukopenia in mice.

Leukopenia caused by radiation hinders the continuous treatment of cancers. Danggui Buxue Decoction (DBD) has been widely used in clinical owing to low toxicity and definite therapeutic effects to increase leukocytes. Meanwhile, icaritin (ICT) has also been proved to have the effect of boosting peripheral blood cells proliferation. However, there is no study to prove the efficacy of MDBD (Modified Danggui Buxue Decoction), a derivative herbal formula composed of DBD and ICT, in the treatment of radiation-induced leukopenia. In this study, we performed a model of 3.5Gy whole-body radiation to induce leukopenia in mice. The results of pharmacodynamic studies demonstrated that MDBD could significantly increase the white blood cells in peripheral blood by improving the activity of bone marrow nuclear cells, reducing bone marrow damage, modulating spleen index, and regulating hematopoietic factors to alleviate leukopenia. We also analyzed the integrated results of metabolomics and transcriptomics and found that MDBD could relieve leukopenia and alleviate bone marrow damage by targeting steroid biosynthesis and IL-17 signaling pathway, in which the key genes are Jun, Cxcl2 and Egr1. Therefore, our study provides a basis for the effectiveness and compatibility in the combination of traditional Chinese medicine formula and small molecule drugs.

Norcantharidin alleviates cyclophosphamide induced immunosuppression and leukopenia in mice through NF κB pathway

Purpose: To investigate the effect of norcantharidin (NCTD) on cyclophosphamide (CY)-induced immunosuppression and leukopenia in mice.
 Methods: Cyclophosphamide (80 mg/kg/day) was intraperitoneally administered to mice from day 1 to 3 and day 9, and from day 4 to 10, NCTD (1, 2 and 4 mg/kg) was intraperitoneally administered. The counts of white blood cells (WBC), red blood cells (RBC) and platelets (PLT) in peripheral blood were determined, followed by the evaluation of mouse spleen and thymus indices. Histopathological examination was performed using hematoxylin-eosin (H & E) staining. In addition, the serum levels of immunoglobulins and cytokines were determined using enzyme-linked immunosorbent assay (ELISA), while plenetic T lymphocyte subsets were assessed by flow cytometry. The expression levels of p65, p-p65, IκBα, and p-IκBα in spleen tissues were determined by immunoblotting.
 Results: Administration of NCTD (1, 2 and 4 mg/kg) increased blood cell counts, and alleviated injuries on splenocytes and thymocytes in CY-treated mice, accompanied by decline in spleen and thymus indices. NCTD ameliorated CY-induced immunosuppression by inhibiting the cytokines but increased the immunoglobulins. CD4+ and CD8+ T lymphocyte expressions were enhanced, especially at the high dose of NCTD (4 mg/kg). Besides, NCTD at high doses (2 and 4 mg/kg) significantly activated NF-κB pathway.
 Conclusion: Administration of NCTD alleviates CY-induced leukopenia and immunosuppression in mice through activation of NF-κB pathway, thus providing a lead for the potential treatment of chemotherapy-induced leukopenia and immunosuppression.

Chemopreventive effects of Xiang Sha Liu Jun Zi Tang on paclitaxel-induced leucopenia and neuropathy in animals.

Paclitaxel frequently induces peripheral neuropathy and myelosuppression during cancer treatment. According to the National Health Insurance Research Database of Taiwan, traditional Chinese medicine doctors widely use Xiang Sha Liu Jun Zi Tang (XSLJZT) to treat breast cancer patients who have received paclitaxel. We explored the combined therapeutic effects of XSLZJT with paclitaxel. XSLJZT did not exhibit significant cytotoxic effects on P388-D1 cells; however, the combination of XSLJZT (100 and 500mg/kg) with paclitaxel prolonged the survival rate in P388-D1 tumor-bearing mice compared to paclitaxel-only. In addition, XSLJZT was found to enhance white blood cells (WBC) counts and promote leukocyte rebound in paclitaxel-induced leukopenia in mice. XSLJZT also reduced paclitaxel-induced mechanical pain and inhibited c-Fos protein expression in the L4-6 spinal cords of Wistar rats. Moreover, paclitaxel-induced shortening of the nerve fibers of dorsal root ganglion cells was ameliorated by pre-treatment with XSLJZT. Therefore, we suggest that XSLJZT could be used as an adjunct for cancer patients, as the formula could decrease paclitaxel-induced neuropathy and myelosuppression.

Therapeutic Effect of Spleen Low Molecular Weight Extracts on Leukopenia Caused by Epirubicin in Mice and Its Mechanism

To investigate the therapeutic effect of spleen low molecular weight extracts on epileptics hydrochloride-induced leukopenia in mice and explore its mechanism. The model of leukopenia in mice was established by the injection of epirubicin hydrochloride (10 mg/kg). After the injection of chemotherapeutic drugs, leukocytopenia mice were treated with different doses of spleen low molecular weight extract, Ganoderma oral solution and recombinant granulocyte colony stimulating factor (rhG-CSF). The general survival status indicators such as body weight, coat color and athletic ability of mice in each group were recorded; the tail vein blood of mice in each group was collected and the white blood cell count in them was calculated; bone marrow of mice was taken and bone marrow smears were observed. In the model group, the weight of the mice gradually decreased in the later period, their coat became dark and rough, and the ability to exercise decreased, while the mice in the treatment groups showed different degrees of improvement in their survival status except for the mice treated by rhG-CSF. There was no significant fluctuation in the white blood cell count of the blank control mice. After injection of epirubicin, the white blood cell count of peripheral blood in the model mice and treated mice were decreased. The white blood cell count was lower in the mice treated with high-dose low molecular weight extract and rhG-CSF than that in other experimental groups. Bone marrow smear showed that the proportion of bone marrow nucleated cells in the mice treated with the low molecular weight extract of the spleen was significantly higher than that of model mice (P<0.05). The low molecular weight spleen extracts can significantly improve the hematopoietic state of mouse bone marrow, promote the proliferation of inhibited bone marrow cells, and thus has the effect of treating leukopenia in mice.

Ziyuglycoside II alleviates cyclophosphamide-induced leukopenia in mice via regulation of HSPC proliferation and differentiation

Ziyuglycoside II (ZGS II) is a major bioactive ingredient of Sanguisorbae officinalis L., which has been widely used for managing myelosuppression or leukopenia induced by chemotherapy or radiotherapy. In the current study, we investigated the pro-hematopoietic effects and underlying mechanisms of ZGS II in cyclophosphamide-induced leukopenia in mice. The results showed that ZGS II significantly increased the number of total white blood cells and neutrophils in the peripheral blood. Flow cytometry analysis also showed a significant increase in the number of nucleated cells and hematopoietic stem and progenitor cells (HSPCs) including ST-HSCs, MPPs, and GMPs, and enhanced HSPC proliferation in ZGS II treated mice. The RNA-sequencing analysis demonstrated that ZGS II effectively regulated cell differentiation, immune system processes, and hematopoietic system-related pathways related to extracellular matrix (ECM)-receptor interaction, focal adhesion, hematopoietic cell lineage, cytokine-cytokine receptor interaction, the NOD-like receptor signaling pathway, and the osteoclast differentiation pathway. Moreover, ZGS II treatment altered the differentially expressed genes (DEGs) with known functions in HSPC differentiation and mobilization (Cxcl12, Col1a2, and Sparc) and the surface markers of neutrophilic precursors or neutrophils (Ngp and CD177). Collectively, these data suggest that ZGS II protected against chemotherapy-induced leukopenia by regulating HSPC proliferation and differentiation.

The protective effects of diallyl sulfide (DAS) on benzene-induced leukopenia in mice

Objective: To study the protective effects of diallyl sulfide (DAS) on leukopenia induced by benzene. Methods: 90 Healthy male ICR mice, adaptive feeding 5 days later, 15 were randomly divided into blank control group、model group、low、middle、high dose DAS intervention groups and DAS control group. Mice in intervention groups and DAS control group were orally given DAS at 40, 80, 160, 160 mg/kg·bw, while mice in the other two groups received an equal volume of corn oil. After 2 hours, model group and the other three intervention groups were given benzene, corn oil suspension (1.3 g/kg·bw) , the two control groups treated with the same volume of corn oil, Benzene and DAS are dissolved in corn oil. one time for each day. 4 weeks later, Anesthesia at 14/29, make blood routine examination and count organ index and observe pathological examinations of spleen and thymus. Results: On day 14, the counts of peripheral blood white blood cells (WBC) , lymphocytes, monocytes in the model group decreased to 68.99%, 71.72%, 53.19% (P<0.05) ; On day 29, the counts of peripheral blood lymphocytes, monocytes, neutrophils in the model group decreased to 83.00%, 81.03%, 89.37%, 20.84%, 19.25% (P<0.05) ; spleen weight, spleen index, white pulp area ratio of spleen, thymus weight, thymus index, thymic cortex area ratio of mice in the model group decreased (P<0.05) . On day 14, the counts of peripheral blood monocytes and lymphocytes in the DAS high dose intervention group increased by 136.36%, 260.00% (P<0.05) ; On day 29, the counts of White blood cells, lymphocytes, red blood cells, platelets, hemoglobin in the DAS low, middle and high dose intervention groups increased (P<0.05) ; spleen weight, spleen index, white pulp area ratio of spleen, thymus weight, thymus index, thymic cortex area ratio of mice in the DAS high dose intervention groups increased (P<0.05) . Conclusion: DAS can effectively suppress benzene-induced leucopenia in mice.

Polysaccharides isolated from &lt;i&gt;Morinda officinalis&lt;/i&gt; How roots inhibits cyclophosphamide-induced leukopenia in mice

Purpose: To investigate the optimum parameters for extracting polysaccharides from Morinda officinalis How (MOP), and explore their inhibitory effects on leukopenia in mice. Methods: Orthogonal design was performed to investigate the optimum parameters for extracting MOP. A leukopenia mouse model was established by injection of cyclophosphamide (CTX) for three days. Thereafter, MOP (100, 200 and 400 mg/kg) was administered orally for 10 days. Furthermore, blood cells (leukocytes, neutrophil, lymphocyte and mononuclear cell) were analyzed, while serum IL-3 and IL- 6 were determined by ELISA. The thymus and spleen of the mice were separated and weighed to determine viscera indices. Results: Orthogonal design showed that the influence order of the four factors was extraction times (C) > ratio of water to raw material (RWM, D) > extraction time (B) > extraction temperature (A). The optimum extraction parameters for MOP were: extraction temperature (80 °C), extraction duration (2 h), no. of extractions (3), and ratio of water to raw material (30 mL/g). Furthermore, the results indicate that MOP (100, 200 and 400 mg/kg) elevated the levels of leukocyte ( p < 0.01), neutrophil ( p < 0.01), lymphocyte ( p < 0.01) and mononuclear cell ( p < 0.01) in leukopenia mice. Besides, MOP (100, 200 and 400 mg/kg) also increased thymus ( p < 0.01) and spleen ( p < 0.05) indices and serum levels of IL-3 ( p < 0.05) and IL-6 ( p < 0.01). Conclusion: Orthogonal design is a good strategy for optimizing extraction parameters of MOP. Furthermore, MOP stimulated synthesis of leukocytes in CTX-induced leukopenia in mice. Thus, MOP is a potential adjunct for the treatment of tumors/cancers. Keywords: Morinda officinalis , Polyscacharide, Orthogonal design, Leukopenia, Thymus index, Spleen index

Deletion of the Dense Core Vesicle Proteins IA-2 and IA-2β Causes Profound Leukopenia in Mice

Abstract Previous studies showed that deletion of the genes encoding the dense core vesicle (DCV)-associated proteins IA-2 and/or IA-2β resulted in a decrease in the secretion of a number of hormones and neurotransmitters and that this, in turn, led to a variety of pathophysiologic and behavioral abnormalities. The effect of the deletion of these genes on the immune system has never been studied. In the present experiments, we found that the knockout of the IA-2 and IA-2β genes (double knockout) resulted in a profound leukopenia characterized by a 78% decrease in WBCs and an 84% decrease in absolute number of lymphocytes. The leukopenia was detected within 1.5 months after birth and persisted for at least 12 months. Both B and T cells were decreased by about 40–50% in bone marrow and thymus and by about 75% in the spleen. LPS-induced cytokine secretion (e.g., IL-6, MCP-1, INF-γ and TNF-α) also was severely impaired. The most likely explanation for these profound immunological abnormalities in single and double knockout mice is that they represent secondary events resulting from impaired secretion of hormones and neurotransmitters that are involved in immunological homeostasis.

ω-3 Fatty Acids Reduce Chemotherapy-Induced Hematological Toxicity by Bone Marrow Stimulation in Mice.

ω-3 Fatty acids exert several benefits during chemotherapy, such as preventing intestinal mucosal damage and improving response to chemotherapy. However, little is known about the effect of ω-3 fatty acids on chemotherapy-induced hematological toxicities. Mice that had consumed either an ω-3-rich or an ω-3-poor diet for 2 weeks were intraperitoneally administered cisplatin. The resultant changes in blood cell count, bone marrow cell count, and cytokine levels in bone marrow supernatant were analyzed. The effect of ω-3 fatty acids on human peripheral blood mononuclear cells (PBMCs) exposed to cisplatin was also examined. Although peripheral blood cell counts decreased after cisplatin treatment in both groups of mice, the decrease in white blood cell count was significantly lower in mice that consumed the ω-3-rich diet. The decrease in bone marrow cells after cisplatin treatment was also reduced in mice that consumed the ω-3-rich diet. Levels of stem cell factor (SCF) and fibroblast growth factor 1 (FGF-1) were significantly higher in bone marrow supernatants from mice that consumed the ω-3-rich diet. The rate of apoptosis in PBMCs (after exposure to cisplatin) cultured in medium containing ω-3 fatty acids was significantly lower than in PBMCs cultured in control medium. ω-3-Rich diets reduced chemotherapy-induced leukopenia in mice. This may be the result of increased numbers of bone marrow cells due to higher levels of SCF and FGF-1 in the bone marrow.

Concomitant treatment with beta-glucan and G-CSF ameliorates altered biochemical indices after cyclophosphamide-induced leukopenia in mice

Cyclophosphamide (CTX) is a DNA-alkylating chemotherapeutic agent and leads to leucopenia and hepatocyte injury. Granulocyte colony-stimulating factor (G-CSF) promotes granulocyte counts and increases leukocytes count. Beta (β)-glucans affect bone marrow cellularity and stem cell mobilization. This study aimed to investigate the effect of β-glucan in combination with G-CSF on leucopenia and hepatocytes injury induced by CTX in mice. Animals were injected with CTX, G-CSF and/or β-glucan for five days, then leukocyte counts and biochemical assays were performed. Our results showed that CTX administration decreased splenocytes and total white blood cells counts. Furthermore, CTX administration led to hepatocyte oxidative stress which was characterized by increased pro-oxidants (thiobarbituric acid reactive substances and xanthine oxidase) and decreased anti-oxidant levels (glutathione peroxidase) and energy disturbance and cell mobilization arrest due to the partial inhibition of adenosine triphosphatase and mannosidase. As a result, hepatocyte injury was observed as indicated by serum aspartate aminotransferase and alanine aminotransferase elevation. β-glucan alone or combined with G-CSF successfully normalized the biochemical parameters and the alteration in cell counts arising from CTX administration. In conclusion, CTX causes leukopenia and liver necrosis through the stimulation of oxidative stress, which leads to inflammation. Both G-CSF and glucan counteract this action through their anti-oxidant and anti-inflammatory properties.

Effect of recombinant Ganoderma lucidum immunoregulatory protein on cyclophosphamide-induced leukopenia in mice

The aim of the study is to investigate the effect of recombinant Ganoderma lucidum immunoregulatory protein (rLZ-8) on mouse models of cyclophosphamide-induced leukopenia, which we have established with both single-phase and multi-phase administration methods. Treatment with rLZ-8 had a strong effect on both models of cyclophosphamide-induced leukopenia. In particular, it increased the number of neutrophils, lymphocytes and monocytes. rLZ-8 treatment also increased the percentage of CD4+ T cells and the levels of secreted IL-3 and IL-4, which contributed to the cyclophosphamide-induced immune dysfunction and immune system imbalance. In conclusion, rLZ-8 treatment benefitted mice with cyclophosphamide-induced leukopenia by improving overall immune function and by specifically increasing the number of white blood cells.

Effect of a structurally modified human granulocyte colony stimulating factor, G-CSFa, on leukopenia in mice and monkeys

BackgroundGranulocyte colony stimulating factor (G-CSF) regulates survival, proliferation, and differentiation of neutrophilic granulocyte precursors, Recombinant G-CSF has been used for the treatment of congenital and therapy-induced neutropenia and stem cell mobilization. Due to its intrinsic instability, recombinant G-CSF needs to be excessively and/or frequently administered to patients in order to maintain a plasma concentration high enough to achieve therapeutic effects. Therefore, there is a need for the development of G-CSF derivatives that are more stable and active in vivo.MethodsUsing site-direct mutagenesis and recombinant DNA technology, a structurally modified derivative of human G-CSF termed G-CSFa was obtained. G-CSFa contains alanine 17 (instead of cysteine 17 as in wild-type G-CSF) as well as four additional amino acids including methionine, arginine, glycine, and serine at the amino-terminus. Purified recombinant G-CSFa was tested for its in vitro activity using cell-based assays and in vivo activity using both murine and primate animal models.ResultsIn vitro studies demonstrated that G-CSFa, expressed in and purified from E. coli, induced a much higher proliferation rate than that of wild-type G-CSF at the same concentrations. In vivo studies showed that G-CSFa significantly increased the number of peripheral blood leukocytes in cesium-137 irradiated mice or monkeys with neutropenia after administration of clyclophosphamide. In addition, G-CSFa increased neutrophil counts to a higher level in monkeys with a concomitant slower declining rate than that of G-CSF, indicating a longer half-life of G-CSFa. Bone marrow smear analysis also confirmed that G-CSFa was more potent than G-CSF in the induction of granulopoiesis in bone marrows of myelo-suppressed monkeys.ConclusionG-CSFa, a structurally modified form of G-CSF, is more potent in stimulating proliferation and differentiation of myeloid cells of the granulocytic lineage than the wild-type counterpart both in vitro and in vivo. G-CSFa can be explored for the development of a new generation of recombinant therapeutic drug for leukopenia.

Two types of C/EBPα mutations play distinct but collaborative roles in leukemogenesis: lessons from clinical data and BMT models

AbstractTwo types of mutations of a transcription factor CCAAT-enhancer binding protein α (C/EBPα) are found in leukemic cells of 5%-14% of acute myeloid leukemia (AML) patients: N-terminal mutations expressing dominant negative p30 and C-terminal mutations in the basic leucine zipper domain. Our results showed that a mutation of C/EBPα in one allele was observed in AML after myelodysplastic syndrome, while the 2 alleles are mutated in de novo AML. Unlike an N-terminal frame-shift mutant (C/EBPα-Nm)–transduced cells, a C-terminal mutant (C/EBPα-Cm)–transduced cells alone induced AML with leukopenia in mice 4-12 months after bone marrow transplantation. Coexpression of both mutants induced AML with marked leukocytosis with shorter latencies. Interestingly, C/EBPα-Cm collaborated with an Flt3-activating mutant Flt3-ITD in inducing AML. Moreover, C/EBPα-Cm strongly blocked myeloid differentiation of 32Dcl3 cells, suggesting its class II mutation-like role in leukemogenesis. Although C/EBPα-Cm failed to inhibit transcriptional activity of wild-type C/EBPα, it suppressed the synergistic effect between C/EBPα and PU.1. On the other hand, C/EBPα-Nm inhibited C/EBPα activation in the absence of PU.1, despite low expression levels of p30 protein generated by C/EBPα-Nm. Thus, 2 types of C/EBPα mutations are implicated in leukemo-genesis, involving different and cooperating molecular mechanisms.

Vitamins C and E prevent AZT-induced leukopenia and loss of cellularity in bone marrow. Studies in mice

A major limitation in the use of AZT for AIDS treatment is the occurrence of side effects, such as leukopenia. The effects of antioxidant vitamins C and E on AZT-induced leukopenia were investigated in mice. Mice were divided into four groups: (1) controls; (2) AZT-treated; (3) treated with AZT plus vitamins C and E; and (4) pre-treated with vitamins and then treated with AZT plus vitamins. Our results demonstrate that AZT causes leukopenia in mice, which was abrogated by administration of vitamins C and E in the pre-treated group. These vitamins prevented the decrease in cellular content induced by AZT in bone marrow and diminished peroxide levels in myeloid precursors in bone marrow. AZT also caused an increase in plasma malondialdehyde and blood oxidized glutathione levels, which was prevented by the administration of antioxidant vitamins. In conclusion, oxidative stress is involved in AZT-induced leukopenia which may be prevented by antioxidant treatment.

Effect of NQO1 induction on the antitumor activity of RH1 in human tumors in vitro and in vivo

NQO1 is a reductive enzyme that is important for the activation of many bioreductive agents and is a target for an enzyme-directed approach to cancer therapy. It can be selectively induced in many tumor types by a number of compounds including dimethyl fumarate and sulforaphane. Mitomycin C is a bioreductive agent that is used clinically for treatment of solid tumors. RH1 (2,5-diaziridinyl-3-(hydroxymethyl)- 6-methyl-1,4-benzoquinone) is a new bioreductive agent currently in clinical trials. We have shown previously that induction of NQO1 can enhance the antitumor activity of mitomycin C in tumor cells in vitro and in vivo. As RH1 is activated selectively by NQO1 while mitomycin C is activated by many reductive enzymes, we investigated whether induction of NQO1 would produce a greater enhancement of the antitumor activity of RH1 compared with mitomycin C. HCT116 human colon cancer cells and T47D human breast cancer cells were incubated with or without dimethyl fumarate or sulforaphane followed by mitomycin C or RH1 treatment, and cytotoxic activity was measured by a clonogenic (HCT116) or MTT assay (T47D). Dimethyl fumarate and sulforaphane treatment increased NQO1 activity by 1.4- to 2.8-fold and resulted in a significant enhancement of the antitumor activity of mitomycin C, but not of RH1. This appeared to be due to the presence of a sufficient constitutive level of NQO1 activity in the tumor cells to fully activate the RH1. Mice were implanted with HL60 human promyelocytic leukemia cells, which have low levels of NQO1 activity. The mice were fed control or dimethyl fumarate-containing diet and were treated with RH1. NQO1 activity in the tumors increased but RH1 produced no antitumor activity in mice fed control or dimethyl fumarate diet. This is consistent with a narrow window of NQO1 activity between no RH1 activation and maximum RH1 activation. This study suggests that selective induction of NQO1 in tumor cells is not likely to be an effective strategy for enhancing the antitumor activity of RH1. In addition, we found that RH1 treatment produced significant leukopenia in mice that may be of concern in the clinic. These results suggest that the ease of reduction of RH1 by NQO1 makes it a poor candidate for an enzyme-directed approach to cancer therapy.

Pharmacological study on the effects of the adenosine uptake inhibitor KF24345 on inflammatory diseases

Adenosine protects against cellular damage and dysfunction under several adverse conditions, including inflammation. We examined the effects of KF24345, a novel adenosine uptake inhibitor, on inflammatory diseases to investigate whether the adenosine uptake inhibition is useful for the treatment of inflammation. KF24345 inhibited adenosine uptake into washed erythrocytes (in vitro) and sampled blood cells from mice after its oral administration (in vivo). KF24345 significantly suppressed lipopolysaccharide-induced tumor necrosis factor-alpha production and leukopenia in mice, and the effects of KF24345 were abolished by the treatment with a non-selective or an A(2A)-selective adenosine receptor antagonist. In the experimental glomerulonephritis induced in mice by anti-glomerular basement membrane antiserum, KF24345 significantly inhibited proteinuria and glomerular damage without exhibiting the side effects observed following the treatment with prednisolone and cyclophosphamide. In addition, KF24345 ameliorated the severity of experimental acute pancreatitis induced by cerulein or choline-deficient and ethionine-supplemented diet in mice, and it decreased mortality accompanying severe acute pancreatitis. The anti-pancreatitis effects of KF24345 were abolished by the treatment with a non-selective or an A(2A)-selective adenosine receptor antagonist. These results suggest that KF24345 and adenosine uptake inhibitors can be a new therapeutic approach for various inflammatory diseases, including glomerulonephritis and acute pancreatitis.

Pathogenesis of pneumococcal pneumonia in cyclophosphamide-induced leukopenia in mice.

Streptococcus pneumoniae pneumonia frequently occurs in leukopenic hosts, and most patients subsequently develop lung injury and septicemia. However, few correlations have been made so far between microbial growth, inflammation, and histopathology of pneumonia in specific leukopenic states. In the present study, the pathogenesis of pneumococcal pneumonia was investigated in mice rendered leukopenic by the immunosuppressor antineoplastic drug cyclophosphamide. Compared to the immunocompetent state, cyclophosphamide-induced leukopenia did not hamper interleukin-1 (IL-1), IL-6, macrophage inflammatory protein-1 (MIP-1), MIP-2, and monocyte chemotactic protein-1 secretion in infected lungs. Leukopenia did not facilitate bacterial dissemination into the bloodstream despite enhanced bacterial proliferation into lung tissues. Pulmonary capillary permeability and edema as well as lung injury were enhanced in leukopenic mice despite the absence of neutrophilic and monocytic infiltration into their lungs, suggesting an important role for bacterial virulence factors and making obvious the fact that neutrophils are ultimately not required for lung injury in this model. Scanning and transmission electron microscopy revealed extensive disruption of alveolar epithelium and a defect in surfactant production, which were associated with alveolar collapse, hemorrhage, and fibrin deposits in alveoli. These results contrast with those observed in immunocompetent animals and indicate that leukopenic hosts suffering from pneumococcal pneumonia are at a higher risk of developing diffuse alveolar damage.

KF24345, an adenosine uptake inhibitor, suppresses lipopolysaccharide-induced tumor necrosis factor-alpha production and leukopenia via endogenous adenosine in mice.

3-[1-(6,7-Diethoxy-2-morpholinoquinazolin-4-yl)piperidin-4-yl]-1,6-dimethyl-2,4(1H,3H)-quinazolinedione hydrochloride (KF24345) is a novel potent adenosine uptake inhibitor. KF24345 inhibited [(3)H]adenosine uptake into erythrocytes from human, mouse, rabbit, and hamster with IC(50) values of 59.5, 130.1, 104.2, and 30.9 nM, respectively. In mice, oral administration of KF24345 at 10 mg/kg almost completely inhibited the [(3)H]adenosine uptake into sampled blood cells at least up to 10 h of the administration. In this study, to examine whether the adenosine uptake inhibition exhibits anti-inflammatory effects, we determined the effects of KF24345 on lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-alpha) production and leukopenia in mice. KF24345 (10 mg/kg p.o.) significantly suppressed the elevation of serum TNF-alpha concentration after the LPS injection, and the suppressing effect of KF24345 was abolished by the treatment with 4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol, a selective adenosine A(2) receptor antagonist, but not with 8-(noradamantan-3-yl)-1,3-dipropylxanthine, a selective adenosine A(1) receptor antagonist. KF24345 (10 mg/kg p.o.) also inhibited the decrease of leukocytes after the LPS injection, and 8-(p-sulfophenyl)theophylline, a nonselective adenosine receptor antagonist, completely reversed the inhibitory effect of KF24345. These results demonstrate that KF24345 inhibits LPS-induced TNF-alpha production and leukopenia via enhancing the effect of endogenous adenosine. It is thus suggested that the adenosine uptake inhibitor has anti-inflammatory effects in vivo and represents a novel therapeutic approach to the treatment of various inflammatory diseases.

Accelerated recovery from cyclophosphamide-induced leukopenia in mice administered a Japanese ethical herbal drug, Hochu-ekki-to

The effect of a Japanese ethical herbal drug, Hochu-ekki-to (HOT), on recovery from leukopenia induced by cyclophosphamide (CY) was investigated. Daily oral administration of 1000 mg/kg HOT into CY-treated mice significantly prevented decrease of leukocyte numbers in the peripheral blood and accelerated recovery from leukopenia. Ginsenoside Rg1 extracted from Ginseng radix, a major herb of HOT, was one of the active ingredients. HOT increased numbers of neutrophils and monocytes in the peripheral blood compared with CY-treated control. Moreover, HOT augmented the resistance against Pseudomonas aeruginosa infection. The number of colony-forming units in the spleen (CFU-S) also increased in HOT-treated mice. The frequencies of IL-3-, GM-CSF- and IFN-γ-producing cells increased in the spleen, bone marrow, liver and IEL on HOT treatment, and HOT clearly augmented the expressions of IL-3, GM-CSF and IFN-γ mRNA in the spleen, bone marrow, liver and IEL except IL-3 and IFN-γ mRNA in the IEL. These results suggest that HOT enhances the production of hematopoietic lymphokines, stimulates the proliferation of hematopoietic progenitor cells and consequently accelerates recovery from leukopenia in CY-treated mice. Additionally, IFN-γ which HOT-augmented the production may contribute the protective effect against the bacterial infection by activating of phagocyte cells.

Effect of orally administered heat-killed Enterococcus faecalis FK-23 preparation on leukopenia in mice treated with cyclophosphamide.

Cyclophosphamide (CY) treated mice were orally administered heat-killed Enterococcus faecalis FK-23 preparation (FK-23) to define the effect of FK-23 on leukopenia associated with chemotherapy. FK-23 had no inhibiting effect on CY induced leukopenia, whereas it augmented leukocyte reconstitution in CY-treated mice. The percentage of neutrophils increased in the mice orally given FK-23. Furthermore, an increase in a myeloid/erythroid ratio and neutrophilic lineages was found in bone marrow of FK-23 treated mice. These findings suggest that FK-23 may have a potent effect on the augmentation of leukocyte reconstituting capacity in patients receiving chemotherapy.