Souza et al, Blood 66 1985 102(a). Sevier et al, Clin Chem 27/u, 1981, pp. 1797-1806. Kipps et al., Handbook of Experimental Immunol, ed Weir et al., 1986, pp. 108.1-108.9 Nature, vol. 319, Jan. 30, 1986, by S. Nagata, "Molecu lar Cloning and Expression of cDNA for Human Gran ulocyte Colony Stimulating Factor" pp. 415-418. Science, vol. 232, No. 4746 Apr. 4, 1986, by Souza et al, "Recombinant Human Granulocyte Colony-Stimulat ing Factor: Effects on Normal and Leukemic Myeloid Cells", pp. 61-65. Primary Examiner-Garnette D. Draper

Recombinant human granulocyte-colony stimulating factor의 토끼에 피부 및 안점막 자극 평가

Emerging evidence suggests that one of the main reasons of chemotherapy treatment failure is the development of multi-drug resistance (MDR) associated with cancer stem cells (CSCs). Our aim is to identify a therapeutic strategy based on MDR-reversing agents. CSC-enriched Ehrlich carcinoma (EC) cell cultures were prepared by drug-resistant selection method using different concentrations of cisplatin (CIS). Cell cultures following drug exposure were analyzed by flow cytometry for CSC surface markers CD44+/CD24−. We isolated murine bone marrow-derived dendritic cells (DCs) and then used them to prepare CSC–DC vaccine by pulsation with CSC-enriched lysate. DCs were examined by flow cytometry for phenotypic markers. Solid Ehrlich carcinoma bearing mice were injected with the CSC–DC vaccine in conjunction with repeated low doses of CIS. Tumor growth inhibition was evaluated and tumor tissues were excised and analyzed by real-time PCR to determine the relative gene expression levels of MDR and Bcl-2. Histopathological features of tumor tissues excised were examined. Co-treatment with CSC–DC and CIS resulted in a significant tumor growth inhibition. Furthermore, the greatest response of downregulation of MDR and Bcl-2 relative gene expression were achieved in the same group. In parallel, the histopathological observations demonstrated enhanced apoptosis and absence of mitotic figures in tumor tissues of the co-treatment group. Dual targeting of resistant cancer cells using CSC–DC vaccine along with cisplatin represents a promising therapeutic strategy that could suppress tumor growth, circumvent MDR, and increase the efficacy of conventional chemotherapies.

Dual-targeted therapeutic strategy combining CSC–DC-based vaccine and cisplatin overcomes chemo-resistance in experimental mice model

A new strategy for enhancing antitumor immune response using dendritic cells loaded with chemo-resistant cancer stem-like cells in experimental mice model

Cyclophosphamide (CTX) has been used to precondition recipient hosts before adoptive T cell therapy to tumor. The aim of the present study was to analyze the host cellularity changes resulting from different doses and timing of CTX administration. To address this aim, Ehrlich ascites carcinoma (EAC)-bearing mice (5×105cells/mouse) were intraperitoneal injected with 2 or 4 mg/mouse CTX either on day 1 or 7. The results revealed a significant decrease in the number of the tumor cells at 4 mg/mouse CTX on day 1 with an induction of 99% (11.6×106 cells/µl) death of the EAC cells. On day 1 post-EAC cells inoculation, there was a significant increase in the absolute number of splenocytes in EAC-bearing mice at 2 and 4 mg/mouse CTX. The absolute number of leukocytes, lymphocytes and monocytes in EAC-bearing mice non-significantly increased at 2 mg/mouse CTX on day 7 and at 4 mg/mouse CTX on day 1 and 7. Additionally, the absolute number of neutrophils was significantly increased at 2 mg/mouse CTX on day 7 and non-significantly increased at 4 mg/mouse CTX on day 7. The results indicate that: 1) the anti-tumor effect of CTX is a dose- and time-dependent, 2) the use of high dose of chemotherapy is preferable to anti-tumor adoptive cell therapy since it induces higher anti-tumor effects with less lymphopenia.

Cytotoxic and Anti - Tumor Effects of the Chemotherapeutic Drug Cyclophosphamide in Tumor Bearing Mice Is Time and Dose Dependent

Background and objective: Cancer is a major health problem that threatens the life of many peoples with an increasing need for anticancer therapy. The present work aimed to investigate the effectiveness of using sonophotodynamic modality (exposure of infra-red laser as photodynamic) and to the ultrasound as sonodynamic modality in combination with nano-chlorophyll, for cancer therapy through histopathological assessment in Ehrlich ascites carcinoma cell implanted to mice as experimental animals.
Methods: A total of 120 male mice were randomly assigned to 12 treatment groups. Ehrlich ascites carcinoma cell was inoculated subcutaneously in all the experimental mice. When the tumor had grown to about 10 mm in diameter at day 10 after inoculation, treatments were started. Experimental groups were investigated histopathologically two weeks after treatments.
Results: The histological evaluation revealed that tumors from various treated groups of mice bearing the tumor showed different percentages of necrosis and tumor proliferation based on the type of treatment. Tumor excised from mice receiving sonophotodynamic with nano-Chlorophyll treatment showed a significant increase of necrosis about 15 – 25% compared to other groups without nano-chlorophyll as a sensitizer. The group of mice subjected to the combination of photodynamic and sonodynamic therapy in the presence of nano-Chlorophyll, showed large foci distinct necrosis areas 95% of large distinct foci of necrosis in the tumor and no tumor proliferation.
Conclusion: The anti-tumor activity of nano-chlorophyll in combination with sonophotodynamic therapy for Ehrlich ascites carcinoma in mice was effective.

https://zjms.hmu.edu.krd/index.php/zjms/article/download/726/624

Anti-tumor synergistic activity of nano-chlorophyll with sonophotodynamic on Ehrlich ascites carcinoma in mice

Background: Dendritic cell (DC)-based vaccination has shown promising application in tumor immunotherapy. However, it is not clear whether the presence of tumor impacts the efficacy of generation and functionality of DCs. Aim : To compare the phenotype of DCs generated from naive or tumor bearing mice and their capability to restore leukopenia-associated chemotherapy. Materials and Methods: DCs were generated from bone marrow (BM) of naive or Ehrlich ascites carcinoma (EAC) bearing mice. EAC is an undifferentiated breast cancer cell line with the high transplantable capability and rapid proliferation. BM cells were cultured in vitro for 7 days with granulocyte macrophage colony-stimulating factor and interleukin-4 (20 ng/ml each), loaded with different concentrations of EAC cell lysate (0.5, 1, 3 and 5 mg/10 6 ) DCs followed by activation with the toll-like receptor 3 ligand poly(I:C). For DC-based vaccination, CD1 mice ( n = 5/group) were inoculated with an intraperitoneal (i.p.) injection of 0.25 × 10 6 EAC cells to form ascites, treated on day 14 with an i.p. injection of cyclophosphamide (4 mg/mouse) and on day 15 with subcutaneous injection of 2 × 10 6 DCs from control or EAC bearing mice. Injected DCs were loaded with or without EAC lysate followed by i.p. injection of 50 μg/mouse poly(I:C). On day 21, mice were bled and sacrificed for peripheral blood count and spleen and BM cellularity. Results : Yield of DCs generated from naIve or EAC bearing mice, as well as their phenotype (CD11c + CD11b + ) and activation (CD40 and CD80) with poly(I:C) were similar. Loading DCs with 1 mg EAC lysate induced better viability and activation phenotype as compared with the other concentrations. Regardless the source of DCs, DCs vaccination restored the total numbers of leukocytes in blood but not in the spleen and BM. The effect on peripheral blood leukocytes was coincided with the restoration of the relative numbers of lymphocytes, monocytes, and granulocytes. Conclusion: These data support the use of allogenic DCs from healthy donors in anticancer DC-based vaccination.

Dendritic cells generated from naïve and tumor-bearing mice uniquely restores different leukocyte subpopulations in chemotherapy-treated tumor-bearing mice

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Dendritic cells generated from naïve and tumor-bearing mice uniquely restores different leukocyte subpopulations in chemotherapy-treated tumor-bearing mice