A novel culture method of canine peripheral blood lymphocytes with concanavalin a and recombinant human interleukin-2 for adoptive immunotherapy   总被引：1，自引：0，他引：1  

Kato M  Watarai S  Nishikawa S  Iwasaki T  Kodama H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(5):481-486

This study was designed to develop a novel culture method for the efficient proliferation of canine peripheral blood lymphocytes (cPBL) for adoptive immunotherapy. When cPBL were cultured in the presence of concanavalin A (Con A), proliferation of cPBL was induced and expression of interleukin-2 receptor (IL-2R) which enables to respond to exogenously added IL-2 was upregulated. And then, when cPBL were cultured with recombinant human interleukin-2 (rhIL-2) in addition to Con A, proliferation was accelerated and increased to about 10-fold after 1 week. The phenotypic analysis showed that the main population of the cultured cPBL was consisted of CD8+ positive lymphocytes. Among them, CD4+CD8+ double positive (DP) lymphocytes had significantly increased, and the ratio of CD4+ single positive (SP) lymphocytes to CD8+ SP lymphocytes (CD4+SP/CD8+SP) was decreased as compared to before culturing. To evaluate the cytotoxic activity of cPBL cultured with Con A and rhIL-2, furthermore, cytotoxic assay was carried out against xenogeneic melanoma cell line (MeWo), which resulted in MHC-unrestricted cytokilling. These results suggest that the culture method of cPBL by the use of Con A and rhIL-2 may be useful for generating lymphokine activated killer cells, and also this may be beneficial for adoptive immunotherapy of tumor-bearing dogs.  相似文献   

Phenotype and function of murine discrete Peyer's patch macrophage derived - dendritic cells     

Makala LH  Reyes JC  Nishikawa Y  Tsushima Y  Xuan X  Huang X  Battsetseg B  Matsuo T  Nagasawa H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(4):491-499

The phenotype and function of peritoneal cavity macrophage-derived dendritic cells (PEC-DC) was previously reported. In this study we have gone further in using our established culture system to generated discrete Peyer's patch dendritic cells (DPP-DC) from murine discrete Peyer's patch macrophages (DPP-M?), following stimulation with granulocyte macrophage colony stimulating factor (GM-CSF) plus interleukin 4 (IL-4) for 7 days. DPP-M? from murine small intestines were obtained by mechanical disruption of discrete Peyer's patches (DPP), followed by metrizamide density gradient centrifugation to remove Peyer's patch resident DC and debri, after which an overnight adherent step in tissue culture medium was carried out for macrophage enrichment. Characterization of the generated DPP-DC was carried out using well-established criteria of morphology, expression of membrane antigens and capacity for antigen presentation. Dendritic cells expressed DEC-205, F4/80 and CD34 at high levels, but exhibited very low CD11c levels. They were shown to present soluble protein antigen to CD3(+) spleen T cells. A comparison of the surface antigen expression in the progenitor DPP-M? population and the generated DPP-DC showed a significant decrease in MHC class II levels and a marked down regulation of the co-stimulatory molecule CD86 (B7-2). High expression of the haemopoietic progenitor marker CD34 indicates that the generated DC, possess a haemopoietic rather than myeloid origin. Taken together, these results may provide a better understanding of the complex network regulating mucosal immune responses.  相似文献   

A role for balance of interferon-gamma and interleukin-4 production in protective immunity against Neospora caninum infection   总被引：3，自引：0，他引：3  

Nishikawa Y  Inoue N  Makala L  Nagasawa H 《Veterinary parasitology》2003,116(3):175-184

A suitable balance in the production of Th1/Th2-type cytokines has a crucial role in the control of microbial infections. We investigated cytokine production patterns and effects during Neospora caninum infection, based on two mouse models and an in vitro system. In the acute infection of N. caninum, BALB/c-background IFN-gamma-deficient mice that were sensitive to the N. caninum infection showed high levels of IL-10 production, whereas significant levels of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production were observed in resistant wild type mice. BALB/c mice vaccinated with recombinant vaccinia virus expressing N. caninum surface protein NcSRS2 resisted parasite spread throughout the body, low levels of IFN-gamma production and high levels of IL-4 production were observed compared to unvaccinated animals. The treatment of N. caninum-infected cells with IFN-gamma or IL-10 decreased the host-cell viability in an in vitro system using mouse macrophage J774A.1 cells. On the other hand, IL-4, but not IL-10 administration, increased the viability of N. caninum-infected and IFN-gamma-treated cells. In the light of the balance of Th1/Th2-type cytokine production, an IFN-gamma/IL-4 balance may have a crucial role for the control of cellular responses against the parasite invasion.  相似文献   

Seedborne fungi detected on stored solanaceous berry seeds and their biological activities     

Junji Nishikawa  Takao Kobayashi  Kazuto Shirata  Takashi Chibana  Keiko T. Natsuaki 《Journal of General Plant Pathology》2006,72(5):305-313

We isolated 629 fungi from 1296 berry seeds of solanaceous plants, including tomato (Lycopersicon esculentum), eggplant (Solanum melongena), bell pepper (Capsicum annuum), and red pepper (Capsicum annuum var. annuum) preserved for long and short terms. The isolates were classified into 22 genera excluding unidentified fungi, and the fungal floras were divided into two types: the tomato–eggplant and pepper groups. The results of cluster analysis with unweighted pair-group method with arithmetic average also supported these groups. Most tomato seeds infested with Geotrichum candidum germinated and grew the same as uninfested seeds. Cladosporium sphaerospermum and Arthrinium sp. isolated from eggplant seeds strongly suppressed germination, and Penicillium variabile suppressed seminal root elongation on eggplant. Alternaria alternata, Botrytis cinerea, and Myrothecium verrucaria detected from red pepper or bell pepper seeds were pathogenic to the fruits and the seedlings after artificial inoculation.  相似文献   

Effects of mithramycin on transplantable canine perianal gland carcinoma (CAC-9) in nude mice: biochemical, histomorphometric, and ultrastructural investigations     

S Nishikawa  T J Rosol  C C Capen 《Veterinary pathology》1986,23(6):698-705

A serially transplantable perianal gland carcinoma (CAC-9) was developed in nude mice from a hypercalcemic dog that has been maintained through passage 20. Tumor doubling rate of CAC-9 was 3.1 +/- 0.4 days. Mithramycin (MMC) injected intraperitoneally (8 mg/kg) into nude mice bearing CAC-9 markedly decreased the tumor volume 2 weeks post-injection. MMC returned the elevated serum and urine calcium levels in mice with CAC-9 back to similar values as controls. The few remaining viable tumor cells after MMC were large and had numerous aggregations of intermediate filaments that displaced cytoplasmic organelles. Histomorphometric evaluation of lumbar vertebrae reveled no significant differences in bone resorption of nude mice bearing CAC-9 compared to saline-treated controls. This rapidly growing tumor line in nude mice associated with mild hypercalcemia will be a useful animal model to evaluate combinations of chemotherapy for cancer-associated hypercalcemia.  相似文献   

The pharmacological effects of the anesthetic alfaxalone after intramuscular administration to dogs     

Jun TAMURA  Tomohito ISHIZUKA  Sho FUKUI  Norihiko OYAMA  Kodai KAWASE  Kenjiro MIYOSHI  Tadashi SANO  Kirby PASLOSKE  Kazuto YAMASHITA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(3):289-296

The pharmacological effects of the anesthetic alfaxalone were evaluated after intramuscular (IM) administration to 6 healthy beagle dogs. The dogs received three IM doses each of alfaxalone at increasing dose rates of 5 mg/kg (IM5), 7.5 mg/kg (IM7.5) and 10 mg/kg (IM10) every other day. Anesthetic effect was subjectively evaluated by using an ordinal scoring system to determine the degree of neuro-depression and the quality of anesthetic induction and recovery from anesthesia. Cardiorespiratory variables were measured using noninvasive methods. Alfaxalone administered IM produced dose-dependent neuro-depression and lateral recumbency (i.e., 36 ± 28 min, 87 ± 26 min and 115 ± 29 min after the IM5, IM7.5 and IM10 treatments, respectively). The endotracheal tube was tolerated in all dogs for 46 ± 20 and 58 ± 21 min after the IM7.5 and IM10 treatments, respectively. It was not possible to place endotracheal tubes in 5 of the 6 dogs after the IM5 treatment. Most cardiorespiratory variables remained within clinically acceptable ranges, but hypoxemia was observed by pulse oximetry for 5 to 10 min in 2 dogs receiving the IM10 treatment. Dose-dependent decreases in rectal temperature, respiratory rate and arterial blood pressure also occurred. The quality of recovery was considered satisfactory in all dogs receiving each treatment; all the dog exhibited transient muscular tremors and staggering gait. In conclusion, IM alfaxalone produced a dose-dependent anesthetic effect with relatively mild cardiorespiratory depression in dogs. However, hypoxemia may occur at higher IM doses of alfaxalone.  相似文献   

Central venous blood gas and acid-base status in conscious dogs and cats     

Jun TAMURA  Takaharu ITAMI  Tomohito ISHIZUKA  Sho FUKUI  Kenjirou MIYOSHI  Tadashi SANO  Kazuto YAMASHITA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(7):865-869

To determine the reference level of central venous oxygen saturation (ScvO₂) and clinical efficacy of central venous blood gas analysis, partial pressures of oxygen and carbon dioxide, pH, oxygen saturation, base excess (B.E.) and HCO₃ concentration were compared between simultaneously obtained central venous and arterial blood samples from conscious healthy 6 dogs and 5 cats. Comparisons between arteriovenous samples were performed by a paired t-test and Bland-Altman analysis. Between arteriovenous samples, B.E. showed good agreement, but there were significant differences in other parameters in the dogs, and no good agreement was detected in cats. The ScvO₂ in dogs and cats were 82.3 ± 3.5 and 62.4 ± 13.5%, respectively. Central venous blood gas analysis is indispensable, especially in cats.  相似文献   

Sparing Effect of Robenacoxib on the Minimum Alveolar Concentration for Blunting Adrenergic Response (MAC-BAR) of Sevoflurane in Dogs     

Jun TAMURA  Takaharu ITAMI  Tomohito ISHIZUKA  Sho FUKUI  Norihiko OOYAMA  Kenjiro MIYOSHI  Tadashi SANO  Kazuto YAMASHITA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(1):113-117

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Growth of the fourspine sculpin <Emphasis Type="Italic">Cottus kazika</Emphasis> in the Gonokawa River,Japan, and effects of water temperature on growth     

Naohiko?TakeshitaEmail author  Itaru?Ikeda  Norio?Onikura  Masato?Nishikawa  Singo?Nagata  Seiichi?Matsui  Seir??Kimura 《Fisheries Science》2005,71(4):784-790

ABSTRACT:   The fourspine sculpin Cottus kazika is indigenous to Japan and found in Honshu except for the waters facing the Seto Inland Sea, and was also found in southern Shikoku and eastern Kyushu. This species has a catadromous lifestyle and migrates as juveniles from the sea to the middle reaches of rivers to grow. The growth pattern of this fish was investigated by a mark-and-recapture method from July 1994 to December 1996, in the Nigorikawa River, a tributary of the Gonokawa River system, Shimane Prefecture. 0-year-old fish of 50–70 mm total length ( TL ) occurred in the study area from June to July, grew to 90–140 mm  TL by the following April, and attained 160–210 mm  TL by December. This fish grew rapidly in September–November and April–July, almost ceasing to grow in July–September. It seems that this stagnant growth phase in summer is a characteristic of the seasonal growth pattern of C. kazika . A rearing experiment indicated that the growth rate of C. kazika was higher at 16–22°C than at 12–14 and 24–26°C. This result supports the field evidence of a stagnant growth phase in summer in the Nigorikawa River.  相似文献   

Production and characterization of alien chromosome addition lines in Allium fistulosum carrying extra chromosomes of Allium roylei using molecular and cytogenetic analyses     

Nur Aeni Ariyanti  Vu Quynh Hoa  Ludmila I. Khrustaleva  Sho Hirata  Mostafa Abdelrahman  Shin-ichi Ito  Naoki Yamauchi  Masayoshi Shigyo 《Euphytica》2015,206(2):343-355

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