MISC - HATABU Toshimitsu

子宮内膜における免疫系は暑熱ストレスにより変化する

酒井 駿介, 畑生 俊光, 山本 ゆき, 木村 康二

The Journal of Reproduction and Development   64 ( Suppl. )   j83 - j83   2018.9

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Eimeria tenella 弱毒株の作出と比較ゲノム解析による弱毒化分子機構の解明

松林 誠, 川原 史也, 山岸 潤也, 八田 岳士, 畑生 俊光, 山口 浩貴, 寺本 勲, 金子 明, 磯部 尚, 北 潔, 辻 尚利, 笹井 和美

日本獣医学会学術集会講演要旨集   161回   316 - 316   2018.8

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ニワトリコクシジウム症 アイメリア・テネラ感染メカニズムの解明に向けて

畑生 俊光

岡山実験動物研究会報   ( 34 )   80 - 80   2018.4

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Eimeria tenella弱毒株の作出および病理組織学的比較解析

山口 浩貴, 松林 誠, 川原 史也, 八田 岳士, 畑生 俊光, 山岸 潤也, 磯部 尚, 寺本 勲, 金子 明, 北 潔, 辻 尚利, 笹井 和美

日本獣医学会学術集会講演要旨集   160回   336 - 336   2017.8

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南米型トリパノソーマ感染細胞におけるオートファゴソーム形成と関連タンパク質の解析

高橋 千由紀, 畑生 俊光, 嶋田 淳子

The Kitakanto Medical Journal   63 ( 3 )   327 - 327   2013.8

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ネズミマラリア感染臓器におけるスカベンジャー受容体の発現解析

宮下 大地, 畑生 俊光, 嶋田 淳子

The Kitakanto Medical Journal   63 ( 3 )   327 - 327   2013.8

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Plasmodium berghei感染マウス臓器におけるスカベンジャー受容体SRCLの発現解析

宮下 大地, 畑生 俊光, 嶋田 淳子

関東甲信地区医学検査学会講演集   48回   145 - 145   2011.10

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Language：Japanese   Publisher：日臨技関甲信支部医学検査学会  

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南米型トリパノソーマ感染細胞におけるオートファジーとアポトーシスとの関連性の解析

高橋 千由紀, 畑生 俊光, 嶋田 淳子

関東甲信地区医学検査学会講演集   48回   144 - 144   2011.10

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群馬県における肺吸虫の分布調査 群馬県内利根川水系産サワガニにおける肺吸虫類メタセルカリア寄生状況について

富澤 将太, 畑生 俊光, 嶋田 淳子

関東甲信地区医学検査学会講演集   48回   145 - 145   2011.10

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Trypanosoma cruzi infection induces nitric oxide production and S-nitrosylation of cellular FLIP in host cell

Junko Nakajima-Shimada, Toshimitsu Hatabu

Medimond S.r.l   2011

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菌類代謝産物の抗マラリア活性(1)

内山 奈穂子, 畑生 俊光, 嶋田 淳子, 川原 信夫, 板橋 武史, 河合 賢一

日本生薬学会年会講演要旨集   56回   208 - 208   2009.9

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高校生における麻疹予防接種歴調査、抗体価測定と予防教育

豊島 幸子, 畑生 俊光, 嶋田 淳子, 大平 孝雄, 八木 秀明

The Kitakanto Medical Journal   59 ( 3 )   325 - 325   2009.8

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Association of molecular markers in Plasmodium falciparum crt and mdr1 with in vitro chloroquine resistance: a Philippine study. Reviewed International journal

Toshimitsu Hatabu, Moritoshi Iwagami, Shin-ichiro Kawazu, Nao Taguchi, Aleyla D Escueta, Elena A Villacorte, Pilarita T Rivera, Shigeyuki Kano

Parasitology international   58 ( 2 )   166 - 70   2009.6

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Specific mutations in the pfcrt and pfmdr1 genes have been reported to be associated with chloroquine-resistant falciparum malaria parasites worldwide. These genetic markers are considered to be useful tools for the elucidation of several aspects of the epidemiology of drug resistant malaria. In this study, Plasmodium falciparum isolates from three distinct areas of the Philippines were analyzed for drug-resistance-associated genetic mutations, and their association with the in vitro chloroquine (CQ) response. Two novel pfcrt 72-76 allelic types, CVMDT and SVMDT, were detected. The frequency of the pfcrt K76T mutation in the isolates that were successfully tested for in vitro CQ susceptibility was found to be 100% in Kalinga, 80% in Palawan, and 87% in Mindanao. The frequency of the pfmdr1 N86Y mutation was 39% in Kalinga, 35% in Palawan, and 93% in Mindanao isolates. No mutations were found at positions 1042 and 1246 of pfmdr1. However, there were no significant associations found between polymorphisms in these genes and in vitro CQ susceptibility. The results of this study indicate that mutations in pfcrt and pfmdr1 are not predictive of in vitro CQ resistance in Philippine isolates and may therefore not be suitable as molecular markers for surveillance.

DOI： 10.1016/j.parint.2009.01.010

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Genetic diversity and population structure of Plasmodium falciparum in the Philippines. Reviewed International journal

Moritoshi Iwagami, Pilarita T Rivera, Elena A Villacorte, Aleyla D Escueta, Toshimitsu Hatabu, Shin-ichiro Kawazu, Toshiyuki Hayakawa, Kazuyuki Tanabe, Shigeyuki Kano

Malaria journal   8   96 - 96   2009.5

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BACKGROUND: In the Philippines, malaria morbidity and mortality have decreased since the 1990s by effective malaria control. Several epidemiological surveys have been performed in the country, but the characteristics of the Plasmodium falciparum populations are not yet fully understood. In this study, the genetic structure of P. falciparum populations in the Philippines was examined. METHODS: Population genetic analyses based on polymorphisms of 10 microsatellite loci of the parasite were conducted on 92 isolates from three provinces (Kalinga, Palawan, and Davao del Norte) with different malaria endemicity. RESULTS: The levels of genetic diversity and the effective population sizes of P. falciparum in the Philippines were similar to those reported in the mainland of Southeast Asia or South America. In the low malaria transmission area (Kalinga), there was a low level of genetic diversity and a strong linkage disequilibrium (LD) when the single-clone haplotype (SCH) was used in the multilocus LD analysis, while in the high malaria transmission areas (Palawan and Davao del Norte), there was a high level of genetic diversity and a weak LD when SCH was used in the multilocus LD analysis. On the other hand, when the unique haplotypes were used in the multilocus LD analysis, no significant LD was observed in the Kalinga and the Palawan populations. The Kalinga and the Palawan populations were, therefore, estimated to have an epidemic population structure. The three populations were moderately differentiated from each other. CONCLUSION: In each area, the level of genetic diversity correlates with the local malaria endemicity. These findings confirm that population genetic analyses using microsatellite loci are a useful tool for evaluating malaria endemicity.

DOI： 10.1186/1475-2875-8-96

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フィリピンの熱帯熱マラリア原虫集団の遺伝的構造と流行度

石上盛敏, リベラ ピラリタ, ビラコルテ エレーナ, エスクエタ アレイラ, 畑生俊光, 河津信一郎, 早川敏之, 田邉和裄, 狩野繁之

Trop Med Health   125   2008.10

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Phylogenetic relationship of Plasmodium falciparum populations in the Philippines

Moritoshi Iwagami, Toshimitsu Hatabu, Shin-ichiro Kawazu, Aleyla S. Escueta, Elena A. Villacorte, Pilarita Tongol-Rivera, Shigeyuki Kano

Southeast Asian Journal of Tropical Medicine and Public Health   39 ( 2 )   200 - 204   2008.3

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Malaria is one of the major infectious diseases in the Philippines. It is being targeted for control through sustained early diagnosis, treatment and mosquito control. It is in this light that understanding the genetic background of the parasite population is important not only for basic biology of the organism but also for epidemiology and control of the disease. In the present study, molecular phylogenetic relationships of the 3 Plasmodium falciparum populations in the Philippines with the other populations in the world were inferred based on polymorphisms of 9 highly polymorphic microsatellite DNA loci in the parasite genome. A total of 92 P. falciparum isolates collected from 3 provinces (Kalinga, Palawan and Davao del Morte) in the Philippines, and 8 from other populations (3 African, 2 South American, 2 Papua New Guinean, and 1 Thai) that were previously reported, were used for the analysis. The phylogenetic tree showed that the 3 Philippine populations were genetically divergent from each other as compared to the other populations. The branching pattern of the tree suggests that the 3 Philippine populations were relatively close to the Thai population, rather than the Papua New Guinean populations, indicating that the ancestor of the 3 Philippines populations were introduced from Indochina peninsula, and not from countries located south of the Philippines such as Papua New Guinea or Indonesia.

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Phylogenetic relationship of Plasmodium falciparum populations in the Philippines International journal

Moritoshi Iwagami, Toshimitsu Hatabu, Shin-ichiro Kawazu, Aleyla S. Escueta, Elena A. Villacorte, Pilarita Tongol-Rivera, Shigeyuki Kano

Southeast Asian Journal of Tropical Medicine and Public Health   39 ( 2 )   200 - 204   2008.3

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Malaria is one of the major infectious diseases in the Philippines. It is being targeted for control through sustained early diagnosis, treatment and mosquito control. It is in this light that understanding the genetic background of the parasite population is important not only for basic biology of the organism but also for epidemiology and control of the disease. In the present study, molecular phylogenetic relationships of the 3 Plasmodium falciparum populations in the Philippines with the other populations in the world were inferred based on polymorphisms of 9 highly polymorphic microsatellite DNA loci in the parasite genome. A total of 92 P. falciparum isolates collected from 3 provinces (Kalinga, Palawan and Davao del Morte) in the Philippines, and 8 from other populations (3 African, 2 South American, 2 Papua New Guinean, and 1 Thai) that were previously reported, were used for the analysis. The phylogenetic tree showed that the 3 Philippine populations were genetically divergent from each other as compared to the other populations. The branching pattern of the tree suggests that the 3 Philippine populations were relatively close to the Thai population, rather than the Papua New Guinean populations, indicating that the ancestor of the 3 Philippines populations were introduced from Indochina peninsula, and not from countries located south of the Philippines such as Papua New Guinea or Indonesia.

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節足動物媒介感染症の効果的な防除等の対策研究 マラリア重症患者の管理および治療体制に関する研究

狩野繁之, 水野泰孝, 石上盛敏, 竹内勤, 奥浩之, 畑生俊光, 谷畑健生, LOOAREESUWAN Sornchai, KRUDSOOD Srivicha

節足動物媒介感染症の効果的な防除等の対策研究 平成19年度総括・分担研究報告書   2008

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熱帯熱マラリア原虫に対するアムホテリシンBの発育阻止機序の解析

畑生俊光, 畑生俊光, 川合覚, 山口武士, 山田圭一, 奥浩之, 片貝良一, 嶋田淳子, 狩野繁之

日本寄生虫学会大会プログラム・抄録集   76th   2007

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Plasmodium falciparum: the fungal metabolite gliotoxin inhibits proteasome proteolytic activity and exerts a plasmodicidal effect on P. falciparum. Reviewed International journal

Toshimitsu Hatabu, Masaki Hagiwara, Nao Taguchi, Masayuki Kiyozawa, Mamoru Suzuki, Shigeyuki Kano, Kumiko Sato

Experimental parasitology   112 ( 3 )   179 - 83   2006.3

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The in vitro antimalarial activity of the fungal metabolite gliotoxin (GTX) was evaluated, and its mechanism of action was studied. GTX showed plasmodicidal activity against both Plasmodium falciparum chloroquine-resistant strain K-1 and chloroquine-susceptible strain FCR-3. GTX cytotoxicity was significantly lower against a normal liver cell line (Chang Liver cells). The intracellular reduced glutathione level of parasitized and of normal red blood cells was not affected by GTX treatment. However, GTX decreased the chymotrypsin-like activity of parasite proteasomes in a time-dependent manner. The results of this study indicate that GTX possesses plasmodicidal activity and that this effect is due to inhibition of parasite proteasome activity, suggesting that GTX may constitute a useful antimalarial therapy.

DOI： 10.1016/j.exppara.2005.11.012

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Pyrimethamine-sulfadoxine treatment of uncomplicated Plasmodium falciparum malaria in Lao PDR. Reviewed International journal

M Kaiissar Mannoor, Viengxay Vanisaveth, Boualy Keokhamphavanh, Hiromu Toma, Hisami Watanabe, Jun Kobayashi, Toshimitsu Hatabu, Nao Taguchi, Bousy Hongvangthong, Rattanaxay Phetsouvanh, Samlane Phompida, Shigeyuki Kano, Yoshiya Sato

The Southeast Asian journal of tropical medicine and public health   36 ( 5 )   1092 - 5   2005.9

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A 28-day in vivo treatment trial to evaluate the efficacy of pyrimethamine/sulfadoxine (Fansidar, PS) was conducted in 21 Lao patients with uncomplicated Plasmodium falciparum malaria. Sixteen patients (76%) were completely cured with PS without any reappearance of asexual stage parasitemia during the follow-up examination. On the other hand, 5 patients (24%) failed to respond to this trial medication, resulting in recrudescence of asexual stage P. falciparum malaria. PS resistance resulted in higher prevalence of post-treatment gametocytemia, 25% gametocyte carriers among PS sensitive cases versus 75% of the resistant cases. These findings suggest that although the level of PS resistance is still valid for treatment of malaria in the study area of Lao PDR, post-treatment induction of gametocytemia among resistant cases may result an increase in transmission rate of PS resistant falciparum malaria.

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Potent plasmodicidal activity of a heat-induced reformulation of deoxycholate-amphotericin B (Fungizone) against Plasmodium falciparum. Reviewed International journal

Toshimitsu Hatabu, Tsuyoshi Takada, Nao Taguchi, Mamoru Suzuki, Kumiko Sato, Shigeyuki Kano

Antimicrobial agents and chemotherapy   49 ( 2 )   493 - 6   2005.2

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The emergence and spread of drug-resistant Plasmodium falciparum continue to pose problems in malaria chemotherapy. Therefore, it is necessary to identify new antimalarial drugs and therapeutic strategies. In the present study, the activity of a heat-treated form of amphotericin B (HT-AMB) against P. falciparum was evaluated. The efficacy and toxicity of HT-AMB were also compared with those of the standard formulation (AMB). HT-AMB showed significant activity against a chloroquine-resistant strain (strain K-1) and a chloroquine-susceptible strain (strain FCR-3) in vitro. The 50% inhibitory concentrations of HT-AMB were 0.32 +/- 0.03 mug/ml for strain K-1 and 0.33 +/- 0.03 mug/ml for strain FCR-3. In the presence of 1.0 mug of HT-AMB per ml, only pyknotic parasites were observed after 24 h of incubation of early trophozoites (ring forms). However, when late trophozoites and schizonts were cultured with 1.0 mug of HT-AMB per ml, those forms multiplied to ring forms but the number of infected erythrocytes did not increase. These results indicate that HT-AMB possesses potent antiplasmodial activity and that the drug is more effective against the ring-form stage than against the late trophozoite and schizont stages. HT-AMB was observed to have little cytotoxic effect against a human liver cell line (Chang liver cells). In conclusion, the results suggest that HT-AMB has promising properties and merits further in vivo investigations as a treatment for falciparum malaria.

DOI： 10.1128/AAC.49.2.493-496.2005

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A study of chloroquine resistance of Plasmodium falciparum using the in-vitro sensitivity test and polymerase chain reaction (PCR).

HATABU, TOSHIMITSU

Acta Medica Philippina   39(2): 7-10   2005

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群馬県における成人のスギ花粉およびダニIgE抗体保有率 10年間における変化および自覚症状との関連について

比田井裕子, 畑生 俊光, 荒木忠晴, 佐藤 久美子, 嶋田 淳子

北関東医学雑誌   Vol.55 307 ( 3 )   307 - 307   2005

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群馬県におけるToxoplasma gondii抗体保有率の推移

川島悟美, 畑生 俊光, 佐藤 久美子, 狩野繁之, 嶋田 淳子

北関東医学会誌   Vol55 307 ( 3 )   307 - 307   2005

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In vitro susceptibility and genetic variations for chloroquine and mefloquine in Plasmodium falciparum isolates from Thai-Myanmar border

HATABU, TOSHIMITSU

Southeast Asian J Trop Med Public Health   Vol 36 suppl 4: 73-78   2005

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A study of chloroquine resistance of Plasmodium falciparum using the in-vitro sensitivity test and polymerase chain reaction (PCR).

HATABU, TOSHIMITSU

Acta Medica Philippina   39(2): 7-10   2005

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A Pilot Field Survey on the in Vitro Drug Susceptibility of Plasmodium Falciparum in Lao PDR

Toshimitsu Hatabu, Nao Taguchi, Shigeyuki Kano, Viengxay Vanisaveth, Samlane Phompida, Jun Kobayashi, M. Kaiissar Mannoor, Hiromu Toma, Hisami Watanabe

Tropical Medicine and Health   33 ( 2 )   103 - 104   2005

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DOI： 10.2149/tmh.33.103

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In vitro susceptibility and genetic variations for chloroquine and mefloquine in Plasmodium falciparum isolates from Thai-Myanmar border. Reviewed International journal

Toshimitsu Hatabu, Shin-ichiro Kawazu, Somei Kojima, Kumiko Sato, Pratap Singhasivanon, Sornchai Looareesuwan, Shigeyuki Kano

The Southeast Asian journal of tropical medicine and public health   36 Suppl 4   73 - 9   2005

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In vitro drug susceptibility to chloroquine (CQ) and mefloquine (MF) were assessed in 39 P. falciparum isolates from the Thai-Myanmar border area. To further characterize CQ- and MF-resistance profiles in this area, we also analyzed pfcrt K76T mutation that is critical for CQ resistance, and pfmdr1 polymorphism that has an association with MF resistance. Eighteen isolates were successfully examined by in vitro tests for CQ, and 17 of them had resistance to the drug. Geometric mean concentration of CQ that inhibited the growth of parasites at 50% (IC50) was 371 +/- 227 nM (105-971 nM). Sixteen isolates were successfully examined by in vitro tests for MF, and 8 of them were resistant to the drug. Geometric mean of IC50 for MF was 41 +/- 31 nM (4-125 nM). Genotypes of drug resistance, such as pfcrt and pfmdr1 mutations, were also analyzed. All the 39 isolates had the same haplotype (CVIET) for PfCRT at its 72-76th amino acids. A pfmdr1 Y86 mutation was found in 95% of isolates. A pfmdr1 D1042 mutation was also present in 7 isolates, while no pfmdr1 Y1246 mutation was observed. These results indicated a correlation between CQ resistance and the pfcrt T76 and pfmdr1 Y86 mutations.

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Plasmodium falciparum: selenium-induced cytotoxicity to P-falciparum

N Taguchi, T Hatabu, H Yamaguchi, M Suzuki, K Sato, S Kano

EXPERIMENTAL PARASITOLOGY   106 ( 1-2 )   50 - 55   2004.1

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The in vitro antimalarial activity of sodium selenite (NaSe) was investigated and the mechanism of its action was studied. NaSe had antimalarial activity against both the chloroquine-susceptible strain FCR-3 and chloroquine-resistant strain K-I of Plasmodium falciparum. The shrunken cytoplasm of the parasite was observed in a smear 12h after treatment with NaSe. Co-treatment with copper sulfate (CuSO4) in culture did not affect the antimalarial activity of NaSe, but NaSe cytotoxicity against the mammalian cell line Alexander was decreased significantly. The intracellular reduced glutathione level of parasitized red blood cells was decreased significantly by treatment with NaSe, and the decrease was consistent with their mortality. Treatment with NaSe had a strong inhibitory effect on plasmodial development, and NaSe cytotoxicity to human cells was decreased by co-treatment with CuSO4. These results suggest that co-treatment with NaSe and CuSO4 may be useful as a new antimalarial therapy. (C) 2004 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.exppara.2004.01.005

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野外調査地における熱帯熱マラリア in vitro 薬剤感受性試験の応用ーラオス人民民主共和国におけるマラリア疫学調査ー

畑生 俊光

熱帯   第37巻221-225 ( 221-292 )   221 - 225   2004

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A Pilot Field Trial of an in Vitro Drug Susceptibility Test Using the Anaeropack® Malaria Culture System on the Thai-Myanmar Border

Toshimitsu Hatabu, Shin-Ichiro Kawazu, Somei Kojima, Pratap Singhasivanon, Srivicha Krudsood, Sornchai Looareesuwan, Shigeyuki Kano

Tropical Medicine and Health   32 ( 4 )   335 - 337   2004

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The AnaeroPack® malaria culture system with a portable thermostat incubator was evaluated in a field laboratory on the Thai-Myanmar border conducting in vitro drug susceptibility tests on blood samples from 5 Karen children infected with P. falciparum. Only one isolate was susceptible to chloroquine
the others were highly resistant. The IC60 value of an isolate was only resistant to mefloquine, whereas the values of the 3 patients who presumably showed recrudescence were slightly elevated in the susceptible ranges. These results suggested that chloroquine should no longer be used for P. falciparum malaria in this geographic area, and that mefloquine should be carefully monitored for its in vivo effectiveness. In this study, the AnaeroPack® malaria culture system with portable thermostatic incubator is a powerful and useful mobile tool, which aids in providing detailed evidence-based distribution data concerning of drug resistant malaria in the field. © 2004, Japanese Society of Tropical Medicine. All rights reserved.

DOI： 10.2149/tmh.32.335

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生薬由来成分による抗マラリア効果の検討

石原亜矢, 畑生俊光, 奥浩之, 鈴木守, 佐藤久美子

日本寄生虫学会大会プログラム・抄録集   73rd   2004

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Binding of Plasmodium falciparum-infected erythrocytes to the membrane-bound form of Fractalkine/CX3CL1. Reviewed International journal

Toshimitsu Hatabu, Shin-Ichiro Kawazu, Masamichi Aikawa, Shigeyuki Kano

Proceedings of the National Academy of Sciences of the United States of America   100 ( 26 )   15942 - 6   2003.12

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Plasmodium falciparum-infected erythrocytes (pRBCs) adhere to the endothelium via receptors expressed on the surface of vascular endothelial cells (EC) and sequester in the microvasculature of several organs and block the blood circulation. The sequestration, which involves receptors, may be related to the severity of malaria. Here, we report that pRBCs bind to the membrane-bound form of Fractalkine/CX3CL1 (FKN), which is expressed on the surface of vascular EC in various organs. pRBCs adhered to FKN on the surface of FKN cDNA-transfected Chinese hamster ovary cells (CHO-FKN cells). Both the recombinant human FKN-chemokine domain (FKN-CD) and anti-FKN-CD antibody efficiently blocked adherence of pRBCs to CHO-FKN cells. Similar to binding between FKN and FKN receptor on blood mononuclear cells, two amino acid residues, Lys-7 and Arg-47 within FKN-CD, were critical for FKN-pRBC binding. Immunohistological analysis revealed the expression of FKN on EC at the site of sequestration in the brain of a patient with cerebral malaria. These results suggest that the membrane-bound form of FKN acts as a receptor for pRBCs, and this may contribute to furthering our present understanding of cytoadherence in the pathology of falciparum malaria.

DOI： 10.1073/pnas.2534560100

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Binding of Plasmodium falciparum-infected erythrocytes to the membrane-bound form of fractalkine/CX(3)CL1

T Hatabu, S Kawazu, M Aikawa, S Kano

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   100 ( 26 )   15942 - 15946   2003.12

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Plasmodium falciparum-infected erythrocytes (pRBCs) adhere to the endothelium via receptors expressed on the surface of vascular endothelial cells (EC) and sequester in the microvasculature of several organs and block the blood circulation. The sequestration, which involves receptors, may be related to the severity of malaria. Here, we report that pRBCs bind to the membrane-bound form of Fractalkine/ CX(3)CL1 (FKN), which is expressed on the surface of vascular EC in various organs. pRBCs adhered to FKN on the surface of FKN cDNA-transfected Chinese hamster ovary cells (CHO-FKN cells). Both the recombinant human FKN-chemokine domain (FKN-CD) and anti-FKN-CD antibody efficiently blocked adherence of pRBCs to CHO-FKN cells. Similar to binding between FKN and FKN receptor on blood mononuclear cells, two amino acid residues, Lys-7 and Arg-47 within FKN-CD, were critical for FKN-pRBC binding. Immunohistological analysis revealed the expression of FKN on EC at the site of sequestration in the brain of a patient with cerebral malaria. These results suggest that the membrane-bound form of FKN acts as a receptor for pRBCs, and this may contribute to furthering our present understanding of cytoadherence in the pathology of falciparum malaria.

DOI： 10.1073/pnas.2534560100

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In vitro susceptibility of Plasmodium falciparum isolates to chloroquine and mefloquine in southeastern Mindanao Island, the Philippines. Reviewed International journal

Toshimitsu Hatabu, Shin-ichiro Kawazu, Jun Suzuki, Reuben F Valenzuela, Elena A Villacorte, Mamoru Suzuki, Pilarita T Rivera, Shigeyuki Kano

The Southeast Asian journal of tropical medicine and public health   34 ( 3 )   546 - 51   2003.9

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Although the presence of multi-drug-resistant falciparum malaria has been reported in the Philippines, the distribution of drug-resistant malaria parasites has not yet been determined in Mindanao Island. In vitro susceptibility of P. falciparum to both chloroquine and mefloquine was assessed to forecast the spread of drug-resistant parasites in various foci in southeastern Mindanao Island. Of the 33 isolates of P. falciparum successfully tested, 10 (30%) were susceptible, 12 (36%) showed decreased susceptibility (80 nM < or = IC50 < 114 nM), and 11 (33%) were resistant (IC50 > or = 114 nM) to chloroquine. Ten (91%) of the resistant isolates and 9 (75%) of those with decreased susceptibility were from northern and northwestern Davao del Norte Province. Chloroquine-susceptible isolates were found among patients in the eastern parts of Davao del Norte and Davao Oriental provinces. Seven isolates from several foci in the study area were all mefloquine- susceptible (IC50 < 10 nM). This is the first report indicating the potential emergence of chloroquine-resistant P. falciparum on Mindanao Island, which is presently regarded as a drug-susceptible area.

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熱帯熱マラリア原虫フィリピン・ミンダナオ島分離株におけるpfcrtおよびpfmdr‐1遺伝子多型の解析

畑生俊光, 河津信一郎, VILLACORTE E A, 佐藤久美子, RIVERA P T, 狩野繁之

日本寄生虫学会大会プログラム・抄録集   72nd   120   2003.3

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タイ・ミャンマー国境地域におけるアネロパックマラリア培養システムを用いた薬剤感受性試験

畑生 俊光, 狩野 繁之, 小島 荘明, シンハシバノン・プラタップ, ルーアリースワン・ソンチャイ

Clinical Parasitology   13 ( 1 )   75 - 77   2003.2

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Language：Japanese   Publisher：日本臨床寄生虫学会  

著者等が開発したアネロパックマラリア原虫培養システムの更なる野外調査地での有用性を確認するため,in vivo多剤耐性熱帯熱マラリアの流行が報告されているタイ・ミャンマー国境地域において,in vitro薬剤感受性試験を試みた.なお,培養システムには改良型冷却機能つき小型恒温槽を使用した.ラジャナガリンドラ・トロピカル・ディジーズ・インターナショナル・センター(RTIC)を来院した5名の患者より分離した熱帯熱マラリア原虫を用いた薬剤感受性試験を行ったところ,80%がクロロキン耐性であった.メフロキンについては,80%の原虫が感受性であったが,再燃を疑わせる患者3名から分離した原虫については,メフロキンに対する感受性の低下が認められた.以上の事から,アネロパックマラリア培養システムと改良携帯型恒温槽を用いたin vitro薬剤感受性試験は,従来報告されているin vivoの結果からだされている薬剤耐性マラリアの流行分布に関する疫学的考察を強化する手法として有用であると考えられた

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[An imported case of falciparum malaria successfully treated with Artemether-Lumefantrine in Japan]. Reviewed

Azumi Ishizaki, Yoshiki Kikuchi, Toshimitsu Hatabu, Shigeyuki Kano, Akira Yasuoka, Shinichi Oka

Kansenshogaku zasshi. The Journal of the Japanese Association for Infectious Diseases   77 ( 1 )   34 - 7   2003.1

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Spread of multi-drug resistant malaria in the endemic areas has made malaria control more difficult. Thus, WHO recommends combination therapy for the treatment of malaria. The aim of combination therapy is to improve efficacy and to reduce the incidence of resistance development to the each component of the combination. Particularly, the combination with artemisinin derivatives shows good outcome in Thailand where high resistance for mefloquine has already been found. We report the first case of falciparum malaria, successfully treated with Artemether-Lumefantrine in Japan. Artemether-Lumefantrine is a newly developed artemisinin-based combination agent for the treatment of uncomplicated multi-drug resistant malaria. This drug has proved highly effective and well tolerated by some clinical trials abroad. This Japanese female case showed a good clinical course without any side effect.

DOI： 10.11150/kansenshogakuzasshi1970.77.34

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An Imported Case of Falciparum Malaria Successfully Treated with Artemether-Lumefantrine in Japan

ISHIZAKI Azumi, KIKUCHI Yoshiki, HATABU Toshimitsu, KANO Shigeyuki, YASUOKA Akira, OKA Shinichi

Journal of the Japanese Association for Infectious Diseases   77(1):34-7 ( 1 )   34 - 37   2003

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Publisher：The Japanese Association for Infectious Diseases  

Spread of multi-drug resistant malaria in the endemic areas has made malaria control more difficult. Thus, WHO recommends combination therapy for the treatment of malaria. The aim of combination therapy is to improve efficacy and to reduce the incidence of resistance development to the each component of the combination. Particularly, the combination with artemisinin derivatives shows good outcome in Thailand where high resistance for mefloquine has already been found. We report the first case of falciparum malaria, successfully treated with Artemether-Lumefantrine in Japan. Artemether-Lumefantrine is a newly developed artemisinin-based combination agent for the treatment of uncomplicated multi-drug resistant malaria. This drug has proved highly effective and well tolerated by some clinical trials abroad. This Japanese female case showed a good clinical course without any side effect.

DOI： 10.11150/kansenshogakuzasshi1970.77.34

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PCR-amplification, sequencing, and comparison of the var/PfEMP-1 gene from the blood of patients with falciparum malaria in the Philippines.

HATABU, TOSHIMITSU

Southeast Asian Journal of Tropical Medicine and Public Health   33 suppl(3):8-13   2003

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群馬大学公開講座実施報告。科学する心を育てようーミクロの世界を覗いてみようー

福田 利夫, 佐藤 久美子, 畑生 俊光, 小河原 はつ江, 佐竹 史明, 吉田 朋美

群馬保健学紀要   24   117 - 122   2003

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Language：Japanese   Publisher：群馬大学医学部保健学科  

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Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2004&ichushi_jid=J03890&link_issn=&doc_id=20040311270015&doc_link_id=http%3A%2F%2Fhdl.handle.net%2F10087%2F1564&url=http%3A%2F%2Fhdl.handle.net%2F10087%2F1564&type=%8CQ%94n%8C%A7%92n%88%E6%8B%A4%93%AF%83%8A%83%7C%83W%83g%83%8A_AKAGI&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F80146_3.gif

薬剤耐性マラリアの分布に関する調査研究

畑生 俊光

北関東医学会誌   53巻3号 ( 3 )   327 - 328   2003

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DOI： 10.2974/kmj.53.327

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亜セレン酸ナトリウムが及ぼすマラリア原虫培養系への影響

田口 直, 畑生 俊光, 澁澤 美奈, 石原 亜矢, 佐藤 久美子

関東甲信地区医学検査学会講演集   39回   121 - 121   2002.11

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Language：Japanese   Publisher：日臨技関甲信支部医学検査学会  

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Severe Falciparum Malaria with Prolonged Hemolytic Anemia after Successful Treatment with Intravenous Artesunate

ITODA Ichiro, YASUNAMI Takeshi, KIKUCHI Ken, YAMAURA Hisashi, TOTSUKA Kyoichi, YOSHINAGA Kentaro, TERAMURA Masanao, MIZOGUCHI Hideaki, HATABU Toshimitsu, KANO Shigeyuki

76 ( 8 )   600 - 603   2002.8

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DOI： 10.11150/kansenshogakuzasshi1970.76.600

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[Severe falciparum malaria with prolonged hemolytic anemia after successful treatment with intravenous artesunate]. Reviewed

Ichiro Itoda, Takeshi Yasunami, Ken Kikuchi, Hisashi Yamaura, Kyoichi Totsuka, Kentaro Yoshinaga, Masanao Teramura, Hideaki Mizoguchi, Toshimitsu Hatabu, Shigeyuki Kano

Kansenshogaku zasshi. The Journal of the Japanese Association for Infectious Diseases   76 ( 8 )   600 - 3   2002.8

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We report a 68-year-old woman with severe falciparum malaria contracted in Tanzania. She presented high parasitemia and was treated successfully with intravenous artesunate, a qinghaosu derivative, and aggressive supportive therapy. She developed hemolytic anemia and jaundice on day 11 and blood transfusion was required. This case illustrates that intravenous artesunate has excellent antimalarial activity with rapid efficacy and that no severe adverse effect but conventional aggressive supportive therapy is still important in the treatment of severe falciparum malaria.

DOI： 10.11150/kansenshogakuzasshi1970.76.600

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ネズミマラリア原虫(Plasmodium yoelii)ペルオキシレドキシンの媒介蚊体内発育ステージにおける発現

河津信一郎, 野崎智義, 坪井敬文, 中野陽子, 畑生俊光, 鳥居本美, 狩野繁之

日本獣医学会学術集会講演要旨集   133rd   72 - 72   2002.3

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ネズミマラリア原虫Plasmodium berghei強毒株(NK65),弱毒株(XAT)間におけるプロテオーム解析の試み

石川浩之, 河津信一郎, 畑生俊光, 駒木加奈子, 中野陽子, 鈴木守, 狩野繁之

日本寄生虫学会大会プログラム・抄録集   71st   114   2002.3

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フィリピン・ミンダナオ島における熱帯熱マラリア原虫クロロキン感受性の検討

畑生俊光, 河津信一郎, 鈴木淳, VALENZUELA R F, VILLACORTE E A, RIVERA P T, 狩野繁之

日本寄生虫学会大会プログラム・抄録集   71st   118   2002.3

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The expression system of biologically active canine interleukin-8 in Leishmania promastigotes

T Hatabu, Y Matsumoto, S Kawazu, Y Nakamura, T Kamio, HG Lu, KP Chang, Y Hashiguchi, S Kano, T Onodera, Y Matsumoto

PARASITOLOGY INTERNATIONAL   51 ( 1 )   63 - 71   2002.3

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Language：English   Publisher：ELSEVIER IRELAND LTD  

It has been reported that Leishmania promastigotes have ability to express foreign genes on drug selectable plasmids. To investigate further abilities of the recently described expression vector, P6.5, in the transfection of Leishmania organisms (Chen D-Q, Kolli BK, Yadava N et al. Episomal expression of specific sense and antisense mRNAs in Leishmania amazonensis: modulation of gp63 levels in promastigotes and their infection of macrophages in vitro. Infect Immun 2000;68:80-86). the constructed expression vector, which contains canine interleukin-8 (cIL-8) coding cDNA, was introduced by electroporation to promastigotes of four species of the genus Leishmania: Leishmania amazonensis, L. equatorensis, L. donovani and L. infantum. Extrachromosomal DNAs and total RNAs from the transfected promastigotes were subjected to polymerase chain reaction (PCR) and reverse transcriptase-PCR, respectively, using cIL-8 gene specific primers, and a predicted product of 330 bp was detected. Western blot analysis using a mouse monoclonal antibody raised against cIL-8 demonstrated the successful expression of cIL-8 in the transfectants and culture supernatants. Culture supernatants of the transfected L. amazonensis and L. equatorensis promastigotes showed a high chemotactic activity to both dog and mouse polymorphonuclear leukocytes. These results indicate that Leishmania promastigotes transfected with the expression vector P6.5 containing cIL-8 cDNA are capable of producing biologically active cIL-8. The Leishmania expression system using the P6.5 vector might be a useful alternative for the production of biologically active recombinant cytokines. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.

DOI： 10.1016/S1383-5769(01)00107-6

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The expression system of biologically active canine interleukin-8 in Leishmania promastigotes. Reviewed International journal

Toshimitsu Hatabu, Yasunobu Matsumoto, Shin-ichiro Kawazu, Yuko Nakamura, Tsugihiko Kamio, Hong Gang Lu, Kwang-Poo Chang, Yoshihisa Hashiguchi, Shigeyuki Kano, Takashi Onodera, Yoshitsugu Matsumoto

Parasitology international   51 ( 1 )   63 - 71   2002.3

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Language：English   Publisher：ELSEVIER IRELAND LTD  

It has been reported that Leishmania promastigotes have ability to express foreign genes on drug selectable plasmids. To investigate further abilities of the recently described expression vector, P6.5, in the transfection of Leishmania organisms (Chen D-Q, Kolli BK, Yadava N et al. Episomal expression of specific sense and antisense mRNAs in Leishmania amazonensis: modulation of gp63 levels in promastigotes and their infection of macrophages in vitro. Infect Immun 2000;68:80--86), the constructed expression vector, which contains canine interleukin-8 (cIL-8) coding cDNA, was introduced by electroporation to promastigotes of four species of the genus Leishmania: Leishmania amazonensis, L. equatorensis, L. donovani and L. infantum. Extrachromosomal DNAs and total RNAs from the transfected promastigotes were subjected to polymerase chain reaction (PCR) and reverse transcriptase-PCR, respectively, using cIL-8 gene specific primers, and a predicted product of 330 bp was detected. Western blot analysis using a mouse monoclonal antibody raised against cIL-8 demonstrated the successful expression of cIL-8 in the transfectants and culture supernatants. Culture supernatants of the transfected L. amazonensis and L. equatorensis promastigotes showed a high chemotactic activity to both dog and mouse polymorphonuclear leukocytes. These results indicate that Leishmania promastigotes transfected with the expression vector P6.5 containing cIL-8 cDNA are capable of producing biologically active cIL-8. The Leishmania expression system using the P6.5 vector might be a useful alternative for the production of biologically active recombinant cytokines.

DOI： 10.1016/S1383-5769(01)00107-6

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PCR-amplification, sequencing, and comparison of the var/PfEMP-1 gene from the blood of patients with falciparum malaria in the Philippines. Reviewed International journal

Nozomu Ikenoue, Shin-ichiro Kawazu, Toshimitsu Hatabu, Elena A Villacorte, Pilarita T Rivera, Shigeyuki Kano

The Southeast Asian journal of tropical medicine and public health   33 Suppl 3   8 - 13   2002

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The adhesion of Plasmodium falciparum-infected erythrocytes to vascular endothelium and to uninfected red blood cells (RBCs) plays a key role in the pathology of severe malaria. Adhesion is known to be mediated in part by the antigenically-variant erythrocyte membrane protein-1 (PfEMP-1), which is encoded by the var-gene family of P. falciparum. It has recently been reported that in vitro a single parasite simultaneously transcribes multiple var-genes but that, through a developmentally regulated process, the parasite selects only one PfEMP-1 that will to reach the surface of the host RBC. Were this to be true in vivo, one would expect a correlation between the type of var/PfEMP-1 that is expressed on the parasite-infected RBC and the severity of clinical disease. In order to test this assumption, we determined the sequence of the var-gene that was expressed by the parasites in patients' blood samples. Seven blood samples were collected from patients with or without severe clinical symptoms (cerebral malaria): two samples were from patients diagnosed as having imported falciparum malaria at the International Medical Center of Japan (IMCJ); the five others were from patients of the Davao Regional Hospital in Davao, the Philippines. The parasites (ring stage) in the blood samples were cultured for 24 hours; the matured trophozoites, in which the var-gene selection had taken place, served as material for mRNA isolation. The cDNA corresponding to the Duffy-binding-like (DBL)-1 domain of the var-gene was amplified by RT-PCR, using a region-specific primer set. The amplified cDNAs were cloned into the plasmid vector; the resultant clones (32) were sequenced on both strands. The results indicated that there was considerable diversity in the sequence of the DBL-1 domain among the clones, even among those from a single patient. In conclusion, it was difficult to demonstrate the correlation between the type of var-gene transcripts found in the RBCs of malaria patients and the severity of their symptoms.

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フィリピン・ミンダナオ島における熱帯熱マラリア原虫の薬剤感受性 酸素吸収・炭酸ガス発生剤(アネロパック)を用いた薬剤感受性試験

畑生俊光, 鈴木淳, VALENZUELA R F, VILLACORTE E A, RIVERA R T, 河津信一郎, 狩野繁之

日本熱帯医学会雑誌   29 ( 増刊 )   160 - 160   2001.8

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Molecular characterization of a 2-Cys peroxiredoxin from the human malaria parasite Plasmodium falciparum

S Kawazu, K Komaki, N Tsuji, S Kawai, N Ikenoue, T Hatabu, H Ishikawa, Y Matsumoto, K Himeno, S Kano

MOLECULAR AND BIOCHEMICAL PARASITOLOGY   116 ( 1 )   73 - 79   2001.8

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We have identified the 2-Cys peroxiredoxin (PfPrx-1) from the human malaria parasite Plasmodium falciparum. The PfPrx-1 showed the highest identity at amino acid level to the type Il Prx among the currently known six subfamilies of mammalian Prx. The sequence identity between the PfPrx-1 and the previously reported I-Cys Prx. of P. falciparum (PfPrx-2), which corresponded to mammalian type VI Prx, was 25%. This suggests that the parasite possesses two Prx subfamilies. The PfPrx-1 showed significant sequence similarities with those of 2-Cys peroxiredoxins of plants in the BLASTX search. This may reflect the consequences of a genetic transfer from an algal endosymbiont to the parasite nucleus during evolution. The recombinant PfPrx-1 protein (rPfPrx-1) was expressed as a histidine fusion protein in Escherichia coli and purified with Ni chromatography. The rPfPrx-1 existed as dimers under non-reducing conditions and dissociated into monomers in the presence of dithiothreitol. The PfPrx-1 protein also exists as a dimer in the parasites themselves. The reduction of the oxidized enzyme by the donation of electrons from E. coli thioredoxin (Trx)/Trx reductase system was demonstrated in its reaction with H2O2, using the rPfPrx-1 protein. These results suggested that the PfPrx-1 can act as a terminal peroxidase of the parasite Trx system. An elevated expression of the PfPrx-1 protein seen in the trophozoite, the stage with active metabolism, suggests an association of the parasite Trx. system with its intracellular redox control. (C) 2001 Elsevier Science B.V. All rights reserved.

DOI： 10.1016/S0166-6851(01)00308-5

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熱帯熱マラリア原虫(Plasmodium falciparum)2‐Cys型ペルオキシレドキシンの性状解析

河津信一郎, 駒木加奈子, 辻尚利, 川合覚, 池ノ上望, 石川浩之, 畑生俊光, 松本芳嗣, 狩野繁之

日本獣医学会学術集会講演要旨集   131st   65 - 65   2001.3

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P181 Immunohistochemical analysis of liver and spleen in beige mutant mice infected with P. berghei XAT

HATABU Toshimitsu, ISHIKAWA Hiroyuki, KOMAKI Kanako, MIZUNO Yasutaka, FUKUDA Toshio, SUZUKI Mamoru, KAWAZU Shinichiro, KANO Shigeyuki

Med. Entomol. Zool.   52 ( 0 )   123 - 123   2001

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Language：Japanese   Publisher：The Japan Society of Medical Entomology and Zoology  

DOI： 10.7601/mez.52.123_1

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A case of falciparum malaria successfully treated with intravenus artesunete.

HATABU, TOSHIMITSU

感染症学雑誌   75(9): 822-5   2001

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DOI： 10.11150/kansenshogakuzasshi1970.75.822

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A case of falciparum malaria successfully treated with intravenus artesunete.

HATABU, TOSHIMITSU

感染症学雑誌   75(9): 822-5   2001

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DOI： 10.11150/kansenshogakuzasshi1970.75.822

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フィリピンミンダナオ島のマラリア流行調査ー携帯型連続培養システムを用いたマラリア原虫薬剤感受性試験ー

畑生 俊光

熱帯   第34巻187-196   2001

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P165 Molecular cloning of 2-Cys peroxiredoxin from the human malaria parasite Plasmodium falciparum

KAWAZU Shinichiro, KOMAKI Kanako, TSUJI Naotoshi, KAWAI Satoru, ISHIKAWA Hiroyuki, HATABU Toshimitsu, MATSUMOTO Yoshitsugu, KANO Shigeyuki

Med. Entomol. Zool.   52 ( 0 )   119 - 119   2001

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Language：Japanese   Publisher：The Japan Society of Medical Entomology and Zoology  

DOI： 10.7601/mez.52.119_1

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P164 Study on vitamin A uptake of Plasmodium falciparum in vitro

MIZUNO Yasutaka, KAWAZU Shin-ichiro, HATABU Toshimitsu, KOMAKI Kanako, ISHIKAWA Hiroyuki, KANO Shigeyuki, OHTOMO Hiroshi

Med. Entomol. Zool.   52 ( 0 )   118 - 118   2001

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Language：Japanese   Publisher：The Japan Society of Medical Entomology and Zoology  

DOI： 10.7601/mez.52.118_4

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P166 Production of transgenic Plasmodium falciparum overexpressing an anti-oxidant enzyme, peroxiredoxin

KOMAKI Kanako, KAWAZU Shinichiro, HATABU Toshimitsu, ISHIKAWA Hiroyuki, KANO Shigeyuki

Med. Entomol. Zool.   52 ( 0 )   119 - 119   2001

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Language：Japanese   Publisher：The Japan Society of Medical Entomology and Zoology  

DOI： 10.7601/mez.52.119_2

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P94W4-4 Growth inhibition of Plasmodium falciparum by treatment with anti-recombinant Pf-enolase IgG

ISHIKAWA Hiroyuki, KAWAZU Shin ichiro, HATABU Toshimitsu, KOMAKI Kanako, IKENOUE Nozomu, SUZUKI Mamoru, KANO Shigeyuki

Med. Entomol. Zool.   52 ( 0 )   101 - 101   2001

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Language：Japanese   Publisher：The Japan Society of Medical Entomology and Zoology  

DOI： 10.7601/mez.52.101_2

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CD4(+) cells are indispensable for ulcer development in murine cutaneous leishmaniasis

M Terabe, T Kuramochi, M Ito, T Hatabu, C Sanjoba, KP Chang, T Onodera, Y Matsumoto

INFECTION AND IMMUNITY   68 ( 8 )   4574 - 4577   2000.8

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One of the most characteristic clinical features in cutaneous leishmaniasis is the development of nodules followed by ulcerations at the site of infection. Leishmania amazonensis-infected mice show similar ulcerative lesions. Leishmania-infected severe combined immunodeficiency (SCID) mice, however, have been shown to develop nonulcerative nodules. In the present study, the roles of T cells in ulceration were examined using SCID mice in cell reconstitution experiments. After development of nonulcerative nodules, SCID mice were inoculated with splenocytes from either Leishmania-infected or naive immunocompetent mice, resulting in ulceration in all mice. When naive splenocytes were depleted of CD4(+), CD8(+), or B220(+) cell populations and the remaining cells were injected into Leishmania-infected SCID mice after the development of nodules, only SCID mice inoculated with splenocytes depleted of CD4(+) cells did not show ulceration. The evidence obtained in this study clearly shows that the CD4(+) cell population is indispensable for ulceration in leishmaniasis lesions of SCID mice.

DOI： 10.1128/IAI.68.8.4574-4577.2000

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Molecular cloning and characterization of a peroxiredoxin from the human malaria parasite Plasmodium falciparum

S Kawazu, N Tsuji, T Hatabu, S Kawai, Y Matsumoto, S Kano

MOLECULAR AND BIOCHEMICAL PARASITOLOGY   109 ( 2 )   165 - 169   2000.7

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Language：English   Publisher：ELSEVIER SCIENCE BV  

DOI： 10.1016/S0166-6851(00)00243-7

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Cultivation of Plasmodium falciparum Isolates Under the Anaeropack Gas Condition in a Portable Thermostatic Incubator.

MIZUNO YASUTAKA, HATABU TOSHIMITSU, KAWAZU SHIN-ICHIRO, MASUDA GOHTA, OHTOMO HIROSHI, SUZUKI MAMORU, KANO SHIGEYUKI

Tropical Medicine and Health   23: 393-399 ( 4 )   383 - 385   2000

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Language：English   Publisher：Japanese Society of Tropical Medicine  

DOI： 10.2149/tmh1973.28.383

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Cultivation of Plasmodium falciparum isolates under the AnaeroPack gas condition in a portable thermostatic incubator.

HATABU, TOSHIMITSU

Japanese Journal of Tropical Medicine and Hygine   23: 393-399 ( 4 )   383 - 385   2000

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Language：English   Publisher：Japanese Society of Tropical Medicine  

DOI： 10.2149/tmh1973.28.383

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改良型ポータブル恒温槽及び酸素吸収・炭酸ガス発生剤(アネロパックTM)を用いた熱帯熱マラリア原虫の培養

畑生 俊光, 水野 泰孝, 河津 信一郎, 鈴木 守, 狩野 繁之

日本獣医学会学術集会講演要旨集   128回   161 - 161   1999.9

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Leishmania amazonensis infection in nude mice

M Terabe, T Hatabu, H Takahashi, M Ito, T Onodera, Y Matsumoto

EXPERIMENTAL ANIMALS   48 ( 2 )   119 - 123   1999.4

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Leishmania amazonensis is an intracellular protozoan parasite of macrophages. Cutaneous leishmaniasis in an immunocompetent host begins as papules or nodules followed by ulceration at the site of promastigote inoculation. In this study, the pathological changes of cutaneous leishmaniasis lesions in T cell deficient nude mice were examined. When infected with L. amazonensis promastigotes, nude mice developed non-ulcerative cutaneous nodules, By histological examination of cutaneous lesions, massive accumulation of vacuolated histiocytes containing amastigotes was observed in all the nude mice. Although infiltration of mononuclear and polymorphonuclear cells was seen in the lesions of immunocompetent mice, few such cells were observed in the lesions of nude mice. These results indicate the importance of T cells on the ulcer formation in cutaneous leishmaniasis.

DOI： 10.1538/expanim.48.119

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LEISHMANIASIS IN CENTRAL EURASIA

MATSUMOTO YOSHITSUGU, HATABU TOSHIMITSU, SANJOBA CHIZU, MATSUMOTO YASUNOBU, SASAKI TETSUHIKO, KAWAZU SHIN-ICHIRO, NAKAI YUTAKA, KATAKURA KEN, ITO MAMORU, NAGAKURA KOICHI, AIKAWA MASAMICHI

Japanese journal of tropical medicine and hygiene   27 ( 1 )   59 - 62   1999.3

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Other Link： http://search.jamas.or.jp/link/ui/1999224312

Non-ulcerative cutaneous lesion in immunodeficient mice with Leishmania amazonensis infection

Takashi Kuramochi, Toshimitsu Hatabu, Mamoru Ito, Yoshihito Ueyama, Shin-Ichiro Kawazu, Takashi Onodera, Yoshitsugu Matsumoto

Parasitology International   48 ( 1 )   47 - 53   1999.3

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Cutaneous leishmaniasis begins as papules or nodules at the site of promastigote inoculation. The next key pathogenic event in this disease is the formation of an ulcer at this site. Leishmania infection in immunodeficient mice, however, showed non-ulcerative cutaneous lesions suggesting the involvement of the immune system in ulcer formation. Severe combined immunodeficient (SCID), recombination-activating gene 2 knockout (RAG-2(-/-)), and immunocompetent mice were inoculated subcutaneously with cultured L. amazonensis promastigotes. Macroscopic nodules appeared at the inoculation site within 2 weeks of infection in all the mice and gradually extended to the surrounding skin tissue. Although nodules of immunocompetent mice ulcerated within 6 weeks, immunodeficient mice did not form ulcers even after 25 weeks of inoculation. These results strongly suggest the importance of functional T and B cells in ulcer formation of cutaneous leishmaniasis and are consistent with clinical features of non-ulcerative cutaneous leishmaniasis in some AIDS patients. The present study also indicates that the L. amazonensis-infected immunodeficient mouse model might be suitable for studying the mechanisms of ulcer formation in cutaneous leishmaniasis.

DOI： 10.1016/S1383-5769(98)00040-3

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リーシュマニア原虫を用いた哺乳動物遺伝子発現系の検討

中村 優子, 畑生 俊光, 松本 安喜, 河津 信一郎, Chang K.-P, 佐伯 圭一, 小野寺 節, 松本 芳嗣

日本獣医学会学術集会講演要旨集   127回   93 - 93   1999.3

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Leishmania spp.とζ-crystallin/NADPH oxidoreductase homologue(P36)遺伝子の比較

河津 信一郎, 片倉 賢, 永倉 貢一, 畑生 俊光, 伊藤 守, 中井 裕, 相川 正道, Chang K.-P, 神尾 次彦, 狩野 繁之, 松本 芳嗣

日本獣医学会学術集会講演要旨集   127回   93 - 93   1999.3

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Leishmania mini-exon genes for molecular epidemiology of leishmaniasis in China and Ecuador

Ken Katakura, Shin-Ichiro Kawazu, Chizu Sanjyoba, Toshimitsu Naya, Yoshitsugu Matsumoto, Mamoru Ito, Koichi Nagakura, Masamichi Aikawa, Yoshihisa Hashiguchi

Tokai Journal of Experimental and Clinical Medicine   23 ( 6 )   393 - 399   1998.12

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The mini-exon gene is unique and is tandemly repeated in the Leishmania genome. The transcribed region is highly conserved, but the non-transcribed spacer region is distinct in length and in sequence among different Leishmania species. The usefulness of PCR amplification of the Leishmania mini-exon gene was examined for molecular epidemiology of visceral and cutaneous leishmaniasis. We previously described a PCR method for amplification of the mini-exon gene and obtained positive amplification in bone marrow aspirates of patients with visceral leishmaniasis in China. In this study, we have cloned and sequenced two PCR products from the patients. The sequences of two products revealed 100% identity and showed more similarity to the mini-exon gene of L. donovani Indian strain than those of L. donovani complex in Africa and South America. We also applied this PCR method to the diagnosis of cutaneous leishmaniasis. We obtained positive PCR amplification in skin biopsy materials taken from patients with cutaneous leishmaniasis in Ecuador. Since this PCR amplification is simple and requires only a pair of primers to detect all Leishmania species distributed in Ecuador, the method may be a useful tool for the detection of parasites, not only from patients, but also from sandflies and reservoir animals in this area of endemicity.

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Diagnosis of kala-azar by nested PCR based on amplification of the Leishmania mini-exon gene

K Katakura, S Kawazu, T Naya, K Nagakura, M Ito, M Aikawa, JQ Qu, LR Guan, XP Zuo, JJ Chai, KP Chang, Y Matsumoto

JOURNAL OF CLINICAL MICROBIOLOGY   36 ( 8 )   2173 - 2177   1998.8

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Language：English   Publisher：AMER SOC MICROBIOLOGY  

To diagnose visceral leishmaniasis (kala-azar), we have developed a nested PCR method based on amplification of the mini-exon gene, which is unique and tandomly repeated in the Leishmania genome. Nested PCR was sufficiently sensitive for the detection of DNA in an amount equivalent to a single Leishmania parasite or less. We examined the usefulness of this PCR method using bone marrow aspirates and buffy coat cells collected from kala-azar patients who had or had not received chemotherapy in northwest China. We obtained PCR positivity for all of the parasitologically positive bone marrow samples from the patients. Some ambiguities with the primary PCR results were eliminated by the subsequent nested PCR. The buffy coat samples from 7 of 12 patients with splenomegaly were positive by the nested PCR, although only 2 of them were positive for parasites by culture. However, buffy coat samples from nine children, whose splenomegaly has been reduced and clinically cured by antimony treatment, were all negative. Thus; this nested PCR method represents a new tool for the diagnosis of kala-azar,vith patient blood samples instead of bone marrow or spleen aspirates obtained by more invasive procedures.

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Membrane knobs of unfixed Babesia bovis-infected erythrocytes: New findings as revealed by atomic force microscopy and surface potential spectroscopy

Masamichi Aikawa, Shin-Ichiro Kawazu, Tujihiko Kamio, Yoshitsugu Matsumoto, Toshimitsu Naya, Motomi Torii, Yoshihiro Ito, Bernard Tandler, Yamaji Nakano, Tsutomu Takeuchi, Soshi Shiraishi, Kiyoshi Kanamura

Parasitology International   46 ( 4 )   241 - 246   1997

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Language：English   Publisher：Elsevier Ireland Ltd  

Atomic force microscopy (AFM) and surface potential microscopy (SPS) were used to examine the structure of the knobs of unfixed Babesia bovis-infected erythrocytes, AFM revealed each knob was found to consist of two subunits, components that have not been observed in chemically fixed knobs by transmission electron microscopy. In addition, SPS revealed the knob surface has a positive electrical charge and that the remainder of the erythrocyte surface has a negative charge. These factors might be central to the phenomenon of cytoadherence in cerebral babesiosis. © 1997 Elsevier Science Ireland Ltd.

DOI： 10.1016/S1383-5769(97)00031-7

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