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Effect of short- and long-term immunization of recombinant disorganized muscle protein-1(rDIM-1) against human filarial parasite Brugia malayi in rodents

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Asian Pacific Journal of Tropical Medicine 2021年第7期14卷 287-298页

作者： Vikas Kushwaha Puvvada Kalpana Murthy Division of Parasitology CSIR-Central Drug Research InstituteNew CampusBS 10/1Sector 10Jankipuram ExtensionSitapur RoadLucknow 226031India

Objective: To evaluate the effect of short-term and long-term immunization of recombinant disorganized muscle protein-1(r DIM-1) in rodents against human filarial parasite Brugia malayi.Methods: Recombinant Brugia malayi DIM-1(rDIM-1 bm) protein was cloned, expressed and purified using a Ni-NTA affinity column. Mastomys coucha were immunized with rDIM-1 bm in three immunization schedules: short-term(3-dose of rDIM-1 bm), and long-term(booster doses till 3-and 6-week) and subsequently challenged with infective third-stage larvae of filarial parasite Brugia malayi(L3). Microfilaraemia was monitored in L3 exposed groups on day 90 post larval inoculation(p.l.i.) and continued till day 205 p.l.i. On day 205 p.l.i. all the infected animals were killed and total worm burden was estimated. Cellular proliferative response, macrophage activity, nitric oxide(NO) release, specific IgG and its subtypes, IgE, IgA and Th1(IFN-γ, TNF-ααand IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release were determined. Results: Of the 3 different immunization schedules, shortterm immunization(3-dose schedule) showed better reduction in microfilarial burden(36%-63%) in the peripheral circulation, adult worm load(52%), whereas long-term immunization(3-and 6-week schedule) exerted less effect on peripheral microfilariae count(9%-58%), and adult worm burden(9%-12.5%). Short-term immunization resulted in upregulation of cellular proliferation, macrophages activity, NO release, specific IgG, IgG1, IgG2 a, Ig G2 b, IgE and IgA levels and both Th1(IFN-γ, TNF-α and IL-2) and Th2(IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release whereas long-term immunization(3-and 6-week schedule) exerted less effect on parasite burden and showed mixed immunological responses. None of the rDIM-1 bm administration schedules induced any pathology in lymphoid tissues, or alteration in mast cell number and granularity. Conclusions: The short-term immunization with rDIM-1 bm(3-dose schedule) induces robust immune responses and protects the host from filar

关键词： Brugia malayi Disorganized muscle protein-1 Th1/Th2 cytokines Macrophage activity

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Inflammatory mediator release by Brugia malayi from macrophages of susceptible host Mastomys coucha and THP-1 and RAW 264.7 cell lines

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Asian Pacific Journal of Tropical Medicine 2011年第2期4卷 92-96页

作者： Shiv Kumar Verma Vikas Kushwaha Vijaya Dubey Kirti Saxena Aakanksha Sharma Puvvada Kalpana Murthy Division of Parasitology Central Drug Research InstituteCSIRLucknow 226001India

Objective:To investigate which life stage of the parasite has the ability to stimulate release of pro- or anti-inflammatory mediators from macrophages.Methods:The human macrophage/ monocyte cell line THP-1,the mouse macrophage cell line RAW 264.7 and naive peritoneal macrophages（PM） from the rodent host Mastomys coucha（M.coucha） were incubated at 37℃in 5%CO2 atmosphere with extracts of microfilariae（Mf）,third stage infective larvae（L3） and adult worms（Ad） of Brugia malayi.After 48 hr post exposure,IL-1β,IL-6,TNF-α,IL-10 and nitric oxide（NO） in cell-free supernatants were estimated.Results:Extracts of all the life stages of the parasite were capable of stimulating pro-（IL-1β,IL-6 and TNF-α） and anti-inflammatory （IL-10） cytokines in both the cell lines and peritoneal macrophages of M.coucha.Mf was the strongest stimulator of pro-inflammatory cytokines followed by L3 and Ad;however,Ad was a strong stimulator of IL-10 release.Mf was found to have potential to modulate LPS-induced NO release in RAW cells.Ad-induced NO release was concentration dependent with maximum at 20μg/mL in both RAW and PMs.Conclusions:The results show that parasites at all life stages were capable of stimulating pro-（IL-1β,IL-6 and TNF-α） and anti-inflammatory（IL-10） cytokines and NO release from macrophages of susceptible host M.coucha,human and mouse macrophage cell lines.Mf can suppress the LPS-induced NO release in RAW cells.The findings also show that the two cell lines may provide a convenient in vitro system for assaying parasite-induced inflammatory mediator release.

关键词： Cytokines Macrophage ThP-1 RAW 264.7 Brugia malayi Infective larva Microfilaria Adult worm Mastomys coucha

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Fasciola hepatica infestation as a very rare cause of extrahepatic cholestasis

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World Journal of Gastroenterology 2004年第20期10卷 3076-3077页

作者： Ahmet Dobrucali Rafet Yigitbasi Yusuf Erzin Oguzhan Sunamak Erdal Polat Hakan Yakar division of Gastroenterology Cerrahpasa Medical FacultyIstanbul UniversityTurkey division of Microbiology and parasitology Cerrahpasa Medical FacultyIstanbul UniversityTurkey

Fasciola hepatica,an endemic parsaite in Furkey,is still a viry rare cause of cholestasis worodwide,Through ingestion of contaminated water plants like watercress,humans can become the definitive host of this parasite,Cholestatic symptoms may be sudden but in some cases they may be preceeded by a long period of fever,eosinophilia and vague gsatronintestinal symptoms,we report a woman with cholangitis symptoms of sudden omset which was proved to be due to Fasciloa hepatica infestation by an endoscopic retrograde cholangiography.

关键词：小束苔类植物感染性肝外胆汁淤积胆管炎

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Needs of exploring the burden of recent onset seizures due to neurocysticercosis and challenges in southeast Asia focusing on scenario in Malaysia

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Asian Pacific Journal of Tropical Medicine 2017年第4期10卷 313-321页

作者： Sahu Priyadarshi S Lim Yvonne AL Mahmud Rohela Somanath Sushela D Tan Chong T Ramachandran CP division of Pathology School of MedicineInternational Medical University Department of parasitology Department of MedicineFaculty of MedicineUniversity of Malaya division of Neurology Department of MedicineFaculty of MedicineUniversity of Malaya Chairperson Programme Review GroupNeglected Tropical Diseases-WHO-Western Pacific Region

Seizures due to neurocysticercosis(NCC) is a neglected human-to-human transmitted disorder and an emerging problem worldwide.A substantial portion of recent onset seizures is known to be attributed to NCC in Taenia solium(T.solium) endemic areas where populations which neither raise pigs nor eat pig meat are also at risk.High prevalence of NCC causing epilepsy has been reported in the underdeveloped areas of Southeast Asia(SEA) however,only fragmentary information on its incidence is available in countries like Malaysia.In Malaysia T.solium infection was previously thought to be infrequent due to Muslim population majority and the religious prohibition of consuming pork,but it is not totally absent There is an evident lack of knowledge and awareness of the actual burden,routes of transmission,and the impact of NCC in this region.The problem is assumed to be more prevalent particularly in cities because of the frequent inflow of possibly T.solium infected individuals or carriers among those who migrate from neighboring endemic countries to Malaysia.The issue of imported cases that are likely to be emerging in Malaysia is highlighted here.An accurate quantification of regional burdens of epilepsy due to NCC in Malaysia is warranted considering the disease emergence in its neighboring countries.It is suggested that the importance of NCC be recognized through quantification of its burden,and also to collect epidemiological data for its subsequent elimination in line of World Health Organization's mission for control of cysticercosis as a neglected tropical disease.In this review the need as well as a strategy for neurc-care center screening of epilepsy cases,and various issues with possible explanations are discussed.It is also proposed that NCC be declared as a reportable disease which is one of the eradicable public health problems in SEA.

关键词： Seizure Recent onset seizure NTD Cysticercosis Neurocysticercosis Southeast Asia Malaysia

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Heterologous expression in transgenic mosquitoes

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Asian Pacific Journal of Tropical Medicine 2010年第3期3卷 244-250页

作者： Santhosh PK Yuhua Deng Weidong Gu Xiaoguang Chen Department of parasitology School of Public Health and Tropical MedicineSouthern Medical University division of Infectious Diseases Department of MedicineUniversity of Alabama

Arthropod-borne diseases such as malaria and dengue virus afflict billions of people worldwide imposing major economic and social burdens.Control of such pathogens is mainly performed by vector management and treatment of affected individuals with drugs.The failure of these conventional approaches due to emergence of insecticide-resistant insects and drug-resistant parasites demonstrate the need of novel and efficacious control strategies to combat these diseases.Genetic modification(GM) of mosquito vectors to impair their ability to be infected and transmit pathogens has emerged as a new strategy to reduce transmission of many vector-borne diseases and deliver public health gains.Several advances in developing transgenic mosquitoes unable to transmit pathogens have gained support,some of them attempt to manipulate the naturally occurring endogenous refractory mechanisms,while others initiate the identification of an exogenous foreign gene which disrupt the pathogen development in insect vectors. Heterologous expression of transgenes under a native or heterologous promoter is important for the screening and effecting of the transgenic mosquitoes.The effect of the transgene on mosquito fitness is a crucial parameter influencing the success of this transgenic approach.This review examines these two aspects and describes the basic research work that has been accomplished towards understanding the complex relation between the parasite and its vector and focuses on recent advances and perspectives towards construction of transgenic mosquitoes refractory to vector-borne disease transmission.

关键词： Anopheles Fitness load Malaria Transgene Transposable element Vector

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OBSERVATIONS ON EXPERIMENTAL INFECTION OF PERIODIC BRUGIA MALAYI IN MAN

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中华医学杂志(英文版) 1991年第9期104卷 736-741页

作者：王培义甄天民谷宗藩陈敬亭王尊哲刘伦华 Shandong 272133 Division of Parasitology Changwei Medical College Shandong

Three volunteers were inoculated with different numbers of infective larvae of periodic Brugia malayi from an artifcially infected Meriones unguiculatus. At different times after inoculation, the volunteers developed clinical manifestations such as chills, fever, cough, asthma, skin itching, edema, adenolymphangitis and eosinophilia. Microfilariemia was first detected at 41 and 46 weeks after inoculation in two subjects. At 11 weeks, the percentage of E-rosett forming lymphocytes in these subjects was below the normal level. Specific antibody was first detected in three volunteers at 2, 3 and 5 weeks after inoculation, respectively, and increased to various extent at 12-16 weeks, but decreased in varying degrees at 44-56 weeks. The results also showed that antibody titres fluctuated at different periods of infection.

关键词： FILARIASIS

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Promoted differentiation of cynomolgus monkey ES cells into hepatocyte-like cells by co-culture with mouse fetal liver-derived cells

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World Journal of Gastroenterology 2006年第42期12卷 6818-6827页

作者： Ko Saito Masahide Yoshikawa Yukiteru Ouji Kei Moriya Mariko Nishiofuku Shigehiko Ueda Noriko Hayashi Shigeaki Ishizaka Hiroshi Fukui Department of Gastroenterology and Hepatology Nara Medical University Kashihara Nara Japan division of Developmental Biology Department of Parasitology Nara Medical University Kashihara Nara Japan

AIM: To explore whether a co-culture of cynomolgus monkey embryonic stem (cES) cells with embryonic liver cells could promote their differentiation into hepatocytes. METHODS: Mouse fetal liver-derived cells (MFLCs) were prepared as adherent cells from mouse embryos on embryonic d (ED) 14, after which undifferentiated cES cells were co-cultured with MFLCs. The induction of cES cells along a hepatic lineage was examined in MFLC- assisted differentiation, spontaneous differentiation, and growth factors (GF) and chemicals-induced differentiations (GF-induced differentiation) using retinoic acid, leukemia inhibitory factor (LIF), FGF2, FGF4, hepatocyte growth factor (HGF), oncostatin M (OSM), and dexamethasone. RESULTS: The mRNA expression of α-fetoprotein, albumin (ALB), α-1-antitrypsin, and hepatocyte nuclear factor 4α was observed earlier in the differentiating cES cells co-cultured with MFLCs, as compared to cES cells undergoing spontaneous differentiation and those subjected to GF-induced differentiation. The expression of cytochrome P450 7a1, a possible marker for embryonic endoderm-derived mature hepatocytes, was only observed in cES cells that had differentiated in a co-culture with MFLCs. Further, the disappearance of Oct3/4, a representative marker of an undifferentiated state, was noted in cells co-cultured with MFLCs, but not in those undergoing spontaneous or GF-induced differentiation. Immunocytochemical analysis revealed an increased ratio of ALB-immunopositive cells among cES cells co-cultured with MFLCs, while glycogen storageand urea synthesis were also demonstrated. CONCLUSION: MFLCs showed an ability to induce cES cells to differentiate toward hepatocytes. The co-culture system with MFLCs is a useful method for induction of hepatocyte-like cells from undifferentiated cES cells.

关键词：细胞分化肝细胞胎儿治疗

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Immunological mechanisms of fecal microbiota transplantation in recurrent Clostridioides difficile infection

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World Journal of Gastroenterology 2022年第33期28卷 4762-4772页

作者： Lucas F Soveral Gabriela G Korczaguin Pedro S Schmidt Isabel S Nunes Camilo Fernandes Carlos R Zarate-Blades Laboratory of Immunoregulation Department of MicrobiologyImmunologyand ParasitologyCenter for Dysbiosis ControlFederal University of Santa CatarinaFlorianopolis 88037-000Brazil division of Infectious Diseases Hospital Nereu RamosFlorianopolis 88025-301Brazil

Fecal microbiota transplantation(FMT)is a successful method for treating recurrent Clostridioides difficile(C.difficile)infection(rCDI)with around 90%efficacy.Due to the relative simplicity of this approach,it is being widely used and currently,thousands of patients have been treated with FMT worldwide.Nonetheless,the mechanisms underlying its effects are just beginning to be understood.Data indicate that FMT effectiveness is due to a combination of microbiological direct mechanisms against C.difficile,but also through indirect mechanisms including the production of microbiota-derived metabolites as secondary bile acids and short chain fatty acids.Moreover,the modulation of the strong inflammatory response triggered by C.difficile after FMT seems to rely on a pivotal role of regulatory T cells,which would be responsible for the reduction of several cells and soluble inflammatory mediators,ensuing normalization of the intestinal mucosal immune system.In this minireview,we analyze recent advances in these immunological aspects associated with the efficacy of FMT.

关键词： Fecal microbiota transplantation Immunity Mechanism Dysbiosis Pseudomembranous colitis Clostridioides difficile

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Pulmonary coinfection by Pneumocystis jiroveci and Cryptococcus neoformans

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Asian Pacific Journal of Tropical Biomedicine 2012年第1期2卷 80-82页

作者： Bava Javier Lloveras Susana Garro Santiago Troncoso Alcides Laboratory of parasitology Infectious Diseases Hospital division of Infectious Diseases Francisco Muniz Hospital Department of Infectious Diseases Tropical MedicineSchool of Medicine.Buenos Aires University

We communicate the diagnosis by microscopy of a pulmonary coinfection produced by Cryptococcus neoformans and Pneumocystis jiroveci,from a respiratory secretion obtained by bronchoalveolar lavage of an AIDS patient.Our review of literature identified this coinfection as unusual presentation.Opportunistic infections associated with HIV infection are increasingly recognized.It may occur at an early stage of HIV-infection.Whereas concurrent opportunistic infections may occur,coexisting Pneumocystis jiroveci pneumonia(PCP)and disseminated cryptococcosis with cryptococcal pneumonia is uncommon.The lungs of individuals infected with HIV are often affected by opportunistic infections and tumours and over two-thirds of patients have at least one respiratory episode during the course of their disease.Pneumonia is the leading HIV-associated infection.We present the case of a man who presented dual Pneumocystis jiroveci and cryptococcal pneumonia in a patient with HIV.Definitive diagnosis of PCP and Cryptococcus requires demonstration of these organisms in pulmonary tissues or fluid.In patients with<200/microliter CD4-lymphocytes,a bronchoalveolar lavage should be performed.This patient was successfully treated with amphotericin B and trimethoprim sulfamethoxazole.After 1 week the patient showed clinical and radiologic improvement and was discharged 3 weeks later.

关键词： Cryptococcus neoformans Pneumocystis jiroveci Pulmonary coinfection Diagnosis test Opportunistic pathogen Pneumonia Definitive diagnosis

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Methane production and small intestinal bacterial overgrowth in children living in a slum

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World Journal of Gastroenterology 2012年第41期18卷 5932-5939页

作者： Carolina Santos Mello Soraia Tahan Lígia Cristina FL Melli Mirian Silva do Carmo Rodrigues Ricardo Martin Pereira de Mello Isabel Cristina Affonso Scaletsky Mauro Batista de Morais Pediatric Gastroenterology division Federal University of Sao Paulo04039-031 Sao PauloBrazil Center Foundation Institute of Education for Osasco 06020-190 Sao PauloBrazil Infectology division Federal University of Sao Paulo04025-010 Sao PauloBrazil Microbiology Immunology and Parasitology DivisionFederal University of Sao Paulo 04023-062 Sao PauloBrazil

AIM:To analyze small intestinal bacterial overgrowth in school-aged children and the relationship between hydrogen and methane production in breath tests.METHODS:This transversal study included 85 children residing in a slum and 43 children from a private school,all aged between 6 and 10 years,in Osasco,Brazil.For characterization of the groups,data regarding the socioeconomic status and basic housing sanitary conditions were collected.Anthropometric data was obtained in children from both groups.All children completed the hydrogen(H 2) and methane(CH 4) breath test in order to assess small intestinal bacterial overgrowth(SIBO).SIBO was diagnosed when there was an increase in H 2 ≥ 20 ppm or CH 4 ≥ 10 ppm with regard to the fasting value until 60 min after lactulose ingestion.RESULTS:Children from the slum group had worse living conditions and lower nutritional indices than children from the private school.SIBO was found in 30.9%(26/84) of the children from the slum group and in 2.4%(1/41) from the private school group(P = 0.0007).Greater hydrogen production in the small intestine was observed in children from the slum group when compared to children from the private school(P = 0.007).A higher concentration of hydrogen in the small intestine(P < 0.001) and in the colon(P < 0.001) was observed among the children from the slum group with SIBO when compared to children from the slum group without SIBO.Methane production was observed in 63.1%(53/84) of the children from the slum group and in 19.5%(8/41) of the children from the private school group(P < 0.0001).Methane production was observed in 38/58(65.5%) of the children without SIBO and in 15/26(57.7%) of the children with SIBO from the slum.Colonic production of hydrogen was lower in methaneproducing children(P = 0.017).CONCLUSION:Children who live in inadequate environmental conditions are at risk of bacterial overgrowth and methane production.Hydrogen is a substrate for methane production in the colon.

关键词：氢气生产贫民窟产甲烷肠细菌儿童生长过度生活

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