Developmental gene expression in Leishmania donovani: differential cloning and analysis of an amastigote-stage-specific gene

1994 ◽  
Vol 14 (5) ◽  
pp. 2975-2984
Author(s):  
H Charest ◽  
G Matlashewski
Keyword(s):  
Life Cycle ◽  
Leishmania Donovani ◽  
Repetitive Sequences ◽  
Protozoan Parasite ◽  
Life Cycle Stage ◽  
Immune Serum ◽  
Sand Fly ◽  
Specific Gene ◽  
Reading Frame ◽  
Developmental Gene Expression

Leishmania protozoans are the causative agents of leishmaniasis, a major parasitic disease in humans. During their life cycle, Leishmania protozoans exist as flagellated promastigotes in the sand fly vector and as nonmotile amastigotes in the mammalian hosts. The promastigote-to-amastigote transformation occurs in the phagolysosomal compartment of the macrophage cell and is a critical step for the establishment of the infection. To study this cytodifferentiation process, we differentially screened an amastigote cDNA library with life cycle stage-specific cDNA probes and isolated seven cDNAs representing amastigote-specific transcripts. Five of these were closely related (A2 series) and recognized, by Northern (RNA) blot analyses, a 3.5-kb transcript in amastigotes and in amastigote-infected macrophages. Expression of the amastigote-specific A2 gene was induced in promastigotes when they were transferred from culture medium at 26 degrees C and pH 7.4 to medium at 37 degrees C and pH 4.5, conditions which mimic the macrophage phagolysosomal environment. A2 genes are clustered in tandem arrays, and a 6-kb fragment corresponding to a unit of the cluster was cloned and partially sequenced. An open reading frame found within the A2-transcribed region potentially encoded a 22-kDa protein containing repetitive sequences. The recombinant A2 protein produced in Escherichia coli cells was specifically recognized by immune serum from a patient with visceral leishmaniasis. The A2 protein repetitive element has strong homology with an S antigen of Plasmodium falciparum, the protozoan parasite responsible for malaria. Both the A2 protein of Leishmania donovani and the S antigen of P. falciparum are stage specific and developmentally expressed in mammalian hosts.

scholarly journals Developmental gene expression in Leishmania donovani: differential cloning and analysis of an amastigote-stage-specific gene.

1994 ◽  
Vol 14 (5) ◽  
pp. 2975-2984 ◽  
Author(s):  
H Charest ◽  
G Matlashewski
Keyword(s):  
Life Cycle ◽  
Leishmania Donovani ◽  
Repetitive Sequences ◽  
Protozoan Parasite ◽  
Life Cycle Stage ◽  
Immune Serum ◽  
Sand Fly ◽  
Specific Gene ◽  
Reading Frame ◽  
Developmental Gene Expression

Leishmania protozoans are the causative agents of leishmaniasis, a major parasitic disease in humans. During their life cycle, Leishmania protozoans exist as flagellated promastigotes in the sand fly vector and as nonmotile amastigotes in the mammalian hosts. The promastigote-to-amastigote transformation occurs in the phagolysosomal compartment of the macrophage cell and is a critical step for the establishment of the infection. To study this cytodifferentiation process, we differentially screened an amastigote cDNA library with life cycle stage-specific cDNA probes and isolated seven cDNAs representing amastigote-specific transcripts. Five of these were closely related (A2 series) and recognized, by Northern (RNA) blot analyses, a 3.5-kb transcript in amastigotes and in amastigote-infected macrophages. Expression of the amastigote-specific A2 gene was induced in promastigotes when they were transferred from culture medium at 26 degrees C and pH 7.4 to medium at 37 degrees C and pH 4.5, conditions which mimic the macrophage phagolysosomal environment. A2 genes are clustered in tandem arrays, and a 6-kb fragment corresponding to a unit of the cluster was cloned and partially sequenced. An open reading frame found within the A2-transcribed region potentially encoded a 22-kDa protein containing repetitive sequences. The recombinant A2 protein produced in Escherichia coli cells was specifically recognized by immune serum from a patient with visceral leishmaniasis. The A2 protein repetitive element has strong homology with an S antigen of Plasmodium falciparum, the protozoan parasite responsible for malaria. Both the A2 protein of Leishmania donovani and the S antigen of P. falciparum are stage specific and developmentally expressed in mammalian hosts.

No stress – Hsp90 and signal transduction inLeishmania

Parasitology ◽  
2014 ◽  
Vol 141 (9) ◽  
pp. 1156-1166 ◽  
Author(s):  
A. HOMBACH ◽  
J. CLOS
Keyword(s):  
Signal Transduction ◽  
Life Cycle ◽  
Protein Kinases ◽  
Leishmania Donovani ◽  
Protozoan Parasite ◽  
Specific Protein ◽  
Signal Transduction Pathways ◽  
Phosphorylation Sites ◽  
Therapeutic Approaches ◽  
Novel Therapeutic

SUMMARYHsp90 (a.k.a. Hsp83) plays a significant role in the life cycle control of the protozoan parasiteLeishmania donovani. Rather than protectingLeishmaniaspp. against adverse and stressful environs, Hsp90 is required for the maintenance of the motile, highly proliferative insect stage, the promastigote. However, Hsp90 is also essential for survival and proliferation of the intracellular mammalian stage, the amastigote. Moreover, recent evidence shows Hsp90 and other components of large multi-chaperone complexes as substrates of stage-specific protein phosphorylation pathways, and thus as likely effectors of the signal transduction pathways inLeishmaniaspp. Future efforts should be directed towards the identification of the protein kinases and the critical phosphorylation sites as targets for novel therapeutic approaches.

Construction and validation of a polycompetitor construct (SWITCH) for use in competitive RT-PCR to assess tachyzoite-bradyzoite interconversion in Toxoplasma gondii

Parasitology ◽  
2001 ◽  
Vol 123 (5) ◽  
pp. 433-439 ◽  
Author(s):  
R. E. LYONS ◽  
K. LYONS ◽  
R. McLEOD ◽  
C. W. ROBERTS
Keyword(s):  
Life Cycle ◽  
Toxoplasma Gondii ◽  
Protozoan Parasite ◽  
Housekeeping Gene ◽  
Life Cycle Stage ◽  
Intermediate Hosts ◽  
Related Family ◽  
Intracellular Protozoan Parasite ◽  
Β Tubulin ◽  
Disease Present

The obligate intracellular protozoan parasite, Toxoplasma gondii exists as 2 life-cycle forms in intermediate hosts. The rapidly dividing tachyzoites responsible for acute disease, present in the first 14 days of infection, give rise to slowly dividing bradyzoites that reside in tissue cysts. Reactivation of disease is associated with conversion of bradyzoites to tachyzoites. A sensitive method for detection and assessment of the number of each life-cycle stage would be useful for following these events. Herein we describe the construction and validation of a plasmid (pSWITCH) containing a polycompetitor construct (SWITCH) for use in competitive reverse transcriptase-PCR (cRT-PCR). pSWITCH contains competitors for SAG2A and LDH2 genes, which are exclusively expressed by tachyzoite and bradyzoite stages respectively, and for β-tubulin, a gene expressed by both stages. Using cRT-PCR, samples can first be accurately normalized for expression of the housekeeping gene, β-tubulin and then the relative levels of SAG2A and LDH2 expression compared to follow stage conversion. The abundance of transcripts for other genes of interest can then be followed during this process as demonstrated here for the SAG2-related family of genes. This technique offers a powerful tool for studying the processes involved in tachyzoite and bradyzoite interconversion.

Natural and synthetic indirubins as potent and selective inhibitors of the protozoan parasite Leishmania donovani

Planta Medica ◽  
2007 ◽  
Vol 73 (09) ◽  
Author(s):  
E Xingi ◽  
D Smirlis ◽  
S Bisti ◽  
V Myrianthopoulos ◽  
P Magiatis ◽  
...  
Keyword(s):  
Leishmania Donovani ◽  
Protozoan Parasite ◽  
Selective Inhibitors ◽  
Parasite Leishmania ◽  
Natural And Synthetic

IN VITRO DESTRUCTION OF LEISHMANIA DONOVANI INFECTED CELLS BY IMMUNE SERUM

1976 ◽  
Vol 4 (3-4) ◽  
pp. 207-211
Author(s):  
SHUN SHINBO ◽  
TAKATOSHI KOBAYAKAWA ◽  
HIROSHI ISHIYAMA ◽  
KAZUSHIGE MASUDA
Keyword(s):  
Leishmania Donovani ◽  
Immune Serum ◽  
Infected Cells

scholarly journals Metabolic reprogramming during the Trypanosoma brucei life cycle

F1000Research ◽  
2017 ◽  
Vol 6 ◽  
pp. 683 ◽  
Author(s):  
Terry K. Smith ◽  
Frédéric Bringaud ◽  
Derek P. Nolan ◽  
Luisa M. Figueiredo
Keyword(s):  
Life Cycle ◽  
Trypanosoma Brucei ◽  
Model Organism ◽  
Protozoan Parasite ◽  
Tsetse Fly ◽  
Metabolic Reprogramming ◽  
Mammalian Host ◽  
Insect Vector ◽  
Environmental Signals ◽  
Cellular Metabolic Activity

Cellular metabolic activity is a highly complex, dynamic, regulated process that is influenced by numerous factors, including extracellular environmental signals, nutrient availability and the physiological and developmental status of the cell. The causative agent of sleeping sickness, Trypanosoma brucei, is an exclusively extracellular protozoan parasite that encounters very different extracellular environments during its life cycle within the mammalian host and tsetse fly insect vector. In order to meet these challenges, there are significant alterations in the major energetic and metabolic pathways of these highly adaptable parasites. This review highlights some of these metabolic changes in this early divergent eukaryotic model organism.

scholarly journals Clinical and laboratory profiles of visceral leishmaniasis among adult patients admitted to Felege Hiwot Hospital, Bahir Dar, Ethiopia

2021 ◽  
Vol 9 ◽  
pp. 205031212110367
Author(s):  
Berhanu Tarekegn ◽  
Ayanaw Tamene
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Donovani ◽  
Clinical Presentation ◽  
Winter Season ◽  
Adult Patients ◽  
Sand Fly ◽  
Timely Initiation ◽  
Vector Borne Disease ◽  
Vector Borne ◽  
Endemic Areas

Background: Visceral leishmaniasis is a vector-borne disease caused by Leishmania donovani transmitted by sand fly species. It is the third most common vector-borne disease globally. Visceral leishmaniasis is endemic in Ethiopia with an estimated annual incidence ranging from 3700 to 7400 cases. This research aimed to assess the clinical presentations and laboratory profiles of visceral leishmaniasis for early diagnosis and timely initiation of management. Objective: To describe the clinical and laboratory manifestation and diagnostic modalities of visceral leishmaniasis among adult patients admitted to Felege Hiwot Hospital, from 1 September 2016 to 30 August 2019. Method: Institution-based retrospective cross-sectional study was conducted among 141 patients admitted to Felege Hiwot Hospital from 1 September 2016 to 30 August 2019. Descriptive statistics were used to describe the clinical presentation and laboratory profiles of patients with visceral leishmaniasis. Results: Among a total of 141 enrolled patients in the study, males were affected 13-fold. Most of them were travelers to endemic areas during the winter season for labor work. The mean duration of illness was 48 days. Common symptoms were fever (96.5%), weightless (82.5%), jaundice (18.4%), vomiting/diarrhea (13.5%), and bleeding episodes (11.3%). Splenomegaly was seen in 98.6%, ascites in 35.5%, and lymphadenopathy in 9.9%. Lymphadenopathy was seen significantly in HIV patients (40%). Anemia was seen in 95%, thrombocytopenia in 90.2%, leukopenia in 86.4%, and pancytopenia in 79.4%. Half of the patients had coinfection. Neutropenic sepsis was seen in 21.3%. The diagnosis was made by tissue aspiration in 65% of patients. Conclusion: The majority of patients who were diagnosed to have visceral leishmaniasis were young male adults who traveled to the endemic areas seasonally. Fever and splenomegaly were seen as the commonest clinical presentation. Lymphadenopathy occurred in high frequency among HIV co-infected patients. Anemia was the commonest hematologic finding.

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