First detection of Cryptosporidium DNA in blood and cerebrospinal fluid of HIV-infected patients

Velásquez, Jorge Néstor; Pantano, María Laura; Vittar, Natalia Belén Rumie; Nigro, Mónica Gabriela; Figueiras, Olga; Astudillo, Osvaldo Germán; Ricart, Javier; Paolera, D. Della; Carnevale, Silvana

doi:10.1007/s00436-018-5766-1

Cited by 7 publications

(6 citation statements)

References 49 publications

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“…In addition, we failed to detect the parasite 18S ribosomal DNA (rDNA) in the exosomes released from IEC4.1 cells following C. parvum infection ( Fig. 5C), using the nested PCR-restriction fragment length polymorphism (PCR-RFLP) of the 18S rDNA, as previously reported (35).…”

Section: Resultssupporting

confidence: 70%

“…DNA molecules are usually not detectable in exosomes from various types of cells (19,20). Accordingly, we failed to detect the parasite 18S rDNA in these exosomes released from infected cells, suggesting that exosome release from infected epithelial cells may not contribute to the systemic circulation of parasite DNA, as reported in HIV-infected patients with intestinal cryptosporidiosis (35).…”

Section: Discussionmentioning

confidence: 52%

“…In a recent report (35), Cryptosporidium 18S rDNA was detected in stool and tissue samples, as well as in blood and cerebrospinal fluid samples, from HIV-infected patients with intestinal cryptosporidiosis. DNA molecules are usually not detectable in exosomes from various types of cells (19,20).…”

Section: Discussionmentioning

confidence: 92%

“…The relative abundance of each RNA was calculated using the ΔΔC T threshold cycle (C T ) method and normalized to the amount of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) RNA (for measurement of whole-cell lysates) or U2 small nuclear (RNU2-1) RNA (for measurement of nuclear extracts). PCR-RFLP was carried out to detect C. parvum 18S rDNA as previously reported (35). The sequences for all the primers are listed in Table S1 in the supplemental material.…”

Section: Methodsmentioning

confidence: 99%

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Induction of Inflammatory Responses in Splenocytes by Exosomes Released from Intestinal Epithelial Cells followingCryptosporidium parvumInfection

et al. 2019

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Cryptosporidium, a protozoan parasite that infects the gastrointestinal epithelium and other mucosal surfaces in humans and animals, is an important opportunistic pathogen in AIDS patients and one of the most common enteric pathogens affecting young children in developing regions. This parasite is referred to as a “minimally invasive” mucosal pathogen, and epithelial cells play a central role in activating and orchestrating host immune responses. We previously demonstrated that Cryptosporidium parvum infection stimulates host epithelial cells to release exosomes, and these released exosomes shuttle several antimicrobial peptides to carry out anti-C. parvum activity. In this study, we detected the upregulation of inflammatory genes in the liver and spleen following C. parvum intestinal infection in neonatal mice. Interestingly, exosomes released from intestinal epithelial cells following C. parvum infection could activate the nuclear factor kappa B signaling pathway and trigger inflammatory gene transcription in isolated primary splenocytes. Several epithelial cell-derived proteins and a subset of parasite RNAs were detected in the exosomes released from C. parvum-infected intestinal epithelial cells. Shuttling of these effector molecules, including the high mobility group box 1 protein, was involved in the induction of inflammatory responses in splenocytes induced by the exosomes released from infected cells. Our data indicate that exosomes released from intestinal epithelial cells upon C. parvum infection can activate immune cells by shuttling various effector molecules, a process that may be relevant to host systemic responses to Cryptosporidium infection.

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Section: Resultssupporting

confidence: 70%

Section: Discussionmentioning

confidence: 52%

Section: Discussionmentioning

confidence: 92%

Section: Methodsmentioning

confidence: 99%

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Induction of Inflammatory Responses in Splenocytes by Exosomes Released from Intestinal Epithelial Cells followingCryptosporidium parvumInfection

et al. 2019

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“…One of these new pathogens is Cryptosporidium parvum, which is currently one of the four most important intestinal pathogens associated with diarrhea in humans (5). Cryptosporidiosis is a parasitic infection caused by Cryptosporidium that occurs in a wide range of vertebrates, including mammals, fish, reptiles, and birds (6,7). Cryptosporidiosis is a gastrointestinal infection caused by Cryptosporidium that is associated with diarrhea.…”

Section: Introductionmentioning

confidence: 99%

Assessment of Cryptosporidium in Patients With Gastroenteritis by Modified Ziehl-Neelsen Staining Method in East Azerbaijan Province of Iran During 2018-2019

Javidmehr

Garedaghi

Sioufi

2020

Int J Med Parasitol Epidemiol Sci

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Introduction: Cryptosporidiosis is a worldwide zoonotic disease that is caused by Cryptosporidium species and leads to acute or chronic diarrhea and vomiting in patients. Due to the suitable conditions of East Azerbaijan province for parasite transmission, the present study was performed to evaluate the prevalence of Cryptosporidium infection in patients with gastroenteritis referred to hospitals and reference laboratories of East Azerbaijan province during 2018-2019. Methods: A descriptive cross-sectional study was performed on patients with diarrhea who referred to hospitals and reference laboratories of East Azerbaijan province during 2018-2019. In this study, 180 fecal samples were collected from patients with gastroenteritis by cluster-random sampling. Stool samples were stored in 10% formalin and transferred to a parasitology laboratory and examined for parasite contamination by modified acid-fast staining (Modified Ziehl-Neelsen) method. Data were analyzed by chi-square test using SPSS. Results: Out of 180 samples, the highest number of diarrhea samples belonged to the age group over 40 years (35.5%) and the lowest belonged to the age group of 10-20 years (7.7%). Additionally, 56% of subjects were male and 44% were female. Of these patients, 57.7% lived in urban areas and 42.3% in rural areas. Cryptosporidium was observed in 1.6% (3 cases) of patients with gastroenteritis. There was a statistically significant relationship between Cryptosporidium infection and the age of patients with diarrhea (P<0.05). Conclusion: In the present study, a lower prevalence of cryptosporidiosis has been determined in comparison with previous studies in East Azerbaijan province. However, Cryptosporidium and other parasitic infections in the feces of patients with diarrhea referred to hospitals or reference laboratories need to be carefully diagnosed by appropriate parasitological methods.

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Calcium-Mediated Biophysical Binding of Cryptosporidium parvum Oocysts to Surfaces Is Sensitive to Oocyst Age

Sarkhosh

Zhang

Jellison

et al. 2019

Appl Environ Microbiol

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Cryptosporidium parvum causes potentially life-threatening gastrointestinal disease in humans and may not be effectively removed from drinking water via conventional methods. Prior research has shown that environmental biofilms immobilize oocysts from the water column, but the biophysical mechanisms driving this attraction are still under investigation. This study investigates the affinity of C. parvum oocysts to silanized surfaces. Surfaces were prepared with hydroxyl, amine, and carboxyl moieties. Binding forces between the oocysts and these engineered substrates were analyzed, with and without divalent ions, using atomic force microscopy. Binding forces were measured over several weeks to investigate the influence of age on adhesion. C. parvum oocysts bind most strongly to carboxylic acid functional groups, with rupture forces greater than that required to break noncovalent molecular bonds, regardless of oocyst age. This adhesion is shown to be due to divalent cation bridging mechanisms. In addition, the binding strength increases over a 5-week period as the oocysts age, followed by a decrease in the binding strength, which may be related to structural or biochemical changes in the outer wall-bound glycosylated proteins. This study sheds new light on the biochemical parameters that influence C. parvum oocyst binding to surfaces. Increased understanding of how age and water chemistry influence the binding strength of oocysts may inform future developments in environmental detection and drinking water treatment, such as with the development of oocyst-specific sensors that allow for more frequent tracking of oocysts in the environment. IMPORTANCE The mechanisms by which pathogens bind to surfaces are of interest to a wide variety of scientific communities, as these mechanisms drive infectivity, fate, and transport of the pathogenic organisms. This study begins to reveal the mechanism of direct binding of Cryptosporidium parvum to surfaces containing both carboxylic acid and amine moieties, in an attempt to understand how much of the binding ability is due to long-range electrostatic forces versus other mechanisms (specific or nonspecific) of bonding. In addition to improving the scientific understanding of fate and transport of oocysts, an expanded understanding of the binding mechanisms may aid in the development of new tools and sensors designed to detect and track oocysts in waterways. Furthermore, the methods used to examine binding in this study could be translated to other waterborne pathogens of interest.

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First detection of Cryptosporidium DNA in blood and cerebrospinal fluid of HIV-infected patients

Cited by 7 publications

References 49 publications

Induction of Inflammatory Responses in Splenocytes by Exosomes Released from Intestinal Epithelial Cells followingCryptosporidium parvumInfection

Induction of Inflammatory Responses in Splenocytes by Exosomes Released from Intestinal Epithelial Cells followingCryptosporidium parvumInfection

Assessment of Cryptosporidium in Patients With Gastroenteritis by Modified Ziehl-Neelsen Staining Method in East Azerbaijan Province of Iran During 2018-2019

Calcium-Mediated Biophysical Binding of Cryptosporidium parvum Oocysts to Surfaces Is Sensitive to Oocyst Age

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