Characterization of Encephalitozoon (Septata) intestinalis Isolates Cultured from Nasal Mucosa and Bronchoalveolar Lavage Fluids of Two AIDS Patients

Abstract

Microsporidia are obligate intracellular protozoan parasites that can cause opportunistic infections in AIDS patients. Species from five genera of microsporidia are presently known to infect man. One species, Septata intestinalis originally was detected in stool specimens of individuals with chronic diarrhea and subsequently was found to disseminate to the kidneys, lungs, and nasal sinuses. This organism has since been reclassified as Encephalitozoon and in this study, we report the culture of Encephalitozoon intestinalis from a bronchoalveolar lavage specimen and a nasal mucus aspirate of two AIDS patients living in the USA. The bronchoalveolar and nasal microsporidian isolates grew in several continuous cell lines including RK-13, MDCK, HT-29, Caco-2, Vero, and I047. Transmission electron microscopy of the clinical and cell culture specimens revealed that the new isolates appeared to be E. intestinalis based on morphology and growth of organisms in septated membrane-bound parasitophorous vacuoles. The new E. intestinalis isolates were characterized and compared with the first isolated E. intestinalis that was cultured from stool to confirm their identity and to determine if there existed any minor differences, as seen in the closely related Encephalitozoon cuniculi strains. By the methods of sodium dodecyl sulfate-polyacrylamide gel electrophoresis staining for proteins and carbohydrates, Western blot immunodetection, and polymerase chain reaction-based methods with restriction endonuclease digestion, double-stranded DNA heteroduplex mobility shift analysis, and DNA sequencing of the ribosomal DNA intergenic spacer region, the new isolates were identical to each other and to the reference isolate of E. intestinalis. In addition, with any of these methods, the E. intestinalis organisms could be distinguished from the three E. cuniculi strains, Encephalitozoon hellem, and Vittaforma corneae, which is important for diagnostics, therapeutic strategies, and epidemiology.

Quantitative detection of the tumor-associated antigen large external antigen in colorectal cancer tissues and cells using quantum dot probe

“…41 and HT29 cells are from a well-differentiated grade I tumor (equivalent to Dukes' type A). 42 This shows that the cell-based QD probe method can reveal the association between LEA expression and tumor characteristics, and remain in agreement with the tissue-based QD method. Furthermore, although nearly identical LEA expression correlation is confirmed using FCM, the cell-based QD probe method offers advantages over FCM, including the observation of cell images, analysis of marker localization, and understanding of tumor heterogeneity.…”

Synergy between expression of fusogenic membrane proteins, chemotherapy and facultative virotherapy in colorectal cancer

“…The permanent human cell lines DLD-1, 44 HT-29, 45 293, A549 and HeLa were obtained from the ATCC (Manassas, VA, USA). The Vero 2-2 cells were kindly provided by Dr Rozanne Sandri-Goldin (University of California, Irvine, CA, USA) via Dr Cornell Fraefel (University of Zurich, Switzerland).…”

Diversity of microsporidia (Fungi: Microsporidia) among captive great apes in European zoos and African sanctuaries: evidence for zoonotic transmission?

“…DNA from E. intestinalis spores originally isolated from an AIDS patient (Didier et al 1996a) and grown in vitro in VERO E6 cells in the Laboratory of Veterinary and Medical Protistology at the institute of Parasitology of Ascr and from E. bieneusi spores of genotype D originally isolated from a pig (Sak et al 2008) was used as positive control.…”