Sequence of the genome RNA of rubella virus: Evidence for genetic rearrangement during togavirus evolution

An aliquot from a continuous fermenter culture of baby hamster kidney cells (BHK-21 Clone PD-4) (Wistar) maintained in Ca free Eagle's Basal Medium containing 2% Kaolin adsorbed fetal calf serum was planted in spinner flasks at 300,000 cells per ml, total volume 600 ml. After equilibration for one day at 35°C to insure that cells were in log phase, the culture was infected with the M-33-AGMK25 BHK-219 strain of rubella at an input multiplicity of about 6 TCID50 per cell. The virus was identified with specific rubella antiserum.Preliminary experiments had shown that such cultures would reach a peak or plateau HA titer of approximately 1:64, 24 hrs after inoculation and would continue to yield virus for 6 to 12 days. One hundred ml aliquot harvests were withdrawn daily and the culture was returned to volume with growth medium and incubation continued. The harvested cells were spun down rapidly at 2500 rpm per 15 mins., fixed in 3.7% gluteraldehyde in Ca free phosphate buffer saline, and post fixed in osmium tetraoxide. After dehydration, the cells were embedded in Epon 812 and cured approximately 20 hrs at 60°C.

Download Full-text

Identification of T-cell epitopes on E2 protein of rubella virus, as recognized by human T-cell lines and clones.

Journal of Virology ◽

10.1128/jvi.66.11.6788-6793.1992 ◽

1992 ◽

Vol 66 (11) ◽

pp. 6788-6793 ◽

Cited By ~ 13

Author(s):

D Ou ◽

P Chong ◽

Y Choi ◽

P McVeigh ◽

W A Jefferies ◽

...

Keyword(s):

T Cell ◽

Cell Lines ◽

Rubella Virus ◽

T Cell Epitopes ◽

E2 Protein ◽

Human T Cell ◽

T Cell Lines

Download Full-text

The rubella virus RNA binding activity of human calreticulin is localized to the N-terminal domain.

Journal of Virology ◽

10.1128/jvi.69.6.3848-3851.1995 ◽

1995 ◽

Vol 69 (6) ◽

pp. 3848-3851 ◽

Cited By ~ 23

Author(s):

C D Atreya ◽

N K Singh ◽

H L Nakhasi

Keyword(s):

Rubella Virus ◽

Rna Binding ◽

Binding Activity ◽

Terminal Domain ◽

Virus Rna

Download Full-text

Unmasking viral sequences by metagenomic next-generation sequencing in adult human blood samples during steroid-refractory/dependent graft-versus-host disease

Microbiome ◽

10.1186/s40168-020-00953-3 ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

M. C. Zanella ◽

S. Cordey ◽

F. Laubscher ◽

M. Docquier ◽

G. Vieille ◽

...

Keyword(s):

Next Generation Sequencing ◽

Graft Versus Host Disease ◽

Rubella Virus ◽

Viral Infections ◽

Next Generation ◽

Host Disease ◽

Graft Versus Host ◽

Viral Sequences ◽

Generation Sequencing ◽

Steroid Refractory

Abstract Background Viral infections are common complications following allogeneic hematopoietic stem cell transplantation (allo-HSCT). Allo-HSCT recipients with steroid-refractory/dependent graft-versus-host disease (GvHD) are highly immunosuppressed and are more vulnerable to infections with weakly pathogenic or commensal viruses. Here, twenty-five adult allo-HSCT recipients from 2016 to 2019 with acute or chronic steroid-refractory/dependent GvHD were enrolled in a prospective cohort at Geneva University Hospitals. We performed metagenomics next-generation sequencing (mNGS) analysis using a validated pipeline and de novo analysis on pooled routine plasma samples collected throughout the period of intensive steroid treatment or second-line GvHD therapy to identify weakly pathogenic, commensal, and unexpected viruses. Results Median duration of intensive immunosuppression was 5.1 months (IQR 5.5). GvHD-related mortality rate was 36%. mNGS analysis detected viral nucleotide sequences in 24/25 patients. Sequences of ≥ 3 distinct viruses were detected in 16/25 patients; Anelloviridae (24/25) and human pegivirus-1 (9/25) were the most prevalent. In 7 patients with fatal outcomes, viral sequences not assessed by routine investigations were identified with mNGS and confirmed by RT-PCR. These cases included Usutu virus (1), rubella virus (1 vaccine strain and 1 wild-type), novel human astrovirus (HAstV) MLB2 (1), classic HAstV (1), human polyomavirus 6 and 7 (2), cutavirus (1), and bufavirus (1). Conclusions Clinically unrecognized viral infections were identified in 28% of highly immunocompromised allo-HSCT recipients with steroid-refractory/dependent GvHD in consecutive samples. These identified viruses have all been previously described in humans, but have poorly understood clinical significance. Rubella virus identification raises the possibility of re-emergence from past infections or vaccinations, or re-infection.

Download Full-text