scholarly journals Mucosal Delivery of a Respiratory Syncytial Virus CTL Peptide with Enterotoxin-Based Adjuvants Elicits Protective, Immunopathogenic, and Immunoregulatory Antiviral CD8+T Cell Responses

2001 ◽  
Vol 166 (2) ◽  
pp. 1106-1113 ◽  
Author(s):  
Cameron P. Simmons ◽  
Tracy Hussell ◽  
Tim Sparer ◽  
Gerhard Walzl ◽  
Peter Openshaw ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
T Cell ◽  
T Cell Responses ◽  
Cd8 T Cell ◽  
Cell Responses ◽  
Mucosal Delivery ◽  
Syncytial Virus

scholarly journals Characterization of the CD8+ T cell responses directed against respiratory syncytial virus during primary and secondary infection in C57BL/6 mice

Virology ◽  
2006 ◽  
Vol 352 (1) ◽  
pp. 157-168 ◽  
Author(s):  
Michaël V. Lukens ◽  
Erwin A.W. Claassen ◽  
Patricia M.A. de Graaff ◽  
Mariska E.A. van Dijk ◽  
Peter Hoogerhout ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
T Cell ◽  
Secondary Infection ◽  
T Cell Responses ◽  
Cd8 T Cell ◽  
Cell Responses ◽  
Syncytial Virus

scholarly journals Gene-gun DNA vaccination aggravates respiratory syncytial virus-induced pneumonitis

2004 ◽  
Vol 85 (10) ◽  
pp. 3017-3026 ◽  
Author(s):  
Christina Bartholdy ◽  
Wieslawa Olszewska ◽  
Anette Stryhn ◽  
Allan Randrup Thomsen ◽  
Peter J. M. Openshaw
Keyword(s):  
Weight Loss ◽  
Respiratory Syncytial Virus ◽  
T Cell ◽  
T Cell Responses ◽  
Dna Vaccination ◽  
T Cell Memory ◽  
Gene Gun ◽  
Cell Responses ◽  
Virus Clearance ◽  
Syncytial Virus

A CD8+ T-cell memory response to respiratory syncytial virus (RSV) was generated by using a DNA vaccine construct encoding the dominant Kd-restricted epitope from the viral transcription anti-terminator protein M2 (M282–90), linked covalently to human β 2-microglobulin (β 2m). Cutaneous gene-gun immunization of BALB/c mice with this construct induced an antigen-specific CD8+ T-cell memory. After intranasal RSV challenge, accelerated CD8+ T-cell responses were observed in pulmonary lymph nodes and virus clearance from the lungs was enhanced. The construct induced weaker CD8+ T-cell responses than those elicited with recombinant vaccinia virus expressing the complete RSV M2 protein, but stronger than those induced by a similar DNA construct without the β 2m gene. DNA vaccination led to enhanced pulmonary disease after RSV challenge, with increased weight loss and cell recruitment to the lung. Depletion of CD8+ T cells reduced, but did not abolish, enhancement of disease. Mice vaccinated with a construct encoding a class I-restricted lymphocytic choriomeningitis virus epitope and β 2m suffered more severe weight loss after RSV infection than unvaccinated RSV-infected mice, although RSV-specific CD8+ T-cell responses were not induced. Thus, in addition to specific CD8+ T cell-mediated immunopathology, gene-gun DNA vaccination causes non-specific enhancement of RSV disease without affecting virus clearance.

scholarly journals Enzyme-Linked Immunospot Assay for Detection of Human Respiratory Syncytial Virus F Protein-Specific Gamma Interferon-Producing T Cells

2014 ◽  
Vol 21 (5) ◽  
pp. 628-635 ◽  
Author(s):  
Kathryn Patton ◽  
Shahin Aslam ◽  
Jim Lin ◽  
Li Yu ◽  
Stacie Lambert ◽  
...  
Keyword(s):  
T Cells ◽  
Respiratory Syncytial Virus ◽  
T Cell ◽  
Elispot Assay ◽  
T Cell Responses ◽  
The Elderly ◽  
Assay Sensitivity ◽  
Cell Responses ◽  
Ifn Γ ◽  
Syncytial Virus

ABSTRACTRespiratory syncytial virus (RSV) causes significant disease in elderly adults, and we have previously reported that individuals 65 years of age and older have reduced RSV F protein-specific gamma interferon (IFN-γ)-producing T cells compared to healthy younger adults. To measure RSV F-specific memory T cell responses in the elderly following infection or vaccination, we optimized and qualified an IFN-γ enzyme-linked immunospot (ELISPOT) assay. Since peripheral blood mononuclear cells (PBMC) from the elderly could be more fragile, we established optimal cryopreservation techniques and minimal viability acceptance criteria. The number of cells per well, types and concentrations of stimulation antigens, and incubation times were evaluated to maximize assay sensitivity and precision. The optimized assay uses 300,000 cells/well, 2 μg/ml of an RSV F peptide pool (RSV Fpp), and incubation for 22 ± 2 h in serum-free CTL-Test medium. The assay was qualified by 3 analysts using 3 RSV F-responding donor PBMC samples (high, medium, and low responders) tested on 5 different assay days. The assay sensitivity or limit of detection (LOD) was determined to be 21 spot-forming cells (SFC) per 106PBMC, and the lower limit of quantitation (LLOQ) was estimated to be 63 SFC/106PBMC. The intra- and interassay percent coefficients of variation (CV) were <10.5% and <31%, respectively. The results of the qualification study demonstrate that a robust, precise, and sensitive IFN-γ ELISPOT assay has been developed that is fit for measuring RSV F-specific IFN-γ T cell responses in subjects enrolled in a vaccine clinical trial or in epidemiology studies.

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