READ ME File For 'Dataset in support of the Southampton Master of Philosophy thesis 'Airway epithelial cell regulation of T cell responses during viral infection'

Dataset DOI: 10.5258/SOTON/D2807 

ReadMe Author: CLAIRE SIMMS, University of Southampton 0000-0003-2937-5079

This dataset supports the thesis entitled 'Airway epithelial cell regulation of T cell responses during viral infection' 
AWARDED BY: Univeristy of Southampton
DATE OF AWARD: 2023

DESCRIPTION OF THE DATA

This dataset contains: RT-PCR data generated from in vitro Air Liquid Interface cultured BCI-NS1.1 cells as well as PBEC derived from two anonymized human lung tissue samples. 
Samples were exposed to live RSV-A2 obtained from Virapur as pre-purified aliquots with an approximate concentration of 7x107 PFU/Ml. Stock virus was prepared into 5 2-fold serial dilutions in HBSS with 1.25ul to 20ul of stock added per sample in addition to uninfected controls.
RNA was collected at 2h and 72h post infection using Qiazol reagent and converted to cDNA as described in the associated thesis, chapter 2.5. RT-PCR was perfomed in singleplex TaqMan experiments with HPRT1 and GAPDH used as housekeeping genes for analysis.
Reactions were performed in duplicate and amplified using the following cycling parameters: 95°C for 10 min and 40 cycles of 95°C for 15s and 60°C for 1 min using a 7900HT Fast Real-Time PCR System. 

Raw Ct values generated in this way are included in the file 'RAW RT-PCR data'. The data and fold for the genes discussed in the associated thesis, RSV-N, CXCL10, HPRT1 and GAPDH, are included in the 'DataSubmission_RT_PCR_ALI. Both .xlsx files are accessible using Microsoft Excel. 
Fold changes are calculated for 72h infection relative to 2h uninfected samples using the DDCT method and presented for replicate BCi ALI samples (N=5) and PBEC ALI (N=2) on seperate labelled sheets.
BCI ALI sample names reflect the replicate assay (BCI-NY1, BCI-NY2 etc) as well as RSV dose with 'P1' indicating 1.25ul of stock, 'P5' 20ul of stock and 'NI' indicating Not infected.
For the PBEC samples, the RSV load was presented as multiplicity of Infection (MOI).  


Date of data collection: December 2020 - August 2022

Information about geographic location of data collection: Collected at the school of Clinical and Experimental Sciences, University of Southampton


Date that the file was created: October 2023

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