Genome-wide epitope mapping reveals significant diversity in antibody responses to Coxiella burnetii vaccination and infection

Coxiella burnetii is an important zoonotic bacterial pathogen of global importance, causing the disease Q fever in a wide range of animal hosts. Ruminant livestock, in particular sheep and goats, are considered the main reservoir of infection. Vaccination is a key control measure and two commercial vaccines based on formalin-inactivated C. burnetii bacterins are currently available. However, their deployment is limited due to significant reactogenicity in individuals previously sensitized to C. burnetii antigens. Furthermore, these vaccines interfere with available serodiagnostic tests which are also based on C. burnetii bacterin preparations. Subunit vaccines based on recombinant proteins offer significant advantages, as they can be designed to reduce reactogenicity and can be co-designed with defined antigen serodiagnostic tests to allow discrimination between vaccinated and infected individuals. This study aimed to investigate the diversity of antibody responses to C. burnetii vaccin..., Peptide microarray design
In this study, peptide microarrays were designed using the genome sequence of the C. burnetii Nine-Mile strain RSA493 (Accession no. NC_002971), which included 2092 proteins. The arrays also contained internal control peptides from Human Herpes Virus 5 with 170 proteins and tetanus toxin protein from Clostridium tetani. These 2263 protein sequences were processed into 168,792 unique 15 amino acid peptide sequences tiling four amino acids (overlapping by 11) in their parent proteins. 960 random 15 amino acid peptide sequences with similar amino acid frequencies were generated from the 2263 protein sequences to assess background control reactivity. C. burnetii-derived, tetanus toxin protein, and control peptide sequences were synthesized in duplicate, leading to a total of 328,984 peptide sequences.
Peptide microarray synthesis and probing
The microarrays were synthesized using a Nimble Therapeutics Maskless Array Synthesizer via light-directed solid-phase peptid..., Aggregated microarray data for all samples:
microarray_data_aggregated.txt
Fluorescence intensity values for each peptide on the microarray microarray are found in an aggregated tab-seperated file format. First column contains the synthesized peptide sequences, second column contains the peptide group:

EXPR_HHV5: HHV5-derived peptides

EXPR_MAIN_REP1/2: C. burnetii and tetanus toxin protein derived peptides
EXPR_NEG_CTRL_REP1/2: The random peptides

The remaining columns contain the fluorescence intensity values extracted for each of the 156 serum samples. The column names correspond to the microarray identifier (ID).
Peptide to protein mapper file:
peptide_map.txt
The peptide parent protein names and their locations are found in a tab-seperated file format. First column contains the synthesized peptide sequences, second column the name of their parent proteins, third column the length of the parent proteins, and fourth and fifth columns the start and coordinates of the peptides in the...,