Of these proteins, 633 proteins (94

Of these proteins, 633 proteins (94.07%) were efficiently expressed as determined by dual positive signals against the amino-terminal HIS and carboxy-terminal hemagglutinin (HA) tags. Identification of antigens and selection of vaccine candidates The antibody response profile to proteins in infected, uninfected, and SPF domestic AF-353 cats is shown as a heatmap in Fig.?1. and information for individual cats. 12014_2018_9218_MOESM5_ESM.pdf (39K) GUID:?35D44528-0C44-4BA8-95BA-A98F2ACFE91F Additional file 6: Supplementary Table?3. Summary of supportive care administered to individual cats. Cats vaccinated with CF-Library received less overall supportive care. 12014_2018_9218_MOESM6_ESM.pdf (39K) GUID:?6C91A8B9-98A7-42A0-9332-B53A5C7057BF Additional file 7: Supplementary Fig.?3. Serological responses to antigens are increased during chronic contamination. Serologic reactivity of antigens are depicted as a heatmap in (A). Antigens are outlined in rows while grouping of individuals are denoted in columns. Average signal intensity of individual antigens against serum from acute and chronic groups are depicted in (B). The majority of antigens that experienced a higher average reactivity against serum from chronically infected cats (n?=?50), but two antigens were more reactive against serum from acutely infected cats (n?=?2, denoted with yellow boxes). These two antigens were incorporated into the CF-Library vaccine as Candidates 32-33. 12014_2018_9218_MOESM7_ESM.pdf (182K) GUID:?B1A3D28D-E26D-44EA-ADD3-2DE79F86262E Additional file 8: Supplementary Fig.?4. Serological profiles of individual vaccinated cats to candidates throughout study. Arrays made up of the 32 vaccine candidates included in CF-Library and in CF-1 were probed with sera samples from individual cats. Heat map shows normalized signal intensity with red strongest, white weakest, and gray intermediate. Rows denote 32 different candidates included in vaccines outlined in descending order of reactivity; candidates 19 and 3 were included in CF-1, while all outlined candidates were included in AF-353 CF-Library. Results are organized by individual cats (recognized by number at top), and survival status of each AF-353 cat is usually indicated as A (alive) or D (lifeless). Individual columns within each cats array symbolize serum samples collected at different time points through study (labeled by the day in the study the sample was collected; refer to Supplemental Fig.?2 for timeline). There was no correlation between immunization protocol, individual reactivity, and survival for most cats, with the exception of Cat 77, who experienced widespread yet poor AF-353 reactivity against all candidates in the CF-Library vaccine prior to infection and subsequently experienced milder disease and did not require supportive care or antiprotozoal therapy. 12014_2018_9218_MOESM8_ESM.pdf (170K) GUID:?C8CBF4B9-AFC1-4D0E-B2DD-0EB926BD6386 Data Availability StatementAll relevant data is included in this manuscript or as figures, furniture, or additional files. Any additional datasets analyzed during the current study are available from your corresponding author on reasonable request. Abstract Background Cytauxzoonosis is a disease of felids in North America caused by the tick-transmitted apicomplexan parasite genome has recognized over 4300 putative protein-encoding genes. From this pool we constructed a protein microarray containing 673 putative proteins. This microarray was probed with sera from antigens that were incorporated into the two expression library vaccines. However, expression library immunization failed to prevent contamination or disease in cats challenged with antigens. These antigens should be considered for development of vaccines, Rabbit Polyclonal to CHRM4 diagnostics, and therapeutics. Electronic supplementary material The online version of this article (10.1186/s12014-018-9218-9) contains supplementary material, which is available to authorized users. is usually a tick-transmitted apicomplexan parasite that is the causative agent of cytauxzoonosis in domestic and wild felids in North and South America [1C8]. A closely related, genetically unique sp. has been recognized in Europe [9C13], but has not been associated with vintage cytauxzoonosis which is usually characterized by high mortality rates and massive proliferation and vascular dissemination of schizont-infected myeloid cells. Although no longer considered uniformly fatal in domestic cats, morbidity and mortality remain high for AF-353 individuals presenting with acute cytauxzoonosis [14C19]. Even with treatment, which can cost thousands of dollars, mortality remains at least 40% [20]; a vaccine is not currently available. Following its initial discovery in Missouri in.