American journal of veterinary research · 2025
Ex vivo coccidioidal antigen chitinase-1 stimulation increases production of interleukin-1β in healthy cats.
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Gordon Danielle, Jaffey Jared A, Shubitz Lisa F, Johnson Michael D L, Bolch Charlotte, Menghani Sanjay V, Benecke Brooke
Аннотация
OBJECTIVE: To evaluate coccidioidal antigen-stimulated production of cytokines in healthy cats and determine optimum testing conditions (ie, incubation time and antigen concentration).
METHODS: 6 client-owned, healthy cats that were seronegative for Coccidioides spp antibodies were included in this prospective experimental study. Whole blood cultures were performed with exposure to PBS (vehicle control) or a coccidioidal antigen (recombinant coccidioidal antigen chitinase-1 [rCTS1]105-310) at 5 final well concentrations (1 µg/mL, 10 µg/mL, 25 µg/mL, 50 µg/mL, and 75 µg/mL) with incubation times of 12 hours and 24 hours. A validated feline-specific, multiplex bead-based assay was used to measure 10 cytokines in cell culture supernatant.
RESULTS: Supernatant concentrations of IL-1β increased when stimulated with rCTS1105-310 for 12 hours at 50 µg/mL (274.4 pg/mL; P = .02) and 75 µg/mL (298.4 pg/mL; P = .003) compared to control (PBS; 102.9 pg/mL). Similarly, IL-1β concentrations increased after exposure to rCTS1105-310 for 24 hours at 50 µg/mL (340.3 pg/mL) and 75 µg/mL (364.8 pg/mL) compared to control (110.5 pg/mL; P = .04). Production of IL-1β was greater when whole blood was stimulated with rCTS1105-310 at 1 µg/mL for 24 hours (237.0 pg/mL) compared with 12 hours (116.8 pg/mL; P = .04). No other differences in cytokine concentrations were identified at either incubation time or rCTS1105-310 concentration compared to control.
CONCLUSIONS: Exposure of whole blood to rCTS1105-310 increases production of IL1β in healthy cats. Incubation time and rCTS1105-310 concentration did not seem to impact production of most cytokines in this study.
CLINICAL RELEVANCE: These results provide information on rCTS1105-310-stimulated cytokine concentrations in healthy cats.
METHODS: 6 client-owned, healthy cats that were seronegative for Coccidioides spp antibodies were included in this prospective experimental study. Whole blood cultures were performed with exposure to PBS (vehicle control) or a coccidioidal antigen (recombinant coccidioidal antigen chitinase-1 [rCTS1]105-310) at 5 final well concentrations (1 µg/mL, 10 µg/mL, 25 µg/mL, 50 µg/mL, and 75 µg/mL) with incubation times of 12 hours and 24 hours. A validated feline-specific, multiplex bead-based assay was used to measure 10 cytokines in cell culture supernatant.
RESULTS: Supernatant concentrations of IL-1β increased when stimulated with rCTS1105-310 for 12 hours at 50 µg/mL (274.4 pg/mL; P = .02) and 75 µg/mL (298.4 pg/mL; P = .003) compared to control (PBS; 102.9 pg/mL). Similarly, IL-1β concentrations increased after exposure to rCTS1105-310 for 24 hours at 50 µg/mL (340.3 pg/mL) and 75 µg/mL (364.8 pg/mL) compared to control (110.5 pg/mL; P = .04). Production of IL-1β was greater when whole blood was stimulated with rCTS1105-310 at 1 µg/mL for 24 hours (237.0 pg/mL) compared with 12 hours (116.8 pg/mL; P = .04). No other differences in cytokine concentrations were identified at either incubation time or rCTS1105-310 concentration compared to control.
CONCLUSIONS: Exposure of whole blood to rCTS1105-310 increases production of IL1β in healthy cats. Incubation time and rCTS1105-310 concentration did not seem to impact production of most cytokines in this study.
CLINICAL RELEVANCE: These results provide information on rCTS1105-310-stimulated cytokine concentrations in healthy cats.