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. 2018 Jul 31;38(4):BSR20180305. doi: 10.1042/BSR20180305

Figure 6. Percentage of B cells (CD45RA+) in spleen, PLN, MLN, and anti-T. canis antibody titration from uninfected and infected rats.

Figure 6

We used five animals per group and two rounds of experiments. Infection was performed by intragastric administration using a type Foley metallic probe at 8 weeks of age inoculating 2000 T. canis eggs per rat. They were performed by Flow cytometry. Briefly, spleen, PLN and MLN were collected at the time of killing. They were disaggregated using a 70-μm sterile nylon mesh. The following primary antibodies were used: PE anti-rat CD45RA. A secondary antibody solution was added AF488 or AF647 anti-Rabbit IgG. Data analysis was performed using the software FlowJo v10.0. The data represent the mean average (± S.D.). *Significant differences due to infection status (**P<0.01, ***P<0.001). TES-Ag were obtained by culturing the T. canis larvae according to the method described by de Savigny [17]. Purity and integrity of ESA was determined by SDS/PAGE and Coomassie staining, protein content was quantitated by Bradford method. The levels of ESA targetting antibodies in serum were measured indirectly by ELISA, Plate reading was done at 492 nm, 15 s agitation in an ELISA plate reader. The data represent the mean average (± S.D.). ***P<0.001. Abbreviations: Ctrl, uninfected rats; HPRL, adenohypophysis implant in renal capsule surgery; Infx, infected rats; Intact, untreated rats; Sh-HPRL, simulated adenohypophysis implant in renal capsule surgery.