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. 2022 May 10;13:868551. doi: 10.3389/fimmu.2022.868551

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Copyright © 2022 Simón, Martín-Martín, Morel, Díaz-Rosales and Tafalla

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Proliferation of IgM⁺ B cells in AT cells after the intraperitoneal immunization of rainbow trout with TNP-LPS. Confocal microscopy images of rainbow trout AT sections obtained from fish intraperitoneally immunized with the antigen TNP-LPS and mock immunized (control) at day 21 post-immunization. Sections were labeled with anti-IgM (green) in combination with the anti-proliferating cell nuclear antigen (PCNA) (red). All sections were also counterstained with DAPI (blue). (A) Representative images for each condition are shown (scale bars = 20µm) along with a graph showing the mean number of IgM⁺ B cells in immunized fish compared to control fish, calculated in 15 digital fields (400 x magnification) from 6 different individuals (mean + SD; n = 6 fish). Asterisks denote significantly different values between indicated groups (**p ≤ 0.01). (B) Representative images for each condition are shown (scale bars = 20µm) along with higher magnification images from control and TNP-LPS immunized tissues (white square; scale bar = 5 µm). Note that in non-proliferating cells, nuclei appear blue whereas they appear violet in proliferating cells. Graphs showing the mean number of total PCNA⁺ proliferating cells as well as the mean percentage of proliferating IgM⁺/PCNA⁺ cells in immunized fish compared to those in control fish are also included, calculated in 15 digital fields (400 x magnification) from 6 different individuals (mean + SD; n = 6).