DNP-KLH Yields Changes in Leukocyte Populations and Immunoglobulin Isotype Use with Different Immunization Routes in Zebrafish

Abstract

Distinct methods are required for inducing mucosal versus systemic immunity in mammals for vaccine protection at the tissues most commonly breached by pathogens. Understanding of mucosal immunization in teleost fish is needed to combat aquaculture disease, understand emerging ecological threats, and know how vertebrate adaptive immunity evolved. Here, we quantitatively measured expression levels of IgM as well as the teleost mucosal immunoglobulin, IgZ/IgT, in zebrafish given an antigen systemically via intraperitoneal (i.p.) injection or mucosally via bath immersion. Both immunoglobulin isotypes and the B cell activating factor gene transcription was induced in fish injected with antigen as compared to saline injected or antigen immersed fish, though these failed to reach statistical significance. Here we provide additional reference hematology for this model species. Differential blood counts revealed a greater lymphocyte percentage in both i.p. and immersed fish, with increase in large lymphocyte counts and decrease in neutrophils. These humoral adaptive gene transcription and cytological data should provide a foundation for more studies connecting immunology in this dominant developmental and genetic fish model to other species where mucosal immunization is of greater commercial importance.

A new genetic strategy for targeting microglia in development and disease

“…In the thymus, P2ry12-CreER; Rosa26 Ai14 recombination partially overlapped with Cx3cr1-EGFP and CD206+. The most prominent non-CNS recombination was in the spleen, where recombination was moderate (27.5 ± 5.8% of Cx3cr1-EGFP+ cells of the marginal zone and white pulp) and primarily restricted to CD206+; Cx3cr1-EGFP+ cells of the marginal zone between the red and white pulp, an area that contains macrophages that surveil the bloodstream, although there were a small number of TdT+ cells in the white pulp as well ( Figure 4B , Figure 4—figure supplement 1A–B ; Borges da Silva et al, 2015 ). In the liver, P2ry12-CreER; Rosa26 Ai14 recombination was excluded from perivascular SMA-adjacent Cx3CR1-EGFP cells, but was seen in Kupffer cells, in line with previous reports that Kupffer cells lack Cx3cr1 expression ( Figure 4—figure supplement 1C ; Yona et al, 2013 ).…”

Loss of runx1 function results in B cell immunodeficiency but not T cell in adult zebrafish

“…Keyhole limpet haemocyanin (KLH) (Sigma-Aldrich, St. Louis, USA) dissolved in sterile PBS fully, then emulsified in CFA (Sigma-Aldrich, St Louis, USA) and in IFA (Sigma-Aldrich, St Louis, USA) at the concentration of 5 µg µl −1 [ 35 , 60 ]. Ultrasonic emulsification on ice with 4 s, 2 s, 350 W was performed until the mixture would stay agglomerated when dropped on the ice-water.…”

TLR4 Agonist Combined with Trivalent Protein JointS of Streptococcus suis Provides Immunological Protection in Animals

“…Reliable animal models are crucial for vaccine testing. It is known that zebrafish are an ideal animal model for vaccine research because of their innate and adaptive immune systems and the similarity of their nonspecific immunity system to that in mammals [ 33 , 34 ]. A large number of zebrafish can be used without the limitation of animal resources [ 25 , 35 ].…”