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

Weir H, Chen PL, Deiss TC, Jacobs N, Nabity MB, Young M, Criscitiello MF - Front Immunol (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Comparative Immunogenetics Laboratory, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University , College Station, TX , USA ; Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University , College Station, TX , USA ; Department of Science, A&M Consolidated High School , College Station, TX , USA.

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.

No MeSH data available.


Related in: MedlinePlus

IgM levels were raised in injected fish compared to control and mucosal, but not to statistical significance. All statistically relevant differences observed via ANOVA and post hoc TukeyHSD indicated by asterisks p < 0.01.
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Figure 3: IgM levels were raised in injected fish compared to control and mucosal, but not to statistical significance. All statistically relevant differences observed via ANOVA and post hoc TukeyHSD indicated by asterisks p < 0.01.

Mentions: Quantitative real-time PCR was used to measure gene expression of the major immunoglobulin isotypes and BAFF. BAFF binding to the BAFF receptor activates both the classical and non-canonical NFκB pathways generating signals critical for B cell formation and maintenance (38). BAFF was chosen as a transcription factor whose expression was indicative of humoral adaptive immune induction as it and IL-4 have been shown to play a critical role in zebrafish IgM responses (39). More BAFF, IgM, IgZ1, and IgZ2 were expressed in the injected fish than the control or mucosally immunized, yet wide individual variation prevented any significance (Figure S1A in Supplementary Material). The expression profiles of products of the two IgZ constant genes appeared to be in lock-step in the three groups. When the mean immunoglobulin expression differences were analyzed, the injection group was statistically greater than that of the control and mucosal groups (Figure S1B in Supplementary Material). We analyzed individual fish at the experimental terminus compared to PBS injected controls. IgZ was too low to detect for most samples but IgM values are shown in Figure 3. DNP-KLH injected fish had generally higher responses than control and mucosally stimulated fish, but individual variation was high and comparisons between groups did not reach statistical significance.


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

Weir H, Chen PL, Deiss TC, Jacobs N, Nabity MB, Young M, Criscitiello MF - Front Immunol (2015)

IgM levels were raised in injected fish compared to control and mucosal, but not to statistical significance. All statistically relevant differences observed via ANOVA and post hoc TukeyHSD indicated by asterisks p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664633&req=5

Figure 3: IgM levels were raised in injected fish compared to control and mucosal, but not to statistical significance. All statistically relevant differences observed via ANOVA and post hoc TukeyHSD indicated by asterisks p < 0.01.
Mentions: Quantitative real-time PCR was used to measure gene expression of the major immunoglobulin isotypes and BAFF. BAFF binding to the BAFF receptor activates both the classical and non-canonical NFκB pathways generating signals critical for B cell formation and maintenance (38). BAFF was chosen as a transcription factor whose expression was indicative of humoral adaptive immune induction as it and IL-4 have been shown to play a critical role in zebrafish IgM responses (39). More BAFF, IgM, IgZ1, and IgZ2 were expressed in the injected fish than the control or mucosally immunized, yet wide individual variation prevented any significance (Figure S1A in Supplementary Material). The expression profiles of products of the two IgZ constant genes appeared to be in lock-step in the three groups. When the mean immunoglobulin expression differences were analyzed, the injection group was statistically greater than that of the control and mucosal groups (Figure S1B in Supplementary Material). We analyzed individual fish at the experimental terminus compared to PBS injected controls. IgZ was too low to detect for most samples but IgM values are shown in Figure 3. DNP-KLH injected fish had generally higher responses than control and mucosally stimulated fish, but individual variation was high and comparisons between groups did not reach statistical significance.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Comparative Immunogenetics Laboratory, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University , College Station, TX , USA ; Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University , College Station, TX , USA ; Department of Science, A&M Consolidated High School , College Station, TX , USA.

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.

No MeSH data available.


Related in: MedlinePlus