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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

Lymphocyte percentages shift upon immunization. (A) Bar graphs displaying percentages of small lymphocytes only and (B) large lymphocytes in peripheral blood at termination of 4-week immunization experiment. (C) Box and whisker plots of all cell type percentages. The box extends from first quartile to third quartile, the line in the box is the median, and whiskers extend to 1.5 times the inter-quartile range, outliers are points outside this range. Asterisks p < 0.01.
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Figure 2: Lymphocyte percentages shift upon immunization. (A) Bar graphs displaying percentages of small lymphocytes only and (B) large lymphocytes in peripheral blood at termination of 4-week immunization experiment. (C) Box and whisker plots of all cell type percentages. The box extends from first quartile to third quartile, the line in the box is the median, and whiskers extend to 1.5 times the inter-quartile range, outliers are points outside this range. Asterisks p < 0.01.

Mentions: Having standardized our differentiation of lymphocytes and granulocytes, we could perform differential blood cell counts on the terminal bleeds of the zebrafish in the three experimental groups (Table 1 and Datasheet S1 in Supplementary Material). Comparing the percentages of small lymphocytes, a slightly higher percentage was found in both the mucosal and i.p. immunization groups compared to the control group, but this did not reach statistical significance (Figure 2A). The percentage of large lymphocytes in peripheral blood was increased significantly in the i.p. injected fish over the mucosal fish (Figure 2B); however, neither of these was significantly increased over control PBS injected. The relative percentages of other cell types remained largely consistent between the three treatments; however, the neutrophil percentage was compromised when lymphocytes expanded (Figure 2C). This dip in neutrophil percentages could simply indicate an increase in lymphocyte percentages, without a corresponding change in absolute neutrophil number. However, it could also reflect an actual decrease in neutrophil number due to localized diapedesis or decreased production.


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)

Lymphocyte percentages shift upon immunization. (A) Bar graphs displaying percentages of small lymphocytes only and (B) large lymphocytes in peripheral blood at termination of 4-week immunization experiment. (C) Box and whisker plots of all cell type percentages. The box extends from first quartile to third quartile, the line in the box is the median, and whiskers extend to 1.5 times the inter-quartile range, outliers are points outside this range. Asterisks p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 2: Lymphocyte percentages shift upon immunization. (A) Bar graphs displaying percentages of small lymphocytes only and (B) large lymphocytes in peripheral blood at termination of 4-week immunization experiment. (C) Box and whisker plots of all cell type percentages. The box extends from first quartile to third quartile, the line in the box is the median, and whiskers extend to 1.5 times the inter-quartile range, outliers are points outside this range. Asterisks p < 0.01.
Mentions: Having standardized our differentiation of lymphocytes and granulocytes, we could perform differential blood cell counts on the terminal bleeds of the zebrafish in the three experimental groups (Table 1 and Datasheet S1 in Supplementary Material). Comparing the percentages of small lymphocytes, a slightly higher percentage was found in both the mucosal and i.p. immunization groups compared to the control group, but this did not reach statistical significance (Figure 2A). The percentage of large lymphocytes in peripheral blood was increased significantly in the i.p. injected fish over the mucosal fish (Figure 2B); however, neither of these was significantly increased over control PBS injected. The relative percentages of other cell types remained largely consistent between the three treatments; however, the neutrophil percentage was compromised when lymphocytes expanded (Figure 2C). This dip in neutrophil percentages could simply indicate an increase in lymphocyte percentages, without a corresponding change in absolute neutrophil number. However, it could also reflect an actual decrease in neutrophil number due to localized diapedesis or decreased production.

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