Immunomodulatory Effects of Saccharomyces cerevisiae Fermentation Product Supplementation on Immune Gene Expression and Lymphocyte Distribution in Immune Organs in Broilers

Abstract

A study was conducted to evaluate the molecular and cellular immunomodulatory effects of a Saccharomyces cerevisiae fermentation product (Original XPC, Diamond V) in broilers. Our lab has previously demonstrated that broilers fed XPC generate faster and stronger antigen-specific humoral immune responses to Newcastle disease virus (NDV) vaccination. This study aims at investigating the mechanism behind this increased immunocompetence. One-day-old broilers were randomly assigned to one of two treatments: 1.25 kg/ton S. cerevisiae fermentation product (XPC treatment group) or control diet. Birds were vaccinated against NDV on day 1 (B1 strain) and day 21 (LaSota strain) post-hatch. Innate and adaptive immune-related gene expression profiles in central (thymus and bursa of Fabricius) and peripheral (spleen) immune organs were investigated at 14 and 28 days of age by qPCR array. Fold changes larger than 1.2 (P < 0.05) between treated and control were considered significant. Lymphocyte subpopulations in central and peripheral immune organs and blood leukocytes were analyzed by flow cytometry at 14, 21, 28, and 42 days of age. In the spleen, Th1 immune responses and antiviral genes, such as IFN-γ, and its downstream genes signal transducer and activator of transcription (STAT4) and NFκB, were significantly upregulated in the treated group by 14 days of age. In the thymus, genes belonging to different functional groups were influenced at different time points. Cytokine genes associated with lymphocyte maturation, differentiation, and proliferation, such as IL-1R, IL-2, and IL-15 were significantly upregulated in the treated group by 28 days of age. Genes preferentially expressed in the medulla of the thymus and mature thymocytes, such as Myxovirus resistance gene 1, interferon regulatory factor-1, interferon regulatory factor-7, and STAT1, were upregulated in the birds supplemented with XPC. Birds supplemented with XPC had significantly higher percentages of CD3⁺, CD4⁺, and CD8⁺ T-cells in the thymus at day 28 of age, indicating production of more mature T-cells, which was consistent with gene expression results. Results suggest that XPC supplementation primes broilers to become more immunocompetent, without compromising growth performance.

Keywords: Saccharomyces cerevisiae fermentation product; immunomodulation; qPCR; spleen; thymus.

Figure 1

Lymphocyte subpopulations in thymus at 14, 21, 28, and 42 days of age. Percentage of (A) CD3⁺; (B) CD4⁺; (C) CD8⁺; (D) Bu-1⁺ lymphocytes in control (dark column) and XPC-supplemented (gray column) broilers (1.25 kg/ton; Diamond V Original XPC, Cedar Rapids, IA, USA). n = 5 per treatment per day. Data are presented as group means ± SEM. Different letters indicate significant differences (P < 0.05).

Figure 2

Lymphocyte subpopulations in blood at 14, 21, 28, and 42 days of age. Percentage of (A) CD3⁺; (B) CD4⁺; (C) CD8⁺; (D) Bu-1⁺ lymphocytes in control (dark column) and XPC-supplemented (gray column) broilers (1.25 kg/ton; Diamond V Original XPC, Cedar Rapids, IA, USA). n = 5 per treatment per day. Data are presented as group means ± SEM. Different letters indicate significant differences (P < 0.05).

Figure 3

Lymphocyte subpopulations in spleen at 14, 21, 28, and 42 days of age. Percentage of (A) CD3⁺; (B) CD4⁺; (C) CD8⁺; (D) Bu-1⁺ lymphocytes in control (dark column) and XPC-supplemented (gray column) broilers (1.25 kg/ton; Diamond V Original XPC, Cedar Rapids, IA, USA). n = 5 per treatment per day. Data are presented as group means ± SEM. Different letters indicate significant differences (P < 0.05).

Figure 4

Lymphocyte subpopulations in bursa at 14, 21, 28, and 42 days of age. Percentage of (A) CD3⁺; (B) CD4⁺; (C) CD8⁺; (D) Bu-1⁺ lymphocytes in control (dark column) and XPC-supplemented (gray column) broilers (1.25 kg/ton; Diamond V Original XPC, Cedar Rapids, IA, USA). n = 5 per treatment per day. Data are presented as group means ± SEM. Different letters indicate significant differences (P < 0.05).

Figure 5

Number of genes significantly regulated between XPC feed additive group and control in immune organs on the host response to a live Newcastle disease virus vaccine strain in vivo. Immune-associated gene expression pattern in spleen, bursa, and thymus between XPC and control groups measure by qPCR array on days 14 and 28 of age. Fold regulation more than 1.2 and P < 0.05 was considered as significantly regulated.