Immune cell counts and signaling in body fluids of cows vaccinated against Clostridium difficile

Christiane Schmautz,Michael W Pfaffl,Ines Ballweg,Nadine Müller,Heike Kliem,Marlene Auer

doi:10.1186/s40709-018-0092-4

Christiane Schmautz, Michael W Pfaffl + Show 4 more

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https://doi.org/10.1186/s40709-018-0092-4

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Abstract

BackgroundNew treatment options are needed to prevent relapses following failed antibiotic therapies of Clostridium difficile infections (CDI) in humans. The concomitant therapy with an anti-C. difficile IgA containing whey protein concentrate can support the sustainable recovery of CDI patients. For 31 weeks, nine dairy cows were continuously vaccinated with several anti-C. difficile vaccines by certain routes of administration to produce anti-C. difficile IgA enriched milk. The study aimed at finding decisive differences between low responder (LR) and high responder (HR) cows (> 8.0 µg ml−1 total milk C. difficile specific IgA) concerning their immune response to vaccination on cellular and molecular biological levels.ResultsThe results of total and differential cell counting (DCC) in blood and milk and the outcomes of the gene expression analysis of selected immune factors were assessed relating to the usage of two vaccine batches for injection (MucoCD-I batch A and B), marking two immunization (IM) periods, and compared to a control group (Ctr). The MucoCD-I batch A caused short-term leukopenia followed by leukocytosis in the blood of LR and HR. The total somatic cell counts in milk were not altered by the treatment. The DCC revealed that the leukocytes of the treated groups were partly impaired by the treatment. The gene expression analysis exposed cumulative and sustainable differences (p < 0.05) between LR and HR for the genes encoding for lactoferrin, CXCL8, IL1β, IL2, IL6, IL12β, IFNγ, CD4 and CD163. The regulation of the epithelial IgA cell receptor PIGR was not impaired by the IM. In contrast to the vaccination with MucoCD-I batch A, the second IM period with MucoCD-I batch B resulted in mitigation and synchronization of the treated groups’ immune responses.ConclusionsThe inversely regulated cytokines in the blood and milk cells of the treated groups led to a variously directed, local T cell response resulting in their different production intensities of C. difficile specific IgA in milk.

Highlights

New treatment options are needed to prevent relapses following failed antibiotic therapies of Clostridium difficile infections (CDI) in humans
All cows designated for immunization (IM) showed a basic level of less than 2 μg of anti-C. difficile IgA per milliliter milk on average owing to the natural dissemination of C. difficile
The gene expression analysis of peripheral blood leukocytes (PBL) spawned sustainable differences between high responder (HR) and low responder (LR). Their expression patterns of the examined cytokines differed significantly during the first IM period. This outcome might have been crucial for the different response of the treated groups to the vaccinations against C. difficile because the intercellular mediators are important to orchestrate the immune cells for thwarting a pathogen attack

Summary

Introduction

New treatment options are needed to prevent relapses following failed antibiotic therapies of Clostridium difficile infections (CDI) in humans. The study aimed at finding decisive differences between low responder (LR) and high responder (HR) cows (> 8.0 μg ml−1 total milk C. difficile specific IgA) concerning their immune response to vaccination on cellular and molecular biological levels. The outcome of their immune reaction to the same immunological stimulus may vary fundamentally Given this fact, high responder (HR) and low responder (LR) cows were identified, as measured by their production of anti-C. difficile specific antibodies in milk [21]. The first encounter with the antigen provokes the innate IS, which includes components like the complement system or antigen-presenting cells (APCs) The latter activate the second arm of the IS, the adaptive IS, which is associated with lymphocytes as effector cells, antibody production and immunological memory [22]. A more fine-tuned cause analysis for the determined different responsiveness of the treated cows was exemplarily achieved by molecular genetic analysis to verify the degree of cellular activity by the expression of genes coding for specific surface determinants and for typical chemokines [26,27,28,29,30]

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