Mycoplasma bovis-Induced Inhibition of Bovine Peripheral Blood Mononuclear Cell Proliferation Is Ameliorated after Blocking the Immune-Inhibitory Programmed Death 1 Receptor.

Mycoplasma bovis-induced immune suppression is a major obstacle faced by the host for controlling infections. M. bovis impairment of antigen-specific T-cell responses is achieved through inhibiting the proliferation of peripheral blood mononuclear cells (PBMCs). This impairment may contribute to the persistence of M. bovis infection in various sites, including lungs, and its systemic spread to various organs such as joints, with the underlying mechanisms remaining elusive. Here, we elucidated the role of the immune-inhibitory receptor programmed death 1 (PD-1) and its ligand (PD-L1) in M. bovis infection. Flow cytometry (FCM) analyses revealed an upregulation of PD-L1 expression on tracheal and lung epithelial cell lines after M. bovis infection. In addition, we found increased PD-L1 expression on purified lung lavage macrophages following M. bovis infection by FCM and determined its localization by immunofluorescence analysis comparing infected and control lung tissue sections. Moreover, M. bovis infection increased the expression of the PD-1 receptor on total PBMCs and in gated CD4+ and CD8+ T-cell subpopulations. We demonstrated that M. bovis infection induced a significant decrease in CD4+ PD-1INT and CD8+ PD-1INT subsets with intermediate PD-1 expression, which functioned as progenitor pools giving rise to CD4+ PD-1HIGH and CD8+ PD-1HIGH subsets with high PD-1 expression levels. We blocked PD-1 receptors on PBMCs using anti-PD-1 antibody at the beginning of infection, leading to a significant restoration of the proliferation of PBMCs. Taken together, our data indicate a significant involvement of the PD-1/PD-L1 inhibitory pathway during M. bovis infection and its associated immune exhaustion, culminating in impaired host immune responses.

Induction of a balanced IgG1/IgG2 immune response to an experimental challenge with Mycoplasma bovis antigens following a vaccine composed of Emulsigen™, IDR peptide1002, and poly I:C.

Prevention and or control of Mycoplasma bovis infections in cattle have relied on the treatment of animals with antibiotics; herd management including separation and or culling infected animals; and the use of vaccines with limited protection. Due to the negative reactions and incomplete protection observed after vaccination with some bacterin-based vaccines, there is a need to put more efforts in the development of recombinant-based vaccines. However, the arsenal of antigens that may be suitable for a fully protective vaccine is rather limited at this point. We have tested a vaccine formulation containing M. bovis proteins formulated with adjuvants that have been shown to aid in the protection against other pathogens. After vaccinations, the animals were challenged using a BHV-1/M. bovis co-infection model. While the PBMC proliferation and cytokine responses to the antigens in the vaccine were negligible, humoral responses reveal that eight antigens elicit a balanced IgG1/IgG2 response although this was not enough to confer protection against M. bovis.

Effect of Mycoplasma bovis on bovine neutrophils.

Mycoplasma bovis (M. bovis) is a small bacterium that lacks a cell wall. M. bovis infection results in chronic pneumonia and polyarthritis syndrome (CPPS), otitis media, conjunctivitis, and meningitis in feedlot cattle and mastitis in dairy cattle. Numerous studies of peripheral mononuclear cells (PBMC) indicate that M. bovis evades host immunity through targeted effects on immune cell activity, including inhibition of effector function and simultaneous aberrant activation of immune cell activity that has no effect on protection against the bacterium. Few studies have addressed the interaction between M. bovis and neutrophils, one of the most important cell subsets of innate immunity. We hypothesized that M. bovis modifies specific neutrophil activities to support its persistence and systemic dissemination. In this study, we demonstrate that M. bovis enhances neutrophil apoptosis, stimulates production of pro-inflammatory cytokines, IL-12 and TNF-α, inhibits production of nitric oxide (NO) but augments elastase release. We also show that IL-17 an inflammatory cytokine produced by Th-17 cells does not enhance the capacity of neutrophils to destroy M. bovis. These findings present novel mechanisms of mycoplasma evasion of host innate immunity and provide potential opportunities for immuno-therapeutic interventions.

Status of the development of a vaccine against Mycoplasma bovis.

Mycoplasma bovis is an important pathogen of cattle and, despite numerous efforts an effective vaccine for control of the disease it causes remains elusive. Although we now know more about the biology of this pathogen, information is lacking about appropriate protective antigens, the type of immune response that confers protection and adjuvants selection. The use of conserved recombinant proteins, selected using in silico approaches, as components of a vaccine may be a better choice over bacterin-based vaccines due to the limited protection afforded by them and adverse reactions caused by them. More studies are needed on the characterization of host-pathogen interactions and to elucidate M. bovis products modulating these interactions. These products could be the basis for development of vaccines to control M. bovis infections in dairy farms and feedlots.

Novel B regs down-regulate TLR9-induced cytokine responses in sheep Peyer's patches.

Regulation of TLR responses is required in the intestine to prevent unnecessary responses to commensal microorganisms and maintain tissue homeostasis. Several mechanisms have been proposed for the regulation of TLR responses in intestinal epithelial and lamina propria cells. However, whether this regulation occurs in Peyer's patches (PP) is not known. While investigating cellular responses to the TLR9 agonist CpG ODN, we observed that PP cells responded poorly to CpG ODN stimulation despite expressing TLR9. We hypothesized that PP cells produced the immunoregulatory cytokine IL-10 which suppressed TLR-induced cytokine responses. In vitro neutralization of IL-10 or depletion of CD21+ B cells from PP resulted in significant increases in IL-12, IFNγ and IFNα responses in PP cells stimulated with CpG ODN. Essentially, our investigation have identified a novel population of IL-10-secreting B cells in PP with regulatory functions (B(regs)). These B(regs) may play an important role in the maintenance of intestinal homeostasis.

Modulation of B cell responses by Toll-like receptors.

B lymphocytes are well known because of their key role in mediating humoral immune responses. Upon encounter with antigen and on cognate interaction with T cells, they differentiate into antibody-secreting plasma cells, which are critical for protection against a variety of pathogens. In addition to their antibody-production function, B cells are efficient antigen-presenting cells and express a variety of pathogen recognition receptors (PRRs). Engagement of these PRRs with their respective ligands results in cytokine and chemokine secretion and the upregulation of co-stimulatory molecules. These events constitute innate immune responses. Toll-like receptor (TLR) activation provides a third signal for B cell activation and is essential for optimal antigen-specific antibody responses. In some situations, TLR activation in B cells can result in autoimmunity. The purpose of this review is to provide some insights into the way that TLRs influence innate and adaptive B cell responses.