Group 2 innate lymphoid cells (ILC2) are now recognized as an important innate source of type-2 effector cytokines. Although initially associated with mucosal tissues, it is clear that ILC2 are present in diverse anatomical locations. The function of ILC2 at these sites is equally varied, and although ILC2 represent a relatively minor population, they are fundamentally important regulators of innate and adaptive immune processes. As such, there is much interest to understand the role of ILC2 in diseases with a type-2 inflammatory component. This review explores the known roles of ILC2 in disease, and the diseases that show associations or other strong evidence for the involvement of ILC2.

/pdf/group-2-innate-lymphoid-cells-in-disease-2qnzb2aai3.pdf

Group 2 innate lymphoid cells in disease.

Anthrax, caused by Bacillus anthracis, a Gram-positive spore-forming bacterium, is initiated by the entry of spores into the host body. There are three types of human infection: cutaneous, inhalational, and gastrointestinal. For each form, B. anthracis spores need to cross the cutaneous, respiratory or digestive epithelial barriers, respectively, as a first obligate step to establish infection. Anthrax is a toxi-infection: an association of toxemia and rapidly spreading infection progressing to septicemia. The pathogenicity of Bacillus anthracis mainly depends on two toxins and a capsule. The capsule protects bacilli from the immune system, thus promoting systemic dissemination. The toxins alter host cell signalling, thereby paralysing the immune response of the host and perturbing the endocrine and endothelial systems. In this review, we will mainly focus on the events and mechanisms leading to crossing of the respiratory epithelial barrier, as the majority of studies have addressed inhalational infection. We will discuss the critical gaps of knowledge that need to be addressed to gain a comprehensive view of the initial steps of inhalational anthrax. We will then discuss the few data available on B. anthracis crossing the cutaneous and digestive epithelia.

/pdf/crossing-of-the-epithelial-barriers-by-bacillus-anthracis-2pic791c85.pdf

Crossing of the epithelial barriers by Bacillus anthracis: the Known and the Unknown

A commercial Bacillus anthracis (Anthrax) whole genome protein microarray has been used to identify immunogenic Anthrax proteins (IAP) using sera from groups of donors with (a) confirmed B. anthracis naturally acquired cutaneous infection, (b) confirmed B. anthracis intravenous drug use-acquired infection, (c) occupational exposure in a wool-sorters factory, (d) humans and rabbits vaccinated with the UK Anthrax protein vaccine and compared to naive unexposed controls. Anti-IAP responses were observed for both IgG and IgA in the challenged groups; however the anti-IAP IgG response was more evident in the vaccinated group and the anti-IAP IgA response more evident in the B. anthracis-infected groups. Infected individuals appeared somewhat suppressed for their general IgG response, compared with other challenged groups. Immunogenic protein antigens were identified in all groups, some of which were shared between groups whilst others were specific for individual groups. The toxin proteins were immunodominant in all vaccinated, infected or other challenged groups. However, a number of other chromosomally-located and plasmid encoded open reading frame proteins were also recognized by infected or exposed groups in comparison to controls. Some of these antigens e.g., BA4182 are not recognized by vaccinated individuals, suggesting that there are proteins more specifically expressed by live Anthrax spores in vivo that are not currently found in the UK licensed Anthrax Vaccine (AVP). These may perhaps be preferentially expressed during infection and represent expression of alternative pathways in the B. anthracis "infectome." These may make highly attractive candidates for diagnostic and vaccine biomarker development as they may be more specifically associated with the infectious phase of the pathogen. A number of B. anthracis small hypothetical protein targets have been synthesized, tested in mouse immunogenicity studies and validated in parallel using human sera from the same study.

/pdf/whole-genome-protein-microarrays-for-serum-profiling-of-3d8tn073dd.pdf

Whole genome protein microarrays for serum profiling of immunodominant antigens of Bacillus anthracis

Mucosa-associated lymphoid tissues contain roughly 80% of all immune cells and produce virtually all of the body’s IgA1–3. Although the majority of IgA-secreting cells educated within a mucosal site home back to the same anatomic region, some cells are also found in distant mucosal tissues2–6. These observations underlie the notion of a common mucosal immune system, which holds that anatomically unrelated mucosal sites are functionally connected by a shared immune system2,3. However, the ontological basis of this separation between site of immune education and functionality has remained elusive. Here we show that mice lacking Peyer’s patches (PPs)—small-intestinal lymphoid tissue covered by antigen-sampling M cells—have no immunologic defect in the small-intestinal lamina propria. Surprisingly, the primary immunological abnormality in PP-deficient mice was a reduction in colonic B cells, including plasmablasts but not plasma cells. Adoptive transfer experiments conclusively demonstrated that PP-derived cells preferentially give rise to colonic—but not small-intestinal—B cells and plasmablasts. Finally, these PP-derived colonic B cells were critical for restraining colonic inflammation. Thus, PPs bridge the small-intestinal and colonic immune systems and provide a clear example of immune education being required in an anatomic compartment distinct from the effector site. Our findings, which highlight that the majority of fecal IgA is produced by colonic plasmablasts that originate from PPs, will help inform design of mucosal vaccines.

/pdf/anatomically-remote-education-of-b-cells-is-required-for-28jwuov0e3.pdf

Anatomically remote education of B cells is required for colonic health

The Effects of Stress Exposure on One Day Aged Broiler Chicken: A pathological Study

Ingestion of Bacillus anthracis spores causes gastrointestinal (GI) anthrax. Humoral immune responses, particularly immunoglobulin A (IgA)-secreting B-1 cells, play a critical role in the clearance of GI pathogens. Here, we investigated whether B. anthracis impacts the function of colonic B-1 cells to establish active infection. GI anthrax led to significant inhibition of immunoglobulins (eg, IgA) and increased expression of program death 1 on B-1 cells. Furthermore, infection also diminished type 2 innate lymphoid cells (ILC2) and their ability to enhance differentiation and immunoglobulin production by secreting interleukin 5 (IL-5). Such B-1-cell and ILC2 dysfunction is potentially due to cleavage of p38 and Erk1/2 mitogen-activated protein kinases in these cells. Conversely, mice that survived infection generated neutralizing antibodies via the formation of robust germinal center B cells in Peyer's patches and had restored B-1-cell and ILC2 function. These data may provide additional insight for designing efficacious vaccines and therapeutics against this deadly pathogen.

/pdf/impaired-colonic-b-cell-responses-by-gastrointestinal-238am5abnu.pdf

Impaired Colonic B-Cell Responses by Gastrointestinal Bacillus anthracis Infection

Canine osteosarcoma is an extremely malignant bone tumor that often arises in the bones of the limbs. It is a highly metastatic disease distinguished by proliferative bone lesions and a tendency for pulmonary metastasis. Overexpression of proliferative proteins are associated with bad prognosis in human osteosarcoma. Here, we tested the expression of the different proliferative proteins (p53, p16, vimentin, and mdm2) in nine archival samples with canine osteosarcoma. Paraffin-embedded tissue sections were confirmed by histopathology and stained by immunohistochemistry for p53, p16, vimentin and mdm2. Positive expression of these proteins was evaluated as the ratio of positive cancer cells and the intensity staining was assessed in several areas. Histopathologically, 95% of samples were grade II and III. All high-grade osteosarcomas were particularly cellular. The cancer cells were generally large spindle-shaped and large nucleus with distribution of osteoid between the cancer cells. Immunohistochemical detection of p53, p16, vimentin and mdm2 was 89%, 56%, 78%, and 89% of samples respectively. The staining intensity for p53, p16, vimentin and mdm2 was particularly nuclear in 81%, 66%, 78%, and 79% of the cancer cells respectively. Our present work suggests that p53, p16, vimentin, and mdm2 were detected in grade III canine osteosarcomas samples. In addition, these proliferative markers are the significant biomarkers in canine osteosarcomas and can be used as a predictor for diagnostic and prognostic value and allowing cancer differentiation. This primary data supports that both canine and human osteosarcomas share same molecular characters which are approved by expression of proliferative genes.

Immunohistochemical expression of proliferation markers in canine osteosarcoma

Article history: Received September 13, 2022 Accepted December 12, 2022 Available online December 12, 2022 Pheochromocytomas (PCs) are tumors originating from the chromaffin cells of the adrenal medulla. In people, there are highly correlated to inherited gene mutations in the succinate dehydrogenase (SDH) pathway; however, to date, little work has been done on the genetic basis of these tumors in animals. Out of the total of 2.203 Gb of canine DNA sequenced, 88.35% of bases mapped to exons and 11.65% mapped to introns. Out of 26 genes of interest containing 404 exons, 278 exons were sequenced 68.81%. Sequencing was considered successful when the average read depth was 3x and the entire exon was covered. Coverage ranged from 30% to 100%. Both SDHA and SDHB had exon mapped 46.6% and 62.5% respectively. Additionally, out of 45 known canine variants, exome technique able to detect 36 variants (80%). We performed SDHA and SDHB immunohistochemistry on 35 canine formalin-fixed, paraffin embedded. Interestingly, we had loss of immunoreactivity for both SDHA and SDHB in four samples, suggesting a mutation in SDHx including SDHA. Out of 35 samples, 6 had immunoreactivity for SDHA and 25 lacked immunoreactivities for SDHB. 29 out of the 35 (82%) may have an SDH family mutation other than SDHA. Exome sequencing and immunohistochemistry are able to predict malignant behaviour and likelihood of reduction of PCC/PGLs in humans. This can be used to determine whether there are similar mutations in the pseudo-hypoxic, kinase signalling, and other genes of interest exist in dogs, as well as finding novel genes involved in canine Pheochromocytomas oncogenesis.

Exome sequencing and SDH (A, B) immunohistochemistry of canine Pheochromocytomas

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Impaired Colonic B-Cell Responses by Gastrointestinal Bacillus anthracis Infection

Immunohistochemical expression of proliferation markers in canine osteosarcoma

Exome sequencing and SDH (A, B) immunohistochemistry of canine Pheochromocytomas