Laryngeal Chondritis in Sheep in Iceland.

Laryngeal chondritis is a chronic disease in sheep with low morbidity, high mortality and unresolved pathogenesis. The disease has been recognized recently in Iceland and affects both ewes and rams. Animals of different ages are affected, but lambs and yearlings predominate. The disease is seen in housed animals and most cases occur during the late winter months. We report the gross and microscopical findings in 45 cases of laryngeal chondritis in Icelandic sheep.

Increased gammadelta T-cell populations in draining lymph nodes of lambs during the elicitation phase of dinitrochlorobenzene-induced contact hypersensitivity.

Ten lambs were sensitised with the hapten DNCB in an acetone/olive oil vehicle. The hapten/vehicle solution was applied onto the skin on the shaved ventral surface of the right ear. Two weeks later these lambs were rechallenged with the DNCB/vehicle solution. Simultaneously, ten non-sensitised lambs were treated with vehicle only, serving as vehicle controls. The 20 lambs were slaughtered 48 h after challenge/vehicle treatment, along with ten untreated animals serving as normal controls. Specimens of draining lymph nodes were collected from the 30 animals. All lambs were between 149 and 187 days old. Lymph node cryosections were stained for several leukocyte markers using monoclonal antibodies with the ABC immunohistochemical method. The stained sections were subsequently assessed in three different cortical compartments in each section, using an image analysis system. The resulting measurements from the three groups were compared. A marked increase of gammadelta T cells was detected in the DNCB group. The number of CD4+ T helper cells was decreased in the DNCB group compared with the normal control group, but not with the vehicle control group. No differences were revealed for CD8+ T cytotoxic cells or B cells. These findings were interpreted to be the consequences of possible downregulatory mechanisms protecting the lymphoid tissue from hypersensitivity. The prominence of gammadelta T-cells could indicate that these cells are involved in downregulation.

Distribution of MHC-II and CD1 molecules in the skin of lambs and changes during experimentally-induced contact hypersensitivity.

The presentation of antigen to specific T-cell populations is a crucial event during the elicitation phase of contact hypersensitivity (CHS). Significant changes in CD4(+) T-cell and gammadelta T-cell populations occur in the skin of sheep 48h after re-exposure to dinitrochlorobenzene but the expression of antigen presentation molecules such as MHC-II and CD1 at this stage of the hypersensitivity response has not been investigated. In the present study, a panel of monoclonal antibodies recognising CD1 and MHC-II subtypes was used in combination with computer assisted morphometric analysis to estimate the distribution of antigen presentation molecules in the superficial and deep dermis of the ears of lambs during the elicitation phase of CHS. The MHC-II molecules showed predominantly a perivascular and peri-appendageal distribution in the dermis and there were scattered MHC-II(+) cells in the basal and suprabasal layers of the epidermis. The CD1w2(+) (CD1b-like) molecules were present on distinct cells that were scattered evenly through the dermis, whereas CD1w3(+) (CD1c-like) molecules were almost exclusively detected on or in close association with the vascular endothelium. There was a significant increase in the presence of MHC-DQ(+) cells in the superficial dermis of dinitrochlorobenzene-treated animals compared with both an untreated control group and a vehicle-treated control group. However, MHC-DQ/DR(+) and CD1w3(+) cells only showed a significant increase compared with the vehicle-treated control group. The present study shows that the distribution of molecules involved in antigen presentation to CD4(+) T-cells and gammadelta T-cells changes during the elicitation phase of CHS in sheep, and suggests a role for MHC-DQ molecules on antigen presenting cells. However, the changes in distribution and expression of MHC-II and CD1 subtypes argue against a prominent role for a CD1-dependent pathway for T-cell recognition in the clinical cutaneous hypersensitivity response in sheep. Based on the expression of MHC-II molecules and CD1c molecules, we also suggest a potential role for endothelial cells in antigen presentation during the clinical dermatitis reaction.

Factor XIIIa positive dendritic cells are a major accessory cell population in the elicitation phase of DNCB-induced contact hypersensitivity.

The phenotypes and distribution of accessory cells in the ear skin of lambs during the elicitation phase of dinitrochlorobenzene (DNCB)-induced contact hypersensitivity (CHS) were examined using indirect immunoperoxidase histochemistry (ABC method), and a panel of antibodies. Thirty lambs, between 21 and 26 weeks of age, were divided into groups of 10. The shaved right ear of one group was treated with DNCB. Two weeks later this group was challenged with DNCB. One group was treated with the vehicle alone and the remaining group was left untreated. The lambs were slaughtered 48 h after challenge, and tissue specimens were collected from the ears of the three groups. Factor XIIIa+ (FXIIIa+) cells were prominent in the superficial dermis and showed predominantly a perivascular and subepidermal distribution. The other markers were less prominent, and whereas CD1+ cells and CD68+ cells showed a reaction pattern similar to the FXIIIa+ cells, CD14+ cells were found scattered predominantly in the deep dermis. There appeared to be an increase in FXIIIa+ cells, CD1+ cells, and CD68+ cells in the dermis of the DNCB-treated lambs 48 h after challenge. Only CD1+ cells were detected in epidermis of normal controls, and these cells appeared to be decreased in number in the two treated groups. Computer-assisted morphometric analysis was used to estimate the relative presence of the accessory cell subpopulations in the superficial and deep dermis and the entire dermis. A statistical analysis of the relative area of immunostaining showed a significantly increased presence of FXIIIa+ cells and CD68+ cells in the dermis of the DNCB-treated lambs 48 h after challenge. Interestingly, FXIIIa+ cells and CD68+ cells were also significantly increased in the vehicle treated group compared with untreated controls. We found no significant difference in the presence of CD1+ cells or CD14+ cells in the DNCB treated group compared with the controls. The study showed that FXIIIa+ DDC are the major accessory cell population in normal ear skin of lambs and the major responsive population during the elicitation phase of CHS. The lack of response in the CD1+ cell population suggests a less prominent role for the LC-related DC in the skin during the elicitation phase.

Accessory cell populations in draining lymph nodes of lambs in the elicitation phase of DNCB-induced contact hypersensitivity.

The effect of experimentally induced contact hypersensitivity on accessory cell populations in draining lymph nodes of lambs was studied. Previous studies of draining lymph nodes of lambs during the elicitation phase of CHS have shown that there are significant changes in T-cell subpopulations, particularly CD4(+) cells and gamma delta T-cells, but the behaviour of accessory (antigen presenting) cell populations was not investigated. The immunohistochemical presence of accessory cell populations was determined using markers for CD68, Pan MHCII, MHCII DQ, MHCII DR, OvCD1w1 (putative human CD1a/c-like) and OvCD1w2 (human CD1b-like). Ten lambs were sensitised, and 14 days later re-challenged, by applying the hapten di-nitro-chloro-benzene (DNCB) together with an acetone and olive oil (AOO) vehicle, onto the skin. Cryosections of the draining lymph nodes were stained immunohistochemically for the accessory cell markers. Using an image analysis system, the areas of staining in the lymph nodes from the challenged animals were compared with measurements in control animals. A significant increase in staining for CD68(+) cells was detected in the cortex of the DNCB-treated group (p=0.003). A significant increase in staining for the Pan MHCII marker was also observed in the DNCB group (p=0. 013). These results show that MHCII(+) cells and CD68(+) cells constitute a prominent cell population in the cortex of the regional lymph nodes of lambs in the late elicitation phase of DNCB-induced contact hypersensitivity.

Accumulation of CD25+ CD4+ T-cells in the draining lymph node during the elicitation phase of DNCB-induced contact hypersensitivity in lambs.

The elicitation phase of DNCB induced contact hypersensitivity in lambs was studied, and the presence of CD25+ cells in the lymph nodes draining the contact site was measured. Confocal laser scanning microscopy was used to capture images of two sets of triple immunofluorescence labellings. One set labelled CD25+, CD4+ and CD3+ cells, while the other labelled CD25+, VPM30+ and CD4+ cells. The CD25+ subpopulation labellings were assessed by area measurements in a morphometric protocol. The CD25+CD4+CD3+ cells were found to be increased in the DNCB treated group. This subpopulation of CD25+ cells comprised 75% of all CD25+ cells measured. The CD25+VPM30+CD4+ cells were also found to be increased in the DNCB group, but comprised only 17% of the total CD25+ cells measured. Since the VPM30 antibody detects an antigen found on activated T-cells, it was concluded that a substantial proportion of the triple CD25+CD4+CD3+ cells could represent a regulatory phenotype that may be active in suppressing the formation of effector immune cells in CHS of sheep.

Tumour gastrin expression and serum gastrin concentrations in dogs with gastric carcinoma are poor diagnostic indicators.

Hypergastrinaemia is observed commonly in human patients with gastric carcinoma and is associated with atrophic gastritis and Helicobacter pylori infection, both of which predispose to development of gastric tumours. Increased expression of gastrin is also described as a prognostic indicator for gastric carcinoma in man. Gastric carcinoma is rare in dogs and generally carries a grave prognosis. In this study, the expression of gastrin was investigated immunohistochemically in gastric biopsy samples from 64 dogs with gastric carcinoma. Serum gastrin concentrations were measured in 15 of these dogs and compared with those of seven healthy control dogs. Tumour tissue expressed gastrin in 8% (5/64) of the dogs with gastric carcinoma. There was no significant difference in serum gastrin concentrations between dogs with gastric carcinoma and healthy controls (P = 0.08). Expression of gastrin in gastric carcinomas is less common in dogs than in man and may therefore not be relied on as a prognostic marker in this species. Serum gastrin concentration alone is also not a useful biomarker for gastric carcinoma in dogs.

Prominence of gammadelta T cells in the elicitation phase of dinitrochlorobenzene-induced contact hypersensitivity in lambs.

Immunohistochemical techniques were used to identify T-cell subpopulations in the ear skin of lambs during the elicitation phase of dinitrochlorobenzene (DCNB)-induced contact hypersensitivity. Thirty lambs (21-26 weeks of age) were divided into groups of 10. The shaved right ear of one group was treated with DNCB. Two weeks later, this group was challenged with DNCB. One group was treated with the vehicle alone, and the remaining group was left untreated. The lambs were killed 48 hours after challenge, and tissue specimens were collected from the ears of the three groups. There was an increase in T-cell populations in the skin of the DNCB-treated lambs 48 hours after challenge. The majority of the T cells were CD8+ and associated predominantly with the blood vessels and adnexa of the superficial dermis. There was also an increased presence of CD4+ cells and gammadelta T cells in the superficial dermis. In the epidermis, clusters of gammadelta T cells and CD4+ cells were associated with microlesions. Computer-assisted morphometric analysis was used to estimate the relative presence of the T-cell subpopulations in the superficial and deep dermis and the entire dermis. Statistical analysis of the relative area of immunostaining showed that the significant increases in all T-cell subpopulations (CD4+, CD8+, and gammadelta T cells) in the entire dermis were accounted for by changes in the superficial dermis. The prominence of gammadelta T cells in delayed-type hypersensitivity in the skin of domestic ruminants has been the subject of conflicting reports. In the present study, CD4+ cell and gammadelta T-cell populations were of similar size in the normal and DNCB-treated lambs, suggesting an equal participation in the elicitation phase of contact hypersensitivity.