Peter J. Felsburg

Selective IgA deficiency in the dog

This study documents the occurrence of selective IgA deficiency in the dog. This is a unique spontaneous animal model with clinical and immunologic findings similar to that of selective IgA deficiency in humans, the most common human primary immunodeficiency. The disease in the dog is characterized by chronic, recurrent respiratory infections and dermatitis, low concentrations of serum IgA, normal concentrations of serum IgG and IgM, normal T-cell function as measured by lymphocyte transformation tests, the presence of autoantibodies, and a defect in the maturation or terminal differentiation of IgA B cells into IgA-secreting plasma cells.

An improved method for the isolation of enriched canine peripheral blood mononuclear cell and peripheral blood lymphocyte preparations

A two-step purification method was developed for obtaining (1) peripheral blood mononuclear cell preparations of greater than 97% purity and (2) peripheral blood lymphocyte preparations of greater than 95% purity from canine whole blood with yields similar to or greater than those obtained by conventional techniques.

X-linked severe combined immunodeficiency in the dog.

This study documents the occurrence of a form of X-linked severe combined immunodeficiency (SCID) in the dog with clinical, immunologic, and pathologic features similar to those of X-linked SCID with B cells in man. The disease in the dog is characterized by growth retardation and increased susceptibility to bacterial and viral infections in young pups. Affected pups have all died or were euthanatized by 5 months with signs of canine distemper, infectious hepatitis, or bacterial pneumonia. Laboratory findings include normal numbers of circulating B lymphocytes and normal concentrations of serum IgM, but low to absent concentrations of serum IgG and IgA, indicating a defect in the terminal differentiation of IgG and IgA B cells into immunoglobulin-secreting plasma cells. This is supported by the failure of peripheral lymphocytes to produce IgG or IgA plaque-forming cells in response to polyclonal activation. There are low-to-normal numbers of circulating T cells, but a severely depressed blastogenic response to T cell mitogens. Postmortem findings include thymic dysplasia and hypoplasia of lymphoid tissue. Family studies and breeding experiments are consistent with an X-linked recessive mode of inheritance.

Experimental T-2 toxicosis in swine following inhalation exposure: effects on pulmonary and systemic immunity, and morphologic changes.

Thirty-four, 9-to 11-week-old, male castrated, crossbred, specific pathogen-free derived pigs were exposed to a T-2 toxin aerosol at a nebulized dose of 0 or 9 mg/kg in pairs, each pair consisting of 1 control and 1 T-2 treated pig which were exposed on the same day. Twenty to 30% of the toxin (1.8 to 2.7 mg/kg) was retained by the pigs. Five pairs were killed on each of 1, 3 and 7 days after dosing. Two pairs of pigs were designated as a 0.33-day group when one T-2 treated pig died and the other was killed in a moribund state at 8 to 10 hours after dosing. The pulmonary and systemic immunity and morphologic changes of the lungs and other organs were examined. Bronchoalveolar lavage was performed to obtain alveolar macrophages (AM) and pulmonary lymphocytes (PL). The phagocytic ability of AM and mitogen-induced blastogenic responses of enriched PL and peripheral blood lymphocytes were evaluated. Clinically, all of the T-2 treated pigs vomited and were cyanotic, anorexic, lethargic and laterally recumbent. In the 0.33-, 1-, and 3-day T-2 treated pigs, there was a marked reduction in AM phagocytosis and mitogen-induced blastogenic responses of PL but not of peripheral blood lymphocytes. Mild to moderate, multifocal interstitial pneumonia was seen in the majority of the T-2 treated pigs. In pigs dying following inhalation of T-2 toxin, there was a more severe pneumonia, as well as marked necrosis of lymphoid tissues, severe necrohemorrhagic gastroenteritis and edema of the gall bladder wall, and multifocal necrosis of the heart and pancreas. Thus, inhalation exposure to T2 toxin can result in clinical signs and morphologic changes resembling those reported previously in pigs given T-2 toxin intravascularly (iv) at a dose of 1.2 mg/kg (approximate LD50) or greater, as well as death. Mild pulmonary injury as well as transient impairment of pulmonary immunity was present in pigs surviving inhalation exposure.

Short-term effect of cyclophosphamide and azathioprine on selected aspects of the canine blastogenic response

The short-term, in vitro responses of canine peripheral blood lymphocytes to mitogenic stimulation and serum immunoglobulin concentrations were evaluated following treatment with currently recommended doses of cyclophosphamide and azathioprine. Cyclophosphamide had no significant effect on either the serum immunoglobulin concentrations or the blastogenic response of lymphocytes to mitogenic stimulation. Serum immunoglobulin concentrations remained unchanged following azathioprine treatment. The blastogenic response was significantly suppressed following one week of azathioprine therapy and returned to baseline values one week following cessation of treatment. The response to phytohemagglutinin was most suppressed, followed, in order, by the response to concanavalin A, and to pokeweed mitogen. These results suggest that the short-term use of azathioprine, but not cyclophosphamide, in clinically used dosages, does suppress selective aspects of the canine immune system, and the T cells appear to be more susceptible than B cells to the immunosuppressive effect of this drug.

Immunohistology of Peyer's patches in the dog.

Canine Peyers patches were examined by light microscopy and immunohistochemistry for possible variations depending on the location within the small intestine and for similarities and dissimilarities to PPs from other species. The duodenal and jejunal PPs were characterized by relatively large domes and interfollicular areas. In contrast, the ileal PP had small domes and poorly developed interfollicular areas and very large follicles. T cells were found in the interfollicular area and corona and in lesser numbers in the dome and germinal centers. The ileal PP contained far fewer T cells than the proximal PPs. Domes of canine PPs contained some cytoplasmic IgA+ (cIgA+) and many cIgG+ cells. Peanut agglutinin (PNA) stained germinal center cells in a selective but not-uniform way and did not stain T cells.

Ultrastructure and alkaline phosphatase activity of the dome epithelium of canine Peyer's patches.

The dome epithelium of Peyers patches from different parts of the intestine of four dogs was examined by scanning and transmission electron-microscopy and by alkaline phosphatase histochemistry on glycol-methacrylate embedded sections. M cells were scattered among more numerous enterocytes in duodenal and jejunal Peyers patches, but constituted the major cellular component of dome epithelia of the ileal Peyers patches. Alkaline phosphatase histochemistry demonstrated low enzyme activity in the brush border of M cells, as compared to enterocytes, in the duodenum and jejunum, allowing identification of M cells at the light microscopic level. Alkaline phosphatase activity was too low in ileal enterocytes to permit visualization of M cells. The presence of intrafollicular invaginations of dome epithelium is a consistent finding in duodenal Peyers patches of the dog and these invaginations were characterized by few M cells, many intraepithelial lymphocytes and strong alkaline phosphatase activity.

Isolation and phenotypic and functional characterization of cells from Peyer's patches in the dog

Cells were isolated from canine Peyers patches (PPs) and their phenotype and capacity to secrete immunoglobulins in vitro were determined. Cells isolated from duodenal and jejunal PPs of adult dogs consisted of 91.4% lymphocytes and 1.6% macrophages with 55.4% mIg(+)-cells and 35.6% Thy-1(+)-cells. In vitro IgA secretion by pokeweed mitogen (PWM)-stimulated PP cells exceeded that by cells from other lymphoid tissues and was specifically increased by concanavalin A, suggesting a role for isotype-specific T-cells. Comparison of duodenal and jejunal (proximal) PPs and the ileal PP revealed that the ileal PP contained fewer T-cells, fewer mIgA(+)-cells and more mIgM(+)-cells. Cells from the ileal PP produced very little IgA and IgG, but abundant IgM in vitro. These data suggest that the proximal PPs of dogs are important in the generation of IgA B-cells, similar to PPs of rodents. The ileal PP of dogs may have a function in the early development of the B-cell system of the dog.

Development and functional characterization of T cell lines from canine Peyer's patches

We have propagated concanavalin A-stimulated cells from canine Peyers patches in vitro in the presence of interleukin-2 (IL-2). The cells were characterized as T cells by determination of their phenotype and by functional assays. They are IL-2 dependent and respond to IL-2 of murine, primate and canine origin. The long-term cultured cells provided help for immunoglobulin production by purified autologous B cells and suppressed IgG production by nonseparated autologous peripheral blood mononuclear cells.

The effect of general anesthesia on canine lymphocyte function

The in vitro response of canine peripheral blood lymphocytes to mitogenic stimulation was evaluated following general anesthesia for a relatively minor diagnostic procedure. A marked suppression in the blastogenic response to phytohemagglutinin and concanavalin A was observed 4 hours postinduction which persisted through the first 24 hours and was normal by 4 days. A mild suppression to stimulation with pokeweed mitogen was observed at 4 hours postinduction but the response was back to normal by 24 hours. These results suggest general anesthesia has a transient immunosuppressive effect on canine lymphocytes and T cells are more susceptible to the immunosuppressive effect than B cells.