Immunohistochemical determination of estrogen receptor-alpha in vaginal and tumor tissues of healthy and TVT-affected bitches and their relation to serum concentrations of estradiol-17beta and progesterone.

One of the most frequent canine neoplasms is the transmissible venereal tumor (TVT), which affects the male and the female genital tract. The objective of this study was to determine (immunohistochemically) estrogen receptor (ER-alpha) expression in vaginal tissue of healthy bitches and in the vaginal and neoplastic tissues of TVT-affected bitches. Fifty-eight bitches were divided into two groups: tumor group (TVT) and control group (healthy). Canine estrous cycle stages were determined by means of exfoliative vaginal cytology, hormone assays, and macroscopic appearance of ovaries. Samples from vaginal and neoplastic tissues were obtained under general anesthesia, fixed in 10% buffered formaldehyde, embedded in paraffin, and sectioned. Anestrus, proestrus and estrus control females had higher ER-alpha expression than diestrus bitches. Within the tumor group, diestrus bitches had significantly higher ER-alpha expression. Although some samples had expression in the endothelium of blood vessels, no ER-alpha expression was observed in neoplastic tissues. In conclusion, vaginal tissue of tumor and control bitches, under different distinct steroid influences, had different ER-alpha expression, whereas ER-alpha expression was not present in neoplastic tissues.

Role of lymphocytes in dogs experimentally re-challenged with canine transmissible sarcoma.

The role of lymphocytes in dogs re-challenged with canine transmissible sarcoma (CTS) was investigated histologically and immunologically. Tumors were detected on the 3rd day, began to regress on the 9th day and disappeared on approximately the 15th day after the secondary transplantation (AST). The CTS cells were surrounded by lymphocytes which had infiltrated from the early stage, degenerated and ultimately disappeared. Most of the lymphocytes expressed thymocyte antigen (Ta) on the cell membrane surface. The numbers of lymphocytes and Ta-positive lymphocytes in peripheral blood increased gradually AST. The blastogenic responses of peripheral blood lymphocytes (PBL) induced by 3 kinds of mitogens were elevated strikingly from the early stage AST. These results suggest that Ta-positive lymphocytes play an important role in vivo in the regression of tumors and that the immunological activity of PBL is stimulated immediately AST.

Regulation of leukocyte glass adherence and tube leukocyte adherence inhibition (LAI) reactivity by serum factors in dogs with progressing or spontaneously regressing canine transmissible venereal sarcoma (CTVS).

We examined the regulation of leukocyte glass adherence and tube leukocyte adherence inhibition (LAI) reactivity by serum factors in dogs with regressing or progressing canine transmissible venereal sarcomas (CTVS). Both regressor and progressor peripheral blood leukocytes (PBL), draining and non-draining lymph node cells (LNC), and splenic leukocytes were significantly responsive to CTVS antigen extract in tube LAI. In contrast, a significant decrease in basal glass adherence of progressor PBL, draining and non-draining LNC, and splenic leukocytes was observed. Normal glass adherence was restored to progressor leukocytes by extensive washing with warm serum-free media, while significant tube LAI responsiveness to CTVS antigen extract was maintained. Preincubation of regressor PBL and LNC with progressor sera in two-stage tube LAI decreased the basal glass adherence of treated leukocytes. This effect of progressor sera was heat labile, a characteristic of CTVS antigen. Collectively, these findings suggest that progressor leukocytes and progressor sera treated regressor leukocytes were activated by interaction with serum CTVS antigen and thus behaved in tube LAI as stimulated cells, even in the absence of CTVS antigen. Regressor but not progressor sera were shown to contain anti-CTVS IgG with specific arming activity for normal dog PBL, but not LNC in two-stage tube LAI. The nonadherent response of peripheral blood neutrophils in two-stage tube LAI was proportional to the concentration of arming IgG, whereas no change was observed in glass adherence of PBL. The results of this study define the role of progressor and regressor serum factors in the mechanism of tube LAI and demonstrate a relationship between leukocyte glass adherence and the clinical course of CTVS. These findings show that tube LAI is a simple and reproducible measure of active factors in the immune response to a tumor.

Canine transmissible venereal sarcoma: distribution of T and B lymphocytes in blood, draining lymph nodes and tumours at different stages of growth.

The levels of T, B and null lymphocytes in the peripheral blood, draining lymph nodes, and tumour masses at different growth stages in dogs transplanted with canine transmissible venereal sarcoma (CTVS) were determined by immunofluorescence techniques. The tumours were classified at excision into "progressor", "steady state", and "regressor" stages of growth. The percentage of B cells in the lymphocytes infiltrating into the progressively growing tumours (n = 10, 37.3 +/- 7.4%) was significantly higher (P less than 0.025) than that in regressing tumours (n = 21, 26.1 +/- 1.9%). In contrast, the percentage of T cells in the lymphocytes infiltrating into the regressing tumours (n = 21, 61.2 +/- 2.6%) was significantly higher (P less than 0.005) than that in the progressively growing tumours (n = 10, 34.0 +/- 5.1%). The tumours at the steady-state growth stage (n = 9) had 50.8 +/- 5.7% infiltrating T-cells, which was significantly higher (P less than 0.005) than the progressors and lower (P less than 0.005) than the regressors. The percentage of null cells of progressors (n = 10, 26.0 +/- 6.9%) was significantly (P less than 0.025) higher than in regressors (n = 21, 13.5 +/- 2.9%). The draining lymph nodes of progressor dogs (n = 5) had significantly fewer (P less than 0.025) B cells (8.2 +/- 2.3%) than in normal (n = 5, 16.1 +/- 3.1%), regressors (n = 12, 19.1 +/- 1.7%) and steady-state dogs (n = 5, 15.8 +/- 2.6%). Although there was slight lymphopenia and fluctuation of null cells, no significant differences in T- and B-lymphocyte levels were noted in the peripheral blood of the tumour dogs (n = 44) studied.