Comparative Expression Analysis of Innate Immune Markers and Phagocytic Activity in Peripheral Blood of Dogs with Mammary Tumors

SIMPLE SUMMARY: The presented study aimed to find out the differences between peripheral blood immune cell markers from healthy bitches and bitches with mammary tumors. Due to the fact that the role of canine innate immune cells in cancer remains poorly understood, the markers of innate cells were chosen for this research. Blood samples from female dogs with mammary tumors of epithelial and mesenchymal origin were investigated by flow cytometry. CD5 and CD11b markers of innate immune cells, phagocytic activity, and cellular killing were assessed. The number of CD11b lymphocytes was increased in tumors with epithelial origin. No significant differences were found between the percentages of phagocytic cells. However, the phagocytes of canine patients with tumors of epithelial origin showed increased phagocytosis compared to the control group. In oxidative burst test, a statistically significant difference between the number of reactive oxygen species (ROS) produced was demonstrated only between the group of bitches with epithelial tumors and the control group. These results may suggest that there are subpopulations of innate immune cells that may be involved in anti-tumor immune mechanisms and have a potential to be supportive diagnostic markers in canine mammary tumors. ABSTRACT: Canine innate immune system role in cancer prevention and progression remains poorly understood. It has been revealed that innate immune cells could play a dual role in cancer immunology promoting or inhibiting tumor development and growth. Current immunotherapies target mainly the adaptive anti-tumor response and that may be a reason why they remain ineffective in a majority of patients. It is important to acquire detailed knowledge about innate immune mechanisms to broaden the diagnostic and therapeutic options and employ innate immune cells in anti-cancer therapies. In the present study, 21 female dogs of different breeds and types of spontaneous mammary tumors were investigated. The study aimed to find simple and cheap markers that can be used for preliminary diagnosis, prior to the surgical resection of the tumor. The differences in innate immune cell quantity and function were investigated between female dogs with malignant mammary tumors of epithelial and mesenchymal origin. Flow cytometry was used to evaluate the percentages of CD5+ lymphocytes including CD5(low) lymphocytes, CD11b integrin expression on leukocytes, phagocytosis, and oxidative burst. The number of CD11b lymphocytes was increased in tumors with epithelial origin compared to the control group. No significant differences were found between the percentages of phagocytic cells neither for granulocytes nor for monocytes. However, the phagocytes of canine patients with tumors of epithelial origin showed increased phagocytosis compared to the control group. The percentages of granulocytes that produced reactive oxygen species (ROS) in response to E.coli and PMA were not altered in patients with malignant tumors compared to control. A statistically significant difference between the number of ROS produced by the single granulocyte was demonstrated only between the group of bitches with epithelial tumors and the control group in case of E. coli stimulation. The obtained results suggest that some innate immune cells may be involved in anti-tumor immune mechanisms and have the potential to be supportive diagnostic markers in canine mammary tumors.