Comparison of cytokine mRNA expression in peripheral CD4+, CD8+ and γδ T cells between healthy Holstein and Japanese Black calves

Japanese Black (JB) calves are more susceptible to infectious diseases compared to Holstein (Hol) calves. To clarify the immunological differences between JB and Hol calves, expression of cytokine messenger RNA (mRNA) was examined using peripheral CD4⁺, CD8⁺ and γδ T cells. Healthy calves, 24 from each species, were examined. Blood samples were obtained from calves at 1 week, 1 month and 3 months old, eight calves for each age of each species. Peripheral blood mononuclear cells were stimulated with phytohemagglutin (PHA), and T cell subsets were isolated by positive selection using magnetic cell sorting (MACS). Levels of interlekin (IL)‐2, IL‐4, IL‐10 and interferon (IFN)‐γ mRNA in three T cell subsets were analyzed. WC1‐N1⁺ γδ T cell percentages were significantly lower in JB calves at 1 week and 1 month of age compared to Hol calves. In addition JB calves had significantly lower IL‐2, IL‐10 and IFN‐γ mRNA in WC1‐N1⁺ γδ T cells at 1 and 3 months of age, whereas there were no significant differences in cytokine mRNA of CD4⁺ and CD8⁺ cells between the two groups. Decreased cytokine mRNA and cell number of peripheral γδ T cells may affect negatively on the immune system of JB calves.     

Relationship between diarrhea and peripheral leukocyte population in neonatal Japanese black calves

Neonatal Japanese Black (JB) calves show a high incidence of diarrhea. The objective of this study was to analyze the immune cell populations of neonatal JB calves in detail and examine its correlation with the incidence of diarrhea immediately after birth. Understanding the immune cell populations is helpful in clinics in order to determine the condition of the immune system for prevention of diseases. Blood samples were obtained from JB calves on the day of birth. The peripheral leukocyte populations were analyzed separately for calves that had diarrhea within 2 weeks after birth (diarrhea group; n = 26) and for calves without diarrhea (control group; n = 74). The numbers of the peripheral blood CD3(+)TcR1-N12(+) and CD8(+) T cells were significantly lower in the diarrhea group compared with the control group. These findings suggest that the congenital lower peripheral γδ and CD8(+) T cells results in a high risk of diarrhea in neonatal JB calves.     

Cattle Infected with Bovine Leukaemia Virus may not only Develop Persistent B‐cell Lymphocytosis but also Persistent B‐cell Lymphopenia

We investigated the distribution of B and T cells in the peripheral blood of haematologically inconspicuous (non‐persistent lymphocytotic, PL^?) cattle infected with the bovine leukaemia virus (BLV). Flow cytometric data were obtained from six PL^? cattle and compared with six age‐matched animals with persistent lymphocytosis (PL⁺) and five non‐infected healthy controls (BLV^?). In the PL^? group, the percentage and number of surface immunoglobulin‐positive (sIg⁺) B cells were significantly reduced. Whereas in BLV^? cattle, about 40% of the peripheral blood lymphocytes (PBL) were sIg⁺ and 24% were sIgM⁺ B cells. In the PL^? group, less than 20% of the PBL were sIg⁺ and sIgM⁺ B cells. Only 5% of the PBL co‐expressed sIgM⁺ and CD5⁺ versus 16% in BLV^?. This decrease was persistent over 3 years and predominantly affected: (i) B cells that did not express sIgM; (ii) sIgM⁺ B cells co‐expressing CD5 and CD11b; and (iii) equally both λ‐ and κ‐type light chain B‐cell subpopulations. In contrast, the number of all circulating lymphocytes, CD5^? and CD11b^? sIgM⁺ B cells and CD2⁺ T cells did not differ. In PL⁺ animals, about 75% of the PBL were sIgM⁺CD5⁺ B cells. These cells were of polyclonal origin, as light chains of the λ‐ and κ‐type were expressed in a ratio of 4:1 (57.7% of PBL λ⁺, 14% κ⁺) as in BLV^? animals (33.6% of PBL λ⁺, 8.7% κ⁺). In PL⁺ cattle the absolute number of B‐cells and, therefore, their relative percentage is significantly increased. For this reason, even in case of absolutely increased T‐cell numbers, the relative percentage of T‐cells could be lower than in normal controls. The cause for the observed B cell decrease in PL^? cattle is unknown, but it can be assumed that cytotoxic T cells are involved in this B‐cell lymphopenia.     

Effect of supplemental vitamin E on the peripheral blood leukocyte population in Japanese Black calves

We investigated the effect of supplemental vitamin E on the peripheral blood leukocyte population in Japanese Black calves. Twenty-six calves kept at the same farm were studied. They were divided into two groups; thirteen calves received 300 IU/day of vitamin E orally from 1 to 3 months of age (VE group), and the other thirteen calves did not receive the vitamin E supplement (control group). The VE group showed a higher serum vitamin E concentration at 2 and 3 months of age compared with the control group (P<0.01). The numbers of CD3⁺ cells and CD4⁺ cells were higher in the VE group than in the control group, and the difference was statistically significant at 3 months of age (P<0.05). The numbers of CD21⁺ cells were higher in the VE group than in the control group, and the difference was statistically significant at 2 months of age (P<0.05). The numbers of CD335⁺ cells tended to be higher in the VE group than in the control group. The numbers of CD8⁺ cells and CD14⁺ cells tended to be higher in the VE group than in the control group at 3 and 4 months of age. This study demonstrated that the supplementation of suckling Japanese Black calves with vitamin E might affect the numbers of some immune cell types in the peripheral blood.     

Anti‐allergic effects of Lactobacillus crispatus KT‐11 strain on ovalbumin‐sensitized BALB/c mice

In this study, we investigated the effects of oral ingestion of Lactobacillus crispatus KT‐11 strain (KT‐11) on the immune response in an allergic rhinitis mouse model, ovalbumin (OVA)‐sensitized BALB/c mice. Sneezing activity in mice that were administered a KT‐11‐supplemented diet was significantly lower than that in mice administered a KT‐11‐free diet (control diet) at age 11 weeks. We found that serum OVA‐specific immunoglobulin E (IgE) levels and total number of interleukin (IL)‐4⁺CD4⁺ spleen cells in mice that were administered a KT‐11‐supplemented diet were significantly lower than in mice administered a control diet. The ratio of spleen interferon‐γ⁺CD4⁺/IL‐4⁺CD4⁺ cells was higher in the mice administered the KT‐11‐supplemented diet compared to that in mice administered the control or L. rhamnosus GG‐supplemented diet. In contrast, the number of CD11b⁺CD80⁺ and FcεRIα⁺CD117⁺ cells was significantly lower in mice administered the KT‐11‐supplemented diet. These results suggested that KT‐11 reduced OVA‐induced allergic symptoms in BALB/c mice via the adjustment of the T helper type 1/T helper type 2 balance, and a decrease in the number of antigen‐presenting cells and high affinity IgE receptor‐positive mast cells.     

Cutaneous infiltrates and peripheral blood immune responses in dogs with immunomodulatory‐responsive lymphocytic–plasmacytic pododermatitis

This study characterizes T‐ and B‐lymphocyte responses in the peripheral blood and lesional skin of dogs with immunomodulatory‐responsive lymphocytic–plasmacytic pododermatitis (ImR‐LPP), a term previously proposed to denote a subpopulation of dogs with idiopathic pododermatitis. T‐cell (CD3⁺, CD4⁺ and CD8⁺) and B‐cell (CD21⁺) counts were significantly increased in both the epidermis and dermis of lesional ImR‐LPP skin compared with that in pedal skin from healthy controls. CD3⁺, CD4⁺, CD8⁺ and CD21⁺ cells were commonly observed in perivascular sites in the superficial dermis, periadnexally, beneath the dermal–epidermal (DE) junction and in the epidermis of lesional ImR‐LPP skin. The CD8⁺/CD3⁺ T‐cell ratio in peripheral blood was significantly increased in the ImR‐LPP group (0.42 versus 0.35 in controls). Serum IgA, IgG and IgM concentrations were all significantly elevated in affected dogs. Lymphocyte stimulation indices in ImR‐LPP dogs were comparable with control levels except for a lower response to ionomycin (6.0 versus 11.1). Dogs with ImR‐LPP had a higher incidence and mean (semi‐quantitative) score for IgA, IgG and IgM deposits in the epidermis, and a significantly increased incidence of dermal IgA⁺, IgG⁺ and IgM⁺ mononuclear inflammatory cells. The results indicate that upregulated T‐ and B‐lymphocyte responses may contribute to the pathogenesis of the skin lesions observed in dogs with ImR‐LPP.     

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions in Large White pigs selected for higher peripheral blood immune capacity

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions were compared between the immunity‐selected Large White line and the non‐selected Large White line. The selected Large White line showed a higher level of pulmonary MPS lesions compared with the non‐selected Large White line. Subsequent to vaccination, the percentage of natural killer cells and T cells (CD3⁺CD4⁺CD8^‐ and CD3⁺CD4^?CD8⁺ T cells) were significantly increased in the non‐selected line but remained unchanged in the immunity‐selected Large White line. Secretion of Mycoplasma hyopneumoniae vaccine‐specific immunoblogulin G and phagocyte activity in peripheral blood were significantly higher in the immunity‐selected Large White line than in the non‐selected line. Expression of interleukin (IL)‐4 and IL‐6 messenger RNA in hilar lymph nodes was significantly lower in the immunity‐selected Large White line than in the non‐selected line. However, expression of IL‐10 in all immune tissues was significantly higher in the immunity‐selected Large White line. These results suggest that the selection for high immunity was not effective in increasing resistance to MPS lung lesions.     

Immunological and angiogenic markers during metronomic temozolomide and cyclophosphamide in canine cancer patients

Metronomic chemotherapy stimulates the immune response via depletion of regulatory T cells (Tregs) and suppresses angiogenesis by modulating the secretion of thrombospondin‐1 (TSP‐1) and vascular endothelial growth factor (VEGF). In this study, blood was collected from 10 healthy dogs and from 30 canine cancer patients before and 2 and 4 weeks after treatment with metronomic temozolomide (6.6 mg m^?2), cyclophosphamide (12.5 mg m^?2) or cyclophosphamide and temozolomide. The percentage of circulating CD25⁺Foxp3⁺CD4⁺ Tregs and the plasma levels of TSP‐1 and VEGF were measured. There was a significant difference in the percentage of Tregs between cancer patients and healthy dogs. A significant decrease in Tregs was noted in patients treated with metronomic cyclophosphamide and the combination. Treatment with temozolomide had no effect on the percentage of Tregs. TSP‐1 and VEGF levels were, respectively, significantly lower and higher in cancer patients than in healthy dogs, but they were not influenced by any of the studied metronomic treatment regimens.     

In vitro effects of dexamethasone on bovine CD25+CD4+ and CD25−CD4+ cells

This paper investigates the in vitro effect of dexamethasone on bovine CD25^highCD4⁺, CD25^lowCD4⁺ and CD25⁻CD4⁺ T cells. Only a small percentage of bovine CD25^highCD4⁺ (2–4%) and CD25^lowCD4⁺ (1–2%) cells expressed Foxp3. Dexamethasone caused considerable loss of CD25⁻CD4⁺ cells, but it increased the relative and absolute numbers of CD25^highCD4⁺ and CD25^lowCD4⁺ lymphocytes, while at the same time reducing the percentage of Foxp3⁺ cells within the latter subpopulations. Considering all these, as well as the intrinsically poor Foxp3 expression in bovine CD25⁺CD4⁺, it can be concluded that the drug most probably increased the number of activated non-regulatory CD4⁺ lymphocytes. It has been found that changes in cell number were at least partly caused by proapoptotic effect of the drug on CD25⁻CD4⁺ cells and antiapoptotic effect on CD25^highCD4⁺ and CD25^lowCD4⁺ cells. The results obtained from this study indicate that the involvement of CD4⁺ lymphocytes in producing the anti-inflammatory and immunosuppressive effect of dexamethasone in cattle results from the fact that the drug had a depressive effect on the production of IFN-γ by CD25⁻CD4⁺ cells. Secretion of TGF-β and IL-10 by CD4⁺ lymphocytes was not involved in producing these pharmacological effects, because the drug did not affect production of TGF-β and, paradoxically, it reduced the percentage of IL-10⁺CD4⁺ cells.     

Chronic pneumonia in calves after experimental infection with Mycoplasma bovis strain 1067: Characterization of lung pathology,persistence of variable surface protein antigens and local immune response

Background

Mycoplasma bovis is associated with pneumonia in calves characterized by the development of chronic caseonecrotic lesions with the agent persisting within the lesion. The purposes of this study were to characterize the morphology of lung lesions, examine the presence of M. bovis variable surface protein (Vsp) antigens and study the local immune responses in calves after infection with M. bovis strain 1067.

Methods

Lung tissue samples from eight calves euthanased three weeks after experimental infection with M. bovis were examined by bacteriology and pathology. Lung lesions were evaluated by immunohistochemical (IHC) staining for wide spectrum cytokeratin and for M. bovis Vsp antigens and pMB67 antigen. IHC identification and quantitative evaluation of CD4⁺ and CD8⁺ T lymphocytes and immunoglobulin (IgG1, IgG2, IgM, IgA)-containing plasma cells was performed. Additionally, expression of major histocompatibility complex class II (MHC class II) was studied by IHC.

Results

Suppurative pneumonic lesions were found in all calves. In two calves with caseonecrotic pneumonia, necrotic foci were surrounded by epithelial cells resembling bronchial or bronchiolar epithelium. In all calves, M. bovis Vsp antigens were constantly present in the cytoplasm of macrophages and were also present extracellularly at the periphery of necrotic foci. There was a considerable increase in numbers of IgG1- and IgG2-positive plasma cells among which IgG1-containing plasma cells clearly predominated. Statistical evaluation of the numbers of CD4⁺ and CD8⁺ T cells, however, did not reveal statistically significant differences between inoculated and control calves. In M. bovis infected calves, hyperplasia of bronchus-associated lymphoid tissue (BALT) was characterized by strong MHC class II expression of lymphoid cells, but only few of the macrophages demarcating the caseonecrotic foci were positive for MHC class II.

Conclusions

The results from this study show that infection of calves with M. bovis results in various lung lesions including caseonecrotic pneumonia originating from bronchioli and bronchi. There is long-term persistence of M. bovis as demonstrated by bacteriology and immunohistochemistry for M. bovis antigens, i.e. Vsp antigens and pMB67. The persistence of the pathogen and its ability to evade the specific immune response may in part result from local downregulation of antigen presenting mechanisms and an ineffective humoral immune response with prevalence of IgG1 antibodies that, compared to IgG2 antibodies, are poor opsonins.     

Phenotypical characterization of peripheral blood leucocytes in the newborn calf

The present study was undertaken to establish reference values for the composition of blood leucocyte populations in neonatal calves by differential leucocyte counts and immunophenotyping. Neonatal calves 1 h post partum (p.p.) were found to have a very high absolute number of granulocytes while the number of peripheral blood mononuclear cells was lower than in calves aged 3-9 weeks. The relative numbers of T cell subpopulations were similar in newborn and older calves, but newborn calves had lower percentages of B cells and MHC class II positive cells. Within the first 4 h of life the relative numbers of CD2+, CD6+, and CD8+ T cells declined in colostrum-fed as well as in colostrum-deprived calves. In contrast, the percentage of MHC class II positive cells and monocytes increased from 1 h to 4 h p.p. particularly in colostrum-fed calves. Although there is some evidence for immaturity of lymphocytes in neonatal calves, the immune system of these animals seems to be fully present at birth.     

Modulatory effects of chitosan adipate on the T and B lymphocyte subsets in mice

This study examined the subsets of T lymphocytes in the thymus, spleen and mesenteric lymph nodes as well as the subsets of B lymphocytes in the spleen and mesenteric lymph nodes in mice administered chitosan adipate (20 mg/kg) intraperitoneally once or four times at 24 h intervals. The results showed that chitosan adipate decreased the percentage of immature CD4⁺CD8⁺ thymic T cells and increased the percentage of mature CD4⁺ and CD8⁺ thymocytes. The most significant stimulating effect was observed after four injections. A single exposure to chitosan adipate increased the percentage of CD4⁺ mesenteric lymph node cells, but four injections of the drug increased the percentage of CD4⁺ and CD8⁺ mesenteric lymph node cells. Chitosan adipate had no effect on the subset of splenic T cells. In contrast, chitosan adipate administered either once or four times increased the percentage of CD19⁺ splenocytes but had no effect on the percentage of CD19⁺ mesenteric lymph node cells. Overall, chitosan adipate induces the maturation and differentiation of thymocytes, and regulates the number of B splenic cells and lymph node T cells irrespective of the number of doses.     

Changes in peripheral leukocyte populations of weak calf syndrome of Japanese black calves

To clarify the cellular immune condition in Japanese Black (JB) calves with a weak syndrome, peripheral leukocyte populations were analyzed by flow cytometry. Twenty JB calves were divided into two groups based on clinical observations; one group of calves was weak, because they had experienced an onset of diarrhea within 3 days after birth and needed treatment (Group 1 ;n=10), and the other group of calves was healthy (Group 2; n=10). With regard to leukocyte populations, CD8(+) cells and gamma delta T cells in Group 1 were markedly lower than those in Group 2 during the experimental periods. It is possible that immune-insufficiency might be based on T lymphocyte function in weak syndrome JB calves during the growth process.     

The effect of feed supplementation with a copper‐glycine chelate and copper sulphate on selected humoral and cell‐mediated immune parameters,plasma superoxide dismutase activity,ceruloplasmin and cytokine concentration in broiler chickens

The varied bioavailability and different effects of organic forms of copper on the immune system of poultry have prompted the search for new feed additives based on copper compounds containing modified chelate complexes. The aim of the study was to determine the effect of inorganic and organic forms of copper on selected parameters of the cellular and humoral immune response in broiler chickens by determining the percentages of CD3⁺CD4⁺, CD3⁺CD8⁺ and CD25⁺ lymphocytes, cells with MHC Class II expression, and BU‐1⁺ cells, as well as the concentrations of SOD, IL‐2, IL‐10 and TNF‐α in the peripheral blood. The experiments were conducted using 500 one‐day‐old Ross 308 roosters divided into five groups. Cu was added in inorganic form (CuSO₄), in inorganic form with the addition of phytase (CuSO4 + F), in organic form in combination with glycine (Cu‐Gly) and in organic form in combination with glycine and a phytase supplement (Cu‐Gly+F). The results of the study indicate an increase in the percentage of CD3⁺CD4⁺ and CD3⁺CD8⁺ T lymphocytes, CD25⁺ T cells, and cells expressing MHC class II molecules, and in the concentration of ceruloplasmin, activity of superoxide dismutase and the concentration of IL‐2 in the groups that received copper, particularly copper‐glycine chelates. Based on the study, we can conclude that supplementation of poultry feed with copper chelates activates mainly the Th1 cellular immune response and the response of peripheral blood T lymphocytes. Furthermore, it promotes secretion of cytokines, which are involved in potentiation and regulation of the immune response in birds.     

Effect of nutritional conditions on changes in leukocyte populations in Japanese black calves

To clarify the effect of nutritive conditions on changes in immune cells in Japanese Black (JB) calves during the growth period, leukocyte populations were analyzed in ten healthy JB calves managed in one herd. The calves were divided into two groups: five calves in Group 1 were given insufficient nutrition, and the other five calves in Group 2 received adequate nutrition. The levels of serum total cholesterol and glucose were significantly lower in Group 1 than in Group 2 at 1 month. The numbers of CD3+, CD4+ and CD8+ cells tended to be lower in Group 1 than in Group 2 at months 1 and 2, and the difference in CD4+ was significant at month 2. The number of MHC class-II(+high) cells was significantly lower in Group 1 than in Group 2 at months 1 and 2. These results suggest that adequate nutrition might stimulate an increase in immune cells in calves during the growth period.     

Changes in Lymphocyte Subsets in the Bronchus‐associated Lymphoid Tissue of Goats Naturally Infected with Different Mycoplasma Species

The distribution of cells containing lysozyme, S‐100 protein, CD3, CD4, CD8, major histocompatibility complex class II antigen and immunoglobulin G (IgG) was analysed in the bronchus‐associated lymphoid tissue (BALT) of goats naturally infected with three Mycoplasma species. This study included the immunohistochemical characterization of the pneumonic lesions of 18 goats (3–5 months old) infected with one of the following Mycoplasma species: M. mycoides ssp. mycoides, Large Colony type (goats no. 1–6), M. mycoides ssp. capri (goats no. 7–12) and M. capricolum ssp. capricolum (goats no. 13–18). Microscopically, infected animals showed a moderate bronchointerstitial pneumonia, characterized by lymphoid hyperplasia of the BALT and infiltration of mononuclear cells in the alveolar walls and airways. The main cellular type in the BALT was represented by CD3⁺ T lymphocytes, and the ratio of CD4⁺:CD8⁺ cells was >2. The BALT showed large germinal centres mainly composed of IgG⁺ B lymphocytes, with numerous S‐100⁺ follicular dendritic cells. The presence of follicular dendritic cells confirmed the high degree of organization of this lymphoid tissue. The immunohistochemical results showed that activated T lymphocytes, particularly in the CD4 subset, and IgG⁺ B cells, play a major role in the immune response of the caprine lung infected with these species of mycoplasmas.     

Histological and immunohistochemical evaluation of duodenal and colonic biopsies after oral bovine lactoferrin supplementation in beagle puppies

Lactoferrin is a natural compound in the milk of mammals and was shown to influence the intestinal micro‐flora and the immune system in mice, calves, dogs and man. The present study was carried out to investigate the effect of orally administered bovine lactoferrin (0, 30, 60 and 120 mg/kg DM feed) on the intestinal morphology and lymphocyte colonization in 36 motherless raised puppies. Endoscopic biopsies from duodenum and colon, taken in week 14, were scored histologically after staining with haematoxylin and eosin (H&E) and lymphocytes (CD3⁺, CD4⁺, CD8⁺) and plasma cells (IgA⁺, IgG⁺, IgM⁺) were enumerated after immunohistochemical staining by computer‐aided quantification. Histological scoring revealed no significant differences amongst the groups. IgG⁺ plasma cells were reduced (p < 0.05) in the lamina propria of the colon of the 30 and the 60 mg group. The number of CD8⁺ lymphocytes was higher (p < 0.05) in the epithelium of the colon of the lactoferrin groups. In conclusion, this study indicated only minimal effects of bovine lactoferrin on the population of selected immune cells in the gut mucosa of puppies. More investigations are needed to describe the impact of lactoferrin on the digestive physiology of puppies.     

Effects of two commercial diets and technical feed treatment on stomach lesions and immune system of fattening pigs

The impact of technical feed treatment and diet on stomach lesions and traits of the local and systemic immune system were investigated in fattening pigs. Feeding groups differed in technical feed treatment (standard ground meal vs. finely ground and pelleted feed) and diet (soya bean meal vs. rapeseed meal/DDGS/soya beans). Pigs were fattened approximately 10 weeks by ad libitum feeding and slaughtered subsequently. Gastric alterations were assessed by a macroscopic scoring system [macroscopic stomach score (MSC) 0 =  normal to 4 =  severe lesions]. For immunological investigations, lymphocytes from blood and jejunal tissues were isolated. T‐cell phenotyping was carried out by staining intestinal lymphocytes with monoclonal antibodies for CD4 and CD8 and flow cytometric measurements. MSC was higher in animals fed finely ground and pelleted feed compared with their counterparts. Significant interactions between diet and feed treatment considering the MSC were observed (p = 0.027). There was no effect of diet or technical feed treatment on T cells of blood, Lymphonodi gastrici or lamina propria (LP) and intraepithelial cells. However, technical feed treatment significantly affected subsets of CD4⁺, CD8⁺, CD8^low, CD4/CD8 double‐positive T cells, the mean fluorescence intensity of CD4⁺ T cells and the ratio of CD8^low/CD8^high T cells in Peyer's patches (PP). All named parameters were reduced in PP of animals fed finely ground and pelleted feed compared with animals fed standard ground meal. Furthermore, significant differences between T cells of lymph nodes and LP were observed between animals with middle MSC (MSC = 1–2.5) and animals with high MSC (MSC = 3–4). Significant alterations in T cells of PP were observed between animals of low (MSC = 0–0.5) and high MSC. The observed effects provide the evidence that the impact of technical feed treatment is not limited on the stomach lesions. Possible stimuli and consequences of the immune system should be studied in more detail.     

Cytometric Analysis of Surface Molecules of Leucocytes and Phagocytic Activity of Granulocytes and Monocytes/Macrophages in Cows with Pyometra

Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non‐specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4⁺CD25⁺ on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p < 0.001). Significantly (p ≤ 0.001) lower percentage of CD4⁺, CD14⁺, CD25⁺ and CD4⁺CD25⁺ phenotype leucocytes was also observed in the peripheral blood of cows from the pyometra group, along with a significantly higher (p < 0.001) percentage of CD8⁺ and CD21⁺ lymphocytes as compared to the healthy group. The results of work indicate that disfunction of cell immunity coexisting with pyometra may be caused by a bacterial infection and the presence of blocking agents (IL‐10), released by the increasing number of CD8⁺ lymphocytes what leads to the advanced inflammation of uterus.