Cytokine gene expression in response to Haemonchus placei infections in Nelore cattle.

This study aimed to evaluate the expression of a subset of cytokine genes in response to Haemonchus placei infections in Nelore cattle presenting different degrees of resistance to natural infections. One hundred weaned bulls, initially 11-12 months old, were evaluated and kept on the same pasture. Faecal and blood samples were collected for parasitological and immunological assays. The seven most resistant and the eight most susceptible animals were selected based on nematode faecal egg counts (FEC) and worm burden. Serum was collected to measure antibody titres, and abomasum and abomasal lymph node tissue samples were collected to analyse the expression of a subset of cytokine genes (IL-2, IL-4, IL-8, IL-12p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, MUC-1) using real-time RT-PCR. Mast cells, eosinophils and globule leukocytes in the abomasal mucosa were enumerated, and IgA levels in the mucus were assessed. Gene expression analysis in the abomasal tissue indicated that IL-4 and IL-13 (TH2 cytokines) were up-regulated in the resistant group, whereas TNF-alpha (TH1/TH2 cytokine) was up-regulated in the susceptible group. In abomasal lymph nodes, IL-4 and IFN-gamma were up-regulated in the resistant and susceptible groups, respectively. In the resistant group, serum IgG1 levels were higher against antigens of H. placei infective larvae on days 14, 42, 70 and 84 and against antigens of H. placei adults on day 84 (P<0.05). The resistant group had higher mast cell counts in the abomasal mucosa than the susceptible group (P<0.05). These results indicate a protective TH2-mediated immune response against H. placei in the resistant group and a less protective TH1 response in the susceptible group.

Immunological responses and cytokine gene expression analysis to Cooperia punctata infections in resistant and susceptible Nelore cattle.

Cellular and humoral immune response, as well as cytokine gene expression, was assessed in Nelore cattle with different degrees of resistance to Cooperia punctata natural infection. One hundred cattle (male, weaned, 11-12 months old), kept together on pasture, were evaluated. Faecal and blood samples were collected for parasitological and immunological assays. Based on nematode faecal egg counts (FEC) and worm burden, the seven most resistant and the eight most susceptible animals were selected. Tissue samples of the small intestine were collected for histological quantification of inflammatory cells and analysis of cytokine gene expression (IL-2, IL-4, IL-8, IL-12p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, and MUC-1) using real-time RT-PCR. Mucus samples were also collected for IgA levels determination. Serum IgG1 mean levels against C. punctata antigens were higher in the resistant group, but significant differences between groups were only observed 14 days after the beginning of the experiment against infective larvae (L3) and 14 and 84 days against adult antigens. The resistant group also presented higher IgA levels against C. punctata (L3 and adult) antigens with significant difference 14 days after the beginning of the trial (P<0.05). In the small-intestine mucosa, levels of IgA anti-L3 and anti-adult C. punctata were higher in the resistant group, compared with the susceptible group (P<0.05). Gene expression of both T(H)2 cytokines (IL-4 and IL-13) in the resistant group and T(H)1 cytokines (IL-2, IL-12p35, IFN-gamma and MCP-1) in the susceptible group was up-regulated. Such results suggested that immune response to C. punctata was probably mediated by T(H)2 cytokines in the resistant group and by T(H)1 cytokines in the susceptible group.

Field study on nematode resistance in Nelore-breed cattle.

The present study evaluated Nelore cattle with different degrees of resistance to natural infections by gastrointestinal nematodes. One hundred weaned male cattle, 11-12 months of age, were kept on the same pasture and evaluated from October 2003 to February 2004. Faecal and blood samples were collected for parasitological, haematological and immunological tests. In February 2004, the 10 most resistant and the 10 most susceptible animals were selected based on individual means of nematode faecal egg counts (FEC). Such animals were slaughtered for worm burden determination and nematode species identification. The repeatability estimates for FEC (+/-S.D.), log-transformed FEC and packed-cell volume (PCV) in all animals were 0.3 (+/-0.05), 0.26 (+/-0.04) and 0.42 (+/-0.05), respectively. The resistant group showed lower FEC and worm burdens than the susceptible group (P<0.05). There were no significant differences between groups regarding mean body weight, weight gain, PCV and total serum protein values (P>0.05). The resistant group showed higher total serum IgE levels (P<0.05) and higher mean eosinophil blood counts. However, the latter was statistically significant only 42 days after the beginning of the study. Nematodes Cooperia punctata and Haemonchus placei were predominant and the correlation between Cooperia and Haemonchus burdens was 0.64 (P<0.05), which indicated that animals presenting increased numbers of one of those genera probably had increased numbers of the other. The current study provides further evidence of IgE active role in nematode immunity and suggests that total serum IgE level might serve as an additional marker to select Nelore cattle that are responsive to H. placei and C. punctata infections.