Anti-IL-12 P70 monoclonal antibody

Postpartum dairy cows experience a heightened inflammatory state coinciding with the time of greatest nutrient deficit. Nutrient availability is sensed on the cellular level by nutrient sensing kinases, such as the PI3K/AKT/mTOR (mTOR) pathway, a key orchestrator of immune cell activation and inflammatory balance. Our objective was to determine the responsiveness of this pathway to inflammatory stimulation with and without nutrient supplementation ex vivo. Blood samples were collected from Holstein cows (n = 14) at -42, -14, 7, 21, and 42 d relative to calving. Control samples and samples pretreated with a mixture of amino acids, glucose, and insulin (AAM) were stimulated with 100 ng/mL E. coli lipopolysaccharide (LPS; LPS, AAMLPS) or left unstimulated (control, AAM). After 1 h, ratios of mean fluorescence intensity for phosphorylated to total protein of AKT and mTORC1 substrates S6RP and 4EBP1 were analyzed in polymorphonuclear cells (PMN), and monocytes by flow cytometry. A separate aliquot was stimulated with LPS for 2 h and relative mRNA abundance of IL10, IL12A, IL12B, and TNFA in whole blood leukocytes from 10 cows was measured by reverse-transcription quantitative PCR. Repeated measures ANOVA was performed with fixed effects of time, treatment, and their interaction. Cells had different ratios of pathway proteins with PMN having the highest phosphorylation of AKT, S6RP, and 4EBP1. Stimulation with LPS consistently activated mTOR signaling in PMN regardless of nutrient supplementation except for postpartum 4EBP1, which increased in response to nutrients alone. In monocytes, AKT baseline phosphorylation was lower and activation could not be induced by either treatment, whereas activation of 4EBP1 responded to nutrient supplementation. Treatment with LPS increased phosphorylation of S6RP in both innate immune cell types. Nutrient supplementation increased baseline IL10 expression and decreased baseline as well as LPS-induced IL12B and TNFA expression. We conclude that the mTOR pathway in bovine innate immune cells can be differentially activated in response to inflammatory stimulation and nutrient supplementation in monocytes versus PMN. Effects of nutrient supplementation on cytokine mRNA abundance are likely specific to immune cell type.

The goal of our study was to evaluate the genetic effects of interleukins genes polymorphisms on the risk of CAD through a case-control study and a meta-analysis. 826 CAD cases and 1153 CAD-free controls in east of China were collected to analyze the association of 28 SNPs in IL1A, IL1B, IL1f7, IL3, IL7Ra, IL17A, IL18, IL18R, IL18RAP, IL28B, IL15RA, IL15, IL9, IL10, IL12A, IL12B, and IL13 genes on the risk of CAD. The distributions of all genotypes were not significantly different between CAD group and control group (P > 0.05), except for IL1f7 rs3811047 G > A and IL10 rs1800872 G > T genetic variants. Our results showed that IL1f7 rs3811047 G > A, IL28B rs8099917 T > G and IL10 rs1800872 G > T polymorphisms were associated with the decreased risk of CAD in various comparison models. No significant association between the other 25 SNPs and the risk of CAD were found in this population. But the meta-analysis on IL10 rs1800872 G > T polymorphism and CAD risk, totaling 3038 CAD patients and 2732 controls, provided no convincing evidence for the genetic association (T vs. G: OR = 1.07, 95% CI 0.88-1.30, P = 0.510), even upon stratified analysis by ethnicity (Asian and Caucasian) or control selection criteria (normal angiography and symptom investigation). The discrepant overall result from previously published studies reflects publication bias or methodological problems. Future well-designed studies with large sample size should be conducted to validate our findings.