Establishment and Expression of Cytokines in a Theileria annulata-Infected Bovine B Cell Line

Abstract

This study aimed to establish a pure single-cell Theileria annulata-infected B cell line for the assessment of cytokine production in transformed and lipopolysaccharide (LPS)-stimulated cells. Several studies have aimed to identify cell surface markers in T. annulata-transformed cells; however, no information on cytokine production in these cells is available. To investigate the potential of the transformed cells to produce cytokines and their potential responses to antigen-stimulation, we purified mature B cells (CD21) from the whole blood of cattle experimentally infected with the T. annulata Kashi strain by magnetic separation. The purity and specificity of the established cell line was assessed by the identification of specific cell surface markers (CD21, IgM, and WC4) by flow cytometry analysis. The transcript levels of the cytokines IL1A, IL1B, IL2, IL4, IL6, IL8, IL10, IL16, LTA, TGFB1, TNFA, IFNA, and IFNB in transformed, buparvaquone (BW720c)-treated cells, and antigen-stimulated cells were analyzed by quantitative polymerase chain reaction (qPCR) using cDNA from these cells. A T. annulata-infected bovine B cell line was successfully established with a purity of ~98.8% (CD21). IL4 and IL12A were significantly (p < 0.01) upregulated in the transformed cells. In BW720c-treated transformed cells, IL12B, TGFB1, and IFNB were significantly (p < 0.01) upregulated. Notably, no significant (p > 0.05) upregulation of cytokines was observed in LPS-stimulated transformed cells. Moreover, IL1A, IL1B, IL8, and IL16 were significantly (p < 0.01) upregulated in LPS-stimulated B cells. Our data signify the potential use of this cell line for cytokine production, observance of immunoglobulins, and production of an attenuated vaccine against tropical theileriosis.

Comparison of the effects of buparvaquone and paromomycin on oocyst excretion and clinical parameters in diarrheal calves naturally infected with cryptosporidiosis

“…A transmission electron microscopic study, in which the effect of BPQ on oxidative phosphorylation in the mitochondria of E. multilocularis cells was measured in mice, revealed that treatment with BPQ prematurely disrupted parasitic mitochondria (Rufener et al, 2018). In studies to explore the survival pathways of infected cells (Guergnon et al, 2003), parasites exposed to the BPQ terminate their proliferation after the death of macroschizonts and trigger apoptosis of host cells within a few days (Rashid et al, 2019, Ram et al, 2021. A research conducted to compare the intestinal permeability of BPQ used for the treatment of Leishmania donovani infections with other drugs emphasized that the active substance that remained in the tissues for the longest time was BPQ (Venkatesh et al, 2007).…”

Bovine Tropical Theileriosis: An Update

“…T. annulata hijacks the host immune response to invade and transform leukocytes [37,38]. Knowledge of the pathogenic mechanisms that underlie tropical theileriosis is scanty but there is evidence that T. annulata schizont-infected cells causes failure of a protective immune response.…”

Expression of TRAF6 in peripheral blood B cells of patients with myasthenia gravis

“…Tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) is a member of the TRAF family that can regulate signaling pathways by acting as an E3 ubiquitin ligase in addition to its role as an adaptor protein [3,4]. TRAF6 acts downstream of CD40 (a member of the TNFR superfamily) and the Toll-like receptor (TLR) superfamily to regulate the activation of the nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways [4][5][6][7], which enhances the activation of B cells and the expression of interleukin (IL)-6 [8], IL-4, IL-1 [9] and other downstream factors. Recently, TRAF6 expression was found to be upregulated in systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), hyperplastic thymuses and MG patient serum.…”