Establishment of canine and feline cells expressing canine signaling lymphocyte activation molecule for canine distemper virus study

Abstract

Signaling lymphocyte activation molecule (SLAM) is one of the receptors for canine distemper virus (CDV). In this study, canine and feline cells expressing canine SLAM, designated A-72/cSLAM and CRFK/cSLAM, were established for the in vitro study of canine distemper. Recent CDV isolates, KDK-1 and 246, which belong to genotypes Asia/H1 and Asia/H2, respectively, rapidly grew and produced distinct syncytia in both the SLAM-expressing cells. The virus-neutralizing (VN) test was successfully performed using these cells, and the results indicated that sera from dogs experimentally infected with KDK-1 had higher VN titers for homologous strain KDK-1 than for heterologous strain 246 and the vaccine Onderstepoort. These newly established cells expressing canine SLAM would help virological and serological analyses of canine distemper.

An Outbreak of Canine Distemper Virus in Tigers (<i>Panthera tigris</i>): Possible Transmission from Wild Animals to Zoo Animals

“…Cells: A72/cSLAM and CRFK/cSLAM cells expressing canine signaling lymphocyte activation molecule (SLAM) [25] were grown (at 37°C in 5% CO 2 ) in Dulbecco's modified Eagle's medium (DMEM: GIBCO, Grand Island, NY, U.S.A.) supplemented with 10% heat-inactivated fetal calf serum (FCS, HyClone®, Ottawa, Canada), 100 U/ml penicillin, and 100 µg/ml streptomycin (GIBCO).…”

Detection of Antibody to Canine Distemper Virus in Wild Raccoons (Procyon lotor) in Japan

“…Total 106 sera were collected from raccoons captured in 2 prefectures, Hyogo and Osaka, under an official control and eradication program of feral raccoons. Virus-neutralizing (VN) test to KDK-1 [10] was carried out by 75% plaque-reduction neutralizing test (PRNT 75 ) using our established cell line, CRFK/cSLAM [11]. For the first screening of CDV-positive raccoons, 10 l of sera was added to 90 l of virus solution containing 100 plaque forming unit (PFU) of KDK-1 diluted with DMEM supplemented with 2% FCS and incubated at 37C for 1 hr (1:10 dilution).…”

“…Sci. 71(12): 1661-1663, 2009 Canine distemper virus (CDV) infects many animal species belonging to the Carnivora and causes lethal disease to them. In Serengenti National Park in Tanzania, an epidemic of CDV spread through lions in 1994, resulting that 30% of lions were dead [13].…”

“…Canine-originated A72/cSLAM and feline-originated CRFK/cSLAM cells expressing canine signaling lymphocyte activation molecule (SLAM) were grown with Dulbecco's modified Eagle's medium (DMEM; GIBCO, U.S.A.) supplemented with 10% FCS and antibiotics at 37C in 5% CO 2 [11]. CDV KDK-1 (genotype Asia-1) [9] and Onderstepoort (a live attenuated vaccine strain) strains were propagated in A72/cSLAM and Vero cells, respectively.…”

Function of Feline Signaling Lymphocyte Activation Molecule as a Receptor of Canine Distemper Virus

“…In order to amplify the corresponding canine gene, the gene encoding canine SLAM without the signal peptide and transmembrane region was amplified from pDisplay/cSLAM [15] with the primers, canineSLAM-95F (5'-CCA GAT CTG AGA GCT TGA TGA ATT GCC C-3') and CDV724R (5'-TCG TCG ACT CTT GGC ACC GAA GAC TCT G-3'). PCR products were digested with restriction endonucleases BglII and SalI and cloned into BglII and SalI sites of the expression plasmid pDisplay (Invitrogen), which has genes for a signal peptide of mouse immunoglobulin kappa-chain V-J2-C, a hemagglutinin A (HA) epitope tag upstream of multiple cloning sites and neomycin-resistance.…”