Hybridomas From Lymphocytes of Normal Mice Produce Monoclonal Autoantibodies

A genetically determined host factor controlling susceptibility to encephalomyocarditis virus-induced diabetes in mice

“…2b). When pancreatic sections were stained with anti-insulin antibody 4 days after EMC-D virus injection as described elsewhere (Yoon et al, 1984;Yoon & Notkins, 1976;Baek& Yoon, 1991), most of the beta cells from C57BL/6J mice stained positively but most of those from SJL/J mice stained negatively. In addition, the concentration of immunoreactive insulin (IRI) in the pancreata was measured as previously described (Yoon et al, 1980;Baek & Yoon, 1990).…”

“…As the first step in virus infection is binding of virus to cells, we attempted to determine the attachment rate of virus to freshly isolated beta cells, which are similar to beta cells in vivo. We previously established a method for isolating beta cells from mice by collagenase treatment (Yoon et al, 1984;Yoon & Notkins, 1976) and for labelling and purifying EMC virus (Bae et al, 1989;Eun et al, 1988;Yoon et al, 1988). Briefly, monolayers of L929 cells were adsorbed with EMC-D virus at an m.o.i, of 10.…”

“…To compare the rate of viral attachment to freshly isolated beta cells from SJL/J mice with that to freshly isolated beta cells from C57BL/6J mice, pancreatic beta cells were prepared from 5-to 6-week-old SJL/J or C57BL/6J male mice (30 mice per group) as described previously (Yoon et al, 1984;Yoon & Notkins, 1976). The isolated beta cells were suspended in MEM with 5 % FBS and kept for 20min at 4°C.…”

“…Each sample was placed on a plastic dish (35 mm) and incubated at 37 °C in a humidified atmosphere containing 5 % CQ. Fourteen hours later, the non-adherent beta cells were collected, thereby eliminating the contaminating fibroblastoid cells that had adhered to the dish (Yoon et al, 1984;Yoon & Notkins, 1976). The collected beta cells were incubated at 37 °C in a humidified atmosphere containing 5 % CO 2 until use.…”