The use of syngeneic cells producing human somatotropic hormone (hSTH) for immunization of mice and development of hybridomas.
We studied the possibility of syngeneic cells expressing heterologous protein being used for sensitization of mice and production of hybridomas. Recombinant retroviral vector containing cloned human somatotropic hormone (hSTH) gene was used to express hSTH in BALB/3T3 cells. BALB/c mice were injected intrasplenically (i/s) or combination of intraperitoneally (i/p) and intrasplenically with hSTH-producing cells. Sensitized splenocytes were fused with myeloma cell P3X63-AgB.653. Screening for anti-hSTH hybridomas was performed by enzyme-linked immunoassay. Both single i/s injection of producer cells as well as combined i/s and i/p injections were effective for sensitization of splenocytes. Combined injection was effective for production of IgG and IgM secreting hybridomas. Single i/s injection led to generation of only IgM producing hybridomas. The results proved that syngeneic cells expressing genes of heterologous proteins can be used for splenocyte sensitization and hybridoma preparation.
Byelorussian Institute of Hematology and Blood Transfusion, Minsk, USSR.
A V
AV
Panyutich
V S
VS
Prassolov
E A
EA
Shydlovskaya
M V
MV
Reznikov
P M
PM
Chumakov
N N
NN
Voitenok
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D005347:Fibroblasts; D013006:Growth Hormone / Q000235:genetics / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007114:Immunization / Q000379:methods*; D007132:Immunoglobulin Isotypes; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology; D012190:Retroviridae / Q000235:genetics; D014162:Transfection
D000911:Antibodies, Monoclonal; D007132:Immunoglobulin Isotypes; D011994:Recombinant Proteins; D013006:Growth Hormone