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Production of antigen-specific hybridomas has been achieved by fusion of HGPRT-negative mutant of Jurkat and antigen-specific TH cell lines. Selection of clones for specificity was best accomplished by the binding of radiolabeled hybrid clones to antigen-pulsed monolayers. Functional lymphokines such as IL-2 and BCGF were induced by antigenic stimulation in the presence of AC. AC function of EBV-transformed autologous B cells was as efficient on a per cell basis as autologous monocytes. Loss of specificity and function was observed in the first 5-6 months of culture which was coincident with chromosome loss. Recovery of specific functional hybrids was accomplished by repeated subcloning during this period. After this time, hybrids have maintained function for at least one year. This methodology allows the study of certain T cell functions without the requirement of periodic restimulation with autologous monocytes. Since EBV-B cell lines can serve as AC to induce lymphokines, large quantities of material can be prepared for biochemical studies.
Production of antigen-specific hybridomas has been achieved by fusion of HGPRT-negative mutant of Jurkat and antigen-specific TH cell lines. Selection of clones for specificity was best accomplished by the binding of radiolabeled hybrid clones to antigen-pulsed monolayers. Functional lymphokines such as IL-2 and BCGF were induced by antigenic stimulation in the presence of AC. AC function of EBV-transformed autologous B cells was as efficient on a per cell basis as autologous monocytes. Loss of specificity and function was observed in the first 5-6 months of culture which was coincident with chromosome loss. Recovery of specific functional hybrids was accomplished by repeated subcloning during this period. After this time, hybrids have maintained function for at least one year. This methodology allows the study of certain T cell functions without the requirement of periodic restimulation with autologous monocytes. Since EBV-B cell lines can serve as AC to induce lymphokines, large quantities of material can be prepared for biochemical studies.