       Document 0513
 DOCN  M9620513
 TI    Influence of antigen presentation and exogenous cytokine activity during
       in vitro primary immunizations employed for the generation of monoclonal
       antibodies.
 DT    9602
 AU    Halabi G; McCullough KC; Institut fur Viruskrankheiten und
       Immunprophylaxe,; Mittelhausern, Switzerland.
 SO    J Immunol Methods. 1995 Oct 26;186(2):205-16. Unique Identifier :
       AIDSLINE MED/96062063
 AB    Hybridomas secreting monoclonal antibodies (MAbs) against African horse
       sickness virus (AHSV) were generated using different AHSV antigen
       preparations (inactivated AHSV, semi-purified virus, and a preparation
       of nonstructural viral proteins) in one of three different in vitro
       primary immunization systems: (i) the Cel-prime kit, a method using
       immunization of splenocytes aided by antigen-primed support cells; (ii)
       a system based on a cytokine soup derived from a mixed lymphocyte
       reaction plus stimulated EL4-IL-2 cells; (iii) a system based on a
       cytokine soup derived from splenocytes stimulated by pokeweed mitogen in
       order to obtain a mixture of cytokines enriched for Th2 lymphokines. The
       viability of immunized BALB/c mouse splenocytes, immunoglobulin
       production by the subsequently generated hybridomas, and the specificity
       of the MAbs were compared. The most efficient in vitro primary
       immunization system was the Cel-prime system employing semi-purified
       antigen. This efficiency was manifest in terms of a greater viability of
       the splenocytes in the immunization, as well as a higher number of
       specific antibody-secreting hybridomas. It seems probable that the
       support cells of the Cel-prime system have an accessory function such as
       that attributed to antigen-presenting cells. Such a function would
       result in impairment of apoptosis, and thus increase the viability of
       the splenocytes in the in vitro primary immunization system, as well as
       enhancing stimulation of the immune response against the antigen used.
       The presence of cytokines at the beginning of the in vitro primary
       immunization did have an influence, but this was secondary to what
       appeared to be the major event of cellular interaction associated with
       the accessory cell function of the support cells.
 DE    African Horsesickness Virus/*IMMUNOLOGY  Animal  Antibodies,
       Monoclonal/*BIOSYNTHESIS/IMMUNOLOGY  Antibodies,
       Viral/*BIOSYNTHESIS/IMMUNOLOGY  *Antigen Presentation  Comparative Study
       Cytokines/*PHARMACOLOGY/SECRETION  Evaluation Studies  Female
       Hybridomas/IMMUNOLOGY  Interleukin-2/PHARMACOLOGY  Lymphocyte Culture
       Test, Mixed  Lymphocyte Transformation/DRUG EFFECTS  Lymphoma/PATHOLOGY
       Mice  Mice, Inbred BALB C  Mice, Inbred C57BL  Multiple
       Myeloma/PATHOLOGY  Pokeweed Mitogens/PHARMACOLOGY
       Spleen/CYTOLOGY/IMMUNOLOGY  Support, Non-U.S. Gov't  Thymus
       Gland/CYTOLOGY/IMMUNOLOGY  Th2 Cells/DRUG EFFECTS/SECRETION  Tumor
       Cells, Cultured  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

