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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).