Heterogeneous profiles of a factor that renders neutrophils cytotoxic obtained from a concanavalin A-stimulated spleen cell culture in partial purification process

Journal Article · Thu May 15 00:00:00 EDT 1986 · J. Immunol.; (United States)

OSTI ID:5007040

Fujii, Y; Inoue, T; Ito, M; Kimura, S; Fuyama, S; Arai, S; Naiki, M; Sendo, F

Concanavalin A (Con A)-stimulated rat spleen cells were cultured in a serum-free conditioned medium. This culture supernatant contained a certain factor(s) that renders neutrophil cytotoxic for various tumor cells. The factor was tentatively termed neutrophil-activating factor (NAF). NAF activity was eluted in broad fractions by the ion exchange chromatography and the gel filtration. Moreover, on the Con A column, some NAF activities were bound to the column, but other activities passed through the column. These results showed the heterogeneity or polydispersity of NAF activity in both molecular size and charge-based separation properties. Monoclonal antibodies were produced by fusing BALB/c myeloma cells (P3-X63 Ag8.653) with spleen cells from syngeneic mice immunized with partially purified NAF (pNAF) obtained from the gel filtration. Absorbent beads which were linked with one monoclonal antibody (ANAF-10) partially absorbed NAF activity from supernatants of a Con A-stimulated spleen cell culture. By further purification of pNAF the NAF activity was concentrated about 10,000-fold. Heterogeneity of NAF activity, however, did not disappear in even this affinity chromatography. On the other hand, /sup 125/I-labeled material of the final product migrated to one major band corresponding with an m.w. of about 20,000 as determined by SDS-PAGE analysis, and NAF activity was detected in the same band.

• Research Organization: Yamagata Univ. School of Medicine, Japan
• OSTI ID: 5007040
• Journal Information: J. Immunol.; (United States), Vol. 136:10
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
LEUKOCYTES
CHEMICAL ACTIVATION
NEUTROPHILS
TOXICITY
TUMOR CELLS
CELL KILLING
BIOLOGICAL PATHWAYS
CELL CULTURES
CONCANAVALIN
IODINE 125
MICE
MONOCLONAL ANTIBODIES
PURIFICATION
RATS
SPLEEN CELLS
TRACER TECHNIQUES
AGGLUTININS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
HEMAGGLUTININS
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LECTINS
MAMMALS
MATERIALS
NUCLEI
ODD-EVEN NUCLEI
RADIOISOTOPES
RODENTS
SOMATIC CELLS
VERTEBRATES
550901* - Pathology- Tracer Techniques