Analysis of HLA-DR from alveolar macrophages and blood monocytes by two-dimensional gel electrophoresis
Human blood monocytes (BM) are more effective than alveolar macrophages (AM) in promoting lymphocyte proliferation to antigen. To further understand these differences, the HLA-DR molecules synthesized by these two cell types were compared. AM were prepared by adherence of cells obtained by bronchoscopic lavage; BM were prepared by adherence of blood mononuclear cells from the same normal volunteer. Cells were cultured for 7 hours with /sup 3/H-leucine and HLA-DR was immunoprecipitated with the murine monoclonal antibody L243. Immunoprecipitates were analyzed by two-dimensional gel electrophoresis. In three experiments, protein synthetic rate was greater and more HLA-DR was immunoprecipitated per cell in BM than in AM. Isoelectric focusing showed identical charge variation for BM and AM. However, molecular weight analysis of AM HLA-DR revealed multiple bands of slightly different molecular weight for each beta-chain peptide, whereas only a single band occurred with BM HLA-DR. Neuraminidase treatment reduced the charge heterogeneity but did not affect the molecular weight differences. These findings may relate to the differential ability of AM and BM to promote lymphocyte proliferation to antigen.
- Research Organization:
- Univ. of Pennsylvania, Philadelphia
- OSTI ID:
- 5388768
- Report Number(s):
- CONF-8604222-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
LYMPHOCYTES
CELL PROLIFERATION
MACROPHAGES
CHEMICAL COMPOSITION
MONOCYTES
BLOOD
CELL CULTURES
ELECTROPHORESIS
LAVAGE
LEUCINE
LUNGS
MAN
MICE
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BIOLOGICAL MATERIALS
BLOOD CELLS
BODY
BODY FLUIDS
CARBOXYLIC ACIDS
CONNECTIVE TISSUE CELLS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LEUKOCYTES
MAMMALS
MATERIALS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PHAGOCYTES
PRIMATES
RESPIRATORY SYSTEM
RODENTS
SOMATIC CELLS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques