Molecular cloning and characterization of a cDNA encoding monoclonal nonspecific suppressor factor.
The monoclonal nonspecific suppressor factor (MNSF) is a lymphokine product of a murine T-cell hybridoma that inhibits the generation of lipopolysaccharide-induced immunoglobulin-secreting cells in an antigen-nonspecific manner. A cDNA clone encoding MNSF beta (an isoform of MNSF) was isolated and expressed in bacteria. The sequence obtained is virtually identical to the Fau protein, a product of the ubiquitously expressed fau gene with unknown function. Northern blot analysis demonstrated a single, 0.6-kb transcript. Specific polyclonal antibodies against synthetic peptides corresponding to the deduced amino acid sequences were elicited in rabbits. Immunoprecipitation experiments with these antibodies showed that MNSF beta is released extracellularly in an aggregate form, albeit it lacks a signal peptide sequence. The anti-MNSF beta affinity eluate from the MNSF-producing murine hybridoma (E17) and concanavalin A-activated splenocyte culture supernatants inhibited the immunoglobulin production by lipopolysaccharide-activated splenocytes. Recombinant MNSF beta also showed a similar biologic activity. Thus, ubiquitin-like protein(s) may be involved in the regulation of the immune responses.
Nakamura
M
M
Department of Biochemistry, Shimane Medical University, Izumo, Japan.
Xavier
R M
RM
Tsunematsu
T
T
Tanigawa
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D018076:DNA, Complementary / Q000235:genetics; D007108:Immune Tolerance; D015151:Immunoblotting; D051379:Mice; D008806:Mice, Inbred AKR; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000494:pharmacology; D012269:Ribosomal Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013491:Suppressor Factors, Immunologic / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology*
D018076:DNA, Complementary; C489668:Fau protein, mouse; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; D013491:Suppressor Factors, Immunologic; C072169:monoclonal nonspecific suppressor factor