Immunoglobulins from scleroderma patients inhibit the muscarinic receptor activation in internal anal sphincter smooth muscle cells.
Systemic sclerosis (SSc) IgGs affecting the M(3)-muscarinic receptor (M(3)-R) have been proposed to be responsible for the gastrointestinal (GI) dysmotility in this disease. However, the effect of SSc IgGs on smooth muscle cell (SMC) function has not been studied. We determined the effect of SSc IgGs on the muscarinic receptor activation by bethanechol (BeCh; methyl derivate of carbachol) in SMC and smooth muscle strips from rat internal anal sphincter. IgGs were purified from GI-symptomatic SSc patients and normal volunteers, with protein G-Sepharose columns. SMC lengths were determined via computerized digital micrometry. The presence of M(3)-R and IgG-M(3)-R complex was determined by Western blot. IgGs from SSc patients but not from normal volunteers caused significant and concentration-dependent inhibition of BeCh response (P < 0.05). The maximal shortening of 22.2 +/- 1.2% caused by 10(-4) M BeCh was significantly attenuated to 8.3 +/- 1.2% by 1 mg/ml of SSc IgGs (P < 0.05). Experiments performed in smooth muscle strips revealed a similar effect of SSc IgG that was fully reversible. In contrast to the effect on BeCh, the SSc IgGs caused no significant effect (P > 0.05) on K(+) depolarization and alpha(1)-adrenoceptor activation by phenylephrine. Western blot studies revealed the specific presence of SSc IgG-M(3)-R complex. SSc IgGs attenuated M(3)-R activation, which was reversible with antibody removal. These data suggest that SSc GI dysmotility may be caused by autoantibodies that inhibit the muscarinic neurotransmission. Future treatment of SSc patients may be directed at the removal or neutralization of these antibodies.
Singh
Jagmohan
J
Department of Medicine, Thomas Jefferson Univ., Philadelphia, PA 19107, USA.
Mehendiratta
Vaibhav
V
Del Galdo
Francesco
F
Jimenez
Sergio A
SA
Cohen
Sidney
S
DiMarino
Anthony J
AJ
Rattan
Satish
S
D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001003:Anal Canal / Q000187:drug effects / Q000276:immunology* / Q000503:physiopathology; D000818:Animals; D001323:Autoantibodies / Q000097:blood*; D001572:Benzofurans / Q000494:pharmacology; D018723:Bethanechol / Q000494:pharmacology; D016022:Case-Control Studies; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D007136:Immunoglobulins / Q000097:blood*; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D018721:Muscarinic Agonists / Q000494:pharmacology; D018727:Muscarinic Antagonists / Q000494:pharmacology; D009119:Muscle Contraction / Q000187:drug effects; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000276:immunology*; D010656:Phenylephrine / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D043587:Receptor, Muscarinic M3 / Q000187:drug effects / Q000276:immunology*; D012595:Scleroderma, Systemic / Q000276:immunology* / Q000503:physiopathology
D000316:Adrenergic alpha-Agonists; D001323:Autoantibodies; D001572:Benzofurans; D007136:Immunoglobulins; D018721:Muscarinic Agonists; D018727:Muscarinic Antagonists; D010880:Piperidines; D011759:Pyrrolidines; D043587:Receptor, Muscarinic M3; D018723:Bethanechol; D010656:Phenylephrine; D011189:Potassium Chloride; C042375:4-diphenylacetoxy-1,1-dimethylpiperidinium; C101207:darifenacin