Role of Muscarinic-3 Receptor in Systemic Sclerosis: Correlation with Disease Duration and Effects of IVIG

1 Disease Duration and Effects of IVIG 2 3 Short Title: SSc duration and cholinergic transmission 4 5 Sumit Kumar1, Jagmohan Singh1, Ramalinga Kedika1, Fabian Mendoza2, Sergio 6 A. Jimenez3, Erik S. Blomain4, Anthony J. DiMarino1, Sidney Cohen1, and Satish 7 Rattan1 8 9 1Department of Medicine, Division of Gastroenterology and Hepatology, 2Division of 10 Rheumatology, 3Jefferson Institute of Molecular Medicine and Scleroderma Center, 11 4Department of Pharmacology & Experimental Therapeutic, Thomas Jefferson University, 12 Sidney Kimmel Medical College, Philadelphia, PA 13 14 Grant Support: The work was supported by Grant Number RO1DK035385 from the National 15 Institutes of Diabetes and Digestive and Kidney Diseases, industrial support from CSL Behring, King 16 of Prussia, PA, and an institutional grant from Thomas Jefferson University. E.S.B. is the recipient of 17 the Ruth L. Kirschstein National Research Service Award for Individual Pre-doctoral MD/PhD Fellows 18 (1 F30 CA180500) from the National Institutes of Health. 19 20 Abbreviations Used in this Paper: Ach, acetylcholine; BeCh, bethanechol; EFS, electrical 21 field stimulation; ELISA, enzyme-linked immunosorbent assay; IFI, immunofluorescence intensity; 22 IHC, immunohistochemistry or immunohistochemical; IgG, immunoglobulin; IVIG, pooled human 23 immunoglobulin for intravenous use; KPS, Krebs physiological solution; NIgG, IgGs from normal 24 subjects; M3-RL2, second extracellular loop peptide of M3-R;F(ab')2, antigen-binding fragment of 25 IVIG; SM, smooth muscle; SMC, smooth muscle cells; MCN, myenteric cholinergic neurons, MP, 26 myenteric neuronal plexus; SSc, systemic sclerosis or scleroderma; SScIgG, IgGs from scleroderma 27 patients;; TMB, 3,3’,5,5’-tetramethylbenzidine 28 29 Corresponding Author: Dr. Satish Rattan, Professor of Medicine; Department of 30 Medicine, Division of Gastroenterology & Hepatology, 1025 Walnut Street, 320 College, 31 Philadelphia, PA 19107; Email # [email protected] ; Tel # (215) 955-5614; Fax # (215) 32 923-7697 33 34 35 Disclosures: The authors have nothing to disclose 36 37 38 Authors’ Contribution to the Manuscript: 39 SK, JS in human blood samples’ collection, data acquisition, analysis and interpretation; and 40 statistical analysis; AD, SC, RK and SAJ in the enrolment and follow up of scleroderma patients; FM, 41 SJ in the isolation and purification of SScIgGs; EB for assistance in immunohistochemistry studies; 42 and SR in funds procurements, concept and design, interpretation of data, and study supervision 43 Articles in PresS. Am J Physiol Gastrointest Liver Physiol (May 12, 2016). doi:10.1152/ajpgi.00034.2016