PMID- 26586612
OWN - NLM
STAT- MEDLINE
DCOM- 20161024
LR  - 20171116
IS  - 1096-0279 (Electronic)
IS  - 1046-5928 (Linking)
VI  - 119
DP  - 2016 Mar
TI  - Cloning and high level expression of the biologically active extracellular domain 
      of Macaca mulatta CD40 in Pichia pastoris.
PG  - 19-26
LID - S1046-5928(15)30101-7 [pii]
LID - 10.1016/j.pep.2015.11.009 [doi]
AB  - The CD40-mediated immune response contributes to a wide variety of chronic 
      inflammatory diseases. CD40 antagonists have potential as novel therapies for 
      immune disorders. However, the CD40 pathway has not been well characterized in 
      the rhesus monkey Macaca mulatta, which is a valuable animal model for human 
      immune disease. An 834 bp transcript was cloned from peripheral blood mononuclear 
      cells (PBMCs) of rhesus monkey using specific primers designed according to the 
      predicted sequence of M. mulatta CD40 (mmCD40) in GenBank. Sequence analysis 
      demonstrated that mmCD40 is highly homologous to human CD40 (hCD40), with an 
      amino acid sequence identity of 94%. Genes encoding the extracellular domain of 
      mmCD40 and the Fc fragment of the hIgG1 were inserted into a pPIC9K plasmid to 
      produce mmCD40Ig by Pichia pastoris. Approximately 15-20 mg of the mmCD40Ig 
      protein with  approximately 90% purity could be recovered from 1 L of culture. The purified 
      mmCD40Ig protein can form dimers and can specifically bind CD40L-positive cells. 
      Additionally, the mmCD40Ig protein can bind hCD40L protein in phosphate buffered 
      saline and form a stable combination in a size-exclusion chromatography assay 
      using a Superdex 200 column. Moreover, mmCD40Ig is as efficient as M. mulatta 
      CTLA4Ig (mmCTLA4Ig) to suppress Con A-stimulated lymphocyte proliferation. 
      Additionally, mmCD40Ig only showed mild immunosuppressive activity in a one-way 
      mixed lymphocyte reaction (MLR) system. These results suggest that mmCD40Ig 
      secreted by P. pastoris was productive and functional, and it could be used as a 
      tool for pathogenesis and therapies for chronic inflammatory diseases in a M. 
      mulatta model.
CI  - Copyright (c) 2015 Elsevier Inc. All rights reserved.
FAU - Zhu, Shengyun
AU  - Zhu S
AD  - Key Lab of Transplant Engineering and Immunology, Ministry of Health, West China 
      Hospital, Sichuan University, Chengdu, 610041, China.
FAU - Wan, Lin
AU  - Wan L
AD  - Key Lab of Transplant Engineering and Immunology, Ministry of Health, West China 
      Hospital, Sichuan University, Chengdu, 610041, China.
FAU - Yang, Hao
AU  - Yang H
AD  - Key Lab of Transplant Engineering and Immunology, Ministry of Health, West China 
      Hospital, Sichuan University, Chengdu, 610041, China.
FAU - Cheng, Jingqiu
AU  - Cheng J
AD  - Key Lab of Transplant Engineering and Immunology, Ministry of Health, West China 
      Hospital, Sichuan University, Chengdu, 610041, China.
FAU - Lu, Xiaofeng
AU  - Lu X
AD  - Key Lab of Transplant Engineering and Immunology, Ministry of Health, West China 
      Hospital, Sichuan University, Chengdu, 610041, China. Electronic address: 
      xiaofenglu@yahoo.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151114
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (CD40 Antigens)
RN  - 0 (Immunosuppressive Agents)
RN  - 0 (Recombinant Proteins)
RN  - 147205-72-9 (CD40 Ligand)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - CD40 Antigens/*biosynthesis/chemistry/genetics/pharmacology
MH  - CD40 Ligand/chemistry
MH  - Cell Line, Tumor
MH  - Cells, Cultured
MH  - Cloning, Molecular
MH  - Gene Expression
MH  - Glycosylation
MH  - Humans
MH  - Immunosuppressive Agents/chemistry/pharmacology
MH  - Leukocytes, Mononuclear/drug effects/physiology
MH  - Macaca mulatta
MH  - Mice
MH  - Molecular Sequence Data
MH  - Pichia
MH  - Protein Binding
MH  - Protein Multimerization
MH  - Protein Processing, Post-Translational
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/biosynthesis/chemistry/genetics/pharmacology
OTO - NOTNLM
OT  - CD40
OT  - Chronic inflammatory disease
OT  - Immunosuppression
OT  - Pichia pastoris
OT  - Rhesus monkey
EDAT- 2015/11/21 06:00
MHDA- 2016/10/25 06:00
CRDT- 2015/11/21 06:00
PHST- 2015/09/17 00:00 [received]
PHST- 2015/10/21 00:00 [revised]
PHST- 2015/11/09 00:00 [accepted]
PHST- 2015/11/21 06:00 [entrez]
PHST- 2015/11/21 06:00 [pubmed]
PHST- 2016/10/25 06:00 [medline]
AID - S1046-5928(15)30101-7 [pii]
AID - 10.1016/j.pep.2015.11.009 [doi]
PST - ppublish
SO  - Protein Expr Purif. 2016 Mar;119:19-26. doi: 10.1016/j.pep.2015.11.009. Epub 2015 
      Nov 14.