Human lymphocyte function-associated antigen 3 (hLFA3) has been identified as an important T cell accessory molecule. Rhesus monkeys (Macaca mulatta) have been widely used as animal models for human immune disorders. Due to the species-specificity of immune system, it is necessary to study M. mulatta LFA3 (mmLFA3). In this study, the gene encoding mmLFA3 CD2-binding domain (mmLFA3Sh) was amplified by polymerase chain reaction (PCR) and genetically fused to human IgG1 Fc fragment in pPIC9K to construct the expression plasmid pPIC9K-mmLFA3Sh-Ig. Approximately 3-4 mg mmLFA3Sh-Ig protein was recovered from 1 L of inductive media, and mmLFA3Sh-Ig produced by the P. pastoris can bind to the CD2 positive cells, and suppress the monkey and human lymphocytes proliferation induced by Con A and alloantigen in a dose-dependent manner. These results suggested that mmLFA3Sh-Ig might be used as a novel tool for pathogenesis and experimental immunotherapy of Rhesus monkey immune disorders.
Animals ; CD58 Antigens ; biosynthesis ; Humans ; Immunoglobulin G ; Lymphocyte Activation ; Macaca mulatta ; Pichia ; Plasmids ; Protein Interaction Domains and Motifs ; Recombinant Fusion Proteins ; biosynthesis ; T-Lymphocytes

Objective:To search for the potential novel mutation site and to discuss related clinical characteristics by collecting detailed information and testing the gene of a family with highly suspected type 7 maturity-onset diabetes of the young (MODY7).Methods:The gene test was conducted in a 28-year-old female patient with a 20-year course of non-ketosis-prone diabetes, with non-effective long-term insulin treatment, and a 3-generation family history of diabetes, and the patient was found to carry KLF11 gene mutation. Thus, the clinical data of family members were collected and investigated, and the pathogenic gene was tested. Firstly, the proband was searched for pathogenic genes by chip-capture high-throughput sequencing method. Then the mutation sites were verified by Sanger sequencing technology, and other family members were searched for the same mutation sites by the Sanger sequencing technology.Results:A total of two members of the family was found to have heterozygous mutation of KLF11 gene: c. 920C>T (No. 920 nucleotide of the coding region mutated from cytosine to thymine), resulting in the change of corresponding amino acid p. P307L (No. 307 amino acid changed from proline to leucine), which was a missense mutation and was consistent with their clinical diagnosis of diabetes.Conclusions:The family in this study had a family history of diabetes caused by the missense mutation of KLF11 gene. This is the first report of the mutation site of c. 920C >T (p.P307l), which may be a new mutation site of MODY7.