Molecular cloning, expression and functional characterization of miniature swine CD86.
CD86 is one of the key molecules involved in the co-stimulation of T cells. The complete cDNA encoding CD86 molecule of miniature swine was cloned and analyzed. A comparison of two CD86 amino acid sequences of miniature swine and domestic swine showed only three amino acid differences suggesting that it is unlikely to affect the major structural features of the miniature swine CD86 (msCD86). In the expression study, constitutive expression of CD86 mRNA was detected in various tissues, and the aberrant expression of the transcriptional variant (putative soluble form) was noted. The cDNA and amino acid sequences for this variant were determined and compared with those for the human soluble CD86, which was previously reported to co-stimulate the T cells. Interestingly, an alignment of the two sequences revealed that 51 amino acids corresponding to the sequence for the boundary of the extracellular and intracellular domains including the transmembrane domain are deleted at almost an identical location within the full form of CD86 from both species. This suggests the possibility of a co-stimulatory function of the putative soluble msCD86. In order to determine if the cloned msCD86 molecules has co-stimulatory activity, the proliferative responses of the human CD4(+) T cells to the msCD86-transfected COS cells were measured in the presence of Con A. The results revealed that CD86/COS, but not the mock/COS, efficiently co-stimulated the proliferation of the Con A-stimulated CD4(+) T cells and this co-stimulatory effect was blocked by CTLA4-Ig. The structural and functional information on the miniature swine CD86 from this study will enable a further genetic manipulation of CD86 as a therapeutic strategy for controlling the xenogeneic T cell immune responses mediated by the CD86-CD28 signal pathway.
Choi
Inho
I
Xenotransplantation Research Center and Clinical Research Institute, Seoul National University Hospital, Seoul 110-744, South Korea.
Cho
Bumrae
B
Kim
Sung Dae
SD
Park
Dongkyoo
D
Kim
Jae Young
JY
Park
Chung-Gyu
CG
Chung
Doo Hyun
DH
Hwang
Woo Suk
WS
Lee
Jung Sang
JS
Ahn
Curie
C
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000276:immunology; D051940:B7-2 Antigen / Q000235:genetics* / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D019556:COS Cells; D060908:CTLA-4 Antigen; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D006801:Humans; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity; D011993:Recombinant Fusion Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics* / Q000276:immunology; D014162:Transfection
D015703:Antigens, CD; D000943:Antigens, Differentiation; D051940:B7-2 Antigen; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C556707:Ctla4 protein, mouse; D011993:Recombinant Fusion Proteins